Home

Manual - VTP UP

1. ccccceceeeeeeeeeeeee ee eeeeaeeeeeeeeeeeeeeneeeeeeeeaaees viii Do s and Don ts oeroceaan eet ES iaa dead yaaealedzeseeansdadecdesaletuespeeataeeenalearels xi CHAPTER 1 INTRODUCTION 1 1 Overview of the Corona CAD Detectors cccccccceseeeeececeeteeeeeeeseeceeeeseeeanaeeneeeess 1 1 1 2 Features of the Corona and Corona Plus DeteCtors cccccceccseeeeeeeeseeeeeeeeeeeanaees 1 2 1 3 Overview of this M nual ccccccccccccccceeccceeeeeeesessueeuseseueeeeeeueeuaeeeueeueeeusanseeeueaaaaes 1 2 1 4 Benefits of the Detectors 2 cA eee eee AAS ek Se aed denen 1 3 1 4 1 The Detector Module INCIUGES cccccecceeecceceeeeeeeeeeeseeteeeeeeetaneeeeetaneeeents 1 3 1 4 2 Corona Detector Organizer Modules ccceeeeeceeeeeeeeeeeeeeeesaeeeeeesaeeeees 1 4 143 Nitrogen Generator nerenin a Gea e A RE 1 4 1 5 COMPONENT Locations na e n a E Ao a E AE a 1 5 1 6 Mode of Operation of the Detector cacaici orcad teseeeeree dice ta tee le scndstcotenrmeadeteeake 1 8 1 7 Additional INfOrMAtION neei arn n TE ROTTO TE 1 10 1 8 SUPPOM FOM ESA e a raa a R aA aE a E a R aai 1 11 1 9 Frequently Asked QUESTIONS cccceccceeeeeeeneeeeteeceeeeeeeaaeeeeseeaeeeeeeteaaaeeeeesaaeeeeeeas 1 12 CHAPTER 2 INSTALLATION 2 1 IMIIFOGUCTION tS Neth E seek ck te et i ad ee Ea Ee te she ae 2 1 2 2 Environmental Conditions 0 cccccccccecceccececeseeeecesueeueeeeueeuseseeeeuseseseauseveeeaaeeeanenes 2 1 2 2 1 Phys
2. Corona Wop Wo Oro Pump n O 6 lt 18 or 46cm _ gt q 72 or 188cm gt Figure 2 1 Approximate Dimensions for a Corona Detector System 2 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation NOTE The detector should be placed in a position so that distance between the end of the column and the mobile phase inlet on the detector is minimized This will reduce post column band broadening effects and will optimize chromatographic resolution 2 2 4 Nebulization Gas A supply of nitrogen is required for operation of the detector The gas must be pure typically 99 99 free from particulates devoid of volatile hydrocarbons solvent vapors e g from a compressor The gas must be supplied at a pressure of 60 psi for use with the Gas Conditioning Module 70 8285 or 35 0 0 1 psi if connecting directly to the detector The use of bottled nitrogen gas may be a quick and easy source of nitrogen during the installation However for longer term use a nitrogen generator Part Number 70 6003 is highly recommended WARNING The nitrogen gas source must not be oil pumped as oil in the gas stream can clog the detectors in line filters and may damage the detector itself WARNING It is required that inline moisture and 0 1um particle traps b
3. Figure 2 13 The Version Copyright Screen Corona CAD and Corona S CAD Detectors 2 17 Operating and Maintenance Manual Chapter 2 c After several seconds the instrument will automatically advance to the MAIN MENU Figure 2 14 MAIN CORONA MENU Select desired operation SYSTEM EDIT RUN Figure 2 14 The Main Corona Menu Screen d Start the delivery of nitrogen to the Gas Conditioning Module at a minimum pressure of 60 psi WARNING Always turn on the gas source first prior to starting the flow of mobile phase Turn on the detector then turn on the gas flow for a few minutes before running the pump e Using the application in preview mode turn on the detector f Confirm the gas pressure is 35 0 psi 0 1 psi If required adjust the gas pressure using the adjustment knob on the ESA External Gas Conditioning Module P N 8285 so that the pressure reads 35 0 psi 0 1 psi Record the pressure and initial current information on the worksheet WARNING The detector is designed to correctly operate at 35 0 0 1 psi If the gas pressure provided is outside of the range specified the detector will not provide consistent results g Allow the gas to flow at least 5 minutes Wait for the reported current to stabilize The current should be 0 05pA gt gt 0 05pA h Start the delivery of the mobile phase see section 0 i Mobile Phase Formulation for instruction on how t
4. Figure 4 1 Typical Corona CAD Detector Baseline Noise Trace Figure 4 2 Typical Chromatogram for Caffeine Standard 5ug on Column 4 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Using the Detector in a HPLC System h A standard curve can be generated from 0 5000ng caffeine using the standards provided A typical standard curve is presented in Figure 4 3 3000000 2500000 2000000 1500000 Peak Area 1000000 500000 0 0 1000 2000 3000 4000 5000 6000 Mass on Column ng Figure 4 3 Typical Caffeine Standard Curve 4 8 Analysis of Data The Corona detector response is not linear typically parabolic in nature and as such an equation such as A a m can be used to describe the response the coefficients a and b are different Mass vs particle diameter has a cube root proportionality while mean charge per particle has a slightly gt 1 proportionality with particle diameter Consequently the response of the Corona detector is approximately proportional to the square root of the analyte mass across the entire dynamic range From a purely theoretical standpoint and in practice calibration is much simpler and ultimately can result in much more accurate quantitation than with ELSD A quadratic plot type present in all or most chromatography data systems provides an easy way to calibrate the Corona detector Alternatively a log log
5. EVENT MARKS Autozero Mark OFF Height 5 Event Width 4 sec Height 5 CANCEL MENU SAVE Figure 3 35 The Event Marks Screen When the screen is accessed the Autozero Mark events entry will blink Use the A or W keys to select the desired value Off or On This new field is accessed via the ENTER key and is used to indicate the height of the Autozero event mark the range is from 5 to 20 and is selected through the keypad After the Mark autozero events field and height field if applicable is edited press ENTER to access the Event width field Select the desired width 1 to 5 seconds of the event marker using the A or Y keys on the key pad press ENTER and then press SAVE 3 20 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware 3 6 5 Date amp Time Setup Screen The Date amp Time Setup screen Figure 3 36 can be accessed through the SYSTEM SETUP menu and then by pressing the A or V keys SYSTEM SETUP MENU Select Date amp Time Set Date and Time CANCEL NEXT Figure 3 36 The System Setup Menu Screen When the NEXT key is selected the DATE amp TIME SETUP screen Figure 3 37 is presented DATE amp TIME SETUP Month Jul Day 7 Year 2004 AM PM PM Hour 3 Min 17 CANCEL MENU SAVE Figure 3 37 The Date amp Time Setup Screen Enter the present date and time by accessing ea
6. defining the communication protocol to a personal computer etc These screens are discussed in Section 3 6 e EDIT accesses a series of screens to select the desired mode of operation and establish a set of analytical parameters for data acquisition A set of analytical parameters is termed a METHOD which can be stored and recalled as desired These screens are discussed in Section 3 4 e RUN accesses the RUN METHOD SELECTION screen which is used to choose a stored method to be run and then execute the selected method Selection and execution of a method is discussed in Section 3 5 Corona CAD and Corona S CAD Detectors 3 3 Operating and Maintenance Manual Chapter 3 3 3 The Keypad The keypad is used to enter data Table 3 1 or initiate an action Table 3 2 Table 3 1 Data Keys Key Function Numeric Keys Used to enter the appropriate value for various parameters e g offset Once the desired values are entered press ENTER to accept Used to change the sign of a numeric entry The sign should be changed before the numeric entry is made Used to accept that the value presented in the display for a given parameter is the desired value for that parameter and stores that value in ENTER the instrument memory If the value that has been entered is outside the range for that parameter the value will not be accepted and a message with the appropriate limits will be presented A Scroll
7. Factory set at 30 C Corona CAD detector only Corona CAD and Corona S CAD Detectors A 1 Operating and Maintenance Manual Appendix A A 2 Timeline Specifications Programmable Changes Programmable Events Program Repeats Program Start External Device Control A 3 External Requirements Power Gas Gas Operating pressure Operating gas flow rate A 4 Physical Specifications Dimensions Corona Module Organizer Module Weights Corona Module Organizer Module Thermal Organizer Module Selectable at times from 0 to 9999 99 minutes in 0 01 minute increments Gas on off Set Output Contact Closures 4 Autozero Filter Marker Current Range Hold Loop Reset Parameters 1 to 9999 using Loop command Via front panel keypad or rear panel contact closure Control of up to 4 external devices via contact closures 100 240 VAC 50 60 Hz 100 VA Nitrogen Gas must not contain volatile hydrocarbons e g compressor oils 60 psi 4 14bar into Gas Conditioning Module approximately 4 L min 20 5 L x 9 W x 11 25 H 52 5 cm x 23 cm x 28 6 cm 18 5 L x 9 W x 5 3 H 47 cm x 23 cm x 13 5 cm 22 lbs 10 kg 7 6 lbs 3 4 kg 9 1 Ibs 4 0 kg A 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Corona and Corona Detector Specifications A 5 Environmental Operating Temperature 10 35 C Humidity Maximum 8
8. S CAD Detectors 3 27 Operating and Maintenance Manual Chapter 3 6 Timeline Parameters a Time 0 00 min Event Hold Input Start Hold for ON Press Add Step soft key after each new timeline event b Time 0 10 min Event Autozero Press Add Step soft key c Time 1 50 min Event Range Range 500pA Press Add Step soft key d Time 3 90 min Event Reset This will reset values to original settings i e Range Press Add Step soft key e Time 4 00 min Event Loop Loop to 0 00 For 9 times In this example 10 injections are to be made therefore the loop must be 9 i e X 1 10 1 9 Press Add Step soft key f Time 4 10 min Event End Stops timeline clock Press Add Step soft key Press Next and Save To start the timeline method Choose Run and then the method that was saved The method will not start until it is triggered It is probably best to initialize the timeline method by a trigger from the autosampler NOTE Initially a blinking cursor will appear on the Range field Press IEE Enter until the cursor is no longer visible A timeline event change cannot take place when a blinking cursor is present on a particular field 3 28 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual CHAPTER 4 USING THE DETECTOR IN AN HPLC SYSTEM 4 1 Introduction The Corona CAD Detector is on
9. These devices can usually be obtained from sources that carry computer supplies If you have any questions concerning this please contact the ESA Service Department or its Representative ce NOTE A larger capacity for the UPS device is needed if the entire HPLC system is to be covered by this UPS device In this situation the total power consumption of your entire HPLC system must be determined and the desired period of time that the system should operate after the power interruption must be determined to calculate the proper capacity of the UPS system In addition it is strongly recommended that the detector does not share a circuit with other electrical equipment that draw significant power from the AC line This equipment includes items such as fume hoods refrigerators ovens centrifuges refrigerated cooling systems vacuum pumps etc viii Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual B Avoid Radio Frequency Interferences RFI Strong RFI may also cause disturbances or artifacts in the chromatogram or with the operation of the detector The detector has been designed to meet various certification agencies stringent requirements for susceptibility to RFI However it is still possible that a strong emitter of RFI could result in interference with the Corona CAD detector Therefore the detector should not be placed close to other instruments or machinery that could emit excessive RFI or magne
10. at Seller s option be terminated D THE WARRANTIES STATED HEREIN ARE THE ONLY WARRANTIES GIVEN BY SELLER WITH RESPECT TO THE PRODUCTS AND THE SOFTWARE PROVIDED WITH THE PRODUCTS AND ARE GIVEN IN LIEU OF ALL OTHER WARRANTIES EXPRESS OR IMPLIED INCLUDING WITHOUT LIMITATION ALL WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE BUYER S EXCLUSIVE REMEDIES AND SELLER S SOLE LIABILITY FOR ANY NON CONFORMITY OR DEFECT IN THE PRODUCTS OR SUCH SOFTWARE WILL BE THOSE EXPRESSED HEREIN Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual E With respect to products sold to Buyer but not manufactured or in the case of software not developed by Seller SELLER MAKES NO WARRANTY OF ANY KIND EXPRESS OR IMPLIED INCLUDING WITHOUT LIMITATION ANY WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE but Seller will make available to Buyer to the extent permitted by law and relevant contracts the warranties of the manufacturer of the relevant product or the developer of the software ii Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual SAFETY OPERATING SYMBOLS The following symbols appearing on the unit or in the manual are defined as follows a bP BP uw This symbol on the instrument indicates that the user should refer to the operating manual before attempting to connect the power interface cables and operate the system This symbol on th
11. i Pressure Corona status view Select Screen f a z 7 Total Flow Ratio Warning eee AE ee i l If TimeLine add event Total flow Flow ratio view __ screen k Corona voltage and current view Time Event 1 Trap voltage view Event conditions N E EE M y ie If Timeline Events view med Save Method screen i screen Time Event Event conditions Some ESS x D 1 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Appendix D This page intentionally left blank D 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual APPENDIX E DRAIN VENT ASSEMBLY OLD STYLES E 1 Drain Cap Assembly Part Number 70 7117 Compression Drain Fitting Components Table E 1 shows the list of items and their part numbers of the components contained in the accessory kit Table E 1 Components Recorded on the Customer Inventory Checklist Qty Description ESA Part Number 1 Corona Detector or Corona Plus Detector 70 6186A or 70 6967 1 Power Cord 70 1164 1 Bottle Assembly 4L Waste 70 7060 1 Organizer Module 70 5499 1 Thermal Organizer Module 70 5499TA 1 Organizer Accessory Kit 70 5651 1 Thermal Organizer Accessory Kit 70 5651TA 1 Corona Corona Plus Accessory Kit including 70 6225A 1 Limited Warranty Registration Card 10 0181 1 Cable BNC conn
12. ppa lt OA A lon K EN I A B N N N 4 sr WYN V V ANA Ati IN VE YVO AN Minutes Figure 5 5 Example of Pump Noise Minimizing the pump noise will be dependent upon the root cause The pump should undergo routine maintenance as recommended by the manufacturer Mobile phase should be degassed prior to use HPLC fittings should be checked routinely 5 18 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting 5 8 2 Mobile Phase Flow Like any other HPLC detector the flow of mobile phase to the Corona detector must be maintained Interruption of mobile phase to the detector e g due to pump issues leaks in the system etc will have adverse effects on performance The flow of mobile phase to the detector must also be within the recommended limits i e gt 0 2 but lt 2 0 mL min Use of flow rates outside the recommended range can have adverse effects on detector performance For example flow rates lt 0 2 mL min can cause detector noise Figure 5 6 due to issues with nebulization This problem is typified by rhythmic noise in the chromatogram that is magnified at the apex of the analytical peak This problem can be avoided by making sure that the flow rate is gt 0 2 mL min Flow rates gt 2 0 mL min may overwhelm the nebulization process ADC1 A ADC1 CHANNEL A CF10 07300409 D mV 37 5
13. the blank injections should be flat and provide a means of measuring base system noise NOTE This is only intended to be a preliminary test that the Corona detector is working properly A more comprehensive Validation Test is available from ESA or can be created using the standards included with your instrument Congratulations your installation of the Corona Detector is complete Corona CAD and Corona S CAD Detectors 2 19 Operating and Maintenance Manual Chapter 2 2 6 ESA Biosciences Corona CAD Detector Test Worksheet GENERAL INFORMATION Organization Address Detector Unit S N Software Version A Startup Information Operator Phone Number Fax Number Email Address Date If there is an error message on the display when the unit is powered up please indicate B Mobile Phase Information Lot Number Expiration Date Value C Tests Initial Conditions Present Value Gas Pressure Expected Value 35 0 0 1 psi Current Measured Value B1 When Nitrogen is initially provided B2 When Mobile Phase is turned on B3 After stabilization Operational Present Value Operational FRatio Reading ICor Reading Total Flow Reading VCor Reading lon Trap Reading Expected Value B2 gt B1 B3 lt B2 Present Value Y N Chromatogram Height Area Retention Time 500 g mL 500 g mL 500 g mL 2
14. with A Peak area m mass of the sample and a b certain factors Can you supply a comparable equitation for the Corona CAD detector Answer The Corona CAD detector s response is non linear and as such an equation such as A am can also be used to describe the response The coefficients a and b would of course be different The difference to ELSD is that CAD response is approximately proportional to the square root of analyte mass across the entire dynamic range The proportionality with ELSD changes from 0 67 to 1 3 to 2 0 through the dynamic range again due to the changes in efficiency of light scattering with particle size From a purely theoretical standpoint and in practice calibration is much simpler and ultimately can result in much more accurate quantitation than ELSD Question Is the Corona CAD detector compatible with columns with a diameter smaller than the conventional 4 6 mm How small Answer In terms of flow rate gt 0 2mL min or columns typically gt 2mm id Question At the rear side of the Corona CAD detector is an USB port can this currently be used Answer Yes The Corona detector can be interfaced to an Agilent Data Station or ESA Elite Data Station using USB Question What are the Corona CAD detector s security features Is it possible to monitor if somebody even with the correct password has changed a method Answer No it is not possible to monitor if someone with the password has changed a method
15. 2 18 Degassing Apparatus b Place mobile phase bottle in the ultrasonic bath c Confirm water level for the ultrasonic bath is at proper operating level about 1 from the top of the ultrasonic bath Figure 2 19 Add more water if necessary Change the water once a week d Turn the timer on the ultrasonic bath to 10 minutes Figure 2 19 2 30 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation Figure 2 19 Water level and timer location e Place the stopper on top of the mobile phase bottle and hold in place until vacuum seal is made when pump is turned on f Turn the vacuum pump on Make sure the stopper is sealed to the bottle Keep pump on for the whole of the degassing process at least 10 minutes Once degassing is completed turn off the vacuum pump Carefully remove the rubber stopper from the mobile phase bottle Remove the mobile phase bottle form the ultrasonic bath Secure cap Wrap in Parafilm around cap and neck of bottle x g h Dry with paper towel Corona CAD and Corona S CAD Detectors 2 31 Operating and Maintenance Manual Chapter 2 2 11 4 Wrap Up a Once degassed place cap tightly on bottle and seal with Parafilm b Label bottle and record the following information on label and in lab log book Methanol Lot Number Date of Preparation Expiration Date Preparers Initials Last Degassed on c Mobile phase not used within 48 hour
16. 24 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware Table 3 6 Thermal Organizer Error Codes Code Message 0 Thermal Organizer not detected 1 Thermal Organizer malfunction 2 Thermal Organizer malfunction logic 3 Temperature out of range under 4 Thermal Organizer sensor malfunction 5 Thermal Organizer malfunction logic 6 Temp sensor or heater not responding 7 Caution excess heating detected es NOTE For proper temperature stabilization the set temperature must be at least 5 C above ambient 3 6 10 Self Test Screen The Self Test screen Figure 3 46 can be accessed through the System Setup menu and by pressing the A or V keys SYSTEM SETUP MENU Select Self Test Self Test Instrument Restart CANCEL NEXT Figure 3 46 The System Setup Screen If NEXT is chosen The Corona detector will restart and proceed through its self test If there are no error messages the user can press the Next soft key to proceed to the Corona detector main menu 3 6 11 Nebulizer Heater Setup Screen The Nebulizer Heater Setup screen is only available when using the Corona S CAD detector The Nebulizer Heater Setup screen Figure 3 47 can be accessed through the System Setup menu and by pressing the A or WV keys Corona CAD and Corona S CAD Detectors 3 25 Operating and Maintenance
17. CAD response is approximately proportional to the square root of analyte mass across the entire dynamic range The proportionality with ELSD changes from 0 67 to 1 3 to 2 0 through the dynamic range again due to the changes in efficiency of light scattering with particle size From a purely theoretical standpoint and in practice calibration of the Corona CAD detector is much simpler and ultimately can result in much more accurate quantitation than ELSD Question We have observed that response factors for the Corona CAD detector is not consistent for the same compound over different concentration ranges Why is this Answer Response factors are not consistent for the same compound over different concentration ranges because the CAD response is nonlinear One would have to calculate the concentrations using a quadratic fit Question Why is the first part of the response curve more parabolic than linear Answer The shape of the response curve is related to the expected square root proportionality between mass and response Mass vs particle diameter has a cube root proportionality Mean charge per particle has a slightly gt 1 proportionality with particle diameter This and perhaps other factors lead to an observed square root proportionality 1 12 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Introduction Question For a UV detector we have Lambert beers law for an ELSD we have the equation A a m
18. Edit the Parameters Screen cece cccceccccceeeceseeeeeceeeeeueueeeuausueeseaueueeneuaeuss 3 7 3 4 5 Creating Editing a CAD TL Method eee eee eeeeneeeeeeeeeeeeeeeeeeaeeeetees 3 8 3 4 5 1 Timeline Screen oo cece ccecccceeeeeeceesueeeseseeeueeseaeueeeeeaeeeesaneeeaes 3 8 3 4 5 2 End Event Screen o ccccccccccccccccsceceecessueeseseeeuseseeeeeeeuseseeeaneass 3 10 3 4 5 3 Set Contact Screen oo cceccccccseeeeessueeesessueeeseseeeeeeeseneneeeeueanes 3 10 3 4 5 4 Autozero SCIEN cccecccccceeeccceeeeeeeessueeeseseueeaseseueeeeeeeneueeeueaaes 3 11 3 4 5 5 Filler SCree n wx cisecte ik Reebok E sock Sasnctaata gaan leks does beaded 3 11 3 4 5 6 Marker SGreenisase deve e eee ek ees kas vaste EE EE EEA SE 3 11 3 4 5 7 Range Screen 25s 242 05 datetene dace repdesestentpusecsiuccaeceneiqercitecdxaataratess 3 12 3 4 5 8 E f SCCM EA E used sheet saved ENT EEES 3 12 ERES Ne E a 01l erea eTe a E EE E T dee tel a E TEE 3 12 3 4 5 10 Lop Sereen nena a E hoo 3 13 g 4 5 1T Roset Ser Oinin ti d a A a rala 3 13 3 4 6 Save Method Screen cccccccccccccccccceceeseeseceseeeueeseeeeuseeeueeueeeeeeeaeeeeuuanseeeneaess 3 13 O 4 7 Method Z6r0ss xii esses alesse secant A sett SPE wie hewisdeei hate he oe ates 3 14 Using a Method to Collect Data s i5 2 5 iaconacicsschtans dened esses de serendaeceaeaaednegaenteaes 3 15 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 3 6 3 7 The System SOFECNIS 2
19. Elution on Response Under isocratic conditions the behavior of the Corona detector is like most other HPLC detectors i e as the elution time increases the peak height decreases peak width increases but area remains the same see Figure 4 6 Mobile Phase 70 MeOH Flow Rate 1mL min Column Shiseido Capcell Pak C18 MG 4 6 x 250mm Column Temperature ambient Injection Volume 10yuL Q Q O Prednisone Hydrocortisone 2 5 5 7 5 10 12 5 15 17 5 Minutes Figure 4 6 Typical Analyte Response under Isocratic Conditions Corona CAD and Corona S CAD Detectors 4 11 Operating and Maintenance Manual Chapter 4 During gradient elution however the response of the Corona detector increases as the organic increase This is purely the consequence of nebulization with an impactor and siphon There is a greater loss of analyte when eluted in aqueous solvent The effect of the gradient on detector response is illustrated in Figure 4 7 high to low organic and Figure 4 8 low to high organic Glucose 2 3 Column Asahipak NH2P 4 6 x 250mm 5um Column Temp 35 C f 5 Injection Volume 10L v a oar her 3 l he a dt LJ mapai a LAU ENTS SA REEE AEE E EERE aaa HPLC Parameters Mobile Phase A acetonitrile Mobile Phase B water Flow Rate 1mL min Gradient Profile 30 B to 70 B from 0 to 40 mins Minutes Fig
20. The only aspect of security is that you can lock the keypad Question Can any nitrogen generator be used with the Corona CAD detector What is the minimum purity required Answer The purity of gas must be 99 and it must be oil free and not contaminated with particles Question How does the ion trap work It must be different than classical ion trap used on a mass spectrometer Answer The ion trap is simply a low voltage negatively charged plate that traps primarily the excess reagent gas ions positive charge and high mobility and allows charged analyte particles lower mobility to reach the electrometer It is therefore nothing like an MS ion trap which consists of an rfide field designed to sort ions based on differences of their m z Question How does CAD differ from APCI Answer CAD has some similarities to APC A main difference is that CAD involves formation of dried particles each presumably consisting of multiple analyte molecules assuming small molecules or even small proteins rather than formation of individual gas phase ions with APCI The charging of aerosol particles then involves collision with reagent gas ions and this process is not significantly dependent on analyte polarity acid base characteristics or eluent pH This is best demonstrated by the fact that one can measure inorganic anions through positive charge measurements with CAD The charge is proportional to particle diameter which is in turn proportional
21. Trouble or flow ratio HPLC plugged into Contact ESA shooting errors v common power Service maintenance source Check source of gas Use new HPLC Ground detector P Contact ESA to HPLC Check drain Servi tubing eIvIice Contact ESA Contact ESA SENCE Service Figure 5 10 Sources of Noise 5 24 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting 5 12 General Troubleshooting Guidelines Although an HPLC system consists of several components troubleshooting can be simplified by consideration of the following guidelines a In almost all cases there is one cause for the problem As an example if an aberrant baseline is observed the problem can be caused by one of the following e The pump e g the pump is not primed e The mobile phase the mobile phase is not suitably degassed or contains contaminants e The column the column is contaminated and strongly eluted compounds are being eluted e The detector there is an electronic problem e The detector may be contaminated e A fitting a fitting may be leaking e Injector not injecting properly e The waste container has overflowed A fundamental knowledge of the role of each component of the system is extremely useful in diagnosing the problem The availability of spare parts to substitute is very useful in diagnosing the problem If a problem is observed run a standard sample to determi
22. Use a quadratic fit for calibration curves DON Ts e Do not use non volatile additives in the mobile phase e Do not use either ammonium carbonate or ammonium bicarbonate in the mobile phase e Do not use a column that has been previously used with nonvolatile mobile phases e Do not empty waste bottle while mobile phase is flowing e Do not flood the detector Severe or prolonged flooding may damage the detector and void the warranty e Do not turn on pump flow without gas flowing through the detector e Do not use columns that bleed e g silica based amino columns e Do not use a linear fit for calibration curves Corona CAD and Corona S CAD Detectors xi Operating and Maintenance Manual Additive Buffer pKa Buffer Range Trifluoroacetic Acid TFA 0 3 Formic Acid 3 75 2 8 4 8 Ammonium Formate 3 75 2 8 4 8 Acetic Acid 4 76 3 8 5 8 Ammonium Acetate 4 76 3 8 5 8 Not all inclusive Some Corona CAD Detector Compatible Volatile Mobile Phase Constituents xii Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual TABLE OF CONTENTS Waray ok ca idee edtee eae he en Meds eee he OAM aA Ie Tue aan i Safety Operating Symbols ccc o5 5 ioc aeses hed eesticeaes neneereiactecernees ekeee ed asta eens ili Warnings and Safety Precautions 0 ccccce cece eee eeneeeeeeeeaeeeeeeeaaeeeeeecaeeeseneeeeeeeneaes iv Important Operating Considerations
23. a a a a E Ea ea EAE aE G 1 INDEX eee eera Eae a Ee EE e Oaea aaa e Aarra eE aaraa Eaa eenia l 1 Corona CAD and Corona CAD Detectors xvii Operating and Maintenance Manual This page intentionally left blank xviii Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual CHAPTER 1 INTRODUCTION 1 1 Overview of the Corona CAD Detectors The ESA Biosciences Corona CAD and Corona S CAD Detectors Figure 1 1 are universal detectors that are designed to measure non volatile and some semi volatile analytes in the eluent from a high performance liquid chromatographic HPLC system The Corona S detector is designed to be used with mobile phases that may adversely affect the performance of the detector due to cooling of the nebulizer e g when using aqueous THF gradients This Operator s Manual will cover many aspects of successfully installing and operating the Corona or Coronas detectors While many areas covered will be similar for the two detectors any differences between them will be highlighted throughout this manual Corona Figure 1 1 The Corona S CAD Detector Optional Thermal Organizer Not Shown Corona CAD and Corona S CAD Detectors 1 1 Operating and Maintenance Manual Chapter 1 1 2 Features of the Corona and Corona S Detectors Reliable and simple to operate Universal detection of nonvolatile and many semi volatile analytes Four order
24. and samples are all run under the same conditions the selection of the filter value will not affect analytical results so long as the peak is clearly observable If the peaks of interest are very sharp and occur soon after injection a short filter time is advised If desired you can also vary the filter during a chromatographic run timeline using smaller filter times for the first few min of a chromatographic run and then using larger filter times later in the run 4 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 4 7 Using the Detector in a HPLC System An Experiment to ensure that the Chromatograph and the Detector are Functioning Properly NOTE Instrument Validation Services Installation and Operational Qualifications and an Instrument Validation Kit are available from ESA Please call your local ESA Biosciences representative for information The experiment described below which involves the detection of caffeine is included so that the analyst can perform an analysis with the Corona detector using a well defined system It might be useful to perform this experiment prior to developing separation detection parameters for the specific needs of the laboratory 1 Materials Needed A mobile phase consisting of 20 80 methanol water HPLC Grade 500 ug mL caffeine standards included in Part Number 70 6565 Caffeine Standards Kit 2 Procedure a Configure system as described i
25. be easier to service the manual injector such as replacing tubing connectors etc with the chassis plate outside of the organizer tray Corona CAD and Corona S CAD Detectors C 17 Operating and Maintenance Manual Appendix C C 6 1 Maintenance on a Daily Basis Check all fluid connections for leaks and fix as required Clean up any spills immediately C 6 2 Maintenance on a Monthly Basis Check all cable connections including the ground strap cable Tighten secure as needed C 6 3 Cleaning Wipe up any spilled mobile phase or other liquids immediately The inside and outside surfaces of the organizer can be cleaned with warm soapy water and a soft cloth Use dilute bleach as a disinfectant Wipe up any excess bleach and dry witha cloth C 18 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual APPENDIX D__ FIRMWARE FLOW CHART I RS 232 gt Del Methods gt 7 Event Marks gt 7 Date amp Time I Remote sae 7 Inputs I Contacts I gt Thermal Org Self test had 4 Nebulizer aag If Plus System select Main Menu taa aaasssssssssma i so System Edit Run Select Method Select Method Security select screen i Range Filter Offset Current Output Nebulizer views Range Filter Offset view modify
26. conditioning duct 26 The detector has a weight of 22lb 10kg and a length of 20 5 52 5 cm During installation consider the combined weight of the HPLC system The lab bench must be strong enough to support the weight of the entire HPLC system It must be level stable and have a depth of at least 23 6 600 mm 27 Ensure that all parts of the HPLC system are properly grounded It is strongly recommended that all parts of the system are connected to a common ground WARNING DO NOT attempt to bypass the earth ground connection A serious shock hazard could result 28 The power consumption of the unit is 100VA Check that the power supply voltage is stable and that its current capacity is sufficient to operate all of the components in the HPLC system If not the instrument s performance may be affected 29 Always filter mobile phase through a 0 2um filter prior to use Always degas the mobile phase as air bubbles may cause the pump to malfunction and cause detector noise 30 Dispose of waste liquid properly 31 If the detector is not going to be used for prolonged periods of time flush the detector with a neat pH neutral solvent for a few minutes to ensure that all buffers etc are removed Turn off the flow of mobile phase Turn off gas flow approximately 5 minutes later Finally turn off the detector DO NOT leave the detector exposed to either acidic or basic mobile phases If the equipment is used in a manner no
27. droplets An impactor is included in the nebulizer to remove large aerosol droplets Large aerosol droplets which would not dry properly and would lead to a larger amount of noise in the detector strike the impactor and are diverted to the drain and then to the waste bottle For the Corona detector the nebulizer is maintained at 30 C in order to prevent freezing of the nebulizer when used with some aqueous organic gradients 1 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Introduction Evaporation of the Solvent from the Aerosol the remaining aerosol droplets are directed into the drying tube where the solvent is completely evaporated Since the droplets are very small they evaporate very quickly The remaining analyte particles are then directed into the corona region of the detector Charging of the Solid Particles before the particles can be detected they must be electrically charged A secondary stream of gas forms positively charged ions as it passes a high voltage corona needle The stream of ions then pass through an orifice into a mixing chamber forming a jet that collides with the opposing jet of analyte particles As the two jets collide and are mixed the charge is diffusionally transferred to the analyte particles After leaving the mixing chamber high mobility excess ions are removed by a negatively charged low voltage ion trap Low mobility singly charged particles greater tha
28. e g CE TUV UL Ie NOTE The detector must be connected to an electrical outlet that shares a common ground with other components of the chromatographic system e g pump autosampler computers etc This will avoid ground loops which can create erratic results e g varying background high noise spiking etc Use of a common power strip is highly recommended to ensure a common ground Although the detector contains a built in line filter to reduce interference at any input voltage connection to an electrical line which also serves units with a large power drain or which may be subject to power surges is not recommended typical units of this type include centrifuges ovens refrigerators and fume hoods In addition a surge suppressor or an uninterruptible power supply UPS may be used Surge suppressors or UPS designed for personal computers are suitable The Corona and Corona S detectors meet the requirements of Underwriters Laboratories UL Canadian Standards Association CSA and European Certification CE see Certification section 2 2 3 Space Requirements The dimensions of the Corona detector are 20 5 L x 9 W x 11 25 H 52 5 x 23 x 28 6 cm The weight of the detector is 22lbs 10kg The space requirements for the entire HPLC system depends on the configuration of the system Figure 2 1 shows the space requirements for a typical ESA HPLC Corona detector system Autosampler Computer
29. electronic issues e g those related to the electrometer Corona needle the ion trap and grounding gas issues e g flow and pressure errors and draining issues Such problems can be manifested either upon start up when the detector goes through its self test and or during use Operational Errors 5 7 1 Internal Electronic Diagnostic Programs Corona detector Self Test The Corona detector includes an internal diagnostic test program that is automatically executed whenever the unit is powered on This program is designed to determine if the electronics are functioning properly If no faults are detected the display will present the standard program screen If the diagnostics program determines that there is an electronic fault a message will appear on the display see Table 5 3 5 12 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting Table 5 3 Self Test Failure Codes Causes and Options Error Test Failure Possible Troubleshooting Code Cause 1 HV Potential Corona wire or high Rerun Self Test If Fails contact ESA Service Max voltage supply 2 HV Potential Min Corona wire or high Rerun Self Test If Fails contact ESA Service voltage supply 4 HV Current Max Corona wire or high Rerun Self Test If Fails contact ESA Service voltage supply 8 HV Current Min Corona wire or high Rerun Self Test If Fails conta
30. esa Corona CAD Detector Corona lus CAD Detector Operating amp Maintenance Manual Corona ESA Biosciences Inc 22 Alpha Road Chelmsford MA 01824 4171 U S A Telephone 978 250 7000 Fax 978 250 7090 www esainc com P N 70 6258 Rev G NOTICES The detector is covered by a limited warranty A copy of the warranty is included with this manual The analyst is required to perform routine maintenance as described herein on a periodic basis to keep the warranty in effect All information in this manual is subject to change without notice and does not represent a commitment on the part of ESA Biosciences Inc The Corona CAD and Corona Plus CAD Detectors and various components in these detectors are covered by the following patents US 5 098 657 5 374 396 6 544 484 and 6 568 245 2004 2005 2006 2007 2008 ESA Biosciences Inc All rights reserved No part of this manual may be reproduced or transmitted in any form or by any means without the written permission of ESA Biosciences Inc ESA CAD and Corona are registered of ESA Biosciences Inc PEEK is a trademark of Victrex plc Kalrez Mylar and Teflon are registered trademarks of E I DuPont deNemours amp Co Inc Kel F is a registered trademark of the 3M Company Printed in the United States of America WARRANTY A ESA Biosciences Inc Seller warrants to Buyer that each product manufactured by Seller will be free from
31. from other components in the system e g the pump can be led into the organizer via the vertical cuts in the cover Section C 4 2 An exploded view of the entire Corona Organizer Module is shown in Figure C 1 Please refer to this figure as you assemble the Organizer Module NOTE The exploded view included the cell brackets and related hardware Corona CAD and Corona S CAD Detectors C 3 Operating and Maintenance Manual Appendix C ORGANIZER COVER 70 5228 PLASTIG THUMB SCREW KNURLED SCREWS 70 5564 BRACKET FOR HOLDING DOWN CELLS 70 5439 KNURLED SCREWS 70 5564 BRACKET FOR MANUAL INJECTOR OVER FLOW LINES 70 5678 COLUMN HOLDER ASSEMBLY 70 5530 ge KNURLED THUMB NUTS FOR PANEL ORGANIZER GROUNDING STRAP 70 5532 70 5625 PANEL WITH HOLE FOR INJECTOR RES CHASSIS PLATE 70 5229 DRAIN TUBE PANEL WITHOUT HOLE FOR AUTOMATED INJECTOR 70 5418 ORGANIZER TRAY ASSEMBLY LATCH TABS Figure C 1 Exploded View of Corona Organizer Module C 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer C 3 2 Removing the Organizer Chassis Plate All components are mounted on the Organizer Chassis Plate Part Number 70 5231 Figure C 2 which is in turn attached to the Organizer Tray It is necessary to remove the chassis plate from the organizer tray to attach the various components Knurled nuts for attaching plat
32. is vapor pressure Compounds with vapor pressures of 1 2 x 10 Torr or greater are susceptible to evaporation at 29 C and may be problematic for detection by the Corona CAD 1 14 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual CHAPTER 2 INSTALLATION 2 1 Introduction This chapter describes e Environmental Conditions Section 2 2 e Unpacking the Detector Section 2 3 e Installing the Detector Section 2 4 e Test Protocol Section 2 5 e ESA Biosciences Corona CAD Detector Test Worksheet Section 2 6 e General Operations Section 2 7 e Other Detector Configurations Section 2 8 e Setting up the Thermal Organizer Section 2 9 e Making Electrical Connections Section 2 10 e Mobile Phase Formulation Section 2 11 e Column Conditioning Procedure Section 2 12 Corona and Corona S detector specifications can be found in Appendix A 2 2 Environmental Conditions 2 2 1 Physical Location of the Corona Detector The detector should be operated in a facility with the following environmental conditions e The temperature range should be maintained between 10 35 C The Corona detector should not be installed near a window ventilation duct or any other device that can cause a significant change in the temperature The humidity should be maintained between 0 85 RH e A suitable ventilation system must be provided Many of the solvents that are used in an HPLC system are toxic Make
33. its use with complex matrices e g human urine waste water etc may require more extensive sample preparation e g solid phase extraction and or gradient separation 5 10 Thermal Organizer Issues If the Thermal Organizer is not functioning properly i e the set temperature is not reached or maintained check to make sure that the interface cable between the organizer and the detector is securely plugged into the Logic Module Power Supply Module and the PC board on the underside of the chassis plate of the organizer Corona CAD and Corona S CAD Detectors 5 23 Operating and Maintenance Manual Chapter 5 In addition make sure that e The remperature probe is placed all the way into the column clamp and secured with the setscrew e The column clamp is flush mounted on the chassis plate and is securely fitted to the plate 5 11 Noise Review A brief synopsis of the potential sources of noise and troubleshooting is presented in Figure 5 10 FLOODING HPLC System GROUNDING DRAINAGE Self Test Error High Background e Narrow spikes in Small dips or Symptom Code 80 currents baseline peaks in baseline High Background Erratic baseline currents noise Liquid in exhaust Mystery peaks or vents Lack of liquid in drain tubing Check gas Remake mobile e Confirm that e Confirm proper pressure 35psi phase HPLC and drain assembly Check total flow Remove column detector are setup
34. l 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual
35. of the detector When Autozero input is in the Active on state as depicted in Figure 3 41 an Autozero event will occur if the autozero terminals on the I O connector block are shorted by an external device It may become useful during Timeline programming to pause two or three times during a Timeline program to await return signals from external devices indicating that they have completed their appointed tasks External devices can include an autosampler a column switching valve manual injectors signal relays etc Normally only one such input is needed and START is reserved for that purpose 3 22 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware This screen also allows the user to set the Autozero and GAS Off inputs to an Inactive off state This allows the user to disable either or both functions without having to remove the connections from the terminals on the I O connector block If the Autozero and GAS On Off states are turned to Inactive off then neither an Autozero event can be triggered nor the gas be turned off via a contact closure in the I O Connector block on the rear panel Under normal circumstances these two devices are always left ON in the System menu 3 6 8 Contacts Screen The Contacts screen is used to activate the individual outputs and view the state of the Inputs related to the I O blocks on the rear panel When the option is selected the screen
36. of the keypad these will allow the user to scroll through the display screens 3 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware The SECURITY LED on the keypad is illuminated when the system is operating in local or remote modes and the security feature is active to prevent accidental changes in the operating parameters In remote secure mode none of the front panel keys are active 3 4 Creating Editing a Corona CAD Detector Method 3 4 1 Overview To create or edit a Corona CAD detector method the user should perform the following steps a e a Edit the Method Select Screen Section 3 4 2 b Edit the Security Screen Section 3 4 3 c Edit the Parameters Screen Section 3 4 4 d Creating Editing a CAD TL Method if Timeline operation has been selected Section 3 4 5 e Save Method Screen Section 3 4 6 f Method Zero Section 3 4 7 3 4 2 Edit the Method Select Screen NOTE The protocol for editing a method is the same as that for generating a new method When EDIT is selected on the Main Corona Menu screen Figures 3 3 and 3 4 the EDIT METHOD SELECT screen Figure 3 5 is presented This screen is used to choose the method that is to be used for the analysis EDIT METHOD SELECT Method 1 Name Default Method Mode CAD D CANCEL NEXT Figure 3 5 The Edit Method Select Screen 1
37. of these may lead to flooding of the detector Severe or prolonged flooding may damage the detector and void the warranty 5 Empty the waste bottle routinely This is a closed system do not allow the detector to become flooded with waste solvent Severe or prolonged flooding may damage the detector and void the warranty 6 Do not start the HPLC pump before the inlet gas is flowing and the Gas On Off valve is open Otherwise solvent can accumulate in the detector and adversely affect performance 7 Never stop the gas flow and never switch the detector off before stopping the HPLC solvent flow and allowing 5 minutes for drainage Failure to do so will cause solvent to accumulate inside the detector and adversely affect performance 8 Wear protective goggles when handling solvents Install a sink as close as possible to the module If any solvents splash on the skin or eyes immediately rinse the affected parts in the sink Ensure that appropriate eye wash and safety shower are available 9 Familiarize yourself with each mobile phase compound s MSDS material safety data sheet Make certain that you understand the toxicological properties safe handling and correct disposal of all chemicals that are used with the detector 10 Only use volatile components buffers solvents etc in the mobile phase Do not use either ammonium carbonate or ammonium bicarbonate in the mobile phase These tend to be dirty and contain particul
38. on the logic module that is used to interface the detector with other components on the system Isocratic Refers to the use of a mobile phase with a constant composition during the chromatographic separation Linearity Refers to the ability of a property of a sample e g concentration to be directly related to an experimentally measurable parameter e g Current in a proportional relationship The linearity of a calibration curve is a factor of critical interest Linear Velocity Average speed of the solvent front though a column usually measured in cm sec Logic Module Component of the detector that contains the microprocessor and associated electronics It includes the input output connections and remote ports so that the detector can interact with other components in the system Longitudinal Diffusion Band spreading in the longitudinal direction of a column as a result of randomized or eddy diffusion of the compound of interest Looping Repetitive operation of a timeline method At the conclusion of the method the method is automatically repeated Mass Sensitive Detector A device that will provide a signal when a particle with the appropriate characteristic is passed through it Mass Transfer Transfer of mass from the mobile phase to the stationary phase or vice versa Stationary phase mass transfer thus refers to the diffusion of molecules into and out of the stationary phase Method A set of analytical parameters
39. plot can be used Table 4 2 shows typical response data for different steroid standards Figures 4 4 and 4 5 presents these data as using a quadratic plot and a log log plot respectively Corona CAD and Corona S CAD Detectors 4 9 Operating and Maintenance Manual Chapter 4 Table 4 2 Response area for Different Steroids a Steroid 1 Steroid 2 Steroid 3 0 0 0 0 10 7741 5 8662 5 10209 20 15361 16932 18731 5 50 45870 33782 5 41635 100 108576 5 78820 5 101639 200 227850 176439 210761 500 605444 5 473053 5 579961 1000 1196883 981872 5 1170350 2000 2147457 1993206 2148938 5000 4622897 4335706 4767831 10000 7504657 7465382 7749171 9000000 8000000 7000000 s 6000000 2 o 5000000 gt Steroid 1 2 OA Steroid 2 a 4000000 Steroid 3 amp 3000000 2000000 1000000 0 0 2000 4000 6000 8000 10000 Mass ng on column Figure 4 4 Response Curves for different Steroids showing Quadratic Fit 4 10 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Using the Detector in a HPLC System 10000000 1000000 g Steroid 1 amp 100000 4 Steroid 2 Steroid 3 10000 4 1000 1 10 100 1000 10000 100000 Figure 4 5 Response Curves for different Steroids showing a Log Log Plot 4 9 Effects of Gradient
40. remove the inner part of the organizer the piece that contains the column s etc to facilitate cleaning in and around the pan and the components see Appendix C The outside of all the column s tubing pulse damper etc may be cleaned according to the above procedure that the cleaning material does not get on the Temperature Probe the Thermostat or the Heater Driver Board located on the underside A CAUTION When cleaning the Thermal Organizer Module take care of the chassis plate as this could damage the organizer Clean the manual injector according to the manufacturer s instructions If it becomes necessary to disinfect the organizer a mild bleach solution can be used on all the surfaces However do not allow the bleach solution to remain on the organizer for more than a few minutes The bleach solution should be wiped off with a damp towel After cleaning the organizer and its contents can be dried using a soft towel Corona CAD and Corona S CAD Detectors 5 9 Operating and Maintenance Manual Chapter 5 5 5 Establishing a System Log A log that includes the usage and maintenance as well as any comments about operation of the system should be maintained This log should include the date time technician s name number of samples any maintenance activities and any relevant user comments about the performance of the system An example of a typical sample log is presented as Table 5 2 If a prescribed sample
41. should refer to the operating manuals for each component of the HPLC system and perform the necessary maintenance activities on a periodic basis ESA provides preventive maintenance services for all ESA components Please contact ESA or your local ESA representative for details 5 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting 5 2 2 Daily Maintenance NOTE The frequency for performing the various activities is dependent on the sample type mobile phase composition sample cleanliness and a number of other factors The frequency indicated below should be considered as a guideline As the user gains experience with the system and the analytical procedure it is recommended that the user creates his or her own maintenance protocols On a daily basis or every time that the unit is started up a Check that the waste bottle is empty and there are no bends kinks or obstructions in the waste lines b Check that the pump is working properly and the solvent bottle s contain sufficient mobile phase for the duration of the expected analysis c Check that there is sufficient pump seal wash solution if equipped Make certain that the pump seal wash system is primed and flowing properly d Check that all connections are leak free e If an autosampler is in the system check that the tray temperature is correctly set the syringe is bubble free and the wash
42. that can cause events such as a gain change autozero event mark etc to occur at designated times during a chromatogram Universal Detector A device that will provide a response for every compound that passes through it USB A standard interface scheme for the transmission of digital information between a computer and a detector Van Deempiter Plot A plot of the height equivalent to a theoretical plate as a function of the linear velocity of the mobile phase A van Deempier plot is used to determine the linear velocity that provides the most efficient separation i e the shortest HETP Void A part of an HPLC column that does not contain packing material A large void causes a dramatic decrease in the efficiency of a column Corona CAD and Corona S CAD Detectors G 9 Operating and Maintenance Manual Glossary Void Volume The available volume of an LC system between the injector and the detector The void volume is the total volume of the system less the volume occupied by the column packing This term is approximated by the peak in the chromatogram that corresponds to the unretained analytes Volt The unit of electrical potential difference W Peak width or band width Wall Effect Band spreading as a result of solvent flow along the column wall being different than the flow through the center of the bed Word The basic unit of information that is transmitted by the serial interface The length of the word i
43. the desired value appears press ENTER After entering all RS232 parameters press SAVE to save them NOTE It is not necessary to scroll through the entire screen Once the appropriate field is edited press SAVE to save the setup 3 6 3 Deleting Methods Screen The Deleting Methods screen Figure 3 32 can be accessed through the SYSTEM SETUP menu and then by pressing the A or Y keys SYSTEM SETUP MENU Select Del methods Delete all user stored methods CANCEL NEXT Figure 3 32 The System Setup Menu Screen When NEXT is selected the METHODS screen Figure 3 33 is presented Corona CAD and Corona S CAD Detectors 3 19 Operating and Maintenance Manual Chapter 3 METHODS Delete all user stored methods CANCEL MENU YES Figure 3 33 The Deletes all user stored methods Screen The DEL key deletes all user entered methods except Method 0 Method 0 contains the parameter values for the last method that was overwritten See Section 3 4 7 for more information about Method Zero 3 6 4 Event Marks Screen The Event Marks screen Figure 3 34 can be accessed through the SYSTEM SETUP menu and by pressing the A or WV keys SYSTEM SETUP MENU Select Event Marks Set Event Mark width height etc CANCEL NEXT Figure 3 34 The System Setup Menu Screen When NEXT is selected the Event Marks screen Figure 3 35 is presented
44. the new assembly Replace the two screws that hold the assembly in place Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual APPENDIX A CORONA AND CORONA DETECTOR SPECIFICATIONS A 1 General Operating Specifications Operating Mode Mobile Phase Flow Rate Wettable Surfaces Full Scale Output Range Filter Time Constants Noise Specification Signal Output Output Resolution Maximum Sampling Rate Output Offset Event Marker Autozero Function Keys RS232 Interface Capability Method Storage Settable Temperature Range Thermal Organizer Option Temperature Stability Thermal Organizer Option Warm up Time Thermal Organizer Option Charged Aerosol Detector Up to 2 mL min 316 stainless steel 1 pA to 500 pA in 1 2 5 sequence None Low Medium High lt 750 fA peak to peak 20 methanol 80 water OV to 1V DC 0 12 uV at 1V full scale 3 5 samples sec 50 of the selected current range in 1 steps Triggered by keypad timed operation or RS232 control width height polarity Triggered by front panel keypad rear panel contact closure timed operation or RS232 control Autozero Event Marker Gas On Off Full parametric instrument control Up to 25 methods any combination of Fixed Mode and Timeline Ambient 5 to 60 C lt 0 5 C lt 30 minutes typically Temperature Accuracy lt 0 5 C Thermal Organizer Option Nebulizer Heater
45. the shipping agent and ESA Biosciences or its representative immediately If any parts are missing call ESA s customer service department and indicate the missing items via the part numbers The shipping carton should be retained as it can be used if it becomes necessary to transport the organizer The module should be placed in an area that is free from drafts or significant temperature changes Avoid placing the organizer near air conditioning vents windows ovens etc Corona CAD and Corona S CAD Detectors C 1 Operating and Maintenance Manual Appendix C The Corona Organizer Module and Thermal Organizer Module are identical to the Coulochem IIl Organizer Module Thermal Organizer Module When the modules are employed with electrochemical cells cell brackets are provided to mount the cells to the housing While these brackets are not required with the Corona Detector they are provided as a convenience to the user in the event that the organizer is used for electrochemical detection with the Coulochem III detector Table C 1 Corona Organizer Module Accessory Kit Quantity Item Part Number 2 8 32 x 3 8 Screw and Star Washer for Manual Injector 50 5213 16 Feet for Cells 70 4336 1 Organizer Ground Strap 70 5625 1 Column Grounding Cable 70 5630 6 feet Tubing for Drain 50 6224 1 Bracket for Manual Injector Overflow Line 70 5678 2 Bracket for Holding Down Cells a 70
46. the yellow wires to the and terminals of the GAS OFF I O Connections respectively c Connect the black wire common to the GND terminal on the I O Connections on the Corona detector Corona CAD and Corona S CAD Detectors 2 27 Operating and Maintenance Manual Chapter 2 To set external events on the pump a While the pump is powered on press the CE key to return to the initial screen b Press FUNC key repeatedly until you come to the following screen c Input 2 and then press ENTER d Press CE to return the initial screen This operation has set the Event 2 of the pump s Relay 2 the Orange and Yellow wires to present a contact closure in the event of a pump error Now when a pump error occurs the detector will turn the gas off In the event that the pump turns the gas off correct the pump error After the pump has been repaired and is running normally the detector can be turned back on Remember to restore gas flow to the detector before starting the flow of mobile phase 2 10 3 Interfacing to Other Solvent Delivery Modules If your Solvent Delivery Module has the capability of providing a contact closure in the event that it experiences an error Types of errors registered by the pump include e g mechanical or electrical problem the back pressure of the HPLC system is outside the range of the upper or lower set pressure limits the pump runs out of mobile phase etc the
47. to the next value in a sequence e g the Current Range value When the desired value is presented press ENTER v Scroll to the previous value in a sequence e g the Current Range value When the desired value is presented press ENTER HELP Presents context sensitive information about the active screen ESCAPE Clears partial entries so you may re enter the correct values Active only when its activity indicator LED is lit Table 3 2 Action Keys Key Function Pe 5 EVENT MARK ie ie a spike on the recorder trace with a deflection 5 to 5 AUTOZERO Sets the output signal to zero at the current range in use GAS This key which is only active whilst running a method is used to open or ON OFF close the valve for the nebulizing gas Itis a toggle switch when the gas valve is opened the LED adjacent to it is illuminated The REMOTE LED on the keypad is illuminated when the detector is under the control of an external device The Corona CAD detector can be controlled by ESA s EZChrom Elite for ESA or Agilent s Chemstation software When configuring the detector for use with EZChrom Elite for ESA software the user has the option to lock the keypad If the user does not choose this option all the keys on the keypad will remain active If the user chooses to lock the keypad or is using Agilent s Chemstation all the keys on the keypad will be inactive except for the two soft keys on the right
48. uncapped end of a ball point pen into the tubing This will slightly widen the opening Take care not to damage the Teflon lining during attachment A CAUTION Because the tubing consists of two layers it may be d Attach the other ends of the Drain Vent tubing to the barbed fittings on the Cap of the Waste Bottle The Vent tubing will be attached to the smaller barbed fitting and the Drain tubing will be attached to the larger barbed fitting Figure E 10 Carefully place the tubing over the barb ensuring that the inner lining is over the barb Grasp the tubing near the barb and push the tubing onto the barb until it is all the way down Drain Tubing Pressure Barbed Fitting Relief Valve Vent Tubing Barbed Fitting Figure E 10 Bottle Cap Connections e Place the Waste bottle in secondary containment in a secure location below the detector Figure E 11 Make sure that the cap is tightened to prevent any gas leaks The waste bottle cap Part Number 70 6915A is a Teflon cap and includes a Teflon gasket Part Number 70 6895 inside to ensure a proper seal Figure E 12 The waste bottle is a closed system under a slight pressure and has a pressure relief valve on the cap in the event that the bottle becomes over pressurized e g if exhaust is blocked Corona CAD and Corona S CAD Detectors E 9 Operating and Maintenance Manual Appendix E Figure E 11 Correct Setup of Drain Tubing Note Secondary Containment Bo
49. underneath so that maximum contact is made As you install the column holder take care that the cable from the sensor will not interfere with placing the column in the holder Figure C 13 Mounting the Column Holder Assembly C 14 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer C 4 8 Manual Sample Injector Overflow Line Bracket The Manual Injector Overflow Line Bracket is an L shaped piece with small holes for tubing Part Number 70 5678 It should be mounted to the underneath side of one of the cell brackets The overflow lines vent tubes from the injector valve should be positioned through the small holes so that any overflow can drip into a Weigh Boat Part Number 40 0172 that is placed on the organizer chassis plate The overflow bracket is designed to keep the outlet of the vent tubes at the same vertical height as that of the needle port to prevent siphoning of the sample Connect the brackets to the left side of the chassis by the lip that fits into the horizontal slit and the knurled screw that fits into the right side of the chassis Figure C 14 The plastic knurled screws Part Number 70 5571 should be threaded into the proper hole over a cell and secured finger tight Figure C 14 Cell Bracket C 4 9 Final Assembly After all the connections are made a Place the chassis plate into the organizer tray so that the three threaded studs protrude t
50. washers Part Number 50 5213 supplied in the accessory kit am NOTE The screws and star washers are provided to ensure appropriate grounding Do not use the screws supplied with the injector to mount the injector e Reattach the injector handle and tighten the two hex setscrews Make sure the setscrews make contact with the flat portions of the valve stem It will be necessary to rotate the handle after tightening one setscrew to gain access to the other setscrew Refer to the Operating Instructions provided with the injector for other information on installation proper use and care of the manual sample injector valve These instructions include information about the wiring for the position sensing switch optional that can be connected to another HPLC component which is used to signal the start of the injection C 12 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer The cabling for the position sensing switch can be directed outside of the organizer at the back of the organizer along with the cell cables If an autosampler is to be used a Place the Organizer Injector Panel without the hole Part Number 70 5418 onto the organizer chassis plate b Affix the organizer injector panel to the organizer chassis plate with the two thumbnuts c Lead in the tubing from the autosampler through one of the ports on the proper side of the organizer C 4 5 Installin
51. will provide an electric signal that can be related to the concentration of the compound flowing through the detector Diffusion The movement of a compound through a phase under the influence of a concentration gradient In the column the compounds of interest will diffuse between the mobile phase and the stationary phase as a part of the separation process Distribution Coefficient D For a chromatographic band that is equilibrated the distribution coefficient is the ratio of the concentration of the compound on the stationary phase Ss to the concentration of the compound in the mobile phase Sm D Ss Sm where Ss is the concentration of the compound in the stationary phase Sm is the concentration of the compound in the mobile phase Drying Tube Device that removes the solvent molecules from the aerosol and thus forms solid particles E Abbreviation for Potential in V or mV Efficiency Chromatographic A measure of the effectiveness of a column for a given separation Efficiency is measured by the number of theoretical plates on the column Efficiency Detector The detector efficiency is the ratio of the amount of analyte actually detected to the total amount of analyte that passes through the detector Electrometer Device that monitors the charge on the particles from the Corona chamber by converting a flux of charged particles into an electrical current Corona CAD and Corona S CAD Detectors
52. with the wrench Pressure Relief Valve Drain Tubing Compression Fitting Vent Tubing Barbed Fitting f Figure E 4 Bottle Cap Connections Place the waste bottle in secondary containment in a secure location below the detector Figure E 5 Make sure that the cap is tightened to prevent any gas leaks The waste bottle cap Part Number 70 7117 is a Teflon cap and includes a Teflon gasket Part Number 70 6895 inside to ensure a proper seal see Figure E 6 The waste bottle is a closed system under a slight pressure and has a pressure relief valve on the cap in the event that the bottle becomes over pressurized e g if exhaust is blocked Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Drain Vent Assembly Old Styles Figure E 5 Correct Setup of Drain Tubing Bottle Cap with Teflon Gasket Teflon Gasket Figure E 6 Bottle Cap and Gasket This section covers the installation of the old style drain vent assembly and bottle cap Corona CAD and Corona S CAD Detectors E 5 Operating and Maintenance Manual Appendix E E 2 Drain Cap Assembly Part Number 70 6915A Barbed Drain Fitting Components Table E 2 shows the list of items and their part numbers of the components contained in the accessory kit Table E 2 Components Recorded on the Customer Inventory Checklist Qty Description ESA Part Num
53. 0 RH 35 C non condensing Storage Temperature 10 to 60 C Specifications are subject to change without notice A 6 Certifications The Corona Detector has the following Laboratory Equipment certifications USA UL 61010A 1 1 Edition Canada CSA Standard C22 2 No 1010 1 92 European Union EN 61326 1997 A1 1998 EN 61010 1 2001 02 FCC Part 15 Subpart B Class A Corona CAD and Corona S CAD Detectors AS Operating and Maintenance Manual Appendix A This page intentionally left blank A 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual APPENDIX B RECOMMENDED SUPPLIES AND SPARE PARTS B 1 Corona Accessories Corona Accessories Part Number Corona Accessory Kit Table 2 1 70 6225A Thermal Organizer Module 70 5499TA Organizer Module 70 5499 Nitrogen Generator 70 6003 B 2 Corona Replacement Parts Corona Replacement Parts Part Number Gas Filter Assembly HEPA Carbon Corona 70 6224 Corona Waste Bottle Cap Assembly Solvent Compatible 70 7117 old style Corona Waste Bottle Cap Assembly Solvent Compatible 70 7754 new style Bottle 5 0 Liter Waste Compatible with 70 7754 only 70 7751 Drain Vent Tubing Assembly Corona old style Drain Vent Tubing Assembly Corona new style Gas Inlet Tubing 4 specify length Tubing Cutter for new style Drain Vent tubing 70 7112 Ya Stainless Nut and Ferrule Set 70 7120 Drain Vent Tubing both old and new
54. 15 Subpart B Class A Specifications subject to change without notice x Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual DO s AND DON Ts This list contains some helpful hints to achieve success with the Corona CAD Detector DO s e Use only volatile mobile phase additives as with MS see a list of suggested additives below e Filter mobile phase through a compatible 0 2 micron filter note pre filtered HPLC grade solvents e g methanol and acetonitrile typically do not require filtration e Avoid contamination of mobile phase with non volatile material e g pH calibrators are a possible source e Thoroughly flush all HPLC lines to remove non volatile materials before connecting e Use mobile phase of pH lt 7 5 whenever possible e Use gas pressure of 35 1 psi as provided by the ESA Gas Conditioning Module 70 8285 e Use an in line filter between the column and detector e Use flow rates from 0 2 2 0 mL min e Allow for proper drainage with appropriate length of waste drain lines e Check level of waste bottle prior to operating the Corona CAD detector e Use the recommended flow splitting technique when using the Corona CAD detector in parallel with other detectors e Always make sure that detector gas flow is on before starting pump flow Ensure that the LED for Gas On Off is illuminated and there are no errors indicated on the Gas Flow screen e
55. 3 4 5 6 Marker Screen The Marker screen Figure 3 15 is used to set an event mark on the output at the indicated time The marker can be set from 10 to 10 in 1 increments TIMELINE Time 0 00 min Event Marker Height 5 CANCEL ADD STEP PREVIOUS NEXT Figure 3 15 The Marker Screen Corona CAD and Corona S CAD Detectors 3 11 Operating and Maintenance Manual Chapter 3 3 4 5 7 Range Screen The Range screen Figure 3 16 is used to select the range for the signal output at the indicated time The range can be set from 1 pA to 500 pA via a 1 2 5 sequence using the A or WV keys TIMELINE Time 0 00 min Event Range Range xx pA CANCEL ADD STEP PREVIOUS NEXT Figure 3 16 The Range Screen lt a NOTE For optimal performance an AUTOZERO should be performed prior to a range change in order to keep the signal on screen If a data station is used set the data station to inhibit integration during the period from 0 1 minutes before and after the gain range change 3 4 5 8 Gas Screen The Gas screen Figure 3 17 is used to turn the gas on off at the indicated time The choices are On or Off and are selected using the A or V keys TIMELINE Time 0 00 min Event Gas Gas valve Off CANCEL ADD STEP PREVIOUS NEXT Figure 3 17 The Gas Screen 3 4 5 9 Hold Screen The Hold screen Figure 3 18 is us
56. 3 Exe rMaliPSQuirG mens viac ches acteccastcg nese sa yeaa and acencenudadan de Saceci edie cp Segeg Sid eat QcvebeensaGiauas enna A 2 A 4 Physical Specifications 5c 55 Be cikeestnecesageshe ds nites ck denakagh has tenho tenes nadageadeaastendaces mat rnent A 2 A 5 Environmental o 2e 2ct0se wets sats Aae tus we Oaa aa ads aa aa Wt eee auntie A 3 A6 COPMPCAT ONS e eree a aaar a e Ana EE ESA S aae genie pleat teceageact es A 3 B 1 Corona Accesories osere a a aa e aan AE E B 1 B 2 Corona Replacement Pamts cc s cccceveeyserzeurssyncepedsauedoeensacstaaextasarueners soli pasoudeeneueseaets B 1 Bast Cale Giese plete Ra etre ona E teas oats eal a a a ed cia cased eta hemes E B 1 B4 WManiialS sie a a SU asa ee tages gS Oe eae Soe ered B 2 B 5 FUSOS a getetselctantdtecrennaieduaateearbiane niet a ARE OE EAEE PUK E tees AARE NAANA EKANA ANANA EEEa En e E B 2 BG Organizer Module Spare Parts ccc ccicis eeccies dtavelnueiecseessieclsceecctiaueeeerdeeeace netics B 2 B 7 Thermal Organizer Module Spare Parts ccccceeeeeeeeeeeeeeeeeeeeeeaeeeeeeseeeeeeeeneeees B 2 xvi Corona CAD and Corona CAD Detectors Operating and Maintenance Manual B 8 Test Standards and Guard Column cccccceeeeeeeeeeeeeneeeeeeecaeeeeeeceaeeeeeeeneeeeeseneeees B 2 B 9 Nitrogen Gas SUPP e ccarenietrn cert tee atcacet de tdetetene eeene teas Meaeeeces eadttceuthaatiartaaacate B 2 APPENDIX C ORGANIZER AND THERMAL ORGANIZER C 1 VOT WCW ees 2 cil cee s
57. 4 32 54 27 575 22 55 Figure 5 6 Low Flow Rate Noise Often the Corona detector is used in parallel with some other detector e g spectrophotometric devices such as UV fluorescence or PDA detectors and others such as MS with the flow of mobile phase from the column being split between the two arms see Section 2 5 7 As the Corona detector exerts a small back pressure it is important that the flow of mobile phase between the arms is balanced such that the Corona detector still receives gt 0 2 mL min This can be achieved by connecting a short piece of PEEK tubing for PEEK compatible mobile phases after the spectrophotometric detector to create a balancing back pressure Corona CAD and Corona S CAD Detectors 5 19 Operating and Maintenance Manual Chapter 5 5 8 3 Column The column and related chemistry see Section 5 8 4 Mobile Phase can have a major impact on the routine performance of the Corona CAD detector Often a problem perceived as being an issue with the Corona CAD detector is in actuality the result of a failing column or chemistry NOTE Generate a caffeine chromatogram using the caffeine standard eS and guard column Chapter 4 Section 4 7 provided with the detector If the Corona detector behaves normally i e like it did upon initial installation then the current problem is due to the column chemistry and not the Corona detector per se e Shifts in retentio
58. 5 yg mL 25 yg mL 25 yg mL Remarks 2 20 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation 2 7 General Operations 2 7 1 Turning Off The Corona Detector a Turn off the pump b c Return to the main menu if in run mode then press the Gas Off switch on the front panel wa Allow the gas to flow for at least 5 minutes d e Turn off the gas source to the detector Turn off the detector using the On OFF switch x 2 7 2 Emptying the Waste Bottle a Turn off the pump b Allow gas to flow for at least 5 minutes c Shut of gas using Gas On Off button d Return Corona detector display to main menu e f Check the FFKM sealing washer Part Number 70 7785 Replace if damaged g Reattach bottle cap Make sure it is securely tightened Undo cap and empty waste bottle NOTE Make sure that the waste bottle cap is tight and that the drain vent tubing connections are tight Failure to do so could result in low or no detector response h Return to Corona detector run screen i Turn on gas flow ensure that the gas pressure is normal and that there are no gas errors e g Flow Ratio error j After a period of 5 minutes the flow of mobile phase can be started NOTE If gas error messages appear check waste bottle cap and tighten if necessary Check tubing connectors Check to ensure
59. 5439 4 Plastic Screws for Cell Bracket a 70 5571 3 Knurled Screws for Cell Bracket a 70 5564 1 Panel with Hole for Manual Injector 70 5229 1 Panel without Hole for Automated Injector 70 5418 2 Knurled Thumbnuts for Panels 70 5532 10 Weigh Boats for Injector Waste 40 0172 2 Column Holder Non Heated Assembly 70 5530 1 Interface Cable Organizer to Corona 70 5595A a Used for mounting electrochemical cells when the organizer is used with the Coulochem III Detector C 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer C 3 Installing the Corona Organizer Module eS NOTE Refer to Section C 4 to install the Corona Thermal Organizer Module C 3 1 General Information Cut the tubing for each connection and prepare the appropriate fittings before installing the various components into the organizer The tubing length should be long enough to allow for a bit of play but excessive tubing length post injector should be avoided It is a good idea to use a heat gun or similar device to heat PEEK tubing in order to bend it without unduly stressing it If PEEK tubing is stressed it becomes weakened and can either burst at lower pressures than expected or leaks or sweating may occur This is especially important when sharp bends are used sharp bends can be found in many configurations or applications used with the Corona Detector and the Organizer Module Tubing to and
60. A D converter is to be used or additional IO connections are required see section 2 10 for further information 2 4 1 9 Setting up Corona Plus Nebulizer Heater a If using a Corona s detector press the System soft key on the main Corona menu Press the A or V key to select the nebulizer heater Select the NEXT soft key to access the Setup Screen Figure 2 11 NEBULIZER HEATER SETUP Status Normal Power OFF Temperature xx xC CANCEL MENU SAVE Figure 2 11 The Nebulizer Heater Setup Screen b Change the Power field to ON with the A or W key Press Enter and then press the SAVE soft key The display will revert back to the Main Corona menu The system will store the nebulizer heater status as ON and can only be changed to OFF if edited and saved in the Nebulizer Heater Setup screen Corona CAD and Corona S CAD Detectors 2 15 Operating and Maintenance Manual Chapter 2 2 5 Test Protocol 2 5 1 Role of the Test Protocol This section provides a procedure that can be used to verify that the Corona detector is functioning properly This procedure is designed for initial system checkout and can also be used for troubleshooting purposes This protocol assumes that the detector is part of either a Chemstation or EzChrom Elite system If it is not the protocol may still be run using the appropriate setup and operational steps for your specific configuration If problems are observed c
61. C 4 5 Installing the In line Filter eee eee eeeseeeeeeeeeeeeeeeeeeeeeeeeeaaeeeeeeeaeeeeeeae C 13 C 4 6 Installing the Temperature Probe cccceceeeeeeeeeeeeeeeeneeeeeseneeeeeeenaaeeeeeee C 13 C 4 7 Installing the Column Holder cccceeeesseceeeeeeeeeeeeeeeaeeeeeeeaaeeeeeeseaeeeeenee C 13 C 4 8 Manual Sample Injector Overflow Line Bracket ceecseeeeeeeeeeteeeeeees C 15 4 9 Final ASSEMDIY inie e hea ia Re ae Ria eas eee Seta C 15 C5 lt TROUDIESNOOUNG ad aisles c tert casa cone ee ene bea hannah maa C 17 C6 Maintenance and Cleaning sisccccicsceivcnnaigesieecneed eae wedi ctavbiianeaeieesbeeeteeeee C 17 C 6 1 Maintenance on a Daily Basis ccceeecceceeeeeeeeeeeeeeeeeeeeeeaaeeeeeeeaeeeeeeee C 18 C 6 2 Maintenance on a Monthly Basis ccccececeeeeeeeeeeeeeeneeeeeeeneeeeteeeaeeeeeeee C 18 CAE CIS ANIC E TESE EA E EAEE ES E EE ETA C 18 APPENDIX D FIRMWARE FLOW CHART 0 0 ccceceeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeenaaeeees D 1 APPENDIX E DRAIN VENT ASSEMBLY OLD STYLES E 1 Drain Cap Assembly Compression Drain Fitting Components ccceeeees E 1 E 1 1 Installing the Drain Vent Assembly eeeeceeccceeeeeeeeeeeeeeeeeeeeeeeeeteeeeeaaaees E 2 E 2 Drain Cap Assembly Barbed Drain Fitting Components ccceeeeeeeteteeeeeneeees E 6 E 2 1 Installing the Drain Vent Assembly ccceeeeeeeeeeeeeeceeeeeeeeeneeeeeneeeeee E 7 GLOSSARY a
62. Chapter 5 changed Gas flow off No gas flow to induce Make sure gas is flowing into nebulization detector pressure should be 35 psi Incorrect gain range Select or move sensitive gain range selection Gas flow reduced Reduced gas flow Increase gas flow pressure at leading to low response regulator but do not exceed 40 psi Check gas filters and replace as necessary Gas leak in waste bottle or Gas leak interrupts Make waste bottle cap on tight and drain vent tubing nebulization Gas flow all drain vent tubing connections are value too low on display tight 5 30 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 5 12 4 Loss of Response Cont Maintenance and Troubleshooting Possible Cause Comments Recommended Solution The corona charger may not be able to maintain the correct corona current Return to ESA for repair Electrometer filter is contaminated Return to ESA for repair 5 12 5 Inability to Autozero the Signal Possible Cause Comments Recommended Solution High background signal ona very sensitive current range background gt 10 x gain Use a mobile phase that provides a lower background lower concentration of impurities Increase the current range Autozeroing on a peak or the void signal Autozero on a relatively flat section of the chromatogram Autozero on a very noisy signal Reduce the noise or i
63. Contaminants leaching from system components Check mobile phase reservoir filters column end frits replace if necessary Passivate system components Waste bottle has been allowed to overflow and backup into the detector May cause permanent damage to electrometer assembly not covered by warranty Empty waste bottle Operate instrument without mobile phase pump off and with gas flowing gas on for several hours Detector flooded with solvent pump on while no gas flow is present May occur if gas tank empties and is not replaced Need to clean and dry all aspects of the flow path Operate the instrument without mobile phase pump off and with gas flowing for several hours The ion trap may be off or have a fault Will be indicated as a power on test failure Contact your ESA service representative a If possible avoid using triethylamine and other organic amines as chromatographic modifiers since organic amines tend to contain impurities b High background currents may be observed when the mobile phase has been stored for a period of time or some other change in the analytical protocol frequently unintentional is made 5 28 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting 5 12 3 Increase in Back Pressure Possible Cause Comments Recommended Solution Accumulation of particulates Start at th
64. D Detectors 1 3 Operating and Maintenance Manual Chapter 1 1 4 2 Corona Detector Organizer Modules A Thermal Organizer Assembly Part Number 70 5499TA and an unheated Organizer Module Part Number 70 5499 are available as options to contain the column manual injector in line filters and related fluidics These modules can be fitted on top of the detector The temperature of the Thermal Organizer Module can be set as described in Appendix C 1 4 3 Nitrogen Generator A nitrogen generator Part Number 70 6003 may also be purchased to convert compressed air into nitrogen for use with the detector The recommended specifications for a nitrogen generator are presented in Table 1 1 Table 1 1 Nitrogen Generator Specifications Nitrogen Purity 99 Nitrogen Flow Up to 4 L min Min Max Pressure 60 125 PSIG 1 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Introduction 1 5 Component Location Figures 1 2 through 1 5 present the location of various components commonly used in the operation of the detector Active Key Indicator Display Manual Gas Valve On Off Key Soft Keys Enter Key Event Mark Key Escape Key Autozero Remote Key Indicator z agii Security Numeric Keypad mgicator and Help Button Scroll Up Down Keys Active Key Indicator Figure 1 2 Front of the Corona CAD and Coronas CAD Detectors Coron
65. ESA Biosciences maintains both applications and service departments to assist in the event of any difficulty with the installation or operation of the instrument Contact Technical Support Phone 1 800 275 0102 or 978 250 7067 Fax 978 250 7065 for assistance The detector includes a number of diagnostic features so that potential problems can be traced to the appropriate sub system When contacting the ESA service department please provide the serial number of your unit and the software version number which is displayed after the system has initialized The software version can also be obtained at any time by pressing the HELP button followed by the ESCAPE button If a diagnostic message or error is presented on the display please be prepared to provide that information to the service representative as well An on line help system is included with the operating program which can be accessed at any time by pressing the HELP button ESA also offers instrument qualification services including Installation Qualification IQ and Operational Qualification OQ Performance Qualification PQ is performed by the customer unless the Corona CAD detector is part of a complete ESA HPLC system For details please contact ESA or your local representative A wide variety of instrumentation and supplies including pumps autosamplers nitrogen generators columns data management software detectors and mobile phases are available from ESA These compon
66. G 3 Operating and Maintenance Manual Glossary Eluent The liquid used as a mobile phase in high performance liquid chromatography Eluate The mobile phase as it leaves the column Elute To remove a band from an LC column through continuous flow of the mobile phase Elution Volume The volume time x flow rate of solvent that is required to elute a given component of the sample using a given HPLC system The time is measured from the instant of sample injection to the time of elution of the center of the band Equilibrium The condition when a chemical system is at rest At equilibrium the rate of the forward reaction and the rate of the reverse reaction are equal and as a result the concentrations of the reactants and products remain constant Evaporation Removal of the mobile phase by vaporization to provide solid sample particles for charging Event Marker A short pulse that is sent to the recorder or other output device that applies a momentary 5 signal It is used to denote some action on the output trace e g an event marker might be used to indicate the injection of the sample via a momentary baseline deflection Filter Signal An electronic device that removes a portion of the noise in the signal output A filter provides a smoother baseline making it easier for peak detection and quantification Flow Rate The rate at which the mobile phase is pumped through a column usually in mL min Fronting The phenom
67. Manual Chapter 3 SYSTEM SETUP MENU Select Nebulizer Nebulizer Heater CANCEL NEXT Figure 3 47 The System Setup Screen Select the NEXT button and the Nebulizer Heater Setup Screen Figure 3 48 is presented NEBULIZER HEATER SETUP Status Normal Power OFF Temperature xx xC CANCEL MENU SAVE Figure 3 48 The Nebulizer Heater Setup Screen This screen is used to turn the nebulizer heater ON or OFF When this screen is accessed the cursor appears in the field to the right of Power After choosing ON press ENTER and then SAVE The choice of ON or OFF is stored in the detectors memory and is automatically recalled when the detector is turned on This is a factory set temperature 30 C and is not editable The Status field indicates the present state of the nebulizer heater e g Normal Board Error Under Range Over Range and is used to indicate error messages Table 3 7 See Chapter 5 for further information Table 3 7 Nebulizer Heater Error Codes Code Message 0 Nebulizer Heater not detected 1 Nebulizer Heater malfunction 2 Nebulizer Heater malfunction logic 3 Temperature out of range under Nebulizer Heater sensor malfunction Nebulizer Heater malfunction logic Temp sensor or heater not responding Caution excess heating detected al NIO ee NOTE For proper equilibration of the nebulizer wait f
68. NIZER C 1 Overview The Corona Organizer Module and the Corona Thermal Organizer Module are optional accessories for the Corona CAD and Corona S CAD Detectors that are designed to accommodate the various components of the fluidics system of the HPLC such as pulse damper column s in line filters a Rneodyne manual sample injector valve tubing and associated cables etc The organizer module is mounted on top of the detector housing as shown in Figure C 1 The Corona Thermal Organizer Module allows the analyst to maintain the temperature of the column between ambient 5 C and 60 C The components that are to be mounted in the module are determined by the application for which the detector will be used A typical organizer configuration might include a manual sample injector a pulse damper an in line filter before the column an HPLC column and an in line filter before the detector This chapter describes how the HPLC components are mounted in the modules as well as maintenance and cleaning instructions C 2 Unpacking the Organizer The organizer module is shipped in a single carton containing the module and an accessory kit Unpack the shipment and inspect the contents to verify receipt of all components A Customer Inventory Checklist for the Corona Organizer Module is presented in Table C 1 Carefully inspect the shipping carton and all components If there is any damage to the carton or to any components contact both
69. Part Number 70 7785 e Perform all of the daily weekly and monthly activities 5 2 6 Additional Periodic Maintenance a The gas filters should be replaced as described in Section 5 3 3 at least once a year v IF the gas pressure falls below 35 psi the external Corona detector gas filters or the Gas Conditioning Module filter may be clogged and require replacement or the Gas Conditioning Module regulator may require adjustement v lf the external gas supply contains volatile hydrocarbons e g compressor oils it may clog the Corona detector s gas filter assembly and they may require more frequent replacement NOTE Volatile hydrocarbons e g compressor oils in the external gas supply to the Corona detector may cause permanent damage to the instrument and void the warranty 5 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting 5 2 7 Nitrogen Generator The nitrogen generator should be serviced every 6 months in order to maintain optimal performance All filter housings requiring routine service and are easily accessible when the filtration access panel is opened Replace the three filter cartridges with those provided in the Maintenance Kit Part Number 70 6230 The maintenance kit contains two coalescing pre filters and one final membrane filter When replacing the pre filter cartridges be sure to install the proper grade filter in the proper housing the housi
70. Somewhat polar side chain e g CN RS232 A standard interface scheme for the transmission of digital information between a computer and a detector This is commonly called the serial mode of data transmission Selectivity Chromatographic The ability of a chromatographic system to separate two compounds Selectivity is measured in terms of the resolution of the chromatogram Selectivity Detector The ability of a detector to provide a distinct signal for the desired compound in a mixture while discriminating against all other materials in the mixture Sensitivity Refers to the minimum detectable quantity of an analyte with a given set of analytical conditions The signal to noise ratio of the measurement should be indicated Serial Port A communication port on the Corona that uses the RS232 protocol or one that uses USB to transmit information A method of transmission of data in which information is transmitted one bit at a time Signal to Noise Ratio S N The signal to noise ratio for a data point is obtained by dividing the signal by the noise level that is associated with the measurement A larger S N ratio is more desirable than a small one As the S N ratio approaches unity the level of certainty that a peak is real falls Signal Output See Output G 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Glossary Solvation The process of dissolving a material by a solvent Solute Th
71. T e If the fixed parameter operation Mode CAD is selected skip Section 3 4 5 and save the method as described in Section 3 4 6 e Ifthe Timeline operation Mode CAD TL is selected enter the timeline as described in Section 3 4 5 3 4 5 Creating Editing a CAD TL Method Timeline 3 4 5 1 Timeline Screen A timeline method permits one or more activities to occur at user specified times during the run To indicate that a timeline is desired select CAD TL in the Mode Field Figure 3 5 The Timeline screen Figure 3 9 is presented when the ENTER key is pressed on the Parameter screen Figure 3 8 if the timeline operation is selected TIMELINE Time 00 00 min Event Set Contact Contact C1 set to Off CANCEL ADD STEP PREVIOUS NEXT Figure 3 9 The Timeline Screen When the Timeline Screen is accessed Figure 3 9 the cursor will be found on the first character of the Time field Now enter the time at which the event is to occur For example to enter in a time of 23 75 minutes press the 2 then the 3 then the 7 and finally the 5 keys in that order The numbers will only be accepted after all of them are entered All times must be entered to two decimal places i e enter 8 00 min not 8 min Press ENTER to accept the number entered and to access the Event field The Event field is used to select the desired action that is to occur at the indicated time Use the A or V key t
72. The Corona CAD detector can store 25 analytical methods which are sequentially numbered from 1 25 When the EDIT METHOD SELECT screen is opened Method 1 is presented displaying the mode of operation used in that method its name xxxx and the date that it was generated or last modified The cursor will be found on the method number Corona CAD and Corona S CAD Detectors 3 5 Operating and Maintenance Manual Chapter 3 lt a NOTE When the detector is first received Method 1 is provided as a default set of conditions that can be used for testing as described in Chapter 2 If desired it can be modified and used as a user method If a different method is to be edited press the A arrow key until the desired method is presented As an example if Method 2 is selected the screen might appear as shown in Figure 3 6 CAD TL indicates a method using the detector in Timeline mode EDIT METHOD SELECT Method 2 Name Default Method Mode CAD TL D CANCEL NEXT Figure 3 6 The Edit Method Select Screen for method 2 When the desired method is accessed it is possible to e Change the mode Press ENTER to move the cursor to the Mode field and use the A or Y key to access the appropriate mode e Access the Security screen Section 3 4 3 by pressing NEXT 3 4 3 Edit the Security Screen The Security screen Figure 3 7 which can be accessed by pressing NEXT while in EDIT SCREEN is use
73. a CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting 5 7 3 Drainage Proper performance of the Corona and Corona detectors require that the waste vent system detector vent drain tubing waste bottle be properly sealed i e under positive pressure Failure to secure the waste bottle cap properly can result in a Flow Ratio Operation error see Table 5 4 As the waste vent system is a closed system make sure that the waste bottle is emptied routinely e g daily prior to the start of an analytical run otherwise there is the possibility for the waste solvent to re enter the detector If this happens error messages such as Self Test Error Code 80 and Operation Error Flow Ratio Error may be presented If this happens please refer to Tables 5 3 and 5 4 for corrective action Severe or prolonged flooding of the Corona detector may require that the unit be returned to ESA for service Damage caused by flooding is not covered by the instrument s warranty Make sure that the Corona CAD detector and its waste bottle and vent drain tubing are installed correctly see Section 2 5 1 Basic Installation and that the vent drain tubing are free from loops bends and obstructions i e areas where waste solvent can accumulate and adversely affect drainage Failure to allow proper drainage of waste solvent from the Corona CAD detector may adversely affect detector performance causing pul
74. a CAD and Corona S CAD Detectors 1 5 Operating and Maintenance Manual Chapter 1 Figure 1 3 Front Left Side of Detector 1 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Introduction Logic Module Gas Module Power Module Thermal Organizer Control Ground Thermal Organizer Power USB Filter Module Brackets Signal Out Filter Module Fuse Connections Power Line On Off Switch Gas Inlet Gas Exhaust I O Connections Figure 1 4 Rear of the Detector Corona CAD and Corona S CAD Detectors 1 7 Operating and Maintenance Manual Chapter 1 ORe CC1 00 Pump Off GND O GND CUON CC2 ff Bee Gas O GND GND On Os F Autozero ccas LOG GND O GND Cy St CC4 eae e GND GND Figure 1 5 Close Up of I O Connections 1 6 Mode of Operation of the Detector The Corona detector is a mass sensitive detector that can respond to essentially all non volatile and some semi volatile compounds in the sample that are eluted from the column The steps in the detection process presented schematically in Figures1 6 and 1 7 include Nebulization of the Eluent the mobile phase containing the analytes from the column is introduced into the nebulizer where pressurized gas e g nitrogen that has been passed through a carbon filter and a high efficiency particulate air HEPA filter is used to form an aerosol of ultra fine
75. al columns to waste before attaching to the detector 5 8 4 Mobile Phase e Use only volatile salts in the mobile phase Exposure of the detector to non volatile salts will lead to increased back ground currents and noise and may adversely affect the nebulizer If the detector is inadvertently exposed to non volatile salts flush with water to remove the salts stop once the background and noise have returned to normal e Do not use either ammonium carbonate or ammonium bicarbonate in the mobile phase These chemicals are typically heavily contaminated and may cause problems with back ground current and noise and may adversely affect the nebulizer If the Corona detector is accidentally exposed to these compounds flush with water as with non volatile electrolytes above e Use only solvents with minimal residue check label on the solvent bottle Ifa particular solvent causes problem due to particulates try a different lot or a different vendor Be aware that some solvents contain stabilizing agents that may cause problems e Usea pH of lt 7 5 Higher pHs may cause increased noise background currents etc Although not harmful detector performance will not be equivalent to conditions that use lower pH ranges e When pH ing make sure that calibrators used in checking the pH meter do not contaminate the mobile phase e Filter mobile phase through a 0 2 um filter prior to use e Mobile phases that can cause freezing of
76. al troubleshooting guidelines that can be used to quickly isolate the cause of a potential problem Subsequent sections cover Corona detector related issues Section 5 7 HPLC related issues Section 5 8 and sample related issues Section 5 9 in greater detail 1 Turn on the Corona detector and observe the results of the automatic power on tests If the power on tests fail note the error message and check the recommended protocol presented in Table 5 3 2 If the Corona detector does not turn on a Check the AC power cord and make sure that power is being supplied to the instrument Check the fuses on the rear of the instrument Replace if necessary Turn on the detector If the fuse blows again contact an ESA Service Representative If still unable to turn on the Corona detector contact your ESA Service Representative 3 Review Chapter 2 Installation a b Check that all gas connections are properly made and that gas is flowing Verify that the input gas pressure is at 35 0 0 1 psi and if not adjust the Gas Conditioning Module as required Check that all fluid connections are properly made Check that the filter assembly on the rear of the Corona detector is properly attached Verify that other HPLC equipment is connected and operating properly according to the instructions for each instrument Verify that all synchronizing I O connections are properly made and functioning correctly Verify that
77. ally the floor beneath the instrument The drain and vent lines should be shortened if necessary and connected to the vented cap and drain bottle provided with the detector If required by your facility please provide a secondary containment vessel for the drain bottle IS A NOTE Ensure that the two drain tubes are not kinked or blocked in any way WARNING If the detector does not drain properly the waste liquid will backup and flood the detector During operation always observe that liquid is draining into the drain bottle Severe or prolonged flooding may permenatatly damage the detector and void the warranty Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation e Only use appropriate mobile phases o If PEEK components are present in the HPLC system DO NOT use dichloroacetic acid acetone tetrahydrofuran THF dichloromethane chloroform dimethylsulfoxide DMSO Although unlikely to be used with the Corona detector PEEK is not compatible with strong acids such as concentrated sulfuric acid and concentrated nitric acid o Evaporation of some organic solvents can cool the nebulizer to such a degree that water as a trace contaminant or as part of an aqueous gradient freezes in the nebulizer leading to adverse detector performance Only use the Coronas detector where nebulizer freezing is an issue e g aqueous THF gradients o If possible keep the pH of the mobile phase
78. and or is stable for a period of 5 to 10 minutes eS NOTE The time that is required to obtain a stable baseline is dependent i j 4 2 upon the gain range used i e sensitivity required as well as other factors such as the nature of the mobile phase type of column being used etc Autozero the detector if necessary The detector is now ready for analytical measurements Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Using the Detector in a HPLC System 4 2 2 Maintaining the Detector between Analyses The ideal situation to minimize downtime is to maintain the mobile phase flow and gas flow in the Run mode when the detector is not being used If the detector is kept ready for operation in this manner it is important to ensure that the mobile phase reservoir s contain sufficient solvent for the maintenance period and the gas supply is sufficient for this period The pump high pressure cut off should be set to approximately 35 bar 500 psi above that observed with the normal flow rate to protect the system in case of a blockage If possible the detector rear panel Gas Off contact closure should be connected to an appropriate device to note that the flow of gas has stopped WARNING Make certain that the volume of the waste bottle is sufficient to allow for the collection of mobile phase during the maintenance period The waste bottle should NEVER be allowed to fill completely S
79. and spikes in the baseline Random baseline perturbations Figure 5 4 can also be caused by poor grounding Always make sure that the Corona detector is plugged into the same power strip or the same common outlet as the other components of the HPLC system Corona CAD and Corona S CAD Detectors 5 17 Operating and Maintenance Manual Chapter 5 5 8 HPLC Related Issues 5 8 1 Pump Noise This common problem is seen as rhythmicity in the baseline Figure 5 5 The frequency is dependent upon on the piston stroke volume i e the mobile phase flow rate The magnitude of the response is typically dependent upon the severity of the issue with the pump The impact of pump noise to the analyst is dependent upon what is to be accomplished For high sensitivity analytical work even slight pump noise can have a major impact on the limit of detection of the assay Factors that can contribute to pump noise include but are not limited to the age and quality of the pump incorrect pump priming sticking check valves scratched pistons damaged seals bubbles in the pump head and mobile phase leaks Pump noise is relatively simple to diagnose The frequency of the noise is directly proportional to the flow rate If the pump flow is halved make sure that the flow rate to the pump is still maintained above the lower recommended flow of 0 2 mL min then the frequency of the noise will halve as will the amplitude Detector Response SS
80. at contains a reservoir of methanol or isopropanol and a membrane that placed in the mobile phase line to diminish pulsations from the pump Corona CAD and Corona S CAD Detectors G 7 Operating and Maintenance Manual Glossary R Abbreviation for current range in A mA pA nA etc Remote Communication The mode of operation in which the control of the detector is via an external device e g a personal computer Resistance Opposition offered by a component to the flow of current in an electrical circuit Resolution The separation of two peaks in a chromatogram The resolution R is defined by R V2 V4 0 5 We2 W where V and V2 are the retention times for peak 1 and peak 2 W and W are the baseline width of peak 1 and peak 2 Response Factor The amount of a compound that was injected divided by the area under the peak Retention Time The time that has elapsed between the injection of a sample and the time when the detector response is maximized i e the top of the peak for a given compound The retention time is usually reported in minutes and fractions of a minute e g 4 62 min Retention Volume The volume of mobile phase that is required to elute a compound from a column Reverse Phase Column A column in which the stationary phase is less polar than the mobile phase used with it Typically a reverse phase column consists of a column packing material with either an alkyl chain e g Cig or a
81. ates 11 Although both detectors are compatible with the entire pH range detector performance decreases under basic conditions If possible keep the pH of the mobile phase lt 7 5 12 Either detector is compatible with all commonly used mobile phase components e g acetonitrile tetrahydrofuran methanol isopropanol Make sure that other components in the HPLC system e g piston seals injector seal PEEK tubing are also compatible with the mobile phase components However only the Corona S CAD detector can be used for aqueous THF gradients see 13 below Corona CAD and Corona S CAD Detectors v Operating and Maintenance Manual 13 Some solvents will cause the nebulizer to freeze during their evaporation e g aqgueous THF gradients and will adversely affect system performance For solvents that have low specific heat capacities use the Corona CAD detector 14 Both detectors are compatible with most HPLC column formats However the detector is very sensitive to particulates Make sure the column does not suffer from bleeding 15 The detector must receive mobile phase at a flow rate of 0 2 2 0 mL min Lower flow rates may lead to aberrant chromatography and noise If the Corona CAD detector is to be used in a system where flow is split between it and other detectors make sure that the flow rate to the Corona CAD detector is between 0 2 2 0 mL min 16 Avoid open flames and sparks Do not use an op
82. attach the Drain Vent Tubing assembly b Connect the Vent tubing to the quick connect fitting located on the bottom front left portion of the detector This is a quick connect fitting which simply latches when the fitting is pushed in Figure E 9 b Connect the Drain tubing end with S S fitting to the compression fitting located on the bottom front left portion of the detector Ensure that the two ferrules inside the nut are properly oriented Figure E 8 The Drain fitting should be tightened with the supplied 9 16 wrench until it is snug Figure E 9 CAUTION Do no over tighten the drain fitting Equipment damage could result Corona CAD and Corona S CAD Detectors E 7 Operating and Maintenance Manual Appendix E Figure E 8 Correct Orientation of Nuts and Ferrules Figure E 9 Location of Fluidics Connections c Using a straight blade or a sharp pair of scissors cut both the Drain and Vent tubing to a length between detector and waste bottle where there are no dips kinks or loops A vertical drop is optimum It may be necessary to cut the tubing depending upon the installation Leave enough excess in case the tubing ends need to be cut and reinstalled The ends must be cut straight to ensure a proper connection to the barbed fittings E 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Drain Vent Assembly Old Styles necessary to flare the tubing by inserting the
83. be performed on a daily weekly monthly and quarterly basis 5 2 1 Overview While the ESA Corona CAD detector requires little day to day maintenance it is recommended that e Samples should be free of particulate matter Samples should be filtered through a 0 22 um Nylon or PVDF membrane filter Filters should be checked to ensure that extractable materials are not present and they are compatible with all constituents of the sample e The mobile phase should contain only volatile components and be free of particulate matter Mobile phase should be filtered through a 0 22 um Nylon or PVDF membrane Make certain that the filter is compatible with all constituents of the mobile phase e In line PEEK filters are used before the detector PEEK filters are acceptable unless the mobile phase contains a solvent that is incompatible with PEEK e g THF If THF is to be used use the stainless steel filter Part Number 70 4538 and appropriate frits Part Number 70 4359 e Columns should be used in accordance with their manufacturers recommendations Avoid columns that show column bleed eS NOTE The Corona detector is just one component of a chromatographic system The performance of the Corona detector will reflect the performance of the overall HPLC system To maintain optimal analytical performance it is important to perform routine maintenance on each of the various HPLC system components e g pump autosampler etc The user
84. ber 1 Corona Detector Including 70 6225 1 Limited Warranty Registration Card 10 0181 p We BNC connector to Black and Red 70 1776 1 Cable I O 2 Conductor 70 4850 1 Small Flat Blade Screwdriver 3 32 x 2 50 0257 1 Corona Detector Operating Manual 70 6258 1 In Line Filter HPLC 70 4538 1 Filter SS Replacement Frits 70 4539 2 Tubing HPLC 10cm 70 6398 1 Filter Gas Holder 70 4379 1 Drain Vent Tubing Assembly 70 6266A 1 Nut 1 16 PEEK Sealtight 10 32 70 5546 8 Ft Tubing Gas Inlet 70 6260 8 Ft Tubing Gas Outlet 70 6261 1 Fitting 1 4 Tube x 1 4 NPT 70 4377 1 ESA Multitool 40 0288 1 Caffeine Standards Kit 70 6565 1 Wrench 1 2 X 9 16 Open End 50 0868 1 Guard Column 20 x 4mm BetaBasic 18 70 7098 Used in detector verification see Chapter 4 E 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Drain Vent Assembly Old Styles E 2 1 Installing the Drain Vent Assembly Drain and Vent Installation The Drain Vent tubing assembly Part Number 70 6266A consists of two pieces of Teflon lined Tygon that must be carefully assembled to promote proper drainage The assembly includes the Drain Tubing Part Number 70 6299 1 4 ID x 3 8 OD x 4 long and the Vent Tubing Part Number 70 6259 1 8 ID x 4 OD x 4 long shown in Figure E 7 Tygon Tubing Tygon Tubing N Teflon Lining gt i _ a 4 Teflon Lining VENT TUBE DRAIN TUBE Figure E 7 Vent and Drain Tubings To
85. c ah seh ce ee eae dee eA dacth athens sah be Mcetndee cnet dened oh gaveae 3 18 3 6 1 Role of the System SCreens cccccceccceeeeeceeeeeeeeeeeeeeeeeeeeeeeeeeaaeeeeeeeaeeeeeee 3 18 3 6 2 R9232 Setup ScregN a aics iencvevecsediaened saad cared EA E ERa Sa eE E Erite 3 18 3 6 3 Deleting Methods Screen cscetsic ceacceevssttecss Weolited gece seedeancesesiadaes Adee 3 19 3 6 4 Event Marks SCreen ski 35 Sosthcea cere tvich iensedeh aloe cetgeetce Noha tease bade seoteenes 3 20 3 6 5 Date amp Time Setup Screen scciticccctstccatotaaetnntind Qin ac ecdighteanntiauee nweless 3 21 3 6 6 ROMOLE Screen iier era e a E ARE EE E E 3 21 3 6 7 INPUTS SCROCM eeen pi areena era raua ae AETA Aa ARE EEEE En a AENEA 3 22 3 6 8 Contacts SCLCCM one 5 ce ances tant se eat cenh ny Penis eaceyies ack teen cad macmienaeaecestehs 3 23 3 6 9 Thermal Organizer Setup Screen ccccceceeeeeeeeeeeeeeeeeeeeeeaeeeeeeneaeeeeeeas 3 24 30 10 Self TEST SOKO CM a ar scale dared Hae tees EE a 3 25 3 6 11 Nebulizer Heater Setup Screen 00 ec ceeceeceeeeeeeeeeeeeeeaceeeeeeaaeeeeeeneaeeeeenee 3 25 How to Create a Timeline Method c ccceeeccceecescceeeeeeccaeeeeeeeneeeeeeeeeaeeeeeseaeeeeeee 3 27 CHAPTER 4 USING THE DETECTOR IN A HPLC SYSTEM 4 1 INMFOGUCTION A EEEE EAEE EEEE EE EEEE OTT E E E A E EE SE A SA 4 1 4 2 Operating the Corona Detector c cccccccceccesssssnneeeeeeeeesseeceneeeeeeeeeeseessesneeeeeeeeess 4 1 4 2 1 Turning On the Dl CO ce
86. certain that the room is equipped with an exhaust system to ventilate vapors safely from the detector Such ventilation MUST be at atmospheric pressure A vacuum or restriction may result in pressure changes within the detector resulting in baseline instability and or other problems Corona CAD and Corona S CAD Detectors 2 1 Operating and Maintenance Manual Chapter 2 e Nitrogen gas must be available A supply of nitrogen gas is required for the operation of the detector The gas must be supplied at 35 0 0 1 psi 2 4 bar and must be 99 99 pure gt gt D WARNING The nitrogen gas source must not be oil pumped as oil in the gas stream can clog the detectors in line filters and may damage the detector itself WARNING The use of open flames in the laboratory must be prohibited Many of the solvents that are used in an HPLC system are flammable Make certain that the room is equipped with an exhaust system to ventilate vapors safely WARNING Do not install the detector where corrosive gas or excessive dust is present These can adversely affect the long term operation of the unit Install on a stable flat surface The detector should be installed on a laboratory bench that is capable of supporting the entire HPLC system e g a complete HPLC system can weigh over 200lbs The bench should have a minimum depth of 24 60cm e The special drain bottle provided with the instrument must be placed on a surface usu
87. cessed the cursor will be found on the Range field CORONA EDIT Range 100pA Filter None Offset 0 CANCEL PREVIOUS NEXT Figure 3 8 The Edit Parameters Screen The Range field is used to select the full scale gain range for the detector It is used to ensure that the chromatographic peak is on scale and is sufficiently large The default value is 100 pA and the range can be set from 1 pA to 500 pA via a 1 2 5 sequence using the A or W key After the desired value is selected press ENTER to move the cursor to the Filter field The Filter field is used to select the desired electronic filter that is used to reduce the noise in the chromatogram The default is None and the filter can be set to None Low Medium and High using A or WV keys A detailed discussion of the use of the filter is presented in Section 4 6 3 After the desired value is selected press ENTER to move the cursor to the Offset field The Offset field is used to change the position of the entire chromatogram with respect to X axis e g so that two or more chromatograms can be presented on a set of common axes The offset can be set to any value within 50 of full scale via the numeric keypad If anon valid value is entered an error message will be presented with the appropriate range Corona CAD and Corona S CAD Detectors 3 7 Operating and Maintenance Manual Chapter 3 After desired settings are selected press NEX
88. ch field and pressing ENTER after the entry is made Press the SAVE key to start the clock at the indicated time and date 3 6 6 Remote Screen The Remote screen Figure 3 38 can be accessed through the SYSTEM SETUP menu and then by pressing the A or WV keys SYSTEM SETUP MENU Select Remote Set Remote mode CANCEL NEXT Figure 3 38 The System Setup Menu Screen Select the NEXT key to present the REMOTE screen Figure 3 39 Corona CAD and Corona S CAD Detectors 3 21 Operating and Maintenance Manual Chapter 3 REMOTE SCREEN MENU will return to Main Menu MENU RUN Figure 3 39 The Remote Screen The Remote LED on the front panel will illuminate Press RUN to enter the Corona Run screen The gas valve will open automatically and the corresponding LED will illuminate 3 6 7 Inputs Screen The nputs screen Figure 3 40 can be accessed through the SYSTEM SETUP menu and then by pressing the A or WV keys SYSTEM SETUP MENU Select Inputs Set Input Active Inactive CANCEL NEXT Figure 3 40 The System Setup Menu Screen Select the NEXT key to present the INPUTS screen Figure 3 41 INPUTS ACTIVE INACTIVE Autozero ACTIVE Gas off ACTIVE CANCEL MENU SAVE Figure 3 41 The Inputs Active Inactive Screen This screen refers to two of the three inputs located on the Logic Module at the rear
89. characteristics Mobile Phase Flow Rate 1 0mL min Column Temperature ambient Injection Volume 10ul Run Time 2 0 minutes 2 5 3 Protocol CAUTION Make certain that the waste bottle is emptied on a routine AN basis It should not be allowed to get full Failure to do this may result in flooding of the detector Severe or prolonged flooding may damage the detector and void the warranty a Turn on the detector using the On Off switch located on the back of the detector Figure 2 2 and allow it to perform a Self Test When the instrument is first powered up the display will present a number of diagnostic screens e g Figure 2 12 The program will sequentially check individual sections of the detector During this time the second line of the display will indicate the sequential progress through the testing procedures CORONA SELF TEST RUNNING Current Test xxxx Figure 2 12 The Corona Self Test Running Screen NOTE If a fault message is presented turn the power off wait a Ce few seconds and power the unit up again If the fault message appears once again check the error code section Chapter 5 and take appropriate action to rectify the problem b After the self test is completed successfully the Version Copyright screen Figure 2 13 will be presented for several seconds Record the version information on the worksheet CORONA Version X xxx Copyright 2004 NEXT
90. controlled using Timeline Being able to change them manually within the system menu permits connections to other devices to be tested 3 6 9 Thermal Organizer Setup Screen The Thermal Organizer Setup screen Figure 3 44 can be accessed through the SYSTEM SETUP menu and by pressing the A or V keys SYSTEM SETUP MENU Select Thermal Org Setup Thermal Organizer CANCEL NEXT Figure 3 44 The System Setup Screen Press the NEXT key to access the Thermal Organizer Setup screen Figure 3 45 T OFF THERMAL ORGANIZER SETUP Status Normal Power OFF Setpoint xx C Temperature xx xC CANCEL MENU SAVE Figure 3 45 The Thermal Organizer Setup Screen This screen is used to set the temperature of a Thermal Organizer Module if present When this screen is accessed the cursor appears in the field to the right of Power After choosing ON and pressing ENTER the cursor will appear to the right of Setpoint Use the A Vor the numeric keypad to set the desired temperature 25 60 C After you have entered the desired value press ENTER then press SAVE The set temperature is stored in non volatile memory and is automatically recalled when the detector is powered up The Status field indicates the present state of the Thermal Organizer e g Normal Not Installed Board Error Under Range Over Range and is used to indicate error messages Table 3 6 See Chapter 5 for further information 3
91. ct ESA Service voltage supply 10 Pressure Failure of transducer 1 Rerun Self Test If Fails contact ESA Service Transducer 1 20 Pressure Failure of transducer 2 Rerun Self Test If Fails contact ESA Service Transducer 2 80 lon Trap Voltage lon Trap Supply Possible flooding of the detector Turn off On HPLC flow Check that bottle is not overfilled Check that drain assembly is free from loops kinks or blockages Check vent and gas lines for liquid Flow gas only through the detector for 1hr and then rerun Self Test If error 80 still present repeat till test passes this may take gt 24hr depending upon the severity of flooding If continues to fail contact ESA Service 100 lon Trap Off lon Trap supply Rerun Self Test If Fails contact ESA Service problem 200 Electrometer Electrometer heater or Wait for instrument to reach room Temp instrument is too cold temperature Rerun Self Test If Fails contact ESA Service 400 Clock Not Set Real time clock Logic Rerun Self Test If Fails contact ESA Service Board 800 Clock Not Real time clock Logic Rerun Self Test If Fails contact ESA Service Running Board 1000 Battery Low Real time clock battery Rerun Self Test If Fails contact ESA Service Logic Board Multiple errors can occur at the same time In this case the error code presented on the display panel of the Corona CAD detector will be a sum of all failed tests oe NOTE Failed tests can be reviewed by selecting the review sof
92. ctor to the splitter output 2 9 Setting up the Thermal Organizer If a thermal organizer is to be used connect it using the Y cable Part Number 70 5595 The double end of the cable is connected to the Thermal Organizer Control socket on the Logic Module and the Thermal Organizer Power connector on the Power Supply Module of the Corona detector The single end is connected to the Thermal Organizer See Appendix C for greater detail a To Enable the Thermal Organizer press the System soft key from the main b Corona menu and then press the A or Y key to select the THERMAL ORGANIZER Select NEXT to access the SETUP Screen Figure 2 15 T Off THERMAL ORGANIZER SETUP Status Normal Power OFF Setpoint xx C Temperature xx xC CANCEL MENU SAVE Figure 2 15 The Thermal Organizer Setup Screen Change the Power Control field to On with the A or W key Press ENTER this will go to the setpoint to enter the desired temperature Use the numerical keypad to enter a temperature 10 C above ambient press ENTER and then press SAVE The display will then revert back to the MAINCORONA MENU and indicate the temperature status The system will store the new temperature and will provide power to the heater to raise the temperature While the temperature is increasing the present temperature will be indicated with an up arrow e g 33TC Indicate on the worksheet that the temperature rose to the des
93. d Corona S CAD Detectors 5 29 Operating and Maintenance Manual Chapter 5 5 12 4 Loss of Response This section describes the abrupt loss of a peak or peaks from the chromatogram when using a set of analytical conditions which is known to provide a useful chromatogram Possible Cause Comments Recommended Solution Accidental change of a Check settings and verify that the parameter on the recorder Gas On Off indicator is lit On chromatograph detector Compounds of interest not Some compounds will Check stability as a function of time sufficiently stable decompose as a and prepare fresh standards If function of time necessary change conditions Use a cooled autosampler Change in pH or mobile Mobile phases should Check mobile phase and prepare phase composition be freshly prepared fresh phases The injector may be Make sure that the Clean the injector partially or fully clogged sample and mobile phase are clean Detector unit malfunction Turn off module and re initialize If error message reappears contact service Perform detector test Chapter 2 If problem remains contact service Column temperature not Temperature variation Maintain column at constant held constant Retention potentially due to temperature times changed problem in organizer or If the Thermal Organizer is included aie orange make sure that it is operating properly see Retention times
94. d at a positive voltage and gas is continuously swept through this chamber This leads to the production of a corona plasma whereby positive ions and electrons are generated from the reagent gas molecules The electrons accelerate toward the Pt wire while the positive reagent ions are swept through an orifice to mix with an opposing stream of analyte particles the aerosol The ion trap removes the high mobility positive ions therefore the excess reagent ions and low mobility positively charged analyte particles are detected The solvent is evaporated and thus does not form particles rather nonvolatile analyte forms particles that are charged and detected Negatively charged particles are essentially not formed and the electron flow to the Pt wire provides the balance of charge Question Is it possible to use a compound s physical characteristics e g molecular weight MW melting point MP or boiling point BP to predict its volatility and therefore detection by the Corona CAD detector Answer The simple answer is no MW MP and BP cannot be used to predict a compound s volatility with any great accuracy For example compounds that have similar MWs may have very different volatilities due to polarity and hydrogen bonding For example glycerol MW 92 BP 182 C was easily detected to lt 1ng o c but propylglycerol propanediol MW 76 BP 188 C was not detected by the Corona CAD detector Perhaps a better indicator of volatility
95. d screen Figure 3 21 is used to store the method in the detector s memory When the screen is opened the cursor will be located on the method number of the next available unused method even if editing a method that was previously created Use the key to return to the method to be overwritten or store the newly edited method as a new method using the unused method number that is presented EDIT METHOD SAVE Method 3 Name Mode ae er eee CANCEL PREVIOUS SAVE Figure 3 21 The Save Method Screen Corona CAD and Corona S CAD Detectors 3 13 Operating and Maintenance Manual Chapter 3 After the method number has been selected press ENTER The cursor will move to the Name field and additional information about the method i e name of the method your initials etc can be entered using the numerical keypad Alphabetic characters can be selected by pressing 9 and then using the A key e g pressing 9 then the arrow key two times will display the letter B Pressing the 9 key again will allow for the next alphanumeric character etc Method names can be up to 16 characters long To name the method CURRENT Press the 9 key and then the A key until C is showing Press the 9 key and then the W key until U is showing Press the 9 key and then the W key until R is showing Press the 9 key and then the V key until R is showing Press the 9 key and then the W key until E is show
96. d through in the same manner as the data screens Once in the Corona CAD detector run mode and all parameters have been verified the Corona CAD detector is now ready to collect data Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 3 17 Chapter 3 3 6 The System Screens 3 6 1 Role of the System Screens The SYSTEM MENU screens are used to edit a variety of parameters that relate to the overall use of the detector rather than being related to a single method and include the following screens RS232 Setup Screen Section 3 6 2 Deleting Methods Screen Section 3 6 3 Event Marks Screen Section 3 6 4 Date amp Time Setup Screen Section 3 6 5 Remote Screen Section 3 6 6 Inputs Screen Section 3 6 7 Contacts Screen Section 3 6 8 Thermal Organizer Setup Screen Section 3 6 9 Self Test Screen Section 3 6 10 Nebulizer Heater Setup Screen Section 3 6 11 The SYSTEM SETUP menu Figure 3 30 is accessed by selecting SYSTEM on the MAIN CORONA MENU Figure 3 3 When the menu is selected the RS232 screen is presented Other screens can be accessed via the A or V keys SYSTEM SETUP MENU Select RS232 setup Setup serial output protocol CANCEL NEXT Figure 3 30 The System Setup Menu Screen To edit the various parameters press NEXT 3 6 2 RS232 Setup Screen The RS232 Setup screen Figure 3 31 is used to select the parameters to be used in communicatio
97. d to indicate if a user selected security code is required in order to use the keypad It allows authorized operators to prohibit unauthorized users from changing parameters CORONA EDIT Run time security Off Run time security code _ 1 CANCEL MENU NEXT Figure 3 7 The Edit Security Screen If the security code is to be activated to lock the keypad of the detector use the A or V keys then press the ENTER key to change the Run time security legend to On This will activate the Run time security code field so that the desired security code four characters can then be entered Press ENTER then NEXT to access the Parameters screen Figure 3 8 3 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware When security is used the operator must enter the correct code to unlock the keypad before performing any actions and then enter the code once more in order to re lock the keypad If the security feature is not to be used or present security code is not to be edited press NEXT as soon as this screen is presented The CANCEL button erases any entries that were made on this screen the MENU button returns to the EDIT METHOD SELECT screen and the NEXT button accesses the Parameters screen Figure 3 8 3 4 4 Edit the Parameters Screen The Parameters screen Figure 3 8 is used to set a variety of detector parameters When the screen is ac
98. defects in material and workmanship in normal use from the date of delivery to Buyer as the original purchaser for the following periods instruments and equipment one year expendable items such as cells sensors and electrodes 90 days This warranty does not cover and no warranty is provided for consumables and parts that by their nature are normally required to be replaced periodically consistent with normal maintenance including without limitation fuses tubing pump piston seals injector rotors check valves and filters If any product covered by this warranty is returned to the original shipping point transportation charges prepaid within the applicable warranty period set forth above and upon examination Seller determines to its satisfaction that such product was defective in material or workmanship Seller will at its option repair or replace the product or the defective part thereof or refund the original purchase price of the product The foregoing notwithstanding Seller will not be responsible for damage to any product resulting from misuse negligence or accident or resulting from repairs alterations or installation made by any person or firm not duly authorized by Seller in writing or for any damage to any cell assembly resulting from the flow being impeded If any cell assembly is used with control modules or potentiostats other than those manufactured by Seller this warranty shall be void B If at any time during the period en
99. ding ninety 90 days after delivery of any product to Buyer Buyer reports and documents any error in any software provided with such product and developed by Seller or any failure of any such software substantially to conform to Seller s software description that limits or prevents use of the software by Buyer Seller at its option will use reasonable efforts to correct any such error or failure will replace such software or will terminate Buyer s license to use the software and refund the price of the related product In connection with any such termination and refund Buyer will return the related product to Seller forthwith upon request This warranty shall apply only to those portions of the software that were developed by Seller and that incorporate all program corrections and modifications if any delivered to Buyer It shall not apply to any error or failure due to machine error or to the misuse by or negligence of any person or entity other than Seller or to any software which is modified by any person or entity other than Seller C If Buyer shall fail to pay when due any portion of the purchase price of any product or any other payment required from Buyer to Seller whether under the contract or otherwise all warranties and remedies granted in this Section and all of Seller s warranties and obligations to service repair replace correct or otherwise remedy defects errors or failures under any other contract between Buyer and Seller may
100. dix C C 4 Installing the Corona Thermal Organizer Module IES NOTE Refer to Section C 3 to install the Corona Organizer Module C 4 1 General Information An exploded view of the entire Corona Thermal Organizer Module is shown in Figure C 8 Please refer to this figure as the Organizer Module is assembled PLASTIC THUMB SCREW 70 5571 J 7 SSENURLED SCREW s 70 5564 s RR KET FOR HOLDING DOWN A Tab 4 SJ BRACKET FOR MANUAL INJECTOR OVER FLOW LINES 70 5678 COLUMN HOLDER ASSEMBLY 70 5367 SHOWN KNURLED SCREW ff 70 5564 KNURLED THUMB NUTS FOR J PANEL Aj 70 5532 PANEL WITH HOLE FOR INJECTOR ORGANIZER sane ST 0 5605 eta CHASSIS TEA 70 5 Bs HEATER MODULE DRAIN TUBE PANEL WITHOUT HOLE FOR AUTOMATED INJECTOR 70 5418 Z ORGANIZER TRAY ASSEMBLY NATCH TAB Figure C 8 Exploded View of Corona Thermal Organizer Module C 10 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer C 4 2 Removing the Organizer Chassis Plate All components are mounted on the Organizer Chassis Plate Part Number 70 5231 Figure C 9 which is attached to the Organizer Tray It is necessary to remove the chassis plate from the organizer tray to attach the various components Figure C 9 Organizer Chassis Plate To remove the Organizer Chassis Plate a Remove the Organizer Cover by lifting it off b Unscrew the three kn
101. e back of the instrument indicates a functional earth terminal The bold WARNING statement used throughout the manual presents dangers that might result in personal injury The bold CAUTION statement used throughout the manual presents hazards on conditions that could cause damage to the instrument or the reporting of erroneous results The bold NOTE statement used throughout the manual highlights important information about the detector and its use Failure to follow these statements may invalidate the warranty Corona CAD and Corona S CAD Detectors iii Operating and Maintenance Manual WARNINGS AND SAFETY PRECAUTIONS The ESA Corona CAD and Corona CAD detectors are general purpose detectors for use with high performance liquid chromatography HPLC They can be used to measure the level of a large number of compounds in a broad variety of samples The following precautions should be followed to minimize the possibility of personal injury and or damage to property while using either detector Make certain that you are familiar with the contents of this manual before using the detector Do not use this instrument before fully understanding the contents of this manual Provide this documentation to the next user in the event that the unit is borrowed To ensure safe operation read the following Safety Precautions before using the instrument This instrument is a detector for use with a high performance liquid chromato
102. e in place for all nitrogen sources Failure to provide adequate nitrogen filtering can potentially cause damage to the detector A qualified trap assembly is included from ESA Gas Conditioning Module Part Number 70 8285 2 3 Unpacking the Detector Carefully unpack your shipment and inspect the contents to verify receipt of all components The detector comes with a Customer Inventory Checklist which lists the parts shipped with the unit This document is found in the Manual Appendix E If an ESA model 584 pump autosampler nitrogen generator Organizer module or Thermal Organizer module are included they will be shipped in separate cartons Ie NOTE See Appendix B for a list of recommended supplies and spare parts Carefully inspect the shipping carton s and all components If any parts are missing call ESA s Customer Service Department and indicate the missing items via the part numbers Corona CAD and Corona S CAD Detectors 2 5 Operating and Maintenance Manual Chapter 2 WARNING If there is any evidence that the Corona detector unit has AN been damaged in shipping do not install or power on the unit Immediately contact ESA 800 275 0102 978 250 7000 or its representative for advice IESE NOTE The shipping carton should be retained as it can be used if it becomes necessary to transport the detector 2 4 Installing the Detector 2 4 1 Basic Installation This section describes how the different connecti
103. e pump and Replace in line filter elements from the mobile phase or check pressures at each Section 5 3 injected samples component between pump and CAD detector Ensure that the mobile phase and or samples are filtered through a 0 22 um Nylon or PVDF membrane filter Use a mobile phase with a substantial fraction of an organic solvent to prevent bacterial growth Use freshly prepared mobile phase Bacterial growth in the mobile phase may lead to clogging of the filter Solubility of analyte Start at the pump and Replace in line filter elements standard or sample in check pressures at each Section 5 3 mobile phase exceeded component between pump and CAD detector oe een using appropriate solvent Make sure standard and or sample are soluble in mobile phase Use diluted sample standard or use a compatible mobile phase Clogged injector or column Isolate suspect component Refer to manufacturer s cleaning directions or replace rotor seal and or stator face on injector Ensure that the mobile phase and sample are filtered before use Plugged tubing Isolate plugged tubing and replace Column temperature has Temperature variation Maintain column at constant fallen potentially due to problem temperature in organizer or f temperature change in If the Thermal Organizer is laboratory included in the system make sure that it is operating properly see Chapter 5 Corona CAD an
104. e substance that is dissolved in a solvent Solvent A liquid that is used as the mobile phase in HPLC or the liquid that dissolves the solute Sparge The process of bubbling He or N through a filtered solvent Sparging is used to displace dissolved gases that have a deleterious effect on the separation and or detection of analytes of interest Sparging serves to displace COs which could change the pH of a buffered solvent and Os which could take part in electrochemical processes Standard A solution containing a known amount of the compound of interest The detector response is measured for a series of standards to obtain a standard curve Stationary Phase The column packing material where adsorbtion and desorbtion of the components of the sample takes place The mechanism of the interaction could be via ion exchange adsorbtion or size exclusion System Menus A selection of instrument parameters for the Corona Detector that are infrequently changed Tailing In theory a chromatographic peak is Gaussian If the longer retention time profile of the peak is not Gaussian the phenomenon is termed tailing and the peak is termed asymmetric Temperature Coefficient The rate of change of a phenomenon that is related to a change in the temperature Theoretical Plate See Plate Time Constant The period of time it takes for the signal to drop to 2 of the signal after the stimulus is removed Timeline A mode in the Corona Detector
105. e to tray Figure C 2 Organizer Chassis Plate To remove the Organizer Chassis Plate a Remove the Organizer Cover by lifting it off b Unscrew the three knurled nuts that attach the organizer chassis plate to the tray One nut is in the front center and the other two are in the corners at the rear of the organizer c Lift the organizer tray from the chassis plate C 3 3 Installing the Pulse Damper To install the Pulse Damper a The pulse damper is mounted on the rear of the front left side of the chassis plate Figure C 3 using 2 of the 4 screws that are supplied with the pulse damper Mount damper here Figure C 3 Mounting the Pulse Damper es NOTE Do not attach the rubber feet to the bottom of the pulse damper when mounting it to the chassis plate Corona CAD and Corona S CAD Detectors C 5 Operating and Maintenance Manual Appendix C C 3 4 Installing a Rheodyne Manual Sample Injector Valve Sa NOTE It may be easier to connect the appropriate tubing to the manual sample injector valve before installing it into the organizer To install the Manual Sample Injector a Place the Organizer Injector Panel with the hole Part Number 70 5229 onto the organizer chassis plate Figure C 4 Figure C 4 Locating the Organizer Injector Panel b Affix the panel to the organizer chassis plate with the two thumbnuts provided c Remove the handle from the injector valve using a hex wrench d Attach the i
106. ection 1 4 ID connection Corona Gas ee Conditioning I J Module dR OOOD I aooo n 39999 S Nitrogen G Soe Source a UTE il b Connect outlet of ESA Gas Conditioning Module to the gas inlet Figure 2 3 using a quick connection 1 2 ID connection Use the PTFE tubing supplied with the system white the smaller diameter single tubing If this tubing needs to be cut use the included tube cutter P N 70 7112 and cut the tubing as neatly as possible Avoid using a pair of wire cutters or scissors as this can lead to tube damage and a leaky fitting bid CE e en S fd Figure 2 3 Nebulization Gas System trap be in place for all nitrogen sources Failure to provide adequate nitrogen filtering can potentially cause damage to the detector A qualified trap assembly is included from ESA Gas Conditioning Module Part Number 70 8285 i WARNING Itis required that inline moisture and a 0 01um particle 2 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation 2 4 1 4 Exhaust Connect the gas outlet located on the back of the detector Figure 2 3 to a fume hood or other venting device with the vent tube supplied Connect one end of the larger diameter single tubing to the Exhaust Outlet quick connect fitting on the back panel of the detector and connect the other end to a fume hood or vent If required this tubing shou
107. ector to Black and Red Wire 70 1776 1 Cable I O 2 Conductor 70 4850 1 Small Flat Blade Screwdriver 3 32 x 2 50 0257 1 Corona Detector Operating Manual 70 6258 1 In Line Filter HPLC 70 4538 1 Filter SS Replacement Frits 70 4539 2 Tubing HPLC 10cm 70 6398 1 Filter Gas Holder 70 4379 1 Drain Vent Tubing Assembly 70 7115 1 Nut 1 16 PEEK Sealtight 10 32 70 5546 1 Nut Ferrule Set 1 4 SS 70 7120 8 Ft Tubing Gas Inlet 70 6260 8 Ft Tubing Gas Outlet 70 6261 1 Fitting 1 4 Tube x 1 4 NPT 70 4377 1 ESA Multitool 40 0288 1 Caffeine Standards Kit 70 6565 2 Wrench 1 2 X 9 16 Open End 50 0868 1 Guard Column 20 x 4mm BetaBasic 18 70 7098 1 Cable USB 70 5713 Used in detector verification see Chapter 4 NOTE See Appendix B for a list of recommended supplies and spare parts Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Appendix E Carefully inspect the shipping carton s and all components If there is any damage to the carton s or to any components contact both the shipping agent and ESA or its representative immediately If any parts are missing call ESA s Customer Service Department and indicate the missing items via the part numbers E 1 1 Installing the Drain Vent Assembly Drain and Vent Installation The Drain Vent tubing assembly Part Number 70 7115 consists of a piece of Teflon lined Tygon tubing and a piece of solid Teflon tubing that must be carefully assembled
108. ed as a single unit Part Number 70 6224 Detach the existing filters by pressing on the ring on the fitting and pulling on the tubing The new filter simply snaps in the clamp and the tubing snaps into the gas in and out connectors Figure 5 1 Gas Filter Unit Figure 5 1 Location of Gas Filter Unit on Rear of the Detector 5 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting 5 4 Cleaning the Detector 5 4 1 Cleaning the Corona Detector Unit The outside of the detector can be cleaned with a soft towel moistened with a mild detergent This should be suitable for removing dust and fingerprints Avoid getting any liquid inside the detector as this could damage the unit Avoid using abrasive cleaners especially on the keypad and the display as this could cause scratches If it becomes necessary to disinfect the detector a mild bleach solution can be used However do not allow the bleach solution to remain on the detector for more than a few minutes The bleach solution should be wiped off with a damp towel After cleaning the detector can be dried using a soft towel 5 4 2 Cleaning the Corona Detector Organizer and Thermal Organizer Follow the general cleaning procedures as described above for the detector unit However it is more likely that the organizer module will become dirty due to spills and leaks of the mobile phase over time It may also be necessary to
109. ed to indicate that the method should wait for an input from an external device and to indicate what the appropriate input should be TIMELINE Time 0 00 min Event Hold Input Gas Off Hold for ON CANCEL ADD STEP PREVIOUS NEXT Figure 3 18 The Hold Screen The Inputs include Gas Off Autozero and Start The Hold for conditions include off to on on to off off or on Options can be selected using the A or Y keys 3 12 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware 3 4 5 10 Loop Screen The Loop screen Figure 3 19 is used to indicate that a looping operation is to occur at the indicated time A detailed discussion of looping is presented in Section 3 7 TIMELINE Time 0 00 min Event Loop Loop to X xx for 5 times CANCEL ADD STEP PREVIOUS NEXT Figure 3 19 The Loop Screen The time to reset the loop and the number of times that the loop should be performed are chosen using the A or W keys 3 4 5 11 Reset Screen The Reset screen Figure 3 20 is used to indicate that the system should be reset to the original values the values at 0 00 min at the indicated time TIMELINE Time 0 00 min Event Reset Resets method to initial values CANCEL ADD STEP PREVIOUS NEXT Figure 3 20 The Reset Screen 3 4 6 Save Method Screen The Save Metho
110. eeneaeeeeenae 5 8 5 4 Cleaning the Detector asrine eee ened cadet AA A ae euep ae TAA EIRE ENE AERES 5 9 5 4 1 Cleaning the Corona Detector Unit sesssseesesseeenernrresserrrnernnnnesrnrrssrrrn ne 5 9 5 4 2 Cleaning the Corona Detector Organizer and Thermal Organizer 5 9 5 5 Establishing a System Logne ssenime a a a dE ENE 5 10 5 6 General Troubleshooting Approach sssssssssesssnerrssernnrrrnnntsstrrnnerrrnnnrnnnrsserer nnet 5 10 5 7 Detector Related ISSUCS si 22 these coeeepenthceasinsccasescsageesedertla necdeeeteeensaueednaeg thane daabe 5 12 5 7 1 Internal Electronic Diagnostic Programs eeeeeesseeeeeeeeeeeeeeenetneeeeeeeeees 5 12 BT MA a AN I EEEE E oe teeta ea dec ne deviate ee eet we cea wae alae tases ava Lae 5 14 DSTO D I e ONE T Ss Hess coss tied oe aed te tee set ob A eect trea end aredes leaded 5 15 574 Gain Ranges e oedaee sted ea AE Eaa a aa eedit 5 16 5 7 5 Grounding ISSUES nennen gensar a r a EEE Ea A 5 17 5 8 HPLG Related ISSUS mrin a a a a A 5 18 5 8 1 Pump NoiSe naianei raei oere enaa EAA AATAKE ae tae 5 18 5 8 2 Mobile Phase FIOW sssssssssssssesseesesesesssserrrrrtrrrnrrnnnnnsenrnnnnnnnssuntnnnnnnneeeeeee 5 19 Geos OMIT EAE E hee ot wen EA E A A A EET 5 20 D0 Os Mobile PNA Se masea a eaa E te Seles hae e aaa e aSa EER 5 21 5 9 Standard and Sample Related Issues ccceceeeeeeeeeeeeeeeeeeeeeeeeeeeeeseeeeeeeeeeaeeees 5 22 5 10 Thermal Organizer Issues 13 scebvess t
111. eer re a a a ea aT ee ee mre ee eer 2 29 ZV 122 SPYOCCOUMC sin dues etestei cts Valaahelegesceth nlp ailag Bes ei atonedZeveanea uaaeeacee tach TN etk Erias e bi 2 29 2 11 2 1 Mobile Phase Preparation ccccceeceeeeeseeneceeeeseeeeeseeeneneees 2 29 2 11 3 Degassing Mobile PHASE sa cccsseas 5 Suen Yass tna tagna casas aac sereseeneecnneetarnananeneee 2 30 raa AA WNDU o EE ceded tece aonde tate at cn ce patatagec te tees tags sear matte nau T 2 32 2 11 5 Mobile Phase Qualification cece ceceeeecceeeeeeeeeeeeeeeeeeeeeeeeeeaaeeeeeeeaeeeeeee 2 32 Column Conditioning Procedure ices dtecccctcrccetarcticts ad viecd ets teteacdir toes nated 2 33 212A UIE 3 seats aes a eaten e ae tek Denard a E tung Masada an 2 33 271222 PIOCCOUIG n sete ate Nica i onus a aa E A E aa 2 33 CHAPTER 3 THE CORONA CAD FIRMWARE 3 1 3 2 3 3 3 4 3 5 xiv mMroductioM eoe a A r a EAT 3 1 Initialization of the Detector oo cece cceccceeeeeceeeeeeseeeeeecueeeeceueeeeeeeeueeeeueueeceueegeueaees 3 2 The TOY AG cb cick ogee Stsnc seine a nestor rA tras sates ie baad Kae nea eras EAERI 3 4 Creating Editing a Corona CAD Detector Method 00 eee eeeeeesseeeeereeeesneeeeeeeeee 3 5 BoA Ah SOV ORVICW seesb ices coie SAM ta rctdokand a a a a 3 5 3 4 2 Edit the Method Select Scree n i cccccccececcceeeceeseeeececeeeueeessaueueeasaneeessaneaes 3 5 3 4 3 Edit the Security SCrEGN cece ee ecceeeeeeeeeeeeeeeeeeeeeeeeeaaaeeeeeeaaaeeeeneaeeennees 3 6 3 4 4
112. en flame and do not use any equipment that can cause sparks in the laboratory 17 Static charges may build up due to use of organic solvents These charges can lead to fire or explosion Take appropriate precautions to eliminate the buildup of static charges e g ground the waste bottle 18 If a leak occurs turn off power to the module and remedy the situation immediately 19 All mobile phase and waste bottles should be placed into a container to provide secondary containment 20 All components of the HPLC system should be plugged into a grounded power line Make certain that all parts of the system are properly connected to a common ground 21 The cover should remain on the detector at all times 22 Replace fuses only with those of proper type and capacity 23 If any water gets onto the instrument wipe it away immediately to prevent rust Do not use organic solvents as these may cause discoloration 24 If the instrument cover or front panel becomes dirty wipe it clean with a soft dry cloth or tissue paper For persistent stains use a soft cloth dipped in dilute neutral detergent Remove detergent by using a soft cloth dipped in water Finally use a dry cloth to remove all moisture vi Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 25 Install the unit in a location where ambient temperature variations are minimized Avoid placing the unit in direct sunlight near a heating or an air
113. enon observed when the front of a peak is less steep than the tail of a peak an asymmetric peak Gain The ratio of the increase in the size of the output signal over the input signal of a detector Ghosting The phenomenon observed when a late eluting peak in one chromatogram is carried over into a second chromatogram Gradient Refers to a programmed change in the composition of the mobile phase during the separation H Height equivalent to a theoretical plate see theoretical plate HETP Height equivalent to a theoretical plate see below G 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Glossary Height Equivalent to a Theoretical Plate H A measure of the efficiency of a chromatographic column H L N where L is the length of the column N is the number of theoretical plates i Abbreviation for electrical current in A mA UA nA etc Injector A device that is used to introduce a specified amount of sample into a LC system Interferent A species that will produce an incorrectly higher signal for the compound of interest due to the fact that it elutes at the same time as the compound of interest and has a detector response that is covered by the conditions used for the compound of interest Integrator A device that can calculate the area under and or the height of a chromatographic peak An integrator is used for quantitative analysis I O Connections A terminal block
114. ent elution on response 4 11 Emptying the Waste Bottle 2 21 End event screen 3 10 Ensure functioning properly 4 7 Environmental A 3 conditions 2 1 Erratic noisy baseline 5 27 ESA Corona Test Worksheet 2 20 Establishing a system log 5 10 Evaporation solvent aerosol 1 9 Event marks screen 3 20 Exhaust 2 9 Exploded view of the Corona C 4 External requirements A 2 Index F Features of the Corona 1 2 Filter screen 3 11 Final assembly C 9 C 15 Firmware flow chart D 1 Flow rate for columns table 4 2 Frequently asked questions 1 12 changing filter elements 5 5 Fuses B 2 G Gain range 5 16 Gas 5 14 screen 3 12 source 2 8 General information C 3 operations 2 21 troubleshooting approach 5 10 guidelines 5 25 Glossary G 1 Grounding issues 5 17 Guard column B 2 H High background currents 5 28 Hold screen 3 12 How to create a timeline method 3 27 HPLC related issues 18 system 2 13 I Important considerations viii Inability to autozero the signal 5 31 Increase in back pressure 5 29 Initialization of the detector 3 2 In line filter elements 5 5 Inputs screen 3 22 Interfacing to other solvent delivery module 2 28 to the solvent delivery module 2 27 Introduction Installing the Detector Installing the column holder corona organizer module corona thermal organizer detector drain vent assembly in line filter pulse damper sample injector valve temperature probe Internal electronic programs I O Connection Bloc
115. ented to confirm that the step has been saved Press the ENTER key to return the cursor to the Time parameter so that the next Timeline event can be created When a time is entered for another step the bottom line returns to that of Figure 3 9 i e if a new unique time is entered the DEL STEP changes back to ADD STEP When the cursor is in the time field the arrow keys can be used to scroll through the entered activities When an entered activity is being displayed the second soft key label will be DEL STEP as shown in Figure 3 10 Pressing this key will delete that entry activity TIMELINE Time 0 00 min Event End Stops time line clock CANCEL DEL STEP PREVIOUS NEXT Figure 3 10 The Event SAVED Screen 3 4 5 2 End Event Screen The End Event screen Figure 3 11 is used to indicate the time at which the timeline will terminate When this screen is accessed the cursor will be found on the Time field which can be edited as described in Section 3 4 5 1 After the desired time is entered press ADD STEP This command is used to prevent the detector from running continuously TIMELINE Time 0 00 min Event End Stops time line clock CANCEL DEL STEP PREVIOUS NEXT Figure 3 11 The End Event Screen 3 4 5 3 Set Contact Screen The Set Contact screen Figure 3 12 is used to change the position of a contact closure at the indicated time When this screen
116. ents have been specifically designed or selected to optimize the performance of your HPLC System A wide variety of Corona detector Application Notes are currently available from ESA Biosciences For additional information please contact ESA Biosciences Inc or your local representative Corona CAD and Corona S CAD Detectors 1 11 Operating and Maintenance Manual Chapter 1 1 9 Frequently Asked Questions Question As the detection principles of CAD and ELSD have two steps in common nebulization and evaporation how can you explain that you can obtain a better consistency of response The lack of consistency of response of the ELSD is mostly due not to the detector itself but to the laws of physics the difference in response is mainly due to volatility and solubility differences Have you for example tested monosaccharides which give very different responses with ELSD Can you do better Answer In direct comparison studies where volatility and solubility are common variables with flow injection analysis yes our data including those with monosaccharides show that charged aerosol response magnitude appears to be much less dependent on analyte nature The effect of analyte volatility appears to be very similar to low temperature ELSD Other variables that reportedly affect light scattering response magnitude include the formation of solid crystals vs solvated particles the former scattering light much more efficiently speci
117. etc However a The Corona detector MUST be the last detector in the sequence b Be aware that other detectors may contribute to band broadening and have an adverse effect on peak shape measured by the Corona 2 8 2 Using the Corona Detector in Parallel with another Detector When installing the Corona detector in parallel with another HPLC detector or in parallel with a mass spec detector care must be taken to assure a balanced distribution of flows to each detector a It is important when using a passive flow splitting device to consider the back pressure from the Corona detector when you are adjusting balancing the flows to each device The Corona contributes a slight back pressure similar to that created by approximately 1 foot of 005 diameter PEEK tubing The Corona detector MUST receive mobile phase flow rates between 0 2 and 2 0 mL min b One possible way to adjust the flows between the devices is to replace the Corona detector with approximately 1 foot of 005 diameter PEEK tubing and to then measure the flow rate emanating from the end of the tubing Adjust the flow balance so that the desired flow rate to the Corona is obtained and then replace the length of tubing with the actual connection to the Corona detector 2 22 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation NOTE Be sure to remove the extra length of tubing before reconnecting the Corona dete
118. etector Ea p Time constant for the Filter filter used to smooth None Low Med Hi None Input baseline and signal Percent offset added to 0 _Bno 0 0 Offset baseline signal 50 to 50 0 Input Indicates current T tomperature otthe TA 10 to 45 C off Output thermal organizer if installed NOTE All values should remain somewhat stable Significant fluctuations within a run may be indicative of a serious issue Review Chapter 5 before contacting ESA Service IS f Press the NEXT button and a second screen of data will be presented Figure 3 27 The parameters on this screen cannot be edited CORONA RUN Gas Pressure xx psi Corona Normal NEXT MENU PREVIOUS Figure 3 27 The Corona Run Screen 2 Gas Pressure refers to the pressure of the incoming gas and should correspond closely to pressures measured at the gas inlet port Corona refers to the status of the Corona discharge and should show normal if the Corona CAD detector is functioning properly If showing Service or Failing see Chapter 5 g Press the NEXT button and a third screen containing diagnostic data will be presented Figure 3 28 The parameters on this screen can not be edited Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 3 16 The Corona CAD Firmware This screen provides current information with regards to both factory set and inter
119. ethanol SuperQ Water or equivalent Guard Cartridge Holder 20mm for 2 0 amp 4 0 mm ID Cartridges P N 88 12414 Guard Cartridges for Capcell Pak C18 MG Columns 3um 20 mm x 4 mm P N 88 12307 0 005 PEEK tubing Red Peek nuts ferrules P N 70 5546 2 12 2 Procedure a b Pour 200mL of HPLC Grade Methanol into cleaned glass Mobile Phase Bottle Perform a purge pump and then initiate flow of methanol at 1mL min through all tubing prior to connecting the column for 5 minutes Stop the mobile phase flow Install the Guard Cartridge in Guard Cartridge Holder in the correct flow orientation Using a permanent marker mark the column holder with an arrow indicating the correct flow direction Using appropriate nuts ferrules P N 70 5546 from the Corona Accessory Kit connect the column holder cartridge to the 0 005 PEEK tubing Red Direct the exit tubing to a waste container Corona CAD and Corona S CAD Detectors 2 33 Operating and Maintenance Manual Chapter 2 Guard Column E Waste a Container h Initiate flow of methanol at 1mL min through the column for 30 minutes i Stop flow of methanol j Switch mobile phase to 20 Methanol 80 H2O Test Mobile Phase prepared as described in section 0 k Flow mobile phase through column for 30 minutes 1 Column is ready for use 2 34 Corona CAD and Corona S CAD Detecto
120. evere or prolonged flooding may damage the detector and void the warranty Maintaining the analytical conditions for lengthy periods may be expensive in terms of solvent and gas consumption and increases the wear and tear on the pump s Two commonly used alternatives are Maintain the flow at a lower flow rate e g 0 1 or 0 2 mL min Increase the flow rate to the desired operating level prior to resuming use then monitor the baseline signal to ensure that it is stable before making an injection Use a nitrogen generator rather than bottled gas Part Number 70 6003 is recommended 4 2 3 Shutting Down the Detector To shutdown the system a Flush the system with a neat neutral solvent for a few minutes to ensure that all buffers etc are removed Do not leave the detector exposed for long periods to either acidic or basic mobile phases oO Turn off the flow of mobile phase c Turn off the gas approximately 5 minutes after the flow of mobile phase was stopped 2 Turn off the detector Corona CAD and Corona S CAD Detectors 4 3 Operating and Maintenance Manual Chapter 4 4 3 Mobile Phase Considerations The detector can be used to measure all non volatile and many semi volatile analytes The basis of the detection is the measurement of the charge on the non volatile particulates formed following the evaporation of the solvent Since the detector will produce a response to essentially all particu
121. ezse teas ens densdecuedoscedeese daateatemeteeengert ada plaoededeageanineg 4 1 4 2 2 Maintaining the Detector between Analyses eeeeeseeeeeeeeeeeeeeeeeeneeeeeeeees 4 3 4 2 3 Shutting Down the Detector gw casch snssdepawiess wb dlbay Rohe evden Soares Ronee 4 3 4 3 Mobile Phase Considerations icc c5 lacs tccdsatenhodedacceevehean God Gand aedhientiend ae ncieeaed Sekt nel 4 4 4 4 Column Considerations erisccon a nal wk ante eran E 4 4 4 5 Sample GOMSIGSrATONS a cescaveledarcageiecunre Lastesess Meresseds beads nesateuebinnensgunligag aeaalabeueedenved 4 5 4 6 Maximizing the Performance of the ASSA Y ccceeceeeeeeeeeennteeeeeeeeeeeesesneeeeeeeeeeees 4 6 ana o R OETA E sce E E bana eun E A peak ens eee T TAN 4 6 4 6 2 Selection of the Proper Output Range cesses eeeeeeeeeeeeeeeeeeeeeeeeeeeeees 4 6 4 6 3 Setting the Fiter reisir eeaeee kear eee EEEa aee EARE 4 6 4 7 An Experiment to ensure that the Chromatograph and the Detector are F n tioning Properly iniae airnean aa ipanaa TE Gari Og ais Mane TA p aa aN 4 7 4 8 Analysis of Datas i os stvee tied Pewaela ct otaazahendhlaciadg dias aa aad aAA ATAARE tinal EEA Seats Vannes 4 9 4 9 Effects of Gradient Elution on ReESpPOnse ccccccceeeeeeseeeeeeeeeeeeeeeeeaaeeeeeesaeeeeeens 4 11 CHAPTER 5 MAINTENANCE AND TROUBLESHOOTING 5 1 trod CtHON Renin ects cen sedis x hireed e aaa oa ct aes a dame tates ud cute daana cies sued cate 5 1 5 2 Det tor Maintenant si
122. f proteins organic extraction etc will be both matrix and analyte dependent As a precaution samples should be passed through a 0 2 um filter prior to injection on the HPLC system As has been mentioned the detector is extremely sensitive to contaminant particles Particles can form if an analyte comes out of solution when injected into the mobile phase Make sure that the analyte is soluble in both preparation solvent and mobile phase and that the preparation solvent and mobile phase are themselves compatible The detector is sometimes used to measure trace contamination down to the 0 01 level Often in order to measure such trace contaminants it is necessary to analyze high concentration of starting material To prevent formation of particles make sure that all analytes in the starting material are soluble at the highest concentration used Corona CAD and Corona S CAD Detectors 4 5 Operating and Maintenance Manual Chapter 4 4 6 Maximizing the Performance of the Assay 4 6 1 Overview Once the appropriate chromatographic conditions for the compound s of interest have been determined a variety of steps can be used to optimize the performance of the assay e Sample cleanup steps are extremely important This is important if using the detector for the analysis of complex biological samples e g urine It is easier to separate and detect a clean sample than one that has undergone only a minimum of sample preparat
123. fic absorption of light probably a minor effect and more importantly the dramatic changes in efficiency of light scattering with particle size Rayleigh vs Mie vs refraction reflection This latter relationship becomes most important with chromatographic separations when considering the actual peak elution volume of each analyte Differences in peak volume between analytes governed by individual analyte chromatographic variables are the basis for differences in the peak concentration seen by the detector Particle size distribution differences that result from peak concentration differences lead to dramatic differences in response with ELSD again because of the changes in efficiency of light scattering The efficiency of particle charging and detection i e Corona CAD detection has been shown to be much more uniform with particle size Also our results suggest that Corona CAD detector response is not as dependent on the physical e g crystalline characteristics of the aerosol Question You state that Corona CAD detector s unique method of detection allows quantitation across a range that exceeds four orders of magnitude On the other hand the calibration curve you show seems to be non linear So what is the difference with ELSD Answer CAD is not linear and a quadratic plot type present in all or most chromatography data systems provides an easy way to calibrate Alternatively a log log plot can be used The difference to ELSD is that
124. g delivered to it Failure to turn the mobile phase off when the gas flow has stopped could damage internal components of the detector Both connections to the drain bottle must always be connected to allow the mobile phase to drain from the nebulizer The drain bottle must be emptied periodically to prevent flooding of the detector Severe or prolonged flooding may damage the detector and void the warranty The GAS OFF terminals on the rear of the detector can accept a signal from an external device e g a pump or data station to turn off the gas flow at the end of a set of analyses or in the event of a system fault e g the pump stops It is intended to save gas for those using nitrogen cylinders A CAUTION Nebulizer gas must be flowing through the detector 2 26 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation Autozeroing is used to return the signal to a zero point regardless of the current being measured If desired the baseline can be autozeroed before injection of a sample by a momentary shorting of the autozero terminal posts The signal should be sent by the autosampler or other external device When autozeroing allow sufficient time between activation and injection to ensure that the autozero command is completed Alternatively an autozero may be performed during a run see Timeline Event Options Chapter 3 If this is the case make sure that autozero occurs at a poin
125. g the In line Filter If an in line filter is employed it should be connected between the injector and the column The filter holder can be allowed to sit on the floor of the chassis plate C 4 6 Installing the Temperature Probe The Temperature Probe Part Number 70 5812 is placed in the access hole on the lower half of the column holder Figure C 12 To install the Temperature Probe a Loosen the clamping screw on the side of the column holder b Slide the probe into the access hole until it is bottoms out c Tighten the clamping screw against the probe d Check to see that the probe is securely in place by slightly pulling back on it C 4 7 Installing the Column Holder The Thermal Organizer is equipped with a 15 cm Column Holder Assembly Part Number 70 5366 that is clamped to the chassis plate on the right side of the chassis Figure C 13 A 5 cm Column Holder Assembly Part Number 70 5365 and a 25 cm Column Holder Assembly Part Number 70 5367 are available as options Corona CAD and Corona S CAD Detectors C 13 Operating and Maintenance Manual Appendix C The lower half of the column holder is mounted on the two short standoffs on the Organizer Chassis Plate If it is necessary to remove the holder e g to replace it or clean the organizer remove the screws that attach the holder to the standoffs When replacing the column holder check that the holder sits flush to the chassis and that nothing is caught
126. g the detector Activation opening closing and or reopening of up to four contact closures Placement of an event mark event mode on the detector output Wait ing for a signal from an external device to perform an action e g autozeroing the detector e Looping the timeline can be made to automatically repeat its programmed sequence for repetitive analyses When a timeline operation is programmed the detector will either perform an operation or wait for an external signal at the selected time If the system includes an optional Thermal Organizer the temperature of the organizer is set via the program as described in Section 3 6 9 and the temperature is displayed on the main menu The temperature cannot be programmed in a timeline method The Corona S detector can be used with the nebulizer heater on for freezing mobile phase mixtures e g aqgueous THF gradients or off The factory set temperature of 30 C prevents the nebulizer from becoming obstructed or blocked by ice which usually presents as erratic baseline and noise Ice may be formed from water in the mobile phase when the nebulizer block is excessively cooled due to the evaporation of THF or some other solvents during nebulization Turning the nebulizer heater on or off is covered in greater detail in Section 3 6 11 3 2 Initialization of the Detector When the Corona detector is turned on the system undergoes an electronic self test If an electr
127. ges Thermal Organizer Code Message Suggested Action If installed check Temperature Probe 0 Thermal Organizer not detected connections 1 Thermal Organizer malfunction Check Temperature Probe connections 2 Thermal Organizer malfunction logic Check Temperature Probe connections 3 Temperature out of range under Check Temperature Probe and set point 4 Organizer sensor malfunction Check Temperature Probe connections 5 Thermal Organizer malfunction logic Check Temperature Probe connections 6 Temp sensor or heater not responding Check Temperature Probe connections CAUTION REMOVE POWER UNPLUG ORGANIZER Excess heating detected and CALL ESA For assistance in troubleshooting and replacing components please call ESA Biosciences Technical Service or its authorized distributor C 6 Maintenance and Cleaning The Corona Organizer Modules are designed to give years of trouble free use however there are some general maintenance items that should be observed as with all HPLC components and systems care that the cleaning material does not get on the Temperature Probe the Thermostat or the Heater Driver Board located on the underside of the chassis plate as this could cause damage A CAUTION When you are cleaning the Thermal Organizer Module take In most cases it is easier if the chassis plate and its contents are removed from the organizer tray when replacing parts It may also
128. graphy system Use this instrument ONLY for this purpose The following precautions should be followed to minimize the possibility of personal injury and or damage to property 1 Maintain a well ventilated laboratory If the mobile phase contains a volatile organic solvent ensure that the laboratory is well ventilated so that a buildup of organic solvent cannot occur 2 The gas port to the instrument must be properly connected The gas should be clean dry devoid of organic solvents and particulates and properly regulated Only nitrogen is recommended DO NOT use helium Use an approved nitrogen generator capable of delivering 99 pure nitrogen 60 125PSI max flow 4 L min 70 6003 All nitrogen gas shall be preconditioned using the ESA Gas Conditioning Module 70 8285 3 Exhaust gases must be vented to an appropriate hood or gas collection device and NEVER vented directly into the laboratory DO NOT use the instrument unless the exhaust vent is connected properly Outlet gas venting should be at atmospheric pressure A vacuum or restriction may result in pressure changes within the detector resulting in baseline instability and or other problems iv Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 4 The drain tubing must be properly connected to both the detector and the waste bottle The drain tubing must be straight without kinks dips or loops i e areas where fluid can accumulate Any
129. he cap in the event that the bottle becomes over pressurized e g if exhaust is blocked NOTE The cap contains a perfluoroelastomer FFKM sealing washer P N 70 7785 to insure a leak free fit to the bottle Use care when emptying the bottle to avoid damage to the washer 3 NOTE The cap contains a pressure relief valve rated at 10 psi to prevent overpressureization of the waste system Figure 2 8 Drain Tubing Installation 2 12 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation Figure 2 9 2 4 1 6 HPLC System Correct Setup of Drain Tubing Ie NOTE In order to achieve maximum detector performance a validated pre conditioned HPLC system should be used Autosampler Hho HEO O Ses 333 HW Of Filter oo e a eg Put the C18 guard cartridge P N 88 12307 in the guard column holder P N 88 12414 Precondition the guard column P N 88 12414 following the procedure outlined in section 2 12 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 2 13 Chapter 2 WARNING Failure to properly condition the guard column can result in varying background high noise or signal spiking Connect the outlet of the autosampler to the inlet side of the guard column holder The autosampler column holder and detector tubing connections should use 0 005 PEEK tubing Red that is a
130. he nebulizer gas to the detector be maintained at all times An additional benefit of maintaining the power to the control module is that the life of the internal battery which protects stored methods will be increased Corona CAD and Corona S CAD Detectors 4 1 Operating and Maintenance Manual Chapter 4 To start the system a Ensure that the waste bottle is empty and check that the waste lines are free from bends kinks or other interferences Make sure that the bottle cap is on tightly Turn on the detector and allow it to proceed through the Self Test Press the Run soft key to access methods Choose the desired method and press the Next soft key This will open the internal gas valve and allow gas to flow Confirm the input gas pressure is 35 0 0 1 psi and adjust the Gas Conditioning Module as required Allow 5 minutes to equilibrate Start the delivery of mobile phase and allow the mobile phase to flow through the system for 3 5 min at the appropriate flow rate see Table 4 1 to expel the air in the system and equilibrate the detector The minimal flow rate to the Corona detector is 0 2 mL min Flow rates lt 0 2 mL min may cause the detector to exhibit aberrant behavior Table 4 1 Flow Rate for Various Columns Column Diameter Typical Flow Rate 4 6mm 1 0 mL min 3 0 mm 0 5 mL min 2 0mm 0 2 mL min The detector is ready for data collection when the baseline falls to 0 5 pA
131. hrough the bottom of the chassis plate b Connect the Ground Strap Assembly Part Number 70 5625 to the protruding threaded stud at the left rear corner of the chassis plate see Figure C 8 Attach the three knurled thumbnuts which were removed in Section C 4 2 and finger tighten c Mount the organizer module on top of the Corona Detector by inserting the latch tabs at the front of the organizer into the two slots at the top back of the detector s bezel Make sure the swivel plate of the latch assembly at the rear of the organizer is underneath the top lip of the detector Then tighten the latch using the knurled thumbnut Corona CAD and Corona S CAD Detectors C 15 Operating and Maintenance Manual Appendix C In most cases the organizer is attached to the top of the Corona Detector but it can also be placed on the bench next to the Corona if so desired Remove the latch mechanism at the rear of the organizer so the organizer will set flat on the bench The organizer must also be set as close as possible to the detector to ensure that the cell cables will readily reach it Connect the loose end of the Ground Strap Assembly to the ground screw on the back of the Corona Detector It is located in the upper right hand corner when facing the back of the detector of the power supply module next to the ground symbol ee NOTE The Ground Strap that is provided will not reach to the Corona h i Detector when the organi
132. ical Location of the Corona Detector c ccceeesecceeeeeeceeeeeseneeeeeenaes 2 1 2 2 2 Power REQUIFOIMECNIS 1 iscd aetete ete tedece ances ted covers daar etee ais dettrnadthe st teeth 2 3 22 3 Space REQUIFOMEING sa nmre eee teta aa uaaa tiaia tilaa 2 4 2 24 Neb lizatioN GaS a a a aa neds 2 5 2 3 Unpacking the Detector erona renne apornas nea aa a aa Aaa ends EAEE ERESSE aN 2 5 2 4 Installing the Det ctor riranin a aeh A a dE a 2 6 241 Basie NSElalON arean E E E TETE EA 2 6 2 4 1 1 Locating the Detector ssssssssssrerrrrrnsssssrrnrnnnnsssstennrnnnneeesreeennne 2 6 2 4 1 2 Power SOUICE E E EE E T ET 2 7 2423 C z CeO D KO 2 E EAE EA OAE EA AN RAPA E T ATT 2 8 2A A EXMAUSES cece TE AE AEN E E AE ETET 2 9 2 4 1 5 Drain and Vent Installation cece cccecceseeecseeeceeeeseeeeeueesenenees 2 9 2A te APEG SYVSIOM aicinn nienia a daetote a naman nay 2 13 2 4 1 7 USB Connections cccccccccccccecceceeeeceeeueeeseseueeeseseeeueeeeseaeeeueanes 2 15 2 4 1 8 Other Connections senesinin nka aaaea a aiao 2 15 2 4 1 9 Setting up Corona Plus Nebulizer Heater c cceeeeeeeeeeseeees 2 15 2 5 DE Aaea a o EEEa I eet eet PEE E AE AA ARN T E ACh oad 2 16 2 5 1 Role of the Test Protocol esonnnnnannnnnonnnnnenennsrenrrnsnrrronsrrresnrnrrnrnnrssnererenn 2 16 22 SOUP E E E AEA E T AATE 2 16 Zas PTO O reau a A a a aT E A AA 2 17 2 6 ESA Biosciences Corona CAD Detector Test Worksheet ccccccc
133. ie E TEE wrst aha hac ease daact aac lge uh cil E wae eae he C 1 G 2 gt Unpackingithe Organizer gg sacs cere co eats Sec oe aue ale etd eed acdc a a EAS C 1 C 3 Installing the Corona Organizer Module cceceeeeeeeeeeeeeeeneeeeeeeenaeeeeeseneeeeeeeeas C 3 Cael General Information ieceri a E k C 3 C 3 2 Removing the Organizer Chassis Plate cccccceeesssceeeeeeeeeeeeeeeeeeeeeeeees C 5 C 3 3 Installing the Pulse Damper 2 c cceeeeseeeeeeeeeeeeeeeesaeeeeeeeaaeeeeeenaaeeeeneaas C 5 C 3 4 Installing a Rheodyne Manual Sample Injector ValVe ceeeeeeeeeeeeeeeees C 6 C 3 5 Installing the In line Filter eee cceceeeeeeeeeeeeeeeeeeeeeeeaaeeeeeenaaeeeeeneaaeeeenees C 7 C 3 6 Installing the Column Holder 0 ecccecceeeeeeeeeeeeeeneeeeeeeeaeeeeeeeneeeeeseeaeeeeneaas C 7 C 3 7 Manual Sample Injector Overflow Line Bracket ceeeceeeteeeeeeeeeeeeeeees C 8 3 8 Fina ASSOMDY ses rr ip eee et ee eee cence eaten a aa cbse teat ected C 9 C 4 Installing the Corona Thermal Organizer Module cceeceeceeeeeeeeteeeeeetteeeeeeee C 10 C41 General Information ei sccactatecttigstet a Gadi head aS E C 10 C 4 2 Removing the Organizer Chassis Plate ccccceceeseeeeeeeeteeeeeetneeeeees C 11 C 4 3 Installing the Pulse Damper cc cceeeeseeeeeeeeeeeeeeeeeaeeeeeeeaeeeeeeeaaeeeeees C 11 C 4 4 Installing a Rheodyne Manual Sample Injector Valve ecceeeeeeeeeee C 12
134. ine Fuses If the unit does not power up when the main power switch is turned on or if the display and LED s are suddenly no longer illuminated it is possible that the fuse has blown WARNING Disconnect the Corona detector from line power before AN removing the cover from the power input module For continued protection against the risk of fire replace only with the same type and rating of fuse To replace a fuse a Turn off the power and remove the power cord from the unit b Open the cover of the power input module using a small screwdriver or similar tool c Open the fuse block assembly d Remove both of the fuses and replace the fuses with new ones The appropriate fuses are indicated in Table 5 1 Corona CAD and Corona S CAD Detectors 5 7 Operating and Maintenance Manual Chapter 5 Table 5 1 Fuses for Corona CAD Detector Voltage Fuse Rating Part Number 250 V TIA 70 6666 e Holding the assembly on the sides slowly slide the assembly in pressing in the front until seated properly f Close the cover to the power input module g If the fuses blow again contact ESA or its representative for service Do NOT continue to replace fuses as this could cause damage to the detector Do NOT remove the covers from the instrument as high voltages may be present and injury could occur 5 3 3 Replacing the Gas Filters The gas filters should be replaced annually The gas filters are replac
135. ing Press the 9 key and then the W key until N is showing Press the 9 key and then the W key until T is showing The screen should now look like that presented in Figure 3 22 EDIT METHOD SAVE Name CURRENT See ee cas PREVIOUS SAVE Figure 3 22 The Save Method Screen with Method Current When the desired alphanumeric characters are entered press ENTER to return the cursor to the Method field Press the ENTER key and then press the SAVE key to complete the save procedure The method will now be stored in memory The date and time will be automatically entered when the method is saved Press NEXT to continue and then press NEXT to return to the Main Menu 3 4 7 Method Zero Method 1 is initially displayed when EDIT is pressed while in the Main Menu Normally the A key would be used to scroll to higher method numbers both saved and unused methods If the key is pressed Method 0 is accessed This method contains the information for the last method that was overwritten IESE NOTE We recommend that you do not deliberately store a method using Method 0 because it will be lost the next time a method is overwritten 3 14 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware 3 5 Using a Method to Collect Data a Create and save the desired method b Return to the MAIN CORONA MENU screen Figure 3 23 MAIN CORONA MENU Select de
136. ing to the compression fitting located on the bottom front left portion of the detector The inner PEEK tubing should extend about 1 2 into the draing fitting on the machine Ensure that the two ferrules inside the nut are properly oriented Figure 2 5 The Drain fitting should be tightened with the supplied 9 16 wrench until it is snug Figure 2 6 CAUTION Do no over tighten the drain fitting Equipment damage could result Figure 2 5 Correct Orientation of Nut and Ferrule Figure 2 6 Location of Fluidics Connections 2 10 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation c Using the tubing cutter Part Number 70 7112 from the Drain Vent kit cut both the drain and vent tubing to a length between the detector and waste bottle where there are no dips kinks or loops Cut the drain tubing 4 longer than the vent tubing A vertical drop is optimum It may or may not be necessary to cut the tubing depending upon the installation Leave enough excess in case the tubing ends need to be cut and reinstalled The inner PEEK tubing does not have to run the entire length of the Teflon Tubing as long as 72 is extended into the fitting on the Corona CAUTION Because the vent tubing consists of two layers it may be AN necessary to flare the tubing This will slightly widen the opening Take care not to damage the Teflon lining during attachment and visually inspect the lining t
137. ion e If the compound of interest can be baseline separated from other components of the mixture detection and quantitation will be far more straightforward e The ideal mobile phase for separation and the ideal mobile phase for detection may be very different It may be necessary to make a compromise to obtain an appropriate level of sensitivity and selectivity 4 6 2 Selection of the Proper Output Range Ensure that the output from the detector agrees with the input of the recording device being used Select the gain range to ensure that the peak is on scale This is usually done by experience and or experimentation typically the scale is set so that the largest peak anticipated is 50 75 full scale If the inappropriate gain range is chosen a peak may overwhelm the detector and produce aberrant data see Chapter 5 Section 5 8 3 4 6 3 Setting the Filter The filter time constant is used to electronically reduce the noise in the chromatogram Four filter values are provided None Low Medium and High Typically none is used for the filter setting e f no filter is used the raw data is presented if the signal is very noisy and the sample concentration is low it is possible that the peak may be lost in the noise e lf the high filter is used and the peak is small it is possible that the peak will not be observed because the high filter setting attenuates the signal The user should note that as long as standards
138. ired level and stabilized Corona CAD and Corona S CAD Detectors 2 23 Operating and Maintenance Manual Chapter 2 2 10 Making Electrical Connections 2 10 1 Description of the Rear Panel of the Detector All electrical connections are made via the rear panel of the detector The rear panel is shown in Figure 2 16 Logic Module Gas Module Power Module Thermal Organizer Control Ground Thermal Organizer USB Power Filter Module Brackets Signal Out Filter Module Fuse Connections Power Line On Off Switch Gas Inlet Gas Exhaust I O Connections Figure 2 16 Rear Panel Corona CAD Detector 2 24 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation The terminals on the rear panel of the Corona detector are used as follows Thermal Organizer Control If a Thermal Organizer Module Part Number 70 5499TA is installed the Y cable Part Number 70 5595 is connected to the Thermal Organizer Control socket on the Logic Module and the Thermal Organizer Power connector on the Power Supply Module of the Corona detector while the single cable end is connected to the Thermal Organizer See Appendix C for greater detail Analog Signal Out Provides a OV to 1V output to the recording device or external A D converter Power Cord Connection The power cord is inserted into the receptacle on the lower right of the rear panel ot CC1 Pu
139. is accessed the cursor will be found on the Contact field The desired contact closure CC1 to CC5 Note CC5 is dedicated to Pump off is selected via the A or W keys Once the contact closure is selected the cursor moves to the Off On field and the position is chosen using the A or V keys After the desired selections are entered press ADD STEP 3 10 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware TIMELINE Time 0 00 min Event Set Contact Contact C1 set to Off CANCEL ADD STEP PREVIOUS NEXT Figure 3 12 The Set Contact Screen for CC1 3 4 5 4 Autozero Screen The Autozero screen Figure 3 13 is used to choose the time at which an autozero event is to occur TIMELINE Time 0 00 min Event Autozero This will perform an Autozero CANCEL ADD STEP PREVIOUS NEXT Figure 3 13 The Autozero Screen See Table 3 3 for further information about autozeroing 3 4 5 5 Filter Screen The Filter screen Figure 3 14 is used to select the level of signal filtering The filter selection None Low Medium and High is chosen using the A or W keys A detailed discussion of the use of the filter is presented in Section 4 6 3 TIMELINE Time 0 00 min Event Filter Filter time None CANCEL ADD STEP PREVIOUS NEXT Figure 3 14 The Filter Screen none selected
140. ither end of the assembly c Remove both end nuts from the filter assembly d Remove the used filter If necessary carefully insert a small wooden dowel or plastic rod to dislodge the filter e Rinse the filter housing with de ionized water 5 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting f Replace one end nut Insert a new ESA filter element into the filter housing Ensure that the element is properly centered and seated against the surface of the end nut AN CAUTION Be careful not to scratch the filter g Replace the second end nut and tighten carefully until contact between the cap and the filter is felt The filter is properly installed if both end nuts are approximately an equal distance from the center of the filter housing The fitting should be finger tight do not use a wrench or pliers and DO NOT over tighten as this can crush the filter rendering it useless h Re install the filter housing in the chromatographic system Ensure that the direction of flow is as indicated on the filter housing NOTE Initially only the upstream end of the filter should be attached eS to the HPLC system Pump about 5 mL of the mobile phase through the filter to waste before attaching the downstream end of the filter to the detector this step will serve to wash the filter and ensure that particulate matter does not enter the detector 5 3 2 Changing the L
141. k Figure K Keypad Keys action data L Locating the Detector Logic board replacement Loop screen Loss of response Maintenance and cleaning troubleshooting Maintaining the detector between analyses Making Electrical Connections Manual injector overflow line Marker screen Maximizing performance Method zero Mobile phase considerations formulation flow preparation qualification 2 11 E 2 E 7 C 7 C 13 C 5 C11 C 6 C 12 C 13 5 12 2 25 3 4 3 4 3 4 2 6 5 32 3 13 5 30 C 17 5 1 4 3 2 24 B 2 C 8 C 15 3 11 4 6 3 14 5 21 4 4 2 29 5 19 2 29 2 32 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Operating and Maintenance Manual Index Mode of operation 1 8 R Monitoring charged particles 1 9 Monthly maintenance 5 4 C 18 Range screen 3 12 Rear of the detector 1 7 N Recommended supplies B 1 Remote screen 3 21 Nebulizationof the eluent 1 8 Removing the chassis plate C 5 C 11 Nebulizer heater Replacing the gas filters 5 8 error codes 3 26 Reset screen 3 13 setup screen 3 25 Role of the test protocol 2 16 Nebulization Gas 2 5 RS232 setup screen 3 18 Nitrogen Run 3 3 gas supply B 2 parameters 3 16 generator 1 4 5 5 Noise review 5 24 S Normal operation 3 1 Safety O precautions iv symbols jii Operating Sample symbols iii considerations 4 5 the detector 4 1 Save method screen 3 13 Organizer Schematic of the detector 1 10 and thermal organi
142. latch tabs at the front of the organizer into the two slots at the top back of the detector s bezel Make sure the swivel plate of the latch assembly at the rear of the organizer is underneath the top lip of the detector Then tighten the latch using the knurled thumbnut In most cases the organizer is attached to the top of the Corona Detector but it can also be placed on the bench next to the unit if so desired Remove the latch mechanism at the rear of the organizer so the organizer will set flat on the bench Connect the loose end of the Ground Strap Assembly to the ground screw on the back of the Corona CAD detector It is located in the upper right hand corner when facing the back of the detector of the power supply module next to the ground symbol see page vii and Figure 2 1 in this manual eS NOTE The Ground Strap that is provided will not reach to the Corona f 9 when the organizer is placed next to it on a bench The user must provide a grounding cable for this configuration Connect the tubing for the drain onto the protruding tube on the right rear of the organizer Place the other end of the drain tubing in a waste container that is situated lower than the organizer Replace the cover Make all the necessary connections to your HPLC system The Corona CAD detector and its organizer module are now ready to be used Corona CAD and Corona S CAD Detectors C 9 Operating and Maintenance Manual Appen
143. late matter contained in the mobile phase care must be taken to ensure that such material does not enter the detector Such extraneous particulate matter can come from the mobile phase itself the column see Section 4 4 and even the sample see Section 4 5 e Make sure the in line filter is installed between the column and the detector e Use only volatile salts in the mobile phase e Do not use either ammonium carbonate or ammonium bicarbonate in the mobile phase e Use only solvents with minimal residue check label on the solvent bottle Ifa particular solvent causes problems due to particulates try a different lot ora different vendor Be aware that some solvents contain stabilizing agents that may cause problems e Use a pH of lt 7 5 Higher pHs may cause increased noise background currents etc e When pH ing make sure that calibrators used in checking the pH meter do not contaminate the mobile phase e Filter mobile phase through a 0 2 um filter prior to use e Mobile phases that can cause freezing of the nebulizer e g aqueous THF gradients should only be used with the Corona detector e Degass prior to use Degass on line if necessary 4 4 Column Considerations The detector can be used with any HPLC compatible column e g normal phase reverse phase gel permeation chiral etc However columns that are unstable e g some diol columns and release particles from the column bed i e column b
144. ld be cut in a similar manner to that of the gas inlet tubing WARNING Make certain that the venting device used does not provide any positive or negative pressure on the detector exhaust stream Excessive pressure can result varying background high noise or signal spiking laboratory The exhaust gas may contain hazardous fumes A WARNING Make certain that the exhaust gas does not enter the 2 4 1 5 Drain and Vent Installation The Drain Vent tubing assembly Part Number 70 7115 consists of a piece of Teflon lined Tygon tubing and a piece of solid Teflon tubing with a piece of PEEK tubing inside that must be carefully assembled to promote proper drainage The assembly includes the Drain Tubing 3 16 ID x 1 4 OD x 6 long Part Number 70 7113 with 015 ID x 1 16 OD x 5 long PEEK Tubing Gray Part Number 70 5996 and the Vent Tubing 1 8 ID x 4 OD x 6 long Part Number 70 6259 Figure 2 4 PEEK Tubing Tygon Tubing a Solid Teflon Tubing Teflon Lining Vent Tube Drain Tube Figure 2 4 Vent and Drain Tubing Corona CAD and Corona S CAD Detectors 2 9 Operating and Maintenance Manual Chapter 2 To attach the Drain Vent Tubing assembly a Connect the Vent tubing to the quick connect fitting located on the bottom front left portion of the detector This is a quick connect fitting which simply latches when the fitting is pushed in Figure 2 6 b Connect the Drain tubing end with SS fitt
145. leed will produce noise in the detector and should not be used If such a problem is encountered try using an equivalent column from a different manufacturer 4 4 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Using the Detector in a HPLC System To minimize the potential effects of column decomposition e Do not use the column at extremes outside of the pH range indicated by the manufacturer e When anew column is installed pump 5 to 10 column volumes of the mobile phase through it to waste before connecting the column to the detector e Make sure that the column is compatible with the mobile phase being used e g some solid phases are not compatible with some organic solvents e g cellulose modified silica with acetone e Use a dedicated column for each application A column with unknown history may have been exposed to non volatile salts or may be contaminated with analytes capable of being eluted under the current experimental conditions e g producing ghost peaks WARNING A column contaminated with non volatile salts will cause noise and spiking in the Corona CAD detector Such issues may continue for several hours after the column is removed 4 5 Sample Considerations The detector can be used with many different sample types As with any HPLC analysis correct sample preparation is essential The type of sample preparation used e g solid phase extraction acid precipitation o
146. lements Corona CAD and Corona S CAD Detectors 5 5 Operating and Maintenance Manual Chapter 5 5 3 1 2 Checking the Pressure Drop Across the Filter To measure the pressure drop across the filter element a Turn off the mobile phase flow Allow system pressure to drop to zero before disconnecting any components on the high pressure side of the column The rapid pressure drop can damage various components in the overall system Always A CAUTION Do not remove the system pressure by opening a fitting allow the system to drop to zero before breaking any connections b Disconnect the tubing on the downstream outlet side of the suspect filter c Run the pump at a flow rate of 1 mL min and record the pressure reading d Disconnect the suspect filter and again record the pressure reading using a flow rate of 1 mL min e Determine the filter pressure drop by subtracting the second reading from the first reading If the pressure drop is significant 100 psi 8 bar or more change the filter 5 3 1 3 Changing a Filter To change a filter a Turn off the mobile phase flow Allow system pressure to drop to zero before disconnecting any components on the high pressure side of the column The rapid pressure drop A CAUTION Do not remove the system pressure by opening a fitting can damage various components in the overall system b Remove the filter assembly from the chromatographic system by removing the nuts on e
147. log format is provided by your organization then that can be used to capture the relevant information Table 5 2 A Typical System Log Back User Number System Maintenance General round Date Name Samples sample Type Pressure f Activities Comments Current 6 3 01 Jones 19 Catechols 220Bar 0 1pA Daily activities OK Leaky fitting 6 6 01 Davis 27 Compd 2319 20Bar 5pA Weekly activities on column repaired 6 6 01 Davis 29 Compd 2301 220Bar 0 1pA Daily activities OK Significant 6 8 01 Jones 21 Compd 2318 220Bar 0 1pA Daily activities Peak tailing replaced column 6 8 01 Smith 33 Compd 2367 220Bar 0 1pA Weekly activities OK 6 9 01 Smith 23 Compd 2400 220Bar 0 1pA Daily activities OK 5 6 General Troubleshooting Approach This section is intended to provide the user with guidance to correctly identify and correct problems that may be experienced with either the Corona or Corona s detectors 5 10 NOTE Ie to bea It is important to remember that although a problem may appear ssociated with the detector itself issues with the HPLC system the analytical method and sample preparation can all adversely affect the detector s performance DO NOT immediately assume that the problem is caused by the detector Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting The following is a set of gener
148. lumn that is not occupied by stationary phase Corona A high voltage electrical discharge that is used to place an electrical charge on a gas Corona Chamber A region of the detector that charges the particles G 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Glossary Current A flow of electric charge usually measured in amperes or amps A mA HA nA pA etc Current Range A setting on the Corona that relates to the current scale used for the signal output For example a Current Range of 100 pA indicates that a signal from 100 pA to 100 pA can be presented as a signal on the output of the detector for display and or quantification Signals higher than 100 pA will be clipped or cutoff Default Setting A setting of the detector parameter that is preselected by the manufacturer e g the default filter response is none Degassing The removal of dissolved gases from the mobile phase Degassing may be affected by helium sparging or vacuum filtration Detection Limit The smallest amount of a compound that will produce an observable signal above the background noise The signal to noise ratio that is selected is an integral part of the definition of the detection limit for a given situation e g a system has a detection limit of 2ug of tricyclyne with a S N of 3 Detector A device that is used to determine the presence or absence of analytes in the mobile phase A detector
149. ly to be used as an integral part of a High Performance Liquid Chromatography HPLC system This chapter discusses the use of the detector in an HPLC system and includes Operating the Corona Detector Section 4 2 Mobile Phase Considerations Section 4 3 Column Considerations Section 4 4 Sample Considerations 4 5 Maximizing the Performance of the Assay Section 4 6 Sample Experiment Section 4 7 Analysis of Data Section 4 8 Effects of Gradient Elution on Response Section 4 9 4 2 Operating the Corona Detector 4 2 1 Turning On the Detector NOTE A review of Cautions Warnings and Safety Procedures are presented in pages iii xii The user should review this information before powering up the detector Like any other HPLC detector when first turned on the Corona detector has to stabilize before use Usually this takes approximately 10 mins With the Corona detector the nebulizer block takes about 30 mins to reach thermal equilibrium The time taken for the thermal organizer to reach equilibrium will be dependent upon the set temperature the ambient temperature and the number and type of components contained within the organizer Usually it should take no more than 30 mins Plus If the Corona CAD detector is turned off and then on again it will be necessary to wait for the detector to stabilize To immediately begin analyzing samples we suggest that the detector is kept on and that the flow of mobile phase and t
150. mp Off O ump GND GND oi cc2 Gas Off L Cree a GND GND am See CC3 O Autozero DO GND GND cca LX t sn GO x GND end Figure 2 17 The I O Connection Block I O Connections The Input Output Connections block contains eight connections for interfacing the detector to external devices such as an autosampler a data station pumps valves etc which can synchronize the operation of these devices Figure 2 17 The inputs are TTL compatible and the outputs are true relays contact closures The two green terminal blocks must be installed before the inputs outputs can be used Make sure they are oriented and aligned properly before pushing or snapping them into place OUTPUTS The terminals labeled CC1 to CC4 provide contact closures that can be used to start an autosampler integrator pump gradient change a valve position etc The terminals are active only when the timeline function is used see Chapter 3 Section 3 4 5 3 Each of the terminals acts in a similar fashion The labeled designations are simply provided for the convenience of the user Corona CAD and Corona S CAD Detectors 2 25 Operating and Maintenance Manual Chapter 2 When the output contact closures are used there is no polarity plus or minus The third wire on the cable the ground wire on Part Number 70 4850 must be attached to ground GND of the Corona detector The PUMP OFF output co
151. n 10nm in diameter pass by the ion trap where they then impinge on a particle screen filter Monitoring the Charged Particles as the charged particles transfer their charge to the screen filter current is measured by a sensitive electrometer which in turn generates a signal that is amplified and placed on the detector output Charging the Particles Monitoring the Charge Figure 1 6 Stages in Detection Corona CAD and Corona S CAD Detectors 1 9 Operating and Maintenance Manual Chapter 1 NEBULIZER DRYING TUBE ae Ore tse GAS INLET EXHAUST L Figure 1 7 Schematic Overview of the Detector 1 7 Additional Information A detailed discussion of the use of aerosol charging detection in HPLC is presented in R W Dixon and D S Peterson Development and Testing of a Detection Method for Liquid Chromatography Based on Aerosol Charging Anal Chem 2002 74 2930 2937 To install and operate the detector in an HPLC system it will be necessary to obtain information from the manuals of other devices that are used in conjunction with the detector These include but are not limited to the pump the analytical column the nitrogen generator autosampler or injector and the data recording processing device recorder integrator personal computer etc 1 10 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Introduction 1 8 Support from ESA
152. n Chapter 2 with pump pulse dampener injection device either autosampler or manual injector with 10 uL injection capacity inline filter and Corona detector Connect the guard column Part Number 88 12414 between the in line filter and the Corona detector Do not use an analytical column Power up the detector and allow nitrogen gas 35 psi to flow through for approximately 10 min Activate the guard column by following the procedure outlined in Chapter 2 if required Prepare a 20 methanol 80 water HPLC grade mobile phase and allow it to pass through the system at 1 mL min Create a method in the Corona detector with the current gain range set to 100 pA full scale the Filter set to none and the offset set to 0 NOTE When the instrument is shipped from ESA this method is provided as method 1 Corona CAD and Corona S CAD Detectors 4 7 Operating and Maintenance Manual Chapter 4 f Wait 30 minutes and monitor the current When it is stable press the Autozero key The baseline noise trace should be similar to that presented in Figure 4 1 g Inject a 10 uL sample of 500 pg mL caffeine solution The chromatogram generated should be similar to that presented in Figure 4 2 Caffeine s retention time should be approximately 0 6 0 7 min typical peak area should be approximately 2 500 000 units 20 Mobile Phase 20 MeOH 80 water Corona Gain Setting 100 pA full scale Corona Filter Setting None
153. n between the detector and personal computer It can be accessed by pressing NEXT on the SYSTEM SETUP MENU when the Select area field indicates RS232 setup The values indicated in Figure 3 31 are the default values set by ESA and may need to be changed to conform to the communication requirements of the computer and or data station 3 18 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware RS232 SETUP Baud 38 4k Parity None Stop bits 1 Data bits 8 Handshaking None CANCEL MENU SAVE Figure 3 31 The RS232 Setup Screen When the screen is accessed The cursor will blink to the right of the Baud entry Use the A or Y keys to select the desired value the system can transmit data at 9 6K 19 2K and 38 4K baud When the desired value appears press ENTER The cursor will blink to the right of the Parity field Use the A or Y keys to select the desired value None Even or Odd When the desired value appears press ENTER The cursor will blink to the right of the Stop Bits entry Use the A or W keys to select the desired value 1 or 2 When the desired value appears press ENTER The cursor will blink to the right of the Data Bits entry Use the A or Y keys to select the desired value 7 or 8 When the desired value appears press ENTER The cursor will blink to the right of the Handshaking entry Use the A or W keys to select None or Hardware When
154. n time or changes in chromatographic peak shape e g Figure 5 7 are unlikely to be due to a problem with the Corona detector but are more likely to be due to column aging changes in temperature and differences in mobile phase composition Good Suspect m Minutes Figure 5 7 The Effect of Column Aging on Analytical Performance The old suspect column was replaced with a new good column and performance returned to normal e Only use the column as recommended by the manufacturer i e make sure the flow rate temperature and mobile phase composition and pH are compatible e Only use stable columns Those that show significant bleed can cause pressure problems e g due to blocking of the in line filter and if the particles enter the Corona detector increased back ground currents and noise 5 20 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting e Not all columns in a particular format are equal For example a diol column from one manufacturer may prove to be unusable e g due to column bleed while one from a different manufacturer may work perfectly well e Make sure that the column format used is compatible with the sample volume and or mass of analyte being separated Overwhelming the column will lead to aberrant chromatography e g odd peak shape shifting retention times etc e Properly flush new guard analytic
155. nally calibrated parameters Keep a log of these values If any of them changes dramatically then maintenance or service may be needed An explanation of these parameters and typical values are presented in Table 3 5 CORONA DIAGNOSTICS Total Flow x xx Flow Ratio x xx Voor X xxxkV lonT x xxV MENU NEXT Icor x xxyA PREVIOUS Figure 3 28 The Corona CAD Detector Diagnostic Screen Table 3 5 Diagnostic Parameters Parameter Definition Value Total Flow The total measured gas flow through the detector gt 0 5 Flow Ratio The ratio of the flows of the the positively charged corona secondary gas stream analyte particle gas stream to 0 2 1 0 VCor Corona Voltage The applied voltage to the corona wire to charge the secondary gas stream lt 3400 Normal 3400 3900 Service gt 3900 Failing ICor Corona Current The current required to charge the secondary gas stream Stable approx 1 0uA lonT required to remove the high mobility charged particles lon Trap Voltage The voltage gt 18V If the method includes a timeline press the NEXT button This will access the first Timeline screen A typical screen is presented in Figure 3 29 TIMELINE Time 0 00 min Gas valve Off Event Gas CANCEL PREVIOUS NEXT Figure 3 29 The Timeline Screen Timeline screens can be scrolle
156. ncrease the current range 5 12 6 Detector Test Procedure Section 2 6 includes a test procedure that is used to verify that the detector unit is operating properly If the operator believes that the detector is not functioning properly the test procedure should be repeated A worksheet is provided with the test procedure that should be completed and sent to the ESA Service Department or your local distributor to assist in diagnosing and solving any problem in the detector unit Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 5 31 Chapter 5 5 13 Power Supply Board and Logic Board Replacement 5 13 1 Replacing the Power Supply Board Part Number 70 5436TB If the AC Power fuses repeatedly blow or if the above procedures indicate that it is necessary change the power supply module as follows Remove the AC power cord Remove the two screws on the rear panel that hold the power supply assembly to the chassis Using the power module removal tool Part Number 70 6476 carefully pull the power assembly away from the chassis Insert the new assembly Replace the two screws that hold the assembly in place 5 13 2 Replacing the Logic Board If it is necessary change the logic module as follows 5 32 Remove the AC power cord Remove the two screws on the rear panel that hold the logic assembly to the chassis Carefully pull the logic assembly away from the chassis Insert
157. ne if the problem is instrument related or analysis related If any aspect of the analytical conditions is to be changed run a before and after to ensure that the effect of the change is well understood Do not consider any change as trivial As an example if you change the supplier of your mobile phase verify that the change has no effect on the analysis To isolate the source of the problem it may be valuable to perform independent checks of each of the components in the HPLC system These tests should be found in the operator s manual for the individual components see Section 2 5 for testing the Corona CAD detector Corona CAD and Corona S CAD Detectors 5 25 Operating and Maintenance Manual Chapter 5 A series of diagnostic procedures is presented below that will assist in pinpointing the cause of the problem CAUTION When you are troubleshooting the system do not relieve the pressure by opening a fitting on the high pressure side of the instrument e g do not open the fitting between the pump and column Instead reduce the flow rate to zero and let the pressure fall slowly Failure to do this could damage various components in the system On rare occasions the Corona display may lock up or present nonsense information If this occurs a Turn off the power to the unit b Wait a minute and then turn the power back on contact ESA Biosciences or your ESA Biosciences representative fo
158. ne must have sequence an ending time This will reset to initial Contact closures will NOT Reset parameters be reset a Typically an autozero operation takes place before an injection make sure that sufficient time is allowed for completion of autozero prior to injection If an autozero is to be performed during a run it should be set at a point where the chromatogram is reasonably flat and the signal is low i e the eluent does not contain an eluted compound For optimal performance allow a period of 15 seconds between an autozero event and the onset of a peak b For optimal performance an AUTOZERO should be performed when a CURRENT RANGE change is performed If a data station is used set the data station to inhibit integration during the period from 0 1 minutes before and after the gain range change The third line of the screen is dependent on the nature of the selected event In most cases additional information will be required to indicate the desired action that is to occur at the indicated time e g for the Filter as event desired filter level can be selected via the A or V keys Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 3 9 Chapter 3 A timeline can include a large number of events and each event should be performed at a separate time Once a timeline step is edited or created press ADD STEP to present Figure 3 10 The DEL STEP label over the second soft key is pres
159. nel to the organizer chassis plate with the two thumbnuts c Lead in the tubing from the autosampler through one of the ports on the proper side of the organizer C 3 5 Installing the In line Filter If an in line filter is employed it should be connected between the injector and the column The filter holder can be allowed to sit on the floor of the chassis plate C 3 6 Installing the Column Holder The column is clamped to the chassis plate using a Column Holder Assembly Part Number 70 5530 that is mounted to the right side of the chassis The holder is assembled as shown in Figure C 5 and is mounted to the side of the chassis via the lower set of screw holes on the right side of the chassis as shown in Figure C 6 Qe Figure C 5 Column Holder Assembly Corona CAD and Corona S CAD Detectors C 7 Operating and Maintenance Manual Appendix C One holder should be used for short columns and two holders can be used for longer columns Figure C 6 Mounting the Column Holder Assembly Connect the brackets to the left side of the chassis by the lip that fits into the horizontal slit and the knurled screw that fits into the right side of the chassis Figure C 6 The plastic knurled screws Part Number 70 5571 should be threaded into the proper hole over a cell and secured finger tight C 3 7 Manual Sample Injector Overflow Line Bracket The Manual Injector Overflow Line Bracket is an L shaped piece with small holes f
160. ngs are labeled To ensure consistent product performance and reliability use only genuine replacement parts and filter cartridges from ESA Biosciences Inc The filter cartridges in the filter assemblies are removed by loosening the collar from the filter assembly lowering the filter bowl away from the filter head and unscrewing the element retainer from the base of the cartridge Insert the new filter cartridge and reassemble the housing in reverse order 5 3 Customer Service of the Detector 5 3 1 In Line Filter Elements 5 3 1 1 Frequency of Changing Filter Elements In typical use the filter elements become clogged and must be replaced on a periodic basis The frequency of replacement is dependent on the level of particulate matter present in the mobile phase pre injector and the sample post injector as well as the production of fine particles from the analytical and or guard column post column If the filter must be replaced frequently e g more than once a month it may be worthwhile to modify the composition of the mobile phase and or switch to a more stable column e g a column from a different manufacturer which might create fewer fines column bleed Microbial growth may occur in mobile phases with low levels lt 3 of organic solvents unless suitable precautions are taken A daily log of system pressure should be kept so that any pressure fluctuations can be monitored as this is a good indication of clogged filter e
161. njector to the chassis so that the valve stem goes through the hole in the Injector Panel and the part number of the injector is facing up Fasten the injector using the two screws and star washers Part Number 50 5213 supplied in the accessory kit NOTE The screws and star washers are provided to ensure appropriate grounding Do not use the screws supplied with the injector to mount the injector e Reattach the injector handle and tighten the two hex setscrews Make sure the setscrews make contact with the flat portions of the valve stem It will be necessary to rotate the handle after tightening one setscrew to gain access to the other setscrew Refer to the Operating Instructions provided with the injector for other information on installation proper use and care of the manual sample injector valve These instructions include information about the wiring for the position sensing switch optional that can be connected to another HPLC component which is used to signal the start of the injection C 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer The cabling for the position sensing switch can be directed outside of the organizer at the back of the organizer along with the cell cables If an autosampler is to be used a Place the Organizer Injector Panel without the hole Part Number 70 5418 onto the organizer chassis plate b Affix the organizer injector pa
162. ntact CC5 is used to send a signal to the pump or other external device to turn the pump off and stop the flow of mobile phase If the pressure of the nebulizing gas falls below 50 of the required gas volume this signal may be used to shut the pump off so that the detector is not flooded with mobile phase INPUTS The connector labeled STRT start is used when a method involving the timeline program is to be started from an external device such as an autosampler A contact closure or shorting of the terminals a signal of at least 0 5 second is required from the external device It is strongly recommended that a true contact closure or relay closure be used for the inputs on the I O connections block GAS OFF AZ and STRT Ifa TTL level change is used the voltage must be between 0 and 5V and the polarity sense of the device must match that of the detector i e the and the of the external device must be connected to the and of the detector respectively The terminal is the top one in each group and the terminal is immediately below it followed by ground across the terminals is set at 5V DC or less When connecting outputs the potential across the terminals must be less than 30V DC and the current must be less than 0 5 A CAUTION When connecting TTL inputs ensure that the potential Gas Off turns off the supply of nitrogen gas to the nebulizer whenever mobile phase is bein
163. o create the mobile phase at a rate of 1mL min Check all fittings to ensure there are no leaks and tighten as appropriate j Monitor the background signal of the detector It will increase initially and then start to fall Wait at least 10 minutes or until the current is stable and not changing by more than 0 01 pA per minute Record the initial and final values on the worksheet Also record the information about the mobile phase used NOTE The initial current when flowing mobile phase can be as much as 50 150 times the initial value without flow 2 18 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation WARNING If the detector does not stabilize after 60 minutes or if the final current is not much lower than the initial value then this may indicate contaminated mobile phase and or solvents column bleed or prior contamination of the HPLC system by mobile phases not compatible with the Corona detector e g ones that contain non volatile components Observe the FRatio and Total flow values and record them on the worksheet Stop the preview mode and now run your sequence of methods and record the results on the worksheet Additionally keep a print out of all chromatograms for future reference NOTE A sharp peak should be observed on all the chromatograms and the retention times for the peaks should be between 1 3 and 1 6 minutes NOTE Other than the initial solvent void peak
164. o cut the tubing depending upon the installation Leave enough excess in case the tubing ends need to be cut and reinstalled The ends must be cut straight to ensure a proper connection to the barbed fittings necessary to flare the tubing by inserting the uncapped end of a ball point pen into the tubing This will slightly widen the opening Take care not to damage the Teflon lining during attachment A CAUTION Because the vent tubing consists of two layers it may be Corona CAD and Corona S CAD Detectors E 3 Operating and Maintenance Manual Appendix E ad 2 Attach the other end of the vent tubing to the barbed fitting on the cap of the Waste Bottle The vent tubing will be attached to the smaller barbed fitting and the drain tubing will be attached to the compression fitting Figure E 4 Carefully place the tubing over the barb ensuring that the inner lining is completely over the barb Grasp the tubing near the barb and push the tubing onto the barb until it is all the way down Install the nut and ferrules onto the drain tubing as shown in Figure E 2 Insert the drain tubing into the drain fitting Figure E 4 until it stops Screw the nut on to the fitting until it is finger tight Using the wrenches supplied in the accessory kit hold the body of the fitting with the 1 2 end of one of the wrenches Tighten the nut with the 9 16 end of the other wrench until it is snug approximately three quarters of a turn
165. o ensure that it does not crimp during installation d Attach the other end of the vent tubing to the barbed fitting on the cap of the Waste Bottle P N 70 7754 The vent tubing will be attached to the smaller barbed fitting and the drain tubing will be attached to the compression fitting Figure 2 7 Carefully place the tubing over the barb ensuring that the inner lining is completely over the barb Grasp the tubing near the barb and push the tubing onto the barb until it is all the way down e Install the nut and ferrule onto the drain tubing as shown in Figure 2 5 Insert the drain tubing into the drain fitting Figure 2 8 until it protrudes approximately 2 from the bottom of the cap Figure 2 8 Screw the nut on to the fitting until it is finger tight Using the wrenches supplied in the accessory kit hold the body of the fitting with the 1 2 end of one of the wrenches Tighten the nut with the 9 16 end of the other wrench until it is snug approximately three quarters of a turn with the wrench Drain Relief Valve Vent Z Figure 2 7 Bottle Cap Connections Corona CAD and Corona S CAD Detectors 2 11 Operating and Maintenance Manual Chapter 2 f Place the waste bottle in secondary containment in a secure location below the detector Figure 2 9 Make sure that the cap is tightened to prevent any gas leaks The waste bottle is a closed system under a slight pressure and has a pressure relief valve on t
166. o select the event and press ENTER The various events that can be used in a timeline are listed in Table 3 3 3 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware Table 3 3 Timeline Event Options Method of Event Data Entry Options Editing Comments Enter time to stop the y This is the final time for data End Numeric keypad method collection A and Y keys External signal set on off Set Select Contact CC1 to Each contact closure can CC4 Select On or Off or be set to on closed or off Contact Toggle open or toggle change state as desired Autozero_ Autozero signal No editing needed See notes a and b Filter None Low Med High A and Y keys Change the smoothing filter on the raw data Marker Height 10 to 10 FS Pioneers Ie any a marker onthe Range Current Range A and V keys The current range for the output See note b Gas On or Off A and keys Opens closes gas valve Input Condition Gas Off A and Y keys Indicates what the system should wait for to perform Autozero Start Hold w i the selected action E g Condition options are off to ae hold for on then perform on on to off off or on autozero Loop to x xx minutes Numeric Keypad Establish a set of repetitive Number of Loops 0 9999 operations See Section 3 7 How to create a timeline Loop method This will stop the Timeline Each Timeli
167. oes ce tcanctcaenas etek tonddete efetebaee dei teless cates seteet 5 23 eM INGISE FRO WIGW seictulea se caughateGeasiecan salou Renee a a ea A eras sides A Ea TAEAE AFE 5 24 5 12 General Troubleshooting Guidelines ccccceeeeseceeeeeesteeeeeeeeeneeeeeeeeeeeeeeeeeeeeees 5 25 5 12 1 Erratic Noisy Baseline iiccc scncciecesiveedvees niichawateeahiicee ace ace eee 5 27 5 12 2 High Background Currents 5 3 0 edie velveteen aan eae 5 28 9 12 3 Increase in Back Pressure sasctoscieatkedeceedsccacceteces daguiencanetesanuedsdan doceecdenpeeds 5 29 5 124 LOSS OF MESDOMSE io rna eaaa ane dao eh aea seeped veel shaatnenaceestincdeacatbceniaiatuee 5 30 5 12 5 Inability to Autozero the Signal cececcceeeeeeeeeeeeeneeeeeeeeaeeeeeesnaeeeeneae 5 31 5 12 6 Detector Test Procedures acc tceciceedsteedocendnsceaasviesonueoeenasexcgecaepiees seoedeness 5 31 5 13 Power Supply Board and Logic Board Replacement ccccccssssssseeeeeeeeeeeeeees 5 32 5 13 1 Replacing the Power Supply Board cececceeeeeeeeeeeeeeneeeeeenseeeeeens 5 32 5 13 2 Replacing the Logic Board csc csscieedudcincitenteathideAidtanboncdenst tact newelees 5 32 APPENDIX A CORONA AND CORONA PLUS DETECTOR SPECIFICATIONS A 1 General Operating Specifications 20 0 2 eeeeceeeeeeeeceeeeeeeeeeeeeeesaaeeeeeeaaeeeeeensaaeeeenaas A 1 A 2 Timeline Specifications sy cssec veg seciedeesencacetesietnszeckscalsnensadendvneetandensniaadlaeetdadeonneesvanan teas A 2 A
168. of the amount of compound adsorbed on the stationary phase to the total amount of compound in the mobile phase The capacity factor for an adsorbed compound on an HPLC column can be determined experimentally by use of the following equation k V1 Vo Vo where V is the retention volume of the compound of interest Vo is the void volume of the column Channel Chromatography A longitudinal void in the column that causes band spreading Channeling usually occurs near the column walls Chromatogram A record of a separation that indicates the detector response as a function of time The chromatogram indicates the elution of electroactive species from the column Chromatography The separation of compounds in a mixture that is effected by exploiting the relative differences between their adsorbtion on a solid phase and their desorbtion into the mobile phase Chromatography involves the use of a dynamic equilibrium of the compounds between the stationary phase and the mobile phase Collapse The settling of the packing material in a column that leads to a column void Column A cylinder that is used to contain the stationary phase The mobile phase is pumped through the column to chromatograph the sample Column Void A portion of the column that is not packed with stationary phase A column void may be due to settling of the stationary phase or dissolution of the stationary phase by the mobile phase Column Volume The volume within a co
169. oi oerien a a a ee ewan TAE EREE p a a 5 2 52l OVE W ense irene a AR a A T E E a E 5 2 5 2 2 Daily Maintenance ssssssssssssssseeeessesesssrrrrttrrrrrrrnnnnsrrnrnnnnnusnnstntnennnneeeeene 5 3 5 2 3 Weekly Maintenance ices cccvss cabs cincsndaseerededeetenenatensndessuhinaeactedeedsPeedeveukenenecrens 5 3 5 2 4 Monthly MGIMIGMANC C22 icant etn ice ene tntdteeealteues pa Pesianc cpbew camncdecadestedeesemiadie 5 4 5 2 5 Quarterly Mamierrance sc ices tc cas ones eee weeh sad oi das sed canese a pactanasneenaveateeneaec 5 4 5 2 6 Additional Periodic Maintenance cccccceeceeeeseeececeeeeeeeeseseseeeeeneeeeeeeeees 5 4 5 2 7 Nitrogen Generator sx 5 en ccas8 ete ewe tacee ces cece thudiotaay cave dadieaadevearasactncereedeneweiad 5 5 5 3 Customer Service of the Detector xt acadetectcohered center tivd aoslse hand heeeneee 5 5 5 3 1 In line Fiter Elements sacasioetecslcsidaasts saree snmvca ts aoeaterbon hones 5 5 5 3 1 1 Frequency of Changing Filter Element 0 ccececeeeeeeeeeeeeeeees 5 5 5 3 1 2 Checking the Pressure Drop across the Filter c cc eeeceeeeeeees 5 6 53 13 Chan ing a PING ace iaer ates cael nat certs aE aae nad a tants eaaa aaRS 5 6 Corona CAD and Corona S CAD Detectors Xv Operating and Maintenance Manual 5 3 2 Changing the Line Fuses ai cei sie is ak Ae ie cectn ee erec hed k gateentemhe 5 7 5 3 3 Replacing the Gas Filters ee eeccceeeeeeeeeeeeeeeeeeeeeeeeaaeeeeeeesaaeee
170. onic fault is found during the self test an error message will be presented on the display see Chapter 5 for an explanation of the error code and possible remedies Upon successful completion of the self test the initial display will appear as shown in Figure 3 2 CORONA Version x xxx Copyright 2004 NEXT Figure 3 2 The Version Copyright Screen 3 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware After several seconds the instrument will automatically advance to the MAIN CORONA MENU Figure 3 3 The temperature will only be indicated if the Thermal Organizer is connected to the detector module Figure 3 4 MAIN CORONA MENU Select desired operation SYSTEM EDIT RUN Figure 3 3 The Main Corona Menu Screen no Thermal Organizer T 23 C MAIN CORONA MENU Select desired operation SYSTEM EDIT RUN Figure 3 4 The Main Corona Menu Screen with Thermal Organizer eS NOTE The role of the four keys immediately below the screen is dependent upon the screen that is presented These keys are termed soft keys since the definition of these keys is context sensitive The soft keys presented in Figures 3 3 and 3 4 are used as follows e SYSTEM accesses a series of screens that are used to select a variety of general operating parameters such as defining default values setting time date
171. ons are made to the Corona detector Make connections in the following order 2 4 1 1 Locating the dectector Position the detector in a location so that the distance between the end of the HPLC column and the detector inlet are minimized Autosampler Corona l ee E III O Ibo O IESE NOTE By minimizing the column to detector distance post column band broadening effects can be minimized and optimal chromatographic resolution can be achieved 2 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation 2 4 1 2 Power Source Connect the power cord to the back of the system Figure 2 2 and into a grounded outlet Fuse Power Line On Off Switch Figure 2 2 Rear Panel Corona CAD Detector NOTE The detector must be connected to an electrical outlet that shares a common ground with other components of the chromatographic system e g pump autosampler computers etc This will avoid ground loops which can create erratic results e g varying background high noise spiking etc Use of a common power strip is highly recommended to ensure a common ground Corona CAD and Corona S CAD Detectors 2 7 Operating and Maintenance Manual Chapter 2 2 4 1 3 Gas Source a Connect the nitrogen source to the gas inlet of the ESA Gas Conditioning Module P N 70 8285 using the quick conn
172. ontact ESA Biosciences or its representative To assist the service engineer please fax the worksheet to ESA Biosciences Service Department Fax 978 250 7092 or your local representative to describe the problem and assist in the resolution of the problem Please retain a copy of the filled in worksheet for future reference If the detector requires service this document will provide critical information as to the original installed state of the detector NOTE ESA Biosciences has developed a Validation Protocol which can be used to demonstrate that the Corona detector and related HPLC equipment is operating in an acceptable manner This protocol includes Installation and Operational Qualifications and is performed by a factory trained service engineer For details please contact ESA Biosciences or your local ESA Biosciences representative for pricing and scheduling 2 5 2 Setup Create a set of methods using your HPLC Application to inject 10ul of each of the following standards included in the installation kit The sequence of the methods should be as follows 1x 500 g mL Used for initial system conditioning 3x water blank 3x 500 g mL 3x 25 ug mL Load the standards into the autosampler to allow them to come to equilibrium with the temperature setting for this instrument 2 16 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation Note a typical method has the following
173. or tubing Part Number 70 5678 It should be mounted to the underneath side of one of the cell brackets The overflow lines vent tubes from the injector valve should be positioned through the small holes so that any overflow can drip into a Weigh Boat Part Number 40 0172 that is placed on the organizer chassis plate The overflow bracket is designed to keep the outlet of the vent tubes at the same vertical height as that of the needle port to prevent siphoning of the sample Connect the brackets to the left side of the chassis by the lip that fits into the horizontal slit and the knurled screw that fits into the right side of the chassis Figure C 7 The plastic knurled screws Part Number 70 5571 should be threaded into the proper hole over a cell and secured finger tight Figure C 7 Cell Bracket C 8 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer C 3 8 Final Assembly After you have made all connections a b Place the chassis plate into the organizer tray so that the three threaded studs protrude through the bottom of the chassis plate Connect the Ground Strap Assembly Part Number 70 5625 to the protruding threaded stud at the left rear corner of the chassis plate see Figure C 1 Attach the three knurled thumbnuts which were removed in Section 5 3 2 and finger tighten Mount the organizer module on top of the Corona Detector by inserting the
174. or 30 minutes 3 26 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual The Corona CAD Firmware 3 7 How to Create a Timeline Method The following example instructs the user on how to create a timeline method It was used to generate data for the Impurity Testing 22 Application Note Part Number 70 6716 see Figure 3 49 Here following an autozero the gain range is automatically changed from 1pA to 500pA at 1 50 mins before being set back to 1pA at 3 90 mins ready for the next run This method is repeated for 10 samples 1 0 re Range 1pA Range 500pA 0 6 Response Solvent Void 0 4 100g on column Acetaminophen 0 2 5ng on column Glucose 0 0 0 0 0 4 0 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 Minutes Figure 3 49 Changing Gain Range During a Timeline Method Typically in the timeline mode the user has to select four parameters 1 Analytical run time 2 Number of samples to be analyzed e g X loop will be X 1 3 Event e g gain range change 4 Time of event Procedure 1 Choose Edit Mode 2 Choose Mode CAD TL press enter to verify and then press Next 3 Choose Security On or Off user s choice then press Next 4 Defaults are as follows Range 100pA Filter None Offset 0 5 In this example the method calls for an initial range of 1pA Choose 1pA Press 5 Enter to verify Press Next Corona CAD and Corona
175. or 50 95 F might lead to artifacts or other interference Therefore the temperature of the laboratory in which the Corona CAD detector is used should be controlled The laboratory temperature should be within the specified operating temperature range and temperature fluctuations should be kept at a minimum Avoid placing the instrument in drafts such as near an air conditioning or heating duct the heat output of a refrigerator or an open window and in direct sunlight Corona CAD and Corona S CAD Detectors ix Operating and Maintenance Manual E If the Optional Thermal Organizer Module is employed to Control the Temperature Allow Sufficient Time for Thermal Equilibrium The time required to reach thermal equilibrium depends on the selected temperature and the ambient temperature The user should ensure that thermal equilibrium has occurred before performing analytical separations F With the Corona S CAD Detector Allow Sufficient Time for the Nebulizer to Reach Thermal Equilibrium The Corna S CAD detector requires approximately 30 minutes for the nebulizer block to reach thermal equilibrium The user should ensure that thermal equilibrium has occurred before performing analytical separations CERTIFICATIONS The Corona CAD and Corona S CAD Detectors have the following certifications USA UL 61010A 1 1 Edition Canada CSA Standard C22 2 No 1010 1 92 EU EN 61326 1997 A1 1998 EN 61010 1 2001 02 FCC Part
176. r A NOTE If the unit does not go through the normal power up procedure assistance 5 26 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 5 12 1 Maintenance and Troubleshooting Erratic Noisy Baseline Possible Cause Comments Recommended Solution Pump problem Check pump seals check valves for wear or leaks Replace seals if worn Replace check valves if necessary Leaks in the system Check for leaks in system Tighten all fittings Detector output voltage not matched to the recorder Review installation procedure Set voltages correctly Mobile phase not properly mixed Stir mobile phase Pulse damper membrane rupture Check for leaks Use a pulse damper known to be good Replace membrane or pulse damper Contaminants eluting from column Trace levels of organic compounds may be tightly retained by the column Remove column and see if problem exists Clean the column and or replace System not grounded All components of the system must be connected to acommon ground Check AC line receptacle verify that you have a true ground Ensure that all HPLC components share the same ground Waste overflowed and waste has re entered the detector May cause permanent damage to electrometer assembly Empty waste bottle Operate instrument without mobile phase pump off and with gas flowing gas on for
177. r Section 5 4 gt Establishing a system log Section 5 5 e Troubleshooting Related Topics General troubleshooting approach Section 5 6 Detector related issues Section 5 7 HPLC related issues Section 5 8 Standard and sample related issues Section 5 9 Thermal organizer issues Section 5 10 Noise review Section 5 11 General troubleshooting guidelines Section 5 12 Logic board and power supply board replacement Section 5 13 VVVVVVVV eS NOTE When the detector is initially installed its performance is verified by flow injection analysis Chapter 2 Section 2 7 ESA Biosciences Corona CAD detector Test Worksheet In Chapter 4 a caffeine chromatogram is generated using the caffeine standard and guard column Section 4 7 These data serve as a benchmark for detector performance and can be used to compare the performance of the system at some time in the future Similarly if problems are observed in the use of a specific analytical procedure these data can be used to ensure that the chromatographic system is functioning properly The user is encouraged to maintain a log of all operations of the detector maintenance activities and all observed problems should be entered into the log A discussion of the log is provided in Section 5 5 Corona CAD and Corona S CAD Detectors 5 1 Operating and Maintenance Manual Chapter 5 5 2 Detector Maintenance This section reviews the maintenance that should
178. re use c Use dedicated glassware Do not clean in soap detergents d Clean 500mL graduated measuring cylinder with 200mL Type 1 108 Mohm cm Make sure internal surface the length of the cylinder is exposed to the water Drain Repeat procedure twice more Drain If not used immediately cover with aluminum foil until use e Clean 2000mL graduated measuring cylinder with 200mL Type 1 1078 Mohm cm Make sure internal surface the length of the cylinder is exposed to the water Drain Repeat the procedure twice more Drain If not used immediately cover with aluminum foil until use f Clean empty 2000mL waste bottle with 200mL Type 1 108 Mohm cm Make sure internal surface the length of the bottle is exposed to the water Drain Repeat twice more Drain Use immediately do not store Corona CAD and Corona S CAD Detectors 2 29 Operating and Maintenance Manual Chapter 2 Measure 400mL Ultra pure HPLC grade Methanol using the 500mL cleaned measuring cylinder and add to uncapped cleaned solvent bottle Record lot number on label and lab note book Measure 1600mL Type 1 108 Mohm cm water using the 2000mL graduated measuring cylinder and add to uncapped cleaned solvent bottle g h x i Cap bottle and invert several times to mix j Place solvent bottle in ultrasonic bath 2 11 3 Degassing Mobile Phase a Setup degassing apparatus as shown in Figure 2 18 Ultrasonic Bath Figure
179. response of the Corona detector for semi volatile compounds is determined by the vapor pressure of the compound of interest An effected semi volatile compound will give a diminished response relative to a non volatile compound when analyzed using flow injection analysis Chromatographically an effected semi volatile compound will show a diminished response Figure 5 9 shows the response of the Corona detector to a number of fatty acids and fatty acid methyl esters The chromatogram illustrates reduced responses for a number of semi volatile analytes including short chain fatty acids and medium chain fatty acid ethyl esters 5 22 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting Palmitic Acid Stearic Response Acid a Palmitic Ethyl Ester Myristic Acid Nonadecanoic Ethyl Ester Lauric Acid nad 3 os 10 15 20 25 30 35 an 45 so Minutes re Figure 5 9 Analysis of Different Fatty Acids and Fatty Acid Ethyl Esters Diminished Response due to Semi Volatility Samples must be processed appropriately and filtered e g through a 0 2 um nylon membrane prior to analysis The analyst must ensure that the final sample extract is soluble not only in the sample diluent but also in the mobile phase being used in order to avoid precipitation The Corona detector is a universal detector for most non volatile analytes so
180. riations in the background noise may arise from a number of sources such as the pump column or detector Band A zone on an HPLC column that contains one or more component s of a mixture that is being separated Band Broadening The phenomenon of diffusion of a band into a greater volume e g post column band broadening occurs in the tubing between the end of the column and the detector Band Spreading See band broadening Baseline Resolution Chromatographic separation of two compounds such that the peaks are totally isolated from each other The tail from the first peak intersects the baseline before the second peak begins Corona CAD and Corona S CAD Detectors G 1 Operating and Maintenance Manual Glossary Baud Rate The rate of transmission of data through the RS232 communications interface from the detector to the computer and vice versa Bed A collection of a chromatographic media usually silica that is used to separate a mixture In HPLC this is usually termed the stationary phase Bonded Phase A stationary phase in which a functional group is chemically bound to the stationary phase e g 5um silica The functional group can be polar e g CN or non polar e g Cis Calibration Curve A plot of the detector response vs concentration of a series of known samples standards It is used to determine the concentration of the analyte in unknowns Capacity Factor The capacity factor is the ratio
181. rols Specifications can be found in Appendix A 1 4 1 The Detector Module Includes The User Interface Power Module Logic Module Nebulizer Drying Tube Corona Chamber Electrometer The detector user interface includes a display panel anda keypad Figure 1 2 The display shows information from the system to the user It provides the operational status of the system assists the user in setting up and editing analytical methods and displays stored methods The keypad is used to enter information from the user to the system and is used for establishing and editing methods The Power Module provides the appropriate power to the various components in the system and the Thermal Organizer if installed Figure 1 4 The Logic Module contains the microprocessor and associated electronics Figure 1 4 The input output connections and the remote ports are present on the Logic Module Analytical methods can be stored in memory i e are saved if the power is turned off and can be modified or deleted as desired Converts the eluent from the HPLC column to an aerosol Removes the solvent molecules from the aerosol to form particles Charges the solute particles Accumulates and measures the charge that has been placed on the solute molecules as they pass through the detector Gas Module Contains the gas filters externally and houses the gas plumbing internally Figure 1 4 Corona CAD and Corona S CA
182. romatographic output that the detector has observed the presence of a compound in the mobile phase The pen response will be above the baseline A peak has an approximately Gaussian shape Peak Area The area underneath a recorder trace and above the extrapolated baseline The peak area is proportional to the amount of material in the sample The peak area can be approximated by the equation A bh where b is the width of the peak at the base h is the maximum peak height Peak Height The measurement of the distance on a chromatogram from the baseline to its highest point for a peak For an EDD generated peak the peak height is usually given in terms of current amps Peak Width The breadth of the peak The peak width is frequently measured at the intensity that is equivalent to 0 5 of the peak maxima The peak width is usually expressed in minutes or seconds Plate In distillation theory a plate is the height required for a single vaporization and one condensation process It is a measure of the efficiency of the column This concept has been transported to HPLC where it is a measure of the efficiency of the column Power Module Component of the detector that provides appropriate power to the various components of the detector Pre column A short column that is placed before the analytical column to trap materials that may be deleterious to the analytical column or interfere with the separation detection Pulse Damper A device th
183. rs Operating and Maintenance Manual CHAPTER 3 _ THE CORONA CAD FIRMWARE 3 1 Introduction All commands for the control and operation of the Corona CAD and Corona s CAD Detectors may be entered via the keypad on the front panel Figure 3 1 which includes a 4 line by 40 character display panel In addition a number of indicator lights LED s adjacent to certain keys on the front panel are used to indicate that the corresponding feature is active e g the GAS ON OFF LED is lit when the gas valve is open and the LED between the A and buttons is lit when the buttons are active The detectors can also be controlled by EZChrom Elite for ESA chromatography data systems A flow chart of the Corona CAD detector firmware is presented in Appendix D CORONA RUN i 0 000pA Output 0 Range 100pA Filter None Offset 0 IMENU PREVIOUS NEXT ENTER AUTO 1 ZERO Figure 3 1 Front Panel of Control Module of ESA Biosciences Corona Detector There are two modes of operation for the Corona detector e Normal Operation All parameters are held constant during a separation e Timeline Based Operation A broad variety of operations can be performed at user specified times Corona CAD and Corona S CAD Detectors 3 1 Operating and Maintenance Manual Chapter 3 The TIMELINE mode feature allows the user to perform the following operations Changing the current range and filter settings Autozeroin
184. s either 7 or 8 bits G 10 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual INDEX A Action keys 3 4 Additional information 1 10 periodic maintenance 5 4 Analysis of data 4 9 Autozero screen 3 11 B Basic installation 2 6 C Cables B 1 Certifications x A 3 Changing a filter 5 6 the line fuses 5 7 Charging solid particles 1 9 Checking the pressure across the filter 5 5 Cleaning C 18 the detectors 5 9 Column 5 20 conditioning procedure 2 33 considerations 4 4 Components location 1 5 Contacts screen 3 23 Corona accessories B 1 firmware 3 1 organizer modules 1 4 replacement parts B 1 Creating a CAD TL method 3 8 Corona method 3 5 Customer service of the detector 5 5 D Daily maintenance 5 3 C 18 Data keys 3 4 Date amp time setup screen 3 21 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Degassing apparatus figure 2 30 mobile phase 2 30 Deleting methods screen 3 19 Description of the detector 1 3 rear panel 2 24 Detector maintenance 5 2 related issues 5 12 test procedure 5 31 Diagnostic parameters 3 17 Do s and don ts xi Drain and vent installation 2 9 E 2 E 7 Drain cap assembly barbed components E 6 compression components E 1 Drainage 5 15 Drain Vent assembly old styles E 1 E Edit 3 3 method select screen 3 5 parameters screen 3 7 security screen 3 6 Editing a CAD TL method 3 8 Corona method 3 5 Effects gradi
185. s must be degassed and mobile phase older than 5 days should be discarded d Clean measuring cylinders with Type 1 1018 Mohm cm water and leave inverted on peg board to dry Do not leave upright open to the air 2 11 5 Mobile Phase Qualification a b c d e Take mobile phase to the HPLC Corona mobile phase test system Place pump inlet line into freshly degassed mobile phase x Open valve on front of pump and purge When purging has stopped automatically close valve and pump at 1 0mL min x Wait 30 minutes for system to stabilize f View Corona response using the software preview screen to ensure baseline is stable reaches a plateau Q Record the background current shown on Corona display panel in the log book If background current is lt or 0 150pA then the mobile phase is usable If background is gt 0 150pA the mobile phase may be contaminated and should be discarded ey Ie NOTE Background current results may vary depending on the state of the HPLC system The background values provided are used internally by ESA on maintained HPLC Systems 2 32 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation 2 12 Column Conditioning Procedure WARNING DO NOT use a column in the system unless it has been conditioned using the following procedure Unconditioned columns will cause erroneous results 2 12 1 Supplies HPLC Grade M
186. s of magnitude dynamic range nonlinear Low ng lower limits of detection Typically lt 2 R S D response variability Response magnitude is minimally dependent on the analyte s chemical properties Broad applicability Isocratic and gradient compatibility 1 3 Overview of this Manual The following topics are covered in this manual Installation Chapter 2 how the user incorporates the detector into the HPLC system Detector Firmware Chapter 3 a step by step discussion of how the user establishes an operating program and prepares the detector for use in the HPLC system Using the Detector in an HPLC System Chapter 4 running the detector as a part of an HPLC system Maintenance and Troubleshooting Chapter 5 includes a series of operations that the user should perform on a routine basis to optimize the performance of the detector and the activities that the operator should use to determine the cause of problems A series of appendices are also included that cover a number of topics including accessories spare parts specifications and related information and installation of the organizer and thermal organizer modules 1 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Introduction 1 4 Description of the Detector The ESA Biosciences Corona and Corona detectors are self contained units Figure 1 1 and include the electronics detector assembly and all cont
187. s short as possible Connect the tubing from the outlet of the guard column holder to the inlet of the 0 5 um stainless steel filter assembly P N 70 4538 Then connect the outlet of the filter assembly to the inlet of the detector Figure 2 10 Ie NOTE Try to minimize the column to detector distance so that post column band broadening effects can be minimized and optimal chromatographic resolution can be achieved WARNING If using TFA based mobile phase is to be used stainless steel tubing must be used between the column filter and detector A CAUTION Do not start the flow of mobile phase at this time Figure 2 10 HPLC Inlet 2 14 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation 2 4 1 7 USB Connections If the HPLC system is using either the Agilent Chemstation or EzChrom Elite Application connect the detector to the workstation using the provided standard USB cable Type A B P N 70 5713 Once the connection is made install the appropriate drivers using the CD s provided Agilent For use with Agilent ChemStation Version B 02 01 SR1 or later using ESA ChemStation Driver P N 70 7102 See driver technical note for Versions B 03 of higher ESA Elite For use with EZChrom Elite for ESA Data Systems Version 3 2 or later using ESA EZChrome Elite Driver included with the application installation media 2 4 1 8 Other Connections If an external
188. sations in the baseline e g see the chromatogram presented in Figure 5 2 and cause flooding ADET CHANNEL A Figure 5 2 Example of Improper Drainage NOTE the periodicity of spiking will be related to slugs of waste solvent leaving the detector Corona CAD and Corona S CAD Detectors 5 15 Operating and Maintenance Manual Chapter 5 5 7 4 Gain Range A given gain range has a finite mass range In order to achieve the highest mass sensitivity the 100pA gain range is typically chosen However although this works well for many applications the measurement of very large masses may overwhelm this gain range and produce aberrant data e g Figure 5 3 This issue can readily be observed when plotting standard mass curves the response may actually starts to decrease as the mass on column increases If this is found to be the case the large mass measurements must be conducted at a less sensitive gain setting i J 0 8 Truncated Response Response J 0 00 0 25 0 50 0 75 1 00 1 25 1 50 1 75 200 225 250 275 3 00 3 25 350 3 75 4 00 Minutes Figure 5 3 Overwhelming a Gain Range 5 16 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Maintenance and Troubleshooting Figure 5 4 Two Examples of Grounding Issues 5 7 5 Grounding Issues Inappropriate grounding of the detector can result in excessive noise
189. se errors cause the pump to stop unexpectedly This contact closure can be connected to the GAS OFF input on the Corona detector so that if an error is registered in the pump the detector can shut off the gas flow to save gas See the manufacturer s User s Manual for the procedure to make this electrical connection to the Corona Detector 2 28 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Installation 2 11 Mobile Phase Formulation WARNING Improper or contaminated mobile phase can result in high background readings as well as extraneous noise Take great care in creating mobile phase and follow the exact steps outilined below 2 11 1 Supplies 500mL Glass graduated measuring cylinder 2000mL 2L Glass graduated measuring cylinder Empty 2000mL 2L glass solvent bottle and cap Type 1 108 Mohm cm water Omnisolve HR GC Ultra pure HPLC grade Methanol EMD MX0480 6 Parafilm Vacuum Pump Ultrasonic bath Stopper with hole Rubber tubing Qualified HPLC Corona test system with guard column Label Lab Safety part 22886 Laboratory note log book 2 11 2 Procedure 2 11 2 1 Mobile Phase Preparation a All glassware must be properly cleaned prior to use in order to remove any particles and extraneous solutes from the mobile phase preparation oO Only use Type 1 108 Mohm cm water Make sure that you let the first 500mL 30 seconds of water drain to waste befo
190. seeeeeeeeeeeeeees 2 20 Corona CAD and Corona S CAD Detectors xiii Operating and Maintenance Manual 2 7 2 8 2 9 2 10 2 11 2 12 General Operations sett o o ee caereehce adh a oo aka comet erahved haan 2 21 2 7 1 Turning off the Corona Detector cceccceeceeeeeeeeeeeeeeeeneeeeeeeeeeeeeennaeeeeeee 2 21 2 7 2 Emptying the Waste Bottle icc cz cceedecped savedosenceaieesonunneeneseecsycaaniocesedtacteats 2 21 2 7 3 Storing the Corona Detector v 04 cacehers ceeiess Weleda ace oder eenchsectnds ie madaes 2 22 Other Detector Configurations xis sctectncersgeiaadench aceite hw Reade edie eae Sones a 2 22 2 8 1 Using the Corona in Series with other Detector cccccccsseeseseeeeeeeeees 2 22 2 8 2 Using the Corona in Parallel with another Detector ceceeeeeeeeees 2 22 Setting up the Thermal OrganiZer ccccceeeesscceeeeeeeeeeeeeeceaeeeeeesaeeeseeeeeeeeeeeeeeaeeee 2 23 Making Electrical CONMections cececcceeeeeseeeeeeeneeeeeeeeeeeeeeeeeeeeeeeeeaaaeeeeeenaeaeeeea 2 24 2 10 1 Description of the Rear Panel of the Detector ccc eeeeeeeteeeeeeeeeeeees 2 24 2 10 2 Interfacing to the ESA Model 584 Solvent Delivery Module 20 2 27 2 10 3 Interfacing to other Solvent Delivery MOdUIES ccceeecteteeeeeeeteeeeeeee 2 28 Mobile Phase Formulation 20 sescetescetsdectedcdcenne niet nt incited lees addhsacnsmendedateueetinwetdes 2 29 Foz Wt UO Mer eee
191. several hours Electronics problem in detector unit Perform detector test Sec 2 5 2 If problem persists contact Service at 800 275 0102 Column temperature not held constant Temperature variation potentially due to problem in organizer or temperature change in laboratory Maintain column at constant temperature If the Thermal Organizer is included in the system make sure that it is operating properly see Chapter 5 See comments in next section Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 5 27 Chapter 5 5 12 2 High Background Currents This section describes situations where the background current has noticeably increased in a short period of time Possible Cause Comments Recommended Solution Impurities in the mobile phase See Notes a b To determine the source of potential impurities use another source for each component of the mobile phase on a sequential basis Reduce the potential if possible Highly retained compounds being eluted from the column This may occur when a new mobile phase or column is used Allow the system to equilibrate for an hour with the new mobile phase or until the baseline is stable and check the signal again Remove column and re establish flow If signal drops clean or replace column Leaking fitting Check for wetness around fittings and tighten fittings
192. shown in Figure 3 42 is presented SYSTEM SETUP MENU Select Contacts Set Contacts amp monitor inputs CANCEL NEXT Figure 3 42 The Contacts Screen If NEXT is selected the SET CONTACTS and VIEW INPUTS Screen Figure 3 43 will be presented SET CONTACTS amp VIEW INPUTS C1 OFF C2 OFF C3 OFF C4 OFF PMP OFF Gas off OFF Autozero OFF Start OFF CANCEL MENU Figure 3 43 The Set Contacts and View Inputs Screen The C1 C4 entries describe the status of the four contact closures The status of each contact closure on to off or vice versa can be changed by pressing ENTER moving the cursor to the desired contact closure pressing the A or W keys and finally pressing the ENTER key If the PMP parameter is set to ON a signal contact closure will be sent to an external device g a pump in order to stop the flow of mobile phase to the system in the event the pressure of nebulizer gas is lt 50 of the nominal gas flow required NOTE This signal can be used to protect the detector from possible damage in the event that the gas flow is stopped or compromised but the mobile phase is still being delivered Corona CAD and Corona S CAD Detectors 3 23 Operating and Maintenance Manual Chapter 3 Changing the initial status of the contact closure outputs may become necessary if the Corona detector is connected to other devices that need to be
193. sired operation SYSTEM EDIT RUN Figure 3 23 The Main Corona Menu Screen c Press the RUN button to present the RUN METHOD SELECT screen Figure 3 24 RUN METHOD SELECT Method 1 Name Default Method Mode CAD CANCEL NEXT Figure 3 24 The Run Method Select Screen d Select the desired method using the A or Y keys and press NEXT e The parameters for that method will be loaded and the first status screen will be presented Figure 3 25 The parameters are summarized in Table 3 4 CORONA RUN i 0 000 pA Output 0 Range 10 pA Filter None Offset 0 MENU PREVIOUS NEXT Figure 3 25 The Corona CAD Run Screen 1 For the Corona detector the Corona run screen will appear as in Figure 3 26 Neb T XX where XX is ON or OFF Use of the nebulizer heater is discussed in Section 3 6 11 CORONA RUN i 0 000 pA Output 0 Neb T XX Range 10 pA Filter None Offset 0 MENU PREVIOUS NEXT Figure 3 26 The Corona CAD Run Screen 1 Corona CAD and Corona S CAD Detectors 3 15 Operating and Maintenance Manual Chapter 3 Table 3 4 Run Parameters Parameter Description Range Default Type i Current 0 pA to 500 pA Output of full scale display Output of signal output sent to 100 to 100 0 Output recording device Current Used to set the Range sensitivity level of the 1 e LAR 100 pA Input R d
194. style B 3 Cables Cables Part Number BNC to Bare Wire Cable Signal out to recording device 70 1776 BNC Male to Double Binding Post Adapter 70 0219 Bare Wire to Bare Wire I O Connections 70 4850 Cable RS232 Corona to Computer 70 1743 Cable Interface Thermal Organizer to Corona 70 5595 Corona CAD and Corona S CAD Detectors B 1 Operating and Maintenance Manual Appendix B B 4 Manuals Part Number Operating and Maintenance Manual 70 6258 B 5 Fuses Part Number Fuse 1 0 amp for 100 240 V operation 2 fuses required 70 0751 B 6 Organizer Module Spare Parts Organizer Module Spare Parts Part Number Corona Organizer Accessory Kit Table C 1 70 5651 B 7 Thermal Organizer Module Spare Parts Thermal Organizer Module Spare Parts Part Number Corona Thermal Organizer Accessory Kit 70 5651T Standard Column Clamp Assembly 15 cm 70 5366 Short Column Clamp Assembly 5 cm 70 5365 Long Column Clamp Assembly 25 cm 70 5367 Cable Interface Organizer to Corona 70 5595A B 8 Test Standards and Guard Column Test Standards and Guard Column Part Number Caffeine Standards Set 1 8 mL 70 6565 Guard Column 70 7098 B 9 Nitrogen Gas Supply Nitrogen Gas Supply Maintenance Kit Nitrogen Generator 70 6230 Filter PVC 0 5um 47mm 5 pk for Gas Filter Holder 70 4507 B 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual APPENDIX C ORGANIZER AND THERMAL ORGA
195. syringe has enough wash solution for the day s analyses f Use fresh filtered degassed mobile phase Check that the solvent filter in the mobile phase reservoir is clean Replace the solvent filter if it is discolored g Perform a test run using a standard or a well defined sample and ensure that the background currents and peak heights areas have not changed appreciably from day to day h Monitor the pump pressure and the temperature of the Thermal Organizer Module if equipped and ensure that they have not changed significantly from expected values 5 2 3 Weekly Maintenance On a weekly basis a Replace the pump washing solution b Perform a flow rate check on the pump c Check all electrical connections to ensure that they are properly seated d Perform all of the daily activities Corona CAD and Corona S CAD Detectors 5 3 Operating and Maintenance Manual Chapter 5 5 2 4 Monthly Maintenance On a monthly basis a Replace the filter elements in any in line filters see Section 5 3 1 b Inspect the PEEK tubing to detect potential problems and replace if necessary c Perform all of the daily and weekly activities 5 2 5 Quarterly Maintenance On a quarterly basis a Inspect and change seals check valves and pistons in the pump if necessary b Replace the mobile phase filter c Replace the filter element between the column and the detector d Inspect and replace waste bottle ffkm sealing gasket
196. t in the chromatogram where the response is expected to be reasonably flat i e 15 secs prior to the elution of a peak When the autozero is triggered via the terminal block connection an event mark can be provided after the output is zeroed 2 10 2 Interfacing to the ESA Biosciences Model 584 Solvent Delivery Module The ESA Biosciences Model 584 Solvent Delivery Module has the capability of providing a contact closure in the event that it experiences an error Types of errors registered by the pump include e g mechanical or electrical problem the back pressure of the HPLC system is outside the range of the upper or lower set pressure limits the pump runs out of mobile phase etc these errors cause the pump to stop unexpectedly This contact closure can be connected to the GAS OFF input on the Corona detector so that if an error is registered in the pump the detector can shut off the gas flow to save gas 3 NOTE If using a pump other than the ESA model 584 solvent delivery module check with the vendor that their pump is capable of providing such contact closures To setup and activate this function in the Model 584 pump it is necessary to connect the pump to the detector using one of the connection cables provided and set the external events function on the pump To connect the pump to the detector a Insert the event cable plug into the pump EXT CONTROL connector see Model 584 Operators Manual b Connect the orange and
197. t key and then pressing the Enter button to make the cursor appear in the test field Use the Up and Down arrows to scroll through the self test results In general if a fault message appears on the display other than an error 80 or a error combination containing error 80 then turn the detector off wait for a minute or two and turn the system on again If the fault message re appears call the ESA Service Department at 800 275 0102 or your ESA Representative Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual 5 13 Chapter 5 Please have the serial number of the instrument the full text of the error message and the software version number available See Table 5 4 for related Operation Errors 5 7 2 Gas Operation errors related to gas issues are presented in Table 5 4 Such errors can be caused by the gas supply under or over pressure the Corona detector and whether the waste drain system is effectively sealed Table 5 4 Operation Errors Causes and Options Error Message Possible Cause Troubleshooting Corona Service High voltage approaching maximum level Turn off and on If detector still fails contact ESA Service Corona Failing High voltage unable to maintain current at 1 0uA Turn off and on If detector still fails contact ESA Service Low ion trap voltage lon trap voltage too low Turn off and on If detector still fails contact ESA Ser
198. t specified by ESA Biosciences the protection provided by the equipment may be impaired Corona CAD and Corona S CAD Detectors vii Operating and Maintenance Manual IMPORTANT OPERATING CONSIDERATIONS This manual contains detailed information about the use of the Corona CAD and Corona S CAD detectors As a convenience to the user a number of points are presented below to assist in optimizing the use of the detector protecting the detector and or minimizing the possibility of erroneous results A Use a Line Voltage Conditioning Device If the input power to the detector is noisy i e fluctuations in the line voltage from the power source such as spikes sags dropouts brownouts etc interruptions in the normal operation of the instrument may be observed Such interruptions may cause the unit to reset this will stop the unit shut off the gas flow and return the display to the initial screen or greeting screen While the detector includes internal AC power filters excessive fluctuations and brownouts may still create problems If line voltage fluctuations or brownouts are a common occurrence in your laboratory or if you would simply like to ensure that power fluctuations do not lead to improper operation loss of analytical methods or default storage parameters the use of a line voltage conditioner uninterruptible power supply UPS or power surge protector with power sag protection is recommended
199. that Gas ON OFF LED is lit CAUTION Make certain that the waste bottle is emptied on a routine AN basis It should not be allowed to get full Failure to do this may result in flooding of the detector Severe or prolonged flooding may damage the detector and void the warranty Corona CAD and Corona S CAD Detectors 2 21 Operating and Maintenance Manual Chapter 2 2 7 3 Storing The Corona Detector If the detector is not to be used for a few days it should remain connected to the HPLC system Keep the gas and mobile phase flows off Remember to check the level of fluid in the waste bottle and empty if needed If the detector is not to be used for extended periods then it is probably best to remove it from the HPLC system Before disconnecting the Corona detector flush it with an appropriate solvent i e devoid of salts that is compatible with the mobile phase Typical mobile phase used for this operation is described in section 0 WARNING DO NOT leave highly acidic or basic mobile phases in the detector as long term sedentary exposure can cause damage to the detector Turn off gas source first and allow the gas to bleed off Then undo the gas quick connect fittings using the tool provided to push in the collet prior to removing the fitting 2 8 Other Detector Configurations 2 8 1 Using the Corona Detector in Series with another Detector The Corona detector can be used in series with other detectors e g UV
200. that defines the operation of the Corona Detector Corona CAD and Corona S CAD Detectors G 5 Operating and Maintenance Manual Glossary Microampere A unit of current equal to 1078 amps abbreviated pA Microgram A unit of mass equal to 10 8 gram abbreviated ug Micromole Amount of material equal to 10 mole abbreviated umole Milliampere A unit of current equal to 1073 amps abbreviated mA Milligram A unit of mass equal to 1073 grams abbreviated mg Millimole Amount of material equal to 1073 moles abbreviated mmole Millivolt A unit of potential equal to 10 3 volts abbreviated mV Mobile Phase The liquid that is used to transport a sample mixture through a chromatographic bed Often referred to as the eluent N Number of theoretical plates for a column as measured by the efficiency of a column N can be calculated by the equation N 16 tr tw where tr is the retention time in seconds tw is the baseline width of the peak in seconds n Number of electrons transferred per molecule or ion in a reduction or oxidation Nanoampere A unit of current equal to 10 9 ampere abbreviated nA Nanogram A unit of mass equal to 10 9 gram abbreviated ng Nanomole A unit of mass equal to 1079 mole abbreviated nmole Nebulization The process of reducing the eluant into a fine mist or spray of liquid droplets The solvent in the mist is then evaporated to provide particles Nebulizer Device that conver
201. the nebulizer e g aqueous THF gradients see Figure 5 8 should only be used with the Corona detector e Degas prior to use Degas on line if necessary This will prevent bubbles from forming in the mobile phase and reduce the chance for pump related issues see above Corona CAD and Corona S CAD Detectors 5 21 Operating and Maintenance Manual Chapter 5 ADC1 CHANNEL A oS Ss 5 2 5385 2 Heat turned OWN Figure 5 8 Aqueous THF Gradients will cause the Nebulizer to Freeze and cause Excessive Noise Only use the Corona CAD detector for this and related mobile phases 5 9 Standard and Sample Related Issues The Corona detector has a dynamic range of four orders of magnitude and can be used to measure trace impurities at the lt 0 1 level With a limit of detection of 10ng on column this requires that the mass of the parent compound analyzed using the Corona detector to be about 100yg on column However not all compounds are soluble in the mobile phase at this concentration leading to precipitation of the analyte on column and in the detector typically manifested by increased system back pressure poor chromatography etc Although a compound may be soluble at high masses in the appropriate diluent e g DMSO the analyst must ensure that the analyte is soluble at high masses in the HPLC mobile phase The Corona detector can detect most non volatile and some semi volatile compounds The
202. the sealed drain assembly is properly connected to the Corona detector and that the waste bottle is empty 4 Review the maintenance section of this chapter Verify that any recent maintenance procedures were properly executed Corona CAD and Corona S CAD Detectors 5 11 Operating and Maintenance Manual Chapter 5 5 Review the troubleshooting section of this chapter For specific problems such as a noisy baseline refer to the appropriate troubleshooting diagnostic chart below eS NOTE If the problem is not corrected using the above approach then please notify ESA or your ESA Trained Service Representative for support You will be asked for a concise description of the problem the appearance of error messages and depending upon the problem information about the HPLC system analytical method sample type and sample preparation Other required information includes the detector serial number and firmware version number eS NOTE When the detector is initially installed its performance is verified Chapter 2 ESA Corona CAD detector Test Worksheet and a caffeine chromatogram is generated e g using the caffeine standard and guard column Chapter 4 Section 4 7 These data serve as a benchmark for detector performance Re validating the detector and re analyzing the caffeine standard may help expedite the troubleshooting process 5 7 Detector Related Issues These problems can be grouped into one of three categories
203. tic fields such as refrigerators fume hoods radio transmitter antennae NMR instruments etc C Avoid Exposure to Electrostatic Discharges The detector has been designed to be immune to electrostatic discharge ESD However strong electrostatic discharges to the instrument could cause a disturbance or artifact to the baseline or could interfere with the operation of the detector ESA strongly recommends that the detector is properly grounded to a good earth ground This should result in most ESD events being harmlessly conducted to ground and thus avoiding changes or damage to the sensitive electronics In addition we recommend that the operator take steps to reduce the generation of static electricity in the vicinity of the detector Measures to reduce the chance or severity of ESD include gt Increasing the relative humidity especially during the winter gt Ensuring that the operator touches a good earth ground e g a metal water pipe just prior to touching the detector gt Installing antistatic mats in front of the detector gt Avoiding the use of carpeting near the detector gt The use of low or antistatic lab coats D Avoid Large Temperature Fluctuations Large temperature fluctuations during a measurement can potentially lead to baseline disturbances or other interference with the operation of the detector Similarly operating the detector at temperatures outside of its intended operating temperature range 10 35 C
204. to 7 5 and always use volatile salts or buffers o Donotuse either ammonium carbonate or ammonium bicarbonate in the mobile phase o Reference section 2 11 Mobile Phase Formulation for the procedure to create the recommended detector mobile phase o Only use volatile mobile phases 2 2 2 Power Requirements includes a ground wire The unit must be connected to a properly grounded three prong power outlet to ensure safe and proper operation If there is any doubt about the power supply a qualified electrician should be contacted A WARNING The Corona detector uses a three prong power cord that The 120 VAC power cable consists of a 3 prong receptacle for attachment to the power inlet on the back of the detector unit and a three prong 15 amp plug configuration for connection to a standard U S grounded outlet The 120 VAC 50 60 Hz power cable consists of a 3 prong receptacle for attachment to the power inlet on the back of the detector unit The other end of the cable has three color coded wires that are used to attach to the appropriate plug The color coding of the wires meets ISO and VDE conventions as follows Earth Ground Green with Yellow Stripe Neutral Blue Line Brown The maximum power consumption of the unit is 100 VA Corona CAD and Corona S CAD Detectors 2 3 Operating and Maintenance Manual Chapter 2 WARNING The power plug should be installed by a qualified electrician and should be an approved plug
205. to peak concentration The negative trap is designed to remove the reagent gas ions The effect of the trap is not a function of analyte MW but of particle size Corona CAD and Corona CAD Detectors 1 13 Operating and Maintenance Manual Chapter 1 Question Why does the Corona CAD detector response increase as the organic increase Answer The dependence of Corona CAD detector s response on organic appears to be purely an effect of nebulization with an impactor and siphon There is a greater loss of analyte when eluted in aqueous solvent The effect appears to be very similar to that of ELSDs e g Alltech ELSD with impactor on Question What occurs when the charge is transferred to particles Is the charge only located at the external surface of the particles Or both inside and outside of the particle Answer It is rational to assume that the majority of charge is toward the outer edges of the particles Question Can the Corona CAD detector be used with SFC Answer We have not tried CAD with SFC at ESA Biosciences but we do have customers that use this approach Question Does the corona description mean that gas is floating in the form of a ring around the platinum wire What sort of high mobility particles are removed by the ion trap Where do the negatively charged particles go If you create a positive charge don t you create a negative charge as well Answer The Pt wire within a small chamber is poise
206. to promote proper drainage The assembly includes the Drain Tubing 3 16 ID x 1 4 OD x 6 long Part Number 70 7113 and the Vent Tubing 1 8 ID x OD x 6 long Part Number 70 6259 Figure E 1 Tygon Tubin yg g eee Olid Teflon Tubing Teflon Lining Vent Tube Drain Tube Figure E 1 Vent and Drain Tubing To attach the Drain Vent Tubing assembly a Connect the Vent tubing to the quick connect fitting located on the bottom front left portion of the detector This is a quick connect fitting which simply latches when the fitting is pushed in Figure E 3 b Connect the Drain tubing end with SS fitting to the compression fitting located on the bottom front left portion of the detector Ensure that the two ferrules inside the nut are properly oriented Figure E 2 The Drain fitting should be tightened with the supplied 9 16 wrench until it is snug Figure E 3 CAUTION Do no over tighten the drain fitting Equipment damage could result E 2 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Drain Vent Assembly Old Styles Figure E 3 Location of Fluidics Connections c Using the tubing cutter Part Number 70 7112 from the Drain Vent kit cut both the drain and vent tubing to a length between the detector and waste bottle where there are no dips kinks or loops Cut the drain tubing 4 longer than the vent tubing A vertical drop is optimum It may or may not be necessary t
207. ts the eluent from the HPLC column into a mist or fine spray of liquid droplets Noise The signal that appears on a chromatographic output due to a variety of events that are not a desired response of the detector Noise can be random or regular e g due to pump pulsations Offset The deflection of a recorder trace baseline so that two or more chromatograms can be overlaid on a single presentation Also the moving of a recorder trace baseline so that the output range of the detector recording device can be altered e g to display for negative dips from the baseline Organic Modifier An organic solvent e g methanol and acetonitrile that is used to alter the elution characteristics of an aqueous solution that is used as the mobile phase G 6 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Glossary Organizer A module that is designed to house the column pulse damper filters and related fluidics The thermal organizer module can be thermostated to maintain the separation at a user selected temperature Output The signal that the detector provides to the recorder or other device e g a data station Packing The stationary phase for HPLC Parity A method of providing a mechanism for the validity of the transmission of data via an RS232 interface Particle Size The average particle diameter of the column packing material usually in microns um Peak An indication on the ch
208. ttle Cap with Teflon Gasket Teflon Gasket Figure E 12 Bottle Cap and Gasket E 10 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual GLOSSARY This glossary provides definitions of a broad variety of terms that are commonly used in HPLC For further information the reader is referred to a standard text in HPLC such as L R Snyder and J J Kirkland Introduction to Modern Liquid Chromatography second edition Wiley Interscience John Wiley and Sons Inc New York 1979 or L R Snyder J J Kirkland and J L Glajch Practical HPLC Method Development 2 Edition Wiley Interscience New York 1997 a Separation factor A measure of the difference in retention of two compounds that takes into account the void volume of a column Q ko ky Vo Vo V1 Vo where k and k are the capacity factors for peaks 1 and 2 respectively V and V2 are the retention volumes for peaks 1 and 2 respectively Vo is the void volume for the column Artifact An unwanted peak in a chromatogram It may be due to sample mobile phase or instrumental contaminants Autozero An instrument function which allows the operator to establish the zero of an output recorder integrator etc so that all chromatograms start at the same baseline Background Noise The recorder or integrator signal that is observed when the mobile phase passing through the detector does not contain any species of interest Va
209. ure 4 7 Effects of Gradient Elution on Analyte Response Measurement of Oligosaccharides using a High to Low Organic Gradient 0 02 4 0 01 4 0 00 4 0 03 4 Mobile Phase A 0 1 TFA P Mobile Phase B 0 1 TFA in ACN T Serine h 11 2 Unknown 3 3 Glutamic Acid 00 4 Arginine a 5 Proline i 6 Valine Injection Volume 10l mares Colum lo MG C18 4 6 x 250 5um 7 Methionine Colum ambient 5 8 Isoleucine 9 Leucine Isocratic 10 Phenylalanine 11 Tryptophan 4 3 5 il Im it AN AY pene a WANI D3 ml MA ALU J ee 2 4 6 8 10 12 14 16 18 20 22 24 26 Minutes 28 Figure 4 8 Effects of Gradient Elution on Analyte Response Measurement of Amino Acids using a Low to High Organic Gradient 4 12 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual CHAPTER 5 MAINTENANCE AND TROUBLESHOOTING 5 1 Introduction Optimal detector performance will be maintained if the user performs a series of routine maintenance activities on a periodic basis Although this chapter primarily relates to the Corona CAD and Corona CAD Detectors optimal analytical performance requires that the complete HPLC system be well maintained This chapter provides information about e Maintenance Related Topics gt Detector maintenance Section 5 2 gt Customer service of the detector Section 5 3 gt Cleaning the detecto
210. urled nuts that attach the organizer chassis plate to the tray One nut is in the front center and the other two are in the corners at the rear of the organizer c Lift the organizer tray from the chassis plate C 4 3 Installing the Pulse Damper The pulse damper is mounted on the rear of the front left side of the chassis plate Figure C 10 using 2 of the 4 screws that are supplied with the pulse damper Mount damper here Figure C 10 Mounting the Pulse Damper I NOTE Do not attach the rubber feet to the bottom of the pulse damper when mounting it to the chassis plate Corona CAD and Corona S CAD Detectors C 11 Operating and Maintenance Manual Appendix C C 4 4 Installing a Rheodyne Manual Sample Injector Valve es NOTE It is recommended that you connect the appropriate tubing to the manual sample injector valve before installing it into the organizer To install the Manual Sample Injector a Place the Organizer Injector Panel with the hole Part Number 70 5229 onto the organizer chassis plate Figure C 11 Figure C 11 Locating the Organizer Injector Panel b Affix the panel to the organizer chassis plate with the two thumbnuts provided c Remove the handle from the injector valve using a hex key d Attach the injector to the chassis so that the valve stem goes through the hole in the Injector Panel and the part number of the injector is facing up Fasten the injector using the two screws and star
211. vice Gas Pressure UNDERpsi Loss of incoming gas from source Check that nitrogen gas is still being delivered to the detector Ensure that the incoming gas pressure is 35psi Flow Ratio Error Gas Flow Ratio is compromised Check gas source Check that waste bottle cap is secured correctly Check that bottle cap gasket is undamaged and sealing correctly Check that Drain Vent tubing is free from loops kinks and potential blockages and that all connections are secure If unit still fails then there may be a blockage within the Corona detector Contact ESA Service Low Gas Flow Ratio Low gas flow ratio error has continued for gt 60 seconds Pump Off contact sent via I O terminal Press any key to continue Check all Flow Ratio variables above If detector still fails contact ESA Service Low Total Gas Flow Total gas flow error has continued for gt 60 seconds Pump Off contact sent via I O terminal Press any key to continue Check all Flow Ratio variables above If detector still fails contact ESA Service High Gas Flow Ratio Gas flow ratio error has continued for gt 60 seconds Pump Off contact sent via I O terminal Press any key to continue Check all Flow Ratio variables above If detector still fails contact ESA Service High Gas Pressure Incoming gas pressure gt 45psi Adjust gas source pressure If detector still fails contact ESA Service 5 14 Coron
212. zer C 1 Self test screen 3 25 module spare parts B 2 Set contact screen 3 10 Other Setting connections 2 15 the filter 4 6 detector configurations 2 22 up the Corona nebulizer heater 2 15 Overview of the Corona detector 1 1 up the thermal organizer 2 23 Setup 2 16 P Shutting down the detector 4 3 Space requirements 2 4 Physical Spare parts B 1 location of the Corona 2 1 Specifications A 1 specifications A 2 Stages in detection 1 9 Power Standard and sample issues 5 22 requirements 2 3 Storing the Corona 2 22 source 2 7 Supplies 2 29 2 33 supply board replacement 5 32 Support from ESA 1 11 procedure 2 29 2 33 System 3 3 Proper output range 4 6 screens 3 18 Protocol 2 17 Pump noise 5 18 Q Quarterly maintenance 5 4 Corona CAD and Corona S CAD Detectors l 3 Index T Test protocol 2 16 standards B 2 Thermal organizer error codes 3 25 issues 5 23 setup screen 3 24 spare parts B 2 Time amp date setup screen 3 21 Timeline based operation 3 1 event options 3 9 screen 3 8 specifications A 2 timer location figure 2 31 Troubleshooting C 17 Turning off the Corona 2 21 on the Corona 4 1 U USB Connections 2 15 Unpacking the detector 2 5 organizer C 1 Using a method to collect data 3 15 the Corona in series w another detector 2 22 parallel w another detector 2 22 the detector in HPLC system 4 1 WwW Warnings iv Warranty i Water level figure 2 31 Weekly maintenance 5 3 Worksheet 2 20 Wrap up 2 32
213. zer is placed next to it on a bench The user must provide a grounding cable for this configuration Connect the Thermal Organizer Cable Part Number 70 5595 between the thermal organizer the Power Input Module and the Logic Module the latter two modules are accessed via the rear of the detector chassis Connect the tubing for the drain onto the protruding tube on the right rear of the organizer Place the other end of the drain tubing in a waste container that is situated lower than the organizer Route the cell cables out the back of the organizer using the slots in the chassis plate located at the rear of the organizer Connect cell cables to the proper connectors on the rear panel of the Corona Detector Replace the cover Make all the necessary connections to your HPLC system Now the Corona CAD detector and its thermal organizer are ready to be used C 16 Corona CAD and Corona S CAD Detectors Operating and Maintenance Manual Organizer and Thermal Organizer C 5 Troubleshooting The left part of the top line of the Corona Main screen presents the temperature of the Thermal Organizer when it is present In the event that a fault is observed in the organizer this field will indicate a negative number as a code to indicate the nature of the fault The Corona help button can be used to present the text associated with the particular error code displayed The codes are indicated in Table C 2 Table C 2 Error Messa

Manual - VTP UP

Contents

Download Pdf Manuals

Related Search

Related Contents