Nanospray Flex Ion Source Getting Connected
Contents
1. 7 Robust XYZ manipulator for accurate emitter positioning Sliding rails for Easy access to fittings and quick retraction adaptors for emitters and of emitter columns Content of Nanospray Flex lon Source Kit The Nanospray Flex Ion Source Kit P N ES071 includes the following Flexible ion source housing with precision XYZ manipulator and fittings DirectJunction adaptor ready for use with a variety of different column emitter configurations for online analysis LiquidJunction MicroCross for one column setup with long packed emitters Dual camera setup including one LCD monitor Nanobore stainless steel emitters fitting the DirectJunction adaptor 2 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 1 Introduction Unpacking the lon Source and Preinstallation All necessary cables and connectors for the ion source Nanospray Flex Ion Source Getting Connected Guide Unpacking the lon Source and Preinstallation The Nanospray source kit is packed in a single box To fully protect the components during transport the source mechanics DirectJunction adaptor and cameras are not assembled see Figure 2 Figure 2 Nanospray ion source components Box with DirectJunction adaptor Dino Lite LCD monitor Screwdrivers cameras LiquidJunction Stainless steel emitters Body of Nanospray Phono BNC Source To unpack the ion source 1 After removing the foam from the top of the box access t2. 2010 Thermo Fisher Scientific Inc All rights reserved Nanospray Flex DirectJunction EASY nLC and EASY Column are trademarks and Xcalibur is a registered trademark of Thermo Fisher Scientifc Inc in the United States The following are registered trademarks in the United States Nanospray is a registered trademark of AB Sciex Pte Ltd Lemo is a registered trademark of Interlemo Holding S A Dino Lite is a registered trademark of Anmo Electronics Corporation Swagelok is a registered trademark of the Crawford Fitting Company Thermo Fisher Scientific San Jose has a non exclusive license with the University of Texas for US Patent 5 572 023 Caprioli Electrospray methods and apparatus for trace analysis All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Thermo Fisher Scientific Inc provides this document to its customers with a product purchase to use in the product operation This document is copyright protected and any reproduction of the whole or any part of this document is strictly prohibited except with the written authorization of Thermo Fisher Scientific Inc The contents of this document are subject to change without notice All technical information in this document is for reference purposes only System configurations and specifications in this document supersede all previous information received by the purchaser Thermo Fisher Scientific Inc makes no representations that this docum
3. ferrule and nut from the venting tee SC601 Figure 15 ZDV union SC600 and pre column SC001 assembly i 7 Tighten the nut while ensuring that the pre column and sleeve are firmly pushed against the center of the venting tee 8 Install the analytical column on the other side of the venting tee using a 1 32 in OD sleeve F385 Check the label for the flow and firmly push the sleeve and column against the center of the venting tee Figure 16 ZDV union SC600 pre column SC001 venting tee SC601 and analytical column SC200 assembly Connecting the Columns to a Stainless Steel Emitter To connect the columns to a stainless steel emitter 1 Remove both red nuts from a second ZDV union SC600 2 Insert the white blind plug into the union 3 Insert the outlet end of the analytical column through a 1 32 in OD sleeve F385 and a red nut 4 Screw the outlet end into the union ensuring that the column and sleeve are firmly pushed against the white blind plug Then remove the white blind plug 5 Insert the sleeved emitter ES542 through a red nut Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 17 3 DirectJunction Adaptor Standard Configuration 6 Screw the sleeved emitter into the other end of the union ensuring that the emitter and sleeve are firmly pressed against the column outlet and sleeve Figure 17 Stainless steel emitter with 1 32 inch sleeve ES542 Mounting
4. Installing the lon Source on the Mass Spectrometer Figure 6 Turning the black handles on top of the ion source Black handles 4 Take the Nanospray source and align the guiding rods of the source with the guiding holes of the mass spectrometer see Figure 7 5 Push the source gently towards the mass spectrometer The top part with the connectors might give some resistance If so push a little harder in the top area of the source 6 Close the two handles by turning them so that they point toward each other you might feel some resistance The mass spectrometer is now in Standby mode and the source is mounted Nanospray Flex lon Source Getting Connected Guide 7 1 Introduction Installing the lon Source on the Mass Spectrometer Figure 7 Guiding holes on the Nanospray source for mounting on the guiding rods of the mass spectrometer Multi pole connector for the Interlock and source recognition HV connector Guiding rods Guiding holes 8 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific ES Electrical Connections For correct operation use these procedures to connect the cameras and monitor electrically and to adjust the focus for sharp pictures Contents e Connecting the Monitor and Cameras e Adjusting the Monitor and Cameras CAUTION When you push the XYZ manipulator onto the mounting frame of the source the HV for the spray emitter switches on For your protection always retract the
5. the optimal distance between the emitter tip and orifice depends on the flow rate as does the optimal potential on the emitter 34 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 6 Operation Maintaining Stable Spray Figure 33 Schematic setup of the ion source and the mass spectrometer interface Sample from Inlet of MS HPLC system Stainless steel or fused silica emitter Optimal flow rate between 50 and 500 nL min Values for stainless steel or fused silica emitters A 3 to 5 mm and the potential on the emitter 1400 to 2400 V Maintaining Stable Spray Thermo Scientific For best results use stainless steel emitters for nano electrospray experiments These emitters ensure straightforward initiation of the spray and help to maintain spray stability over extended periods up to 1000 hours Unstable spray is detrimental to most analyses Table 1 lists the most common reasons for unstable spray and ways to correct it Table 1 Correcting unstable electrospray Cause Possible correction The HV contact is interrupted Check clean the gold contact in the HV clamp where the emitter is held in the DirectJunction adaptor Check the red cable and connectors Check if the mass spectrometer is supplying the correct high voltage Air bubbles in the emitter if glass Check if the solvents are degassed properly might cause the emitter to spit The emitter is blocked because of Stainl
6. drawer of the source before working with the source head You must use the supplied HV cables and connectors in the Nanospray Flex ion source Connecting the Monitor and Cameras After installing the Thermo Nanospray Flex ion source you must set up the LCD monitor and Dino Lite cameras To connect and adjust the LCD monitor and Dino Lite cameras 1 In the ion source kit remove the monitor from the brown box and place it on top of the mass spectrometer by using the support arm on the back of the monitor see Figure 8 Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 9 2 Electrical Connections Connecting the Monitor and Cameras Figure 8 Back of LCD monitor Video connectors for the Dino Lite cameras BNC adaptors Video IN for the cameras 12 V power supply connector for the LCD monitor 2 Connect the 12 V power supply connector to the monitor 3 Locate the two BNC adaptors in the ion source kit and using a quarter turn insert them into the two Video IN ports on the back of the monitor Figure 8 4 Connect the two yellow video connectors from the Dino Lite cameras onto the two BNC adaptors 5 Choose and insert the appropriate connector to the power supply a Locate the power supply for the cameras in the camera box The power supply comes equipped with an EU standard net connector The kit box contains one U K and one U S connector b Connect the low voltage connectors of the
7. emitter contains the stationary phase and has dual roles as the analytical column and the emitter The packed glass emitter is inserted through the ZDV union which only serves as positional support Note EASY nLC software version 2 5 and later require use of the Waste in line connected to the vent ES257 upstream of the analytical column in one column configurations In earlier software versions or with other LC systems you can replace the Liquid Junction Cross ES257 with a Liquid Junction Tee ES258 Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 23 4 Alternative Configurations for DirectJunction Packed Glass Emitter with Liquid Junction in Two Column Setup Packed Glass Emitter with Liquid Junction in Two Column Setup Figure 26 Packed glass emitter with a liquid junction and two columns Packed glass emitter To waste ES257 SC600 Transfer line SCO01 Note The packed glass emitter contains the stationary phase and has dual roles as the analytical column and the emitter The packed glass emitter is inserted through the ZDV union which only serves as positional support 24 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 4 Alternative Configurations for DirectJunction Setup with a Long Analytical Column Setup with a Long Analytical Column Figure 27 Long analytical column Long analytical column To waste Glass emitter gt 20 cm Transfer line ES257
8. versions or with other LC systems you can replace the venting tee with a ZDV union SC600 Glass Emitter with Liquid Junction and One Column Setup Figure 23 Glass emitter with a liquid junction and one column Glass emitter Analytical column To waste ES257 Transfer line Note EASY nLC software version 2 5 and later require use of the Waste in line connected to the vent ES257 upstream of the analytical column in one column configurations For earlier software versions or with other LC systems you can replace the Liquid Junction Cross ES257 with a Liquid Junction Tee ES258 Use small ID emitter tips lt 20 um to prevent out gassing in the tip and subsequent spray instability Glass Emitter with Liquid Junction and Two Column Setup Figure 24 Glass emitter with a liquid junction and two columns Glass emitter Analytical column To waste ES257 SC600 Transfer line El gt El SC001 22 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 4 Alternative Configurations for DirectJunction Packed Glass Emitter with Liquid Junction in One Column Setup Note Use small ID emitter tips lt 20 pm to prevent out gassing in the tip and subsequent spray instability Packed Glass Emitter with Liquid Junction in One Column Setup Figure 25 Packed glass emitter with a liquid junction and one column Packed glass emitter To waste Transfer line ES257 Note The packed glass
9. 1 and adjust the focus with the focusing wheel as above 4 If the picture on the LCD monitor is too dark use the camera s internal light to make adjustments a To turn on the light gently press the small button on the back of either camera Figure 4 b Adjust the picture to achieve the sharpest image by using either camera light or both lights together 12 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific ee I DirectJunction Adaptor DirectJunction is a configurable adaptor for the Thermo Nanospray Flex ion source see Figure 12 It enables the flexible interchange between most online column and emitter configurations and uses stainless steel or glass emitters and a one column or two column setup with or without a liquid junction Contents e Choosing a One Column or Two Column Configuration e Choosing Stainless Steel or Glass Emitters e Standard Configuration e Connecting the DirectJunction to HV Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 13 3 DirectJunction Adaptor Choosing a One Column or Two Column Configuration Figure 12 DirectJunction adaptor HV clamp for stainless steel or coated glass emitters ZDV union for connecting the column and emitter HV protection Mounting bracket for the Multipurpose adaptor for shield Nanospray ion source ZDV unions liquid junctions and connecting tees Choosing a One Column or Two Column Configuration The DirectJuncti
10. Figure 4 Small screw to fix Dino Lite camera placed on camera in place top of the source body 7 Remove the cameras and install the first one in the top of the source body Note that the camera can only be placed one way ensuring optimal alignment Slowly tighten the screw until it touches the camera housing Tighten by another half turn 8 Repeat the procedure for the second camera which is located on the left side of the source body You are now ready to install the Nanospray source Installing the lon Source on the Mass Spectrometer After assembling the body and accessories you are ready to mount the completed source on the mass spectrometer Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 5 1 Introduction Installing the lon Source on the Mass Spectrometer Figure 5 Thermo Nanospray Flex ion source with DirectJunction To install the ion source on the mass spectrometer 1 Before removing the previous ion source disconnect any columns and flow lines from the HPLC system 2 Refer to the mass spectrometer user manual for guidance on how to remove the existing ion source 3 Remove the existing ion source as follows a Turn the two black handles on top of the source in the direction indicated in Figure 6 The handles might give some resistance b Carefully pull out the source 6 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific Thermo Scientific 1 Introduction
11. State and these companies should dispose of or recycle this product See www thermo com WEEERoHS for further information on Thermo Fisher Scientific s compliance with these Directives and the recyclers in your country WEEE Konformitat Dieses Produkt muss die EU Waste Electrical 82 Electronic Equipment WEEE Richtlinie 2002 96 EC erf llen Das Produkt ist durch folgendes Symbol gekennzeichnet Thermo Fisher Scientific hat Vereinbarungen mit Verwertungs Entsorgungsfirmen in allen EU Mitgliedsstaaten getroffen damit dieses Produkt durch diese Firmen wiederverwertet oder entsorgt werden kann Mehr Information ber die Einhaltung dieser Anweisungen durch Thermo Fisher Scientific ber die Verwerter und weitere Hinweise die n tzlich sind um die Produkte zu identifizieren die unter diese RoHS Anweisung fallen finden sie unter www thermo com WEEERoHS Thermo SCIENTIFIC Conformit DEEE Ce produit doit tre conforme la directive europ enne 2002 96 EC des D chets d Equipements Electriques et Electroniques DEEE Il est marqu par le symbole suivant Thermo Fisher Scientific s est associ avec une ou plusieurs compagnies de recyclage dans chaque tat membre de l union europ enne et ce produit devrait tre collect ou recycl par celles ci Davantage d informations sur la conformit de Thermo Fisher Scientific ces directives les recycleurs dans votre pays et les informations sur les produits Thermo Fisher Scie
12. Transfer line Note You can easily fit long columns to the DirectJunction adaptor by coiling them between the ZDV union and the vent ES257 Figure 27 shows a glass emitter and a liquid junction but combinations with a steel emitter a two column configuration or both of these are also possible Note EASY nLC software version 2 5 and later require use of the Waste in line connected to the vent ES257 upstream of the analytical column in one column configurations In earlier software versions or with other LC systems you can replace the Liquid Junction Cross ES257 with a Liquid Junction Tee ES258 Use small ID emitter tips lt 20 um to prevent out gassing in the tip and subsequent spray instability Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 25 ES 1 Nano Flex lon Source Configuration When analyzing with nanospray sources you only need to change a few mass spectrometer settings in the Xcalibur data sytem The MS software will automatically recognize the ion source type To specify configuration options for the ion source 1 Open the Instrument Configuration window from the computer desktop as follows e For Xcalibur 2 0 7 or earlier versions choose Start gt All Programs gt Xcalibur gt Instrument Configuration Or double click the Instrument Configuration icon e For Xcalibur 2 1 0 or later versions choose Start gt All Programs gt Thermo Foundation 1 0 gt Instrument C
13. al Equipment for Measurement Control and Laboratory Use Manufacturers Name Proxeon Biosystems A S Manufacturers Address Edisonsvej 4 DK 5000 Odense Denmark Type of Equipment Laboratory Instrumentation Model Name Nanospray Flex ion source Model Numbers ES071 Serial Number ES 001000 and later Year of Manufacture 2010 L the undersigned hereby declare that the equipment specified above conforms to the above Directive s and Standard s We Ole Vorm Site Manager October 11 2010 xii Nanospray Flex lon Source Getting Connected Guide Thermo Scientific ES Introduction Nanospray Flex ion sources minimize the liquid flow rate nL min and maintain excellent spray stability to ensure evaporation and ionization of liquid samples the key to achieving the highest sensitivity Key benefits of the Nanospray Flex ion source include e User friendly design e Single setup for all online nanoflow applications e Ability to interface with online nanoscale LC separation techniques Contents e Content of Nanospray Flex Ion Source Kit e Unpacking the Ion Source and Preinstallation e Installing the Ion Source on the Mass Spectrometer Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 1 1 Introduction Content of Nanospray Flex lon Source Kit Figure 1 Nanospray Flex ion source Camera with integrated light source and one LCD monitor to visualize emitter position
14. als must be ordered from Thermo Fisher Scientific or one of its authorized representatives FCC Compliance Statement THIS DEVICE COMPLIES WITH PART 15 OF THE FCC RULES OPERATION IS SUBJECT TO THE FOLLOWING TWO CONDITIONS 1 THIS DEVICE MAY NOT CAUSE HARMFUL INTERFERENCE AND 2 THIS DEVICE MUST ACCEPT ANY INTERFERENCE RECEIVED INCLUDING INTERFERENCE THAT MAY CAUSE UNDESIRED OPERATION CAUTION Read and understand the various precautionary notes signs and symbols contained inside this manual pertaining to the safe use and operation of this product before using the device Notice on the Proper Use of Thermo Scientific Instruments In compliance with international regulations Use of this instrument in a manner not specified by Thermo Fisher Scientific could impair any protection provided by the instrument Notice on the Susceptibility to Electromagnetic Transmissions Your instrument is designed to work in a controlled electromagnetic environment Do not use radio frequency transmitters such as mobile phones in close proximity to the instrument For manufacturing location see the label on the instrument WEEE Compliance This product is required to comply with the European Unions Waste Electrical 82 Electronic Equipment WEEE Directive 2002 96 EC It is marked with the following symbol Thermo Fisher Scientific has contracted with one or more recycling or disposal companies in each European Union EU Member
15. copy manuals from the Internet Go to mssupport thermo com agree to the Terms and Conditions and then click Customer Manuals in the left margin of the window To suggest changes to documentation or to Help e Fill out a reader survey online at www surveymonkey com s PQM6P62 e Send an e mail message to the Technical Publications Editor at techpubs lcms thermofisher com This equipment including spray capillaries is designed specifically for creating ions in a mass spectrometer No other use is recommended This equipment is for laboratory use only and is not a medical device This equipment must not be connected to any power supply delivering more than 8 kV and 100 pA Do not allow unauthorized or untrained operators to use this equipment Nanospray Flex lon Source Getting Connected Guide xi Preface e Any misuse will be the sole responsibility of the user owner Thermo Fisher Scientific assumes no implied or inferred liability for direct or consequential damages or injuries from this instrumentation if it is operated or used in any way other than that for which it is designed e Any legal disagreements must be settled before a Danish court of law e Only use this equipment if you agree with the conditions above Declaration of Conformity European Safety Standards Declaration of Conformity Standard s to which IEC 61010 1 EN61010 1 2001 Second edition Safety conformity is declared Requirements for Electric
16. e recommended configuration for the EASY nLC nano flow liquid chromatography system and EASY Columns with a stainless steel emitter installed on the DirectJunction prior to mounting onto the Nanospray Flex ion source Figure 13 Configuration for the EASY nLC and EASY Columns on DirectJunction using a stainless steel emitter Stainless steel EASY nLC Venting tee for two column setup emitter Waste in line for 1 32 in OD tubing ES542 1 32 in 0D SC601 EASY Column SC200 EASY Column SC001 EASY nLC 10 cm 75 um ID 2 cm 100 um ID Column out line 3 um C18 media 3 um C18 media 1 32 in 0D Zero dead volume union for 1 32 in OD tubing SC600 Assembling the Columns for Standard Configuration To assemble the pre column and analytical column 1 Remove both red nuts from the ZDV union SC600 2 Screw the white blind plug into the union Figure 14 ZDV union with white blind plug SC600 3 Insert the pre column SC001 through a 2 cm 0 79 in 1 32 in OD sleeve F 385 with arrows on the label pointing away from the sleeve 16 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 3 DirectJunction Adaptor Standard Configuration 4 Insert the sleeved pre column through a red nut 5 Screw in the red nut while ensuring that the pre column and sleeve are firmly pushed against the white blind plug 6 Insert the other end of the pre column through a 1 32 in OD sleeve F 385
17. ent is complete accurate or error free and assumes no responsibility and will not be liable for any errors omissions damage or loss that might result from any use of this document even if the information in the document is followed properly This document is not part of any sales contract between Thermo Fisher Scientific Inc and a purchaser This document shall in no way govern or modify any Terms and Conditions of Sale which Terms and Conditions of Sale shall govern all conflicting information between the two documents Release history Revision A November 2010 Software version EASY nLC 2 5 and later Xcalibur 1 3 and later For Research Use Only Not for use in diagnostic procedures Regulatory Compliance Thermo Fisher Scientific performs complete testing and evaluation of its products to ensure full compliance with applicable domestic and international regulations When the system is delivered to you it meets all pertinent electromagnetic compatibility EMC and safety standards as described in the next section or sections by product name Changes that you make to your system may void compliance with one or more of these EMC and safety standards Changes to your system include replacing a part or adding components options or peripherals not specifically authorized and qualified by Thermo Fisher Scientific To ensure continued compliance with EMC and safety standards replacement parts and additional components options and peripher
18. ess steel emitter only If sonicating the emitter particulates in the sample other does not remove the blockage replace the emitter small particles from the flow lines valves and so on There is a leak at some point in the Check if the correct flow is being delivered liquid path Nanospray Flex lon Source Getting Connected Guide 35 6 Operation Using the Optional Gas Connection Using the Optional Gas Connection The drawer of the Nanospray Flex ion source is equipped with a gas connector a Swagelok fitting for tubing with 1 8 OD Figure 34 Swagelok 1 8 gas connector option for drying or curtain gas In research analyses use this optional gas connection for drying gas when you want to work in a controlled environment around the spray area 36 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific ES i Accessories and Spare Parts This appendix provides ordering information for the Nano ES Spray Kit for offline analysis and spare parts and accessories for the Thermo Nanospray Flex ion source For detailed descriptions and ordering information on the full range of spare parts and accessories for the Nanospray Flex ion source visit www proxeon com Contents e Nano ES Spray Kit for Offline Analysis e Accessories List e Limited Warranty e Additional Support Nano ES Spray Kit for Offline Analysis Through offline analysis you can extract the maximum amount of information fro
19. he accessories for the Nanospray source 2 Carefully lift up the next foam bar including the accessories to access the body of the Nanospray source 3 Carefully take out the body of the Nanospray source and place it on a table to complete the assembly of the Nanospray source Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 3 4 1 Introduction Unpacking the lon Source and Preinstallation 4 Pull back the drawer of the Nanospray source to access the arm on the XYZ manipulator see Figure 3 The DirectJunction adaptor has to be mounted to this arm using the 5 mm screw in the holder of the DirectJunction Figure 3 Assembly of the DirectJunction to the XYZ manipulator arm DirectJunction adaptor Drawer of the 5 mm screw Arm of the XYZ Nanospray source manipulator 5 Use the supplied screwdriver to mount the DirectJunction adaptor on the XYZ manipulator arm Before tightening the 5 mm screw position the DirectJunction with the back of the DirectJunction elevated about 20 above horizontal Tighten the screw firmly The two white boxes in the box contain Dino Lite cameras which are located in the opening on the top and left side of the source body 6 Loosen the small screw which is used to secure the cameras in place with the supplied screwdriver see Figure 4 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 1 Introduction Installing the lon Source on the Mass Spectrometer
20. ion to HV 19 Chapter4 Alternative Configurations for DirectJunction 21 Stainless Steel Emitter and One Column Setup dire 21 Glass Emitter with Liquid Junction and One Column Setup 22 Glass Emitter with Liquid Junction and Two Column Setup 22 Packed Glass Emitter with Liquid Junction in One Column Setup 23 Packed Glass Emitter with Liquid Junction in Two Column Setup 24 Setup with a Long Analytical Colin ann ne kee eee aes 25 Thermo Scientific Nanospray Flex lon Source Getting Connected Guide vii Contents viii Chapter 5 Chapter 6 Appendix A Nano Flex lon Source Configuration 27 Operation soseda ie a 33 Positioning the Emitter Tipo ais a er 33 Maintaining Stable Pray ea au tench e 35 Using the Optional Gas Connection 2 au anne ana dames 36 Accessories and Spare Parts 37 Nano ES Spray Kit for Offline Analysis 37 Accessories List ua ee nr A 38 Limited Warranty eh 38 Additional Support ri sn Wei ie aia ore ti ate o a A 38 INDE RSR ES ER ser ra een oe 39 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific CC A Preface This guide provides information on the Thermo Nanospray Flex ion source including setup installation and operation The Nanospray Flex ion source works in conjunc
21. m very limited amounts of sample You can also average data extensively to improve the S N ratio and conditions optimized for MS MS experiments Using the Nano ES Spray Kit see Figure 35 you can do the following e Work at low flow rates of 10 40 nL min e Utilize nearly 100 percent of sample e Work effectively with sample volumes down to 300 nL e Avoid cross contamination by using disposable emitters e Spray from purely aqueous as well as purely organic solvents Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 37 Accessories and Spare Parts Accessories List The Nano ES Spray Kit includes all necessary items for offline analysis The kit is not included with the Nanospray Flex ion source but can be ordered separately Figure 35 ES259 offline Nano ES Spray Kit Accessories List This section lists the part numbers for the followingNanospray Flex ion source accessories DireetJuneuon ida do ES256 LiquidJunction with Cross 1 32 16 a tr e id de ES257 LiquidJunction with Tee 1 32 in sarah ee ES258 Off line source head kit 0 ES259 One Dino Lite video camera ES216 One LCD Monitor 8 in 42828088 a an aan an ea ES217 Limited Warranty e Thermo Fisher Scientific limits the warranty on repair of supplied Nano ES equipment and replacement provided that instrument operation has been in accordance with the instruction
22. ment recognizes the source automatically If the source is not correct follow the instructions in the Xcalibur manual d Click OK to close the window 4 From your desktop double click the LXQ Tune icon or the correct system name of your MS The Thermo Tune Plus window opens where you can analyze and choose specific settings see Figure 30 Nanospray Flex lon Source Getting Connected Guide 29 5 Nano Flex lon Source Configuration Figure 30 Thermo Tune Plus window Thermo Tune Plus Untitled moewa eo zul ei Lex 54220 IT 10 000 ST 060 uS 3 NL 1 00 Label F ITMS p NSI Full ms 150 00 2000 00 NSI Source X Spray Voltage KVI X Spray Current nA w Sheath Gas Flow Ra Aux Gas Flow Rate 100 Y 90 Shoah Gas Flow Rate ab i 3 Y Sweep Gas Flow Ral RE Aux Gas Flow Rate arb 0 y Capillay Temp OK Pa Sweep Gas Flow Rate at 0 y o Cane Vote NE Y Capila Temp CL se ISpray Vohage VIE 30 Tube Lens 70 Spray Curent A s Capillary Temp C 20000 19996 se fe Capillay Votege v 3200 3205 Tube Lens vy 500 11508 0 as ww Corel to 0 Ca Y lon Gauge Pressure Y lonGauge lon Gawe E 5 Ton Y Convection Pressure Convection Gauge Relative Intensity o Turbo Pump Status Life hours Speed Ha Power Watts Temperature Cp lon Optics AF Lens Offset V Lens 0V Mukipole 0 Olfset V Len
23. nectors 9 installation ion source on MS 5 instrument configuration window Xcalibur 27 ion source removing existing 6 ion source MS schematic 35 ion source kit contents 2 mass spectrometers ion source compatible ix monitor LCD adjusting 11 connecting to cameras 9 0 offline analysis 37 online offline analysis 31 Nanospray Flex lon Source Getting Connected Guide 39 Index P P positioning emitter tip 33 power supply and connectors 10 preinstallation ion source 3 regulatory compliance iii S safety standards ii spare parts 37 spray capillary 33 spray stability maintaining 35 U unpacking the ion source See preinstallation ion source W warranty information 38 WEEE compliance v X Xcalibur settings for ion source 27 XYZ manipulator 2 33 XYZ manipulator caution 9 40 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific
24. ntific qui peuvent aider la d tection des substances sujettes la directive RoHS sont disponibles sur www thermo com WEEEROHS A A Contents Prelate eC ee ere er er ee ee ix Related Documentations sus peated AA ix Satermand Special NOUS essaie ane sank her side ix Contacting US rasen Rive durs Rhea xi Deiner see beats Re ER ER O xi Declaration of Corona eier xii Chapter Introduction 1 Content of Nanospray Flex Ion Source Kit 2 Unpacking the lon Source and Preinstallation ua an nen anne 3 Installing the Ion Source on the Mass Spectrometer 5 Chapter2 Electrical Connections 9 Connecting the Monitor and Cameras 9 Adjusting the Monitor and Cameras a en ne en 11 Chapter3 DirectJunction Adaptor 13 Choosing a One Column or Two Column Configuration 14 Choosing Stainless Steel or Glass Emitters 4 sue 0 00 0000 sees 15 Standard COMA AR RAS ARA Bere 16 Assembling the Columns for Standard Configuration 16 Connecting the Columns to a Stainless Steel Emitter 17 Mounting the Column and Emitter Setup onto the DirectJunction o Ae dan da eho We uk wea eres ot 18 Connecting the LC System to the Column Setup 19 Connecting the DirectJunct
25. oltage across the entire emitter you can maintain a constant optimal electrical contact with the liquid The result is superior spray stability A cross section of scientists consider glass emitters to be more bio inert than stainless steel emitters This state means reducing the risk of non specific adsorption of biomolecules and leads to slightly improved sensitivity However the degree of this adsorption depends on the chemical characteristics of the sample You can pull glass emitters to produce very small ID emitter openings However the very small opening at the tip often results in stability problems associated with blockage of the tip and means that glass emitters rarely last as long as their steel counterparts Glass emitters usually have HV applied through a liquid junction This means that only a small percentage of the bulk liquid flow is exposed to the electrode carrying the HV Tip When using glass emitters do the following e Place the liquid junction at the high pressure side of the column Electrochemical processes occurring at the electrode can otherwise create gas that leads to spray instability e Use glass emitters with small ID emitter tips lt 20 um to create back pressure in the emitter and avoid outgassing and consequent spray instability Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 15 3 DirectJunction Adaptor Standard Configuration Standard Configuration Figure 13 shows th
26. on adaptor supports both a one and two column configuration A one column configuration has the following advantages e It has fewer connections which minimize any potential peak broadening that results from dead volumes It enables MS analysis of compounds that elute during sample loading as they might not bind to the column material because the fluid path leads directly to the MS 14 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 3 DirectJunction Adaptor Choosing Stainless Steel or Glass Emitters A two column setup typically a short pre trap column with a large inner diameter and a longer analytical column with a narrower inner diameter has these advantages e A pre trap column provides an increased loading capacity and an increased loading flow rate when compared to loading directly onto the analytical column e It directs the loading solvent to waste so that sample contaminants do not enter the MS reducing the need to clean the MS inlet e A pre trap column acts as a guard column by protecting the analytical column from particulate matter Choosing Stainless Steel or Glass Emitters Stainless steel emitters are more robust than glass emitters and help to maintain a stable and consistent spray for longer periods of time The DirectJunction adaptor enables high voltage HV to be applied directly to the stainless steel emitter through the small HV clamp at the front of the adaptor By applying v
27. onfiguration The Thermo Foundation Instrument Configuration window appears with a list of the installed available devices see Figure 28 Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 27 5 Nano Flex lon Source Configuration Figure 28 Thermo Foundation Instrument Configuration window 2 Thermo Foundation Instrument Configuration Device Types Al Available Devices Configured Devices us y Surveyor MS Pump Thermo EASY nLC Plus Thermo EASY nLC 2 Double click the icon that represents your MS system for example LXQ MS A copy of the icon appears in the Configured Devices pane 3 Complete the configuration of the mass spectrometer as follows a In the Configured Devices pane double click the LXQ MS icon or the icon for your system The LXQ Configuration dialog box appears see Figure 29 28 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific Thermo Scientific 5 Nano Flex lon Source Configuration Figure 29 LXQ Configuration dialog box LXO Configuration MS Detector Inlet lon Source Display Tune Plus Default source Nanospray Isolation FAIMS V Allow zero sheath gas flow with ESI source Analog Inputs Ethernet ee AP MALDI Application a Instrument Warnings Service Cancel Help b Select Ion Source in the left pane if it is not already selected c Ensure that the default source is Nanospray the instru
28. or optimal performance Contents e Positioning the Emitter Tip e Maintaining Stable Spray e Using the Optional Gas Connection Positioning the Emitter Tip The correct position of the emitter tip is an important parameter in nano ES analysis RL lt To optimize the position of the emitter tip 1 Position the spray capillary tip almost on axis with the orifice by using the four knobs on the XYZ manipulator see Figure 32 2 As you make adjustments observe the magnified inlet orifice on the monitor Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 33 6 Operation Positioning the Emitter Tip Figure 32 Adjustment knobs on the XYZ manipulator Left right adjustment sideward movement of the emitter Vertical adjustment of the emitter Adjustment toward the orifice Fine adjustment toward the orifice CAUTION Emitter tips are extremely sharp Never touch the tips as this may cause injury CAUTION e Although the ion source is shielded there is easy access to the source head e Always turn off the high voltage before touching the source head Do not leave the source unattended while the spray voltage is on e Always mount the high voltage insulator correctly before turning on the high voltage e The interface region of the mass spectrometer can be hot Do not touch this area Figure 33 illustrates some important settings whose values can vary depending on the mass spectrometer in use Also
29. s 1 Vi Gate Lens Vp GEES 200 400 600 800 1000 1200 1400 1600 1800 2000 maz 5 From the main menu choose Setup gt NSI Source The NSI source dialog box opens Figure 31 NSI Source dialog box NSI Source Actual Sheath Gas Flow Rate ab EJ am Aux Gas Flow Rate arb 0 4 002 Sweep Gas Flow Rate arb iz 0 00 Spray Voltage kV eo 0 02 Spray Current pA 0 04 Capillary Temp C 200 00 200 02 Capillary Voltage 32 00 32 03 Tube Lens V 11500 4 115 10 Apply Cancel Help 30 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific Thermo Scientific 5 Nano Flex lon Source Configuration 6 Set the ion spray voltage for nanospray analysis Pay attention to these nanospray specific settings e For online analysis I Spray Voltage kV 1 40 to 2 40 Start with 1 80 kV e For offline analysis I Spray Voltage kV 0 70 to 1 20 Start with 0 90 kV 7 Click OK Note Before you begin nanospray analysis check that the drawer of the Nanospray Flex ion source is pushed completely toward the frame otherwise a message that the source is open appears and the MS will not scan The Nanospray Flex ion source is now be ready for analysis Nanospray Flex lon Source Getting Connected Guide 31 SH Operation Both for intial setup and regular maintenance use these procedures to monitor the position of the emitter tip and the spray voltage current f
30. s are extremely sharp Never touch the tips as this may cause injury e Always depressurize the LC system with transfer line before removing emitters from the source head Failure to do so may result in the emitter being ejected at high speed CAUTION e Although the ion source is shielded in certain cases there might be access to the emitter which is on high voltage HV Always turn off the HV before touching the source head e Never connect a power supply able to deliver more than 8kV and 100 pA to this Nanospray source e Do not leave the source unattended while the spray voltage is on CAUTION The interface region especially the cone of the mass spectrometer can be hot Do not touch this area x Nanospray Flex lon Source Getting Connected Guide Thermo Scientific Contacting Us Disclaimer Thermo Scientific Preface There are several ways to contact Thermo Fisher Scientific for the information you need RL eo To contact Technical Support Phone 800 532 4752 Fax 561 688 8736 E mail us techsupport analyze thermofisher com Knowledge base www thermokb com Find software updates and utilities to download at mssupport thermo com To contact Customer Service for ordering information Phone 800 532 4752 Fax 561 688 8731 E mail us customer support analyze thermofisher com Web site www thermo com ms To get local contact information for sales or service Go to www thermoscientific com wps portal ts contactus To
31. s outlined in the instruction manual Abuse or misuse of the Nano ES equipment or unauthorized repairs void this warranty e Limited warranty work is performed only at the factory and the user bears the cost of shipment to and from the factory The limited warranty is as stated above and no implied or inferred liability for direct or consequential damage is intended Additional Support For further assistance go to www proxeon com and click Support and Service in the left margin of the window 38 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific ES A Index A accessories 38 adaptor See DirectJunction analysis offline 37 BNC adaptor 10 C camera adjusting 11 Dino Lite 9 column and emitter setup 18 column setup long analytical 25 compliance FCC iii regulatory iii WEEE v components ion source 3 configuring the ion source 27 declaration of conformity xii DirectJunction alternative configurations 21 column and emitter setup 18 description 13 HV connection 19 mounting 4 Disclaimer xi electromagnetic compatibility iii EMC compliance iii Tem Scientific emitter glass liquid junction one column setup 22 liquid junction two column setup 22 packed glass liquid junction one column setup 23 liquid junction two column setup 24 stainless steel and one column setup 21 emitter tip caution 34 F FCC compliance iii G gas connector 36 H HV cables and con
32. that the high voltage connector of the DirectJunction adaptor is inserted into the Lemo HV socket on the underside of the Nanospray ion source drawer Figure 21 a hi 1 m DirectJunction adaptor Nanospray ion source drawer DirectJunction high voltage connector Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 19 ee B Alternative Configurations for DirectJunction Depending on the specific experimental requirements on columns emitters and connections the following configurations might work well in different situations Contents e Stainless Steel Emitter and One Column Setup e Glass Emitter with Liquid Junction and One Column Setup e Glass Emitter with Liquid Junction and Two Column Setup e Packed Glass Emitter with Liquid Junction in One Column Setup e Packed Glass Emitter with Liquid Junction in Two Column Setup e Setup with a Long Analytical Column Stainless Steel Emitter and One Column Setup Figure 22 Stainless steel emitter with one column ES542 Analytical column To waste SC601 Transfer line Note EASY nLC software version 2 5 and later require use of the Waste in line connected to the vent SC601 upstream of the analytical column in one column configurations Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 21 4 Alternative Configurations for DirectJunction Glass Emitter with Liquid Junction and One Column Setup For earlier software
33. the Column and Emitter Setup onto the DirectJunction Adaptor To mount the column and emitter setup onto the DirectJunction adaptor 1 Mount the DirectJunction adaptor onto the Nanospray ion source 2 Connect the red HV cable to the banana plug on the DirectJunction adaptor Open the front HV clamp on the DirectJunction adaptor A Mount the ZDV union connecting the emitter and analytical column in the black holder onto the DirectJunction adaptor and close the clamp over the emitter 5 Adjust the position of the multipurpose adaptor on the steel rod and mount the venting tee in the adaptor Figure 19 DirectJunction adaptor with two column setup and steel emitter mounted on a Nanospray Flex ion source 18 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 3 DirectJunction Adaptor Standard Configuration Connecting the LC System to the Column Setup To connect the LC system to the column setup 1 Connect the Column out line from the EASY nLC system to the pre column through the ZDV union ensuring the line is firmly pushed against the pre column and sleeve 2 Connect the Waste in line from the EASY nLC system to the third port on the venting tec Figure 20 DirectJunction adaptor as in Figure 19 but with Column out and Waste in flow lines connected from the EASY nLC system Ss ES 3 a e e D wi Connecting the DirectJunction to HV Make sure
34. tion with these Thermo Scientific mass spectrometers e LTQ Series LTQ Orbitrap FT Velos e Exactive e TSQ Series Quantum Quantum Access Quantum Discovery MAX Quantum Ultra Vantage LCQ Deca XP MAX Contents e Related Documentation e Safety and Special Notices Contacting Us e Disclaimer e Declaration of Conformity Related Documentation In addition to this guide Thermo Fisher Scientific provides the following documents for the Nanospray Flex ion source available as PDF files e Thermo Xcalibur manual suite Jon Max Source and Ion Max S API Source Hardware Manual The software also provides Help Safety and Special Notices Make sure you follow the precautionary statements presented in this guide The safety and other special notices appear in boxes Thermo Scientific Nanospray Flex lon Source Getting Connected Guide ix Preface Safety and special notices include the following CAUTION Highlights hazards to humans property or the environment Each CAUTION notice is accompanied by an appropriate CAUTION symbol IMPORTANT Highlights information necessary to prevent damage to software loss of data or invalid test results or might contain information that is critical for optimal performance of the system Note Highlights information of general interest Tip Highlights helpful information that can make a task easier CAUTION e Emitter tip
35. two power supplies to the two Dino Lite camera power connectors see Figure 9 10 Nanospray Flex lon Source Getting Connected Guide Thermo Scientific 2 Electrical Connections Adjusting the Monitor and Cameras Figure 9 Connection of the low voltage connectors to the camera power connectors Dino Lite camera power connectors Low voltage connectors on the power supply for the cameras 6 Connect the power adaptors for the two Dino Lite cameras and the power adaptor for the monitor to your net power sockets The cameras and the monitor are now ready for use Adjusting the Monitor and Cameras To adjust the LCD monitor and Dino Lite cameras 1 Turn on the LCD monitor by pressing the POWER button on the front of the monitor see Figure 10 Figure 10 LCD Monitor setup and control panel Power button SOURCE POWER Switch between cameras A After a few seconds the picture from one of the two cameras appears The picture is likely to be very blurred Thermo Scientific Nanospray Flex lon Source Getting Connected Guide 1 2 Electrical Connections Adjusting the Monitor and Cameras 2 Adjust the focus with the focusing wheel Figure 4 Figure 11 Camera light source switch and focusing wheel Switch for the light source in the camera Focusing wheel The adjustments produce a sharp image on the monitor 3 Switch to the second camera by pressing the SOURCE button see Figure 10 on page 1
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