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Addendum to the PAL User Manual PAL DLW Option Installation

1. Holder Assembly is flush with the lower surface of the Needle Adapter Block as in Figure 30 Repeat the needle penetration test and verity the needle penetration depth An example of the correct and incorrect positioning of the syringe needle tip in the valve inlet port is shown in Figure 34 Figure 34 Correct and incorrect Valve Needle penetration depth The ball spring loaded Needle Holder presses the square cut needle tip firmly against the bottom of the valve body This ensures a constant seal during the injection process 38 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual A SYSTEM DLW Option G Operating Instructions 1 Priming the Solvent Lines For trouble free DLW operation make sure the two solvent lines are free of alr bubbles at all times If the solvent lines are being connected for the first time or during a solvent change it is necessary to prime the solvent lines properly until air bubbles are no longer visible Solvent degassing and filtering is recommended The priming of the PAL DLW Option Is conveniently done directly trom the local terminal There are several tasks which can be executed from the Utilities section It is recommended to use the function of the Wash station first followed by Clean Syringe Please follow this description step by step to prime the solvent lines and the entire DLW system Solvent dega
2. Wash1 after sample pickup and before moving to the Injection Valve e The wash steps after injection are reduced to Valve Clean with Wash Solvent 1 and Wash Solvent 2 The DLW needle is flushed in the DLW Wash Station with Wash Solvent 1 only e Stator Wash Valve Toggle is not available Macro Name LC Inj DLW Fast Macro Description Active The system pauses according to the delay time Pre Inject Delay The loop is filled with the sample volume as specified in the sample List The Injection Valve is switched to position Standby the loop contents are injected The plunger of the DLW Syringe is pushed down to dispense the Rear Sample and Air Segment to Waste The Holding Loop is still filled with Wash Solvent 1 Macro Variable The PAL System waits for the Sync Signal Ready Remark before the injection cycle is started Sync Signal setting Start Fig 58 The Injection Valve is brought to a defined Inject to position Standby Standby The Rear air segment is pulled into the Holding Airgap Volume Loop Filling Speed Fig 59 The combined volumes of Rear Sample List Front Volume Fig 60 and Front Volume are aspirated into the Rear Volume Holding Loop SL volume Airgap Volume The Front air segment is aspirated Filling Speed Pullup Delay Fig 61 The Injection Unit moves to the specified Inject to Injection Valve Front Volume
3. DLW Plunger Holder DLW Syringe 100 uL DLW Actuator Control PCB Syringe Holder DLW Actuator Solenoid DLW Manifold ith consisting of DLW Solvent Selector O ring Polyfluor DLW Syringe Adapter DLW Flow Diverter DLW Holding Loop DLW Needle Adapter Block DLW Needle Holder Assembly DLW Syringe Needle Retaining Nut Figure 14 Front view of the DLW Syringe Holder assembly 21 PAL is a registered trademark of CTC Analytics AG Switzerland note note note Addendum to PAL System User Manual a SYSTEM DLW Option 2 5 1 Preparing and Installing the DLW Syringe The description on how to correctly install the Plunger Holder is given for field upgrade situations or when a plunger is replaced Newly shipped PAL xt Systems which include the PAL DLW Option contains a precontigured syringe with the Plunger Holder installed 1 Prepare the DLW Syringe by inserting the DLW Plunger Holder First manually move the plunger down to the stop position release a slight amount of pressure from the plunger tip by pulling a fraction of a mm backwards Install the DLW Plunger Holder and tighten the Allen screw Allen Key 6 firmly en er i nn T a i E i pee bbe cs brecsd bbe ceta bbe cas besa dee sd ee ey Figure15 Inserting DLW Plunger Holder on DLW Syringe It is critical that syringes are primed before beginning sample preparation
4. The Front and Airgap Volume is ejected to Airgap Volume Waste Injection Speed Fig 62 The PAL System waits for a signal from the data Wait for DS system The Injection Valve is switched to position Inject to Pre Injection Delay SL volume Fig 63 Injection Speed Post Inject Delay Fig 64 Injection Speed Fig 65 The DLW Actuator Solenoid is activated to deliver Inject to Wash Solvent 2 into the Holding Loop to clean Wash2 the Injection Valve from Port 1 to Port Waste Needle Gap Valve Clean For this step the needle tip is lifted releasing the sealing pressure to enable rinsing around the tip sealing point Remark The Atom Rinse Inj is new available starting with FW 4 1X The DLW Actuator Solenoid ts activated the Wash Solvent pump the Needle Gap and the Rinse Time are selectable Valve Clean Time Solvent 2 Needle Gap is a parameter from Rinse Inj Atom The variable in this macro is Needle Gap Valve Clean The function of this parameter is to lift the needle in the injection port to allow rinsing around the needle tip The pressure of the spring loaded balls in the DLW Syringe Holder Assembly is released by moving approx 3 mm up default This leaves a gap between the needle tip and the valve bottom of approx 1 mm to enable a flush at this contact point Fig 66 PAL is a registered trademark of CTC Analytics AG Switzerland 41 Addend
5. speed with optimized washing possibilities and the second for a Fast cycle with optimized throughput and less focus on carry over See Table 2 below The two macros or cycles are explained in detail see Tables 3 5 below Macro Name Macro Description Standard injection cycle using all possibilities of the DLW Option The LC Inj injection valve inlet port and the needle are washed with both wash solvents DLW Standard inside and out An extra Stator Wash can be added for intensive washing of the injection valve valve toggle The macro is tuned for speed and high throughput application The difference with regard to the Standard DLW macro is that some steps are left out to shorten the cycle time Details are explained below LC Inj DLW Fast Table 3 The DLW Macro definitions 4 1 Installing the Cycle Composer Macros or ICC Cycles The PAL DLW Option Is shipped with a CD ROM PAL System containing various cycles macros for the DLW option Copy these macros to your Cycle Composer Method folder or the corres ponding folder for applications within the integrated system Navigate in Explorer to the Cycle Composer folder This is usually installed in the following path C Program Files PAL Cycle Composer It is advisable to create a separate Method Folder for the DLW Option Copy the folder DLW Option from the CD to the Cycle Composer folder In Cycle Composer use the menu File Options Choose Method Fo
6. 1 0TFA e HPLC System Binary Solvent Delivery Pump UV Detector Wavelength 273 nm HPLC Column Zorbax SB C18 2 1 x50 mm spherical 1 8 um or equivalent Column Temperature Ambient Mobile Phase Water Acetonitrile 90 10 0 1 TFA Gradient isocratic for 2 Min e Wash Solvents Wash1 Water 0 1 TFA Wash2 Acetonitrile 0 1 TFA e Loop Size 2 uL Loop overfill 7 uL Loop ID 0 13 mm 20 uL partial Loop 10 uL Loop ID 0 25 mm e Injection Valve VICI Valco C72VX 1696D CTC bore size 0 25 mm e Cycle Composer Macro or Cycle ICC CE DLW Standard e Cycle Composer Method Parameter Settings Parameter Value Parameter Value Syringe 100 uL DLW Pullup Del 500 ms Sample Volume OIL Inject to Specified Inj Valve Airgap Volume 3 ul Inject Speed 5 L S Front Volume 5 ul Pre Inj Del 500 ms Rear Volume 5 uL Pst Inj Del 500 ms Needle Gap Valve Clean 3mm Valve Clean Time Solvent2 25 Fill Speed 5 uL Post Clean Time Solvent 2 25 Fill Strokes 0 Valve Clean Time Solvent 3s Stator Wash 0 Post Clean Time solvent 1 2s Table 8 Repeatability Test with UV Detection 19 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 3 3 Carry over and Repeatability Tests with UV Detection as described for the OQ PQ PV Test e Test Sample Benzophenone 50 g mL dissolved in Water Methanol 50 50 e Blank Solution Water Methanol 20 80 e HP
7. 3 to 0 5 rel standard deviation Carry over lt 0 030 If the values are significantly higher it is necessary to troubleshoot the system For hints see the section on Troubleshooting 80 PAL is a registered trademark of CTC Analytics AG Switzerland note note Addendum to PAL System User Manual A SYSTEM DLW Option Replacing Parts Safety Warning A Before beginning any maintenance work always disconnect the power cord s from the power supply or from the various power supplies if optional devices are installed Capacitors inside the instrument can remain charged even if the instrument Is turned off To avoid damaging electrical parts do not disconnect an electrical assembly while the power to the PAL system is on Once the power Is turned off wait approximately 30 seconds before you disconnect an assembly This section provides a short description on parts that can be replaced in the field Only basic instructions are given Additional details will be provided with an update of the addendum The parts along with ordering information are listed in the Appendices point 3 1 DLW Pump Module The following parts keywords from the DLW Pump Module can be individually exchanged Single Pump Module incl electrical connection Solvent Tubing Kit Electrical cable from Pump Module to PAL System Pump control PCB DLW Tubing Kit 2 DLW Wash Station The following parts keywords from
8. PAL Object List If a PAL Module hardware is added to an instrument several Objects have to be loaded into the firmware These Objects are collected in an Object List and stored in a file with the extension pol Object Lists are delivered together with Object Manager Software and are grouped into folders for the different kind of Modules e g Syringes Tray Holders Valve Drives The name of an Object List starts with the Module part number with variants added e g first or second Stack The name of the root folder includes the revision version which Is dependent on the firmware version e g PAL xt Object Lists Rev A for firmware 4 1 X or higher Object Class Each Object belongs to an Object Class e g Syringes Trays Injectors The Object Class defines the Items of an Object 86 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Object Item An Object contains several Items which can be numerical values with a physical unit e g X Y Z Position Penetration Syringe Scale Length Syringe Volume or references to other objects Note that the term Parameter is reserved for ATOM Parameter PAL Firmware commands to be used for a PAL Cycle or Macro Objects Objects are data structures describing the properties of physical modules Certain modules e g a Stack require several objects Module PAL hardware module eith
9. Solution Water Acetonitrile 0 1 0TFA e HPLC System Binary Solvent Delivery Pump UV Detector Wavelength 257 nm HPLC Column C18 Halo 2 1 x50 mm spherical 2 7 um or equivalent Column Temperature Ambient Mobile Phase A Water 0 1 TFA B Acetonitrile 0 1 TFA Gradient 10 A hold for 0 5 Min to 90 B in 7 5 Min hold for 1 Min e Wash Solvents Wash1 Water 0 1 TFA Wash2 Acetonitrile 0 1 TFA e Loop Size 2 uL Loop ID 0 13 mm e Injection Valve VICI Valco C72VX 1696D CTC bore size 0 25 mm e Cycle Composer Macro or Cycle ICC CE DLW Standard e Cycle Composer Method Parameter Settings Parameter Value Syringe 100 uL DLW Sample Volume 7 uL Airgap Volume 3 uL Front Volume 5 ul Rear Volume 5 uL Needle Gap Valve Clean 3 mm Fill Speed 5 L s Fill Strokes 0 Stator Wash 0 Parameter Pullup Del Inject to Inject Speed Pre Inj Del Pst Inj Del Valve Clean Time Solvent 2 Post Clean Time Solvent 2 Valve Clean Time Solvent 1 Post Clean Time solvent 1 Table 7 Carry over Test with UV Detection PAL is a registered trademark of CTC Analytics AG Switzerland Value 500 MS Specified Inj Valve 5 uL s 500 ms 500 ms IES 25 35 25 78 Addendum to PAL System User Manual A SYSTEM DLW Option 3 2 Repeatability Test with UV Detection e Test Sample Loop Overfill Caffeine 520 mg L Partial Loop Caffeine 52 mg L dissolved in Water Acetonitrile 90 10 0
10. Upper Needle Guide Attaching Pump Holder Bracket to X Axis Installing the Solvent Tube Assembly to Pump Holder Electrical Connections at the Pump Holder Bracket Electrical Connection from DLW Pump Holder to PAL xt Board Mounting the DLW Option Modules in combination with the Injection Valve Attaching the DLW Wash Station to the X Axis Front View of the DLW Syringe Holder Assembly Inserting DLW Plunger Holder on DLW Syringe Inserting DLW Syringe into Holder Inserting the DLW Syringe Holder Assembly into the Injection Unit Fixing Syringe Holder to Syringe Slider Connecting the DLW Syringe Plunger Holder Inserting the DLW Syringe Needle Installing the standard Valve Needle Guide Solvent Line Plumbing on Cheminert Valve Types Solvent Line Plumbing on Valco W type Valve Solvent Line Plumbing on Rheodyne Valve PD Series Electrical Connection DLW option on a PAL xt DLW Option in Combination with 4 Injection Valves Electrical Connections to 4 Injection Valves Daisy Chaining with PAL xt System PAL Object Manager choosing Object List Folder Wash Stations DLW Wash Station Wash1 Wash2 Reference Point Waste Reference Position LC Valve Reference Point Positioning Z Axis Needle Guide on Valve Needle Guide Teaching the Needle Penetration Valve Needle Penetration Depth DLW Manifold and Actuator Solenoid Inserting DLW Flow Diverter Standard Cycle Descriptions Timing for Stator Wash Step Additional Cleaning Ste
11. be fixed to the syringe in a leak tight manner A Teflon seal is provided When installing the seal first ensure that the hole Is open and that no fuzzy lint polymer material which could clog the needle is in the path When slipping the Teflon seal over the needle pay attention to the sharp edges of the needle which could cut into the polymer Again this could clog the needle 7 Move the lower needle guide carefully up and down to make sure that the needle tip does not catch on the guide Figure 20 Inserting the DLW Syringe Needle 26 PAL is a registered trademark of CTC Analytics AG Switzerland note note Addendum to PAL System User Manual A SYSTEM DLW Option 2 6 Installing the Valve Needle Guide Assembly For a field upgrade when first installing a DLW option the standard valve needle guide can continue to be used If the current Wash Station is the Active Wash type replace the Needle Guide with the standard Needle Guide provided In all cases unscrew the existing Needle Guide remove the old needle seal inside the valve port and insert a new seal Figure 21 Installing the standard Valve Needle Guide 2 7 Injection Valve Plumbing and Connection to the DLW Wash Station 1 Connect the injection valve Waste Line from port 2 example standard valve Cheminert type as shown in Figure 21 to the front Waste Port of the Wash Station If two injection valves are used in the PAL configuration th
12. free movement of the syringe slider Lower the syringe slider as shown in Figure 17 in order to gain space for installing the DLW Syringe assembly Match the magnetic pins of the syringe holder using the counter positions on the syringe slider Figure 17 Inserting the DLW Syringe Holder Assembly into the Injection Unit 2 Press the syringe holder firmly against the Z axis slider to ensure that the holder clicks into place Do not press against the DLW Actuator Solenoid Its mounting to the DLW Manifold ts fragile 3 Tighten the knurled screw to fix the Holder Assembly to the syringe slider see Figure 18 24 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Figure 18 Fixing the Syringe Holder to the Syringe Slider 4 Move the plunger up plunger holder until the thread of the screw catches the thread of plunger bushing Tighten the screw to fix the plunger holder see Figure 19 Figure 19 Connecting the DLW Syringe Plunger Holder 5 Tighten the holding screw to secure the syringe holder position 6 Insert the syringe needle tip into the Z axis lower needle guide then move the needle back up to the DLW Needle Holder Assembly and tighten firmly 25 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option note The removable needle must
13. included Replacement Plunger for Syringe SYRC DLW R pkg of 10 PAL DLW Plunger Holder Needle Kit for PAL DLW Option Gauge 22 PST 3 length 51 mm 3 pcs per pack incl Needle Retaining Nut DLW Syringe Holder Assembly PAL DLWHolder PAL DLWLoop PAL DLW Manifold PAL DLWFlwDiv PAL DLWAct PAL DLW Syringe Holder Assembly complete Assembly with 100 uL syringe and one syringe needle installed Kit PAL DLW Holding Loop with Needle Adapter mounted PAL DLW Manifold Kit complete assembled PAL DLW Flow Diverter Pkg of 1 PAL DLW Actuator Solenoid Kit incl screws without cable 9 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option PAL DLWActPCB PAL DLW Actuator Control PCB Kit incl cable and screw set PAL DLW Screw Syringe Holder Fixation Screw pkg of 1 Illustration DLW Syringe Holder Assembly contd PAL DLW NdlTool PAL DLW Needle Length Guide Tool pkg of 1 PAL DLWNHA PAL DLW Needle Holder Kit l Table 12 DLW Spare Parts 3 2 PM Kit Order Information A dedicated PM Kit Preventative Maintenance Kit is available for PAL xt Systems including a PAL DLW Option This kit contains all parts from the standard PM Kit for HPLC technique PNo PAL PM Kit1 and the DLW specific parts Packing List PAL PM Kit for HPLC Technique including PAL DLW Option PNo PAL PMKitDLW PNo D
14. left side of the X axis Such a configuration also has the advantage that the waste tube from the injection valve can be connected to the front Waste Inlet of the Wash Station module This waste tube is part of the DLW Option Kit E Figure 12 Mounting the DLW Option Modules in combination with the Injection Valve PAL is a registered trademark of CTC Analytics AG Switzerland note note Addendum to PAL System User Manual oY SYSTEM DLW Option 1 Locate the DLW Wash Station module 2 Attach the holder to the X axis at selected position as shown above 3 Connect the Waste Tube from the Injection Valve to the DLW Wash Station 4 Connect the Waste Tube to the Wash Station Waste Adapter Replace the dummy plug s on the front if used to connect the waste line coming from the injection valve This sequence of waste lines simplifies the system avoiding several waste lines running to the waste container The other two dummy plugs underneath the waste block are used for the DLW option The DLW tubing trom the DLW Pump outlet could be connected at these wash station inlet ports The functionality of the Fast Wash Station would then be configured using this order of connections Figure 13 Attaching the DLW Wash Station to the X Axis The waste tube must be positioned below the injection valve Make sure that the waste liquid flows into the waste container
15. system changes The test is designed for PAL standard valves with a bore size of 0 25 mm which allows a flow rate range of 10 to 500 uL min For both the carry over and repeatability tests conditions will be described for UV detection Details are listed below Note that the given levels to reach for carry over and repeatability do not reflect the specification values The tests described here should give the operator a confirmation whether the system is working as expected These tests are comparable to an OQ Operational Qualification Test The test procedure Is written for standard equipment such as a standard syringe of 100 uL and a loop size of 20 uL The procedure does allow configuring the system with different equipment but the user has to bear in mind certain limitations For example if a 100 uL syringe is installed alongside a loop with a volume lt 10 uL the reliability of a partial loop injection Is questionable The relative standard deviation will be above the expected limit of 1 20 Example 2 uL loop 1 uL partial loop filling yields a relative standard deviation of 2 to 4 Under such conditions a full loop injection Is the only valid test 71 PAL is a registered trademark of CTC Analytics AG Switzerland system Addendum to PAL System User Manual DLW Option 3 1 Carry Over Test with UV Detection e Test Sample Chlorhexidine 0 6 mg mL dissolved in Water Acetonitrile 90 10 0 1 TFA e Blank
16. to orient the guide wire Use the M3x5 screw provided and the corresponding serrated lock washer to connect 3 Test if the wire tension Is enough to keep the tubing in an upright position but low enough to move the syringe holder manually along the X axis The design of the DLW Manifold and the concept of washing by active pumping and closing of the lines by the DLW Actuator does not require a predefined solvent line position in theory Problems could occur if solvent is refilled or exchanged while the tubing still contains solvent from the previous setup Always keep the lines at the same position and carefully prime the entire system to prevent unnecessary contusion See also section G Operation 4 Connect solvent line 1 to the upper and solvent line 2 to the lower port at the left side of the DLW Manifold The order of the tube connections upper and lower positions should be consistent For certain applications it is crucial not to mix types of solvents e g biofluid sample solutions should not come in contact with highly concentrated organic solvents 25 PAL is a registered trademark of CTC Analytics AG Switzerland note note Addendum to PAL System User Manual A SYSTEM DLW Option 2 5 3 Inserting the DLW Syringe Holder Assembly into the Injection Unit It is recommended to insert the DLW Syringe Holder assembly when the PAL System is powered off 1 Move the PAL Injection Unit aside manually to allow
17. 39 1 2 Priming with Utlitities Function Syringe 39 1 3 Other Priming functions or Troubleshooting 40 2 Location of Solvent and Waste Bottles 40 3 Functionality of the DLW Option 41 3 1 DLW Pumps 4 3 2 DLW Actuator Solenoid 4 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Table of Contents contd 4 Installing Cycle Composer Macros or ICC Cycles used for the DLW Option 43 4 1 Installing the Cycle Composer Macros or ICC Cycles 43 4 2 General Considerations 44 4 3 Standard DLW Injection Cycle 45 4 4 Fast DLW Injection Cycle 47 5 PAL DLW Cycle Step by Step 49 5 1 Standard Cycle 49 5 2 Additional Valve Toggle Step to the DLW Standard Cycle 56 5 3 Fast Injection Cycle Step by Step 61 H Troubleshooting 67 1 Troubleshooting based on observed Symptoms or Error Messages 67 2 Special Considerations for Troubleshooting 69 2 1 Tubing Connections 69 2 2 Solvent Delivery Pumps are not priming 74 2 3 Solvent Delivery Pump Is not functioning 74 2 4 Verify the Functionality of the DLW Actuator Solenoid 75 3 Standard Chromatographic Tests 77 3 1 Carry Over Test with UV Detection 78 3 2 Repeatability Test with UV Detection 79 3 3 Carry over and Repeatability Tests with UV Detection as described for the OQ PQ PV Test 80 Replacing Parts 81 1 DLW Pump Module 81 2 DLW Wash Station 81 3 DLW Syringe 82 4 DLW Syringe Holder Assembly 82
18. 5 mm must be considered as well as the tubing and injection loop internal diameter When in doubt about the delivered volume of a pump a simple test can provide the answer Execute the test first and only if the result is negative is it necessary to check the flow from the pump directly e Test 1 Pump flow delivered throughout the entire system 1 Prepare a graduated cylinder with a volume of 10 or 20 mL 2 Test solvent Water 3 Select the following path from the local Terminal Menu Utilities Wash Station select the desired Wash Station 4 The delivery pump and the DLW Actuator are activated the delivered flow can be collected at the DLW syringe needle 5 Expected result A volume of approximately 8 mL should collect within 30 seconds Water DLW syringe needle Gauge 22 74 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option e Test 2 Pump flow delivered at pump outlet 1 Disconnect the outlet tube from the pump module 2 Connect PEEK tubing with the following dimensions to the outlet of the pump module Length 85 mm ID 0 25 mm 0 010 3 The wash solvent in the reservoir bottle should be water If necessary replace the solvent 4 Activate the pump by selecting following path Menu F3 Setup Objects Events Pwr Out1 or Pwr Out2 5 Collect the pumped water in a graduated cylinder 6 Expected Result Within 30 seconds the pump should d
19. 8 Dispense Front Air Segment and Front Sample Volume to Waste Figure 42 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oa SYSTEM DLW Option Standard Steps 9 10 Valve is switched to LOAD position Loop Is filled with Inject Volume Figure 43 Standard Step 11 Valve is switched to INJECT position start chromatographic process Figure 44 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Standard Step 12 Rear Sample Volume and Air Segment are dispensed to Waste Figure 45 Standard Steps 13 14 Valve Clean with Wash Solvent 2 Figure 46 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oY SYSTEM DLW Option Standard Steps 15 16 Active Syringe Needle wash with Wash Solvent 2 Column Figure 47 Standard Steps 17 18 Valve Clean with Wash Solvent 1 2 Column Figure 48 54 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Standard Steps 19 20 Active Syringe Needle Wash with Wash Solvent 1 Figure 49 Standard Cycle End Figure 50 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 5 2 Additional
20. Addendum to PAL System User Manual A SYSTEM DLW Option Addendum to the PAL User Manual PAL DLW Option Installation and Operation DLW Dynamic Load amp Wash Printing History Edition 1 November 2009 Edition2 June 2010 Original Instructions CTC Analytics AG reserves the right to make improvements and or changes to the product s described in this Addendum at any time without prior notice Under no circumstances shall CTC Analytics AG be held liable for any coincidental damage or damages arising as a consequence of or from the use of this document CTC Analytics AG makes no warranty of any kind pertaining to this product including but not limited to the implied warranties of merchantability and suitability for a particular purpose 2010 CTC Analytics AG All rights reserved Neither this publication nor any part hereof may be copied photocopied reproduced translated distributed or reduced to electronic medium or machine readable form without the prior written permission from CTC Analytics AG except as permitted under copyright laws CTC Analytics AG acknowledges all trade names and trademarks used as the property of their respective owners PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option A Safety Information Declaration of Conformity Please refer to the Declaration of Conformity sheet enclosed with the instrument A General Con
21. Inappropriate wash solvent composition Tube of Holding Loop at end not cut square or nut ferrule fixation not correct DLW Actuator Solenoid questionable defect Solvent Frits in wash solvent reservoir blocked Table 6 Troubleshooting Reduce sample volume in sample vial Use setting under F3 Setup System PrePressureVial Check recommended Method Parameters in Flow Chart Firmware Overview Appendices point 4 The following parameters are crucial and should be verified first Fill speed Pullup delay Injection Speed Post Inj Delay Remark The parameters mentioned above are listed in the standard injection cycle LC Inj Please note that this cycle cannot be used for the DLW operation However the values of these parameters can be used as a good starting point Check all connections within the DLW system such as needle Holding Loop injection loop connections etc Use appropriate wash solvent s Replace the waste tubing with larger ID tubing Verify the DLW method parameter for Airgap Volume Prime the entire system Increase Pst Inj Slv1 Slv2 values in method cleaning time Use PreClnSlv1 Slv2 and PreClnSpl Use appropriate wash solvent s Replace the waste tubing with larger ID tubing Replace valve rotor see Section H point 4 Change valve needle seal see Section F point 6 2 5 Use appropriate wash solvent Observe also the order of use for biologi
22. J Maintenance 83 K Limitations 84 1 PAL System 84 2 Sample Volume 84 3 Wetted Parts and Material 84 4 Tubing Internal Diameter ID and System Backpressure 84 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Table of Contents contd L Appendices 85 1 Definition of Terms 85 2 Naming Convention 88 3 DLW Option Spare Parts and PM Kit 89 3 1 Spare Parts Order Information 89 3 2 PM Kit Order Information 92 PAL is a registered trademark of CTC Analytics AG Switzerland syste Addendum to PAL System User Manual DLW Option C List of Figures and Tables Figure Figure Figure Figure Figure Figure Figure Figure Figure 9 Figure 10 Figure 11 Figure 12 CON DUO A N N c Figure 13 Figure 14 Figure 15 Figure 16 Figure 17 Figure 18 Figure 19 Figure 20 Figure 21 Figure 22 Figure 23 Figure 24 Figure 25 Figure 26 Figure 27 Figure 28 Figure 29 Figure 30 Figure 31 Figure 32 Figure 33 Figure 34 Figure 35 Figure 36 Figs 37 50 Figure 51 Figs 52 57 Figs 58 69 Figure 70 Figure 71 Figure 72 Figure 73 Figure 74 PAL DLW Option General System Overview PAL Active Wash Station Components New Upper Needle Guide touching electronic Components New Upper Needle Guide installed with modified PCB Preparing the Injection Unit for DLW Option Adjusting Needle Guide to Needle Length for DLW Option Adjusting the
23. LC System Binary or isocratic Solvent Delivery Pump UV Detector Wavelength 258 nm 10 mm path length HPLC Column C18 2 1 x50 100 mm spherical 2 7 to 5 0 um or equivalent Column Temperature Ambient to 30 C Mobile Phase A 20 Water B 80 Methanol socratic 0 5 to 1 0 mL min e Wash Solvents Wash1 Water Methanol 50 50 Wash2 Water Methanol 50 50 e Loop Size 20 uL Loop ID 0 25 mm e Injection Valve 6 port Injection valve with bore size 0 25 mm or 0 40 mm Example VICI Valco C72VX 1696D CTC e Cycle Composer Macro or Cycle ICC CE DLW Standard e Cycle Composer Method Parameter Settings Parameter Value Parameter Value Syringe 100 uL DLW Pullup Del 5000 ms Sample Volume 60 uL Inject to Specified Inj Valve Airgap Volume 3 uL Inject Speed 5 uL s Front Volume 5 uL Pre Inj Del 500 ms Rear Volume 5 uL Pst Inj Del 500 ms Needle Gap Valve Clean 3 mm Valve Clean Time Solvent 2 6S Fill Speed 2 uL s Post Clean Time Solvent 2 6S Fill Strokes 0 Valve Clean Time Solvent 1 6S Stator Wash 0 Post Clean Time solvent 1 6S Table 9 Test with Benzophenone and UV detection Expected results for the Benzophenone test are given below The OQ PQ PV procedure defines the following limits Repeatability 0 8 rel standard deviation Carry over lt 0 10 These limits can easily be reached with an intact system using the described Benzophenone test Typical values may be Repeatability 0
24. Prime every liquid syringe manually before inserting into the PAL system 2 Screw the prepared DLW syringe into the holder Hold the syringe at the lower metal mount while tightening the syringe Figure 16 Inserting the DLW Syringe into the Holder Holding the glass barrel while tightening can damage the seal where the glass meets the metal 22 PAL is a registered trademark of CTC Analytics AG Switzerland note note note note Addendum to PAL System User Manual A SYSTEM DLW Option Do not mount the syringe needle at this stage 2 5 3 Connecting the DLW Tubing to the Syringe Holder It is recommended to connect the DLW Tubing from the kit to the Syringe Holder while the assembly ts not yet inserted in the X axis The wetted parts of the DLW Actuator Solenoid are composed of PEEK for the body and FFKM Simriz for the seal material PEEK exhibits excellent resistance to most commonly used chemicals However the following solvents are not recommended for use with PEEK DMSO THF methylene chloride dichloromethane nitric acid sulfuric acid For more details see the compatibility tables provided by the manufacturer of PEEK materials or components 1 As described under point 2 3 Installing Pump Module the end of the the DLW kit sleeved tubing with the short tubes has to be connected to the DLW Syringe Holder Assembly 2 Use the groove in the block at the back side of the DLW Manifold
25. Valve Toggle Step to the DLW Standard Cycle 5 2 1 Considerations for additional Stator Wash Cleaning Step The DLW Standard Cycle has the built in option for the user to toggle the injection valve at the end of the chromatographic run before equilibration of the column to the start conditions If the Method variable Stator Wash is set to 1 the extra cleaning process for the valve with Valve Toggle becomes part of the standard cycle If the Method variable is deactivated setting O the DLW Standard cycle ends as shown in Figure 50 The macro cycle is written such that the optional valve toggle steps can be executed before re equilibration of the column The exact time the valve needs to be switched on has to be synchronized with the chromatographic method by the user Method variable Delay Stator Wash The two wash solvents are timed by the Method variables Stator Wash Time Solvent 1 and Stator Wash Time Solvent 2 After these wash times have elapsed the valve is Switched back to the start position Figure 51 illustrates the recommended retention time for Stator Wash or Valve Toggle times B Valve Toggle Absorbance Gradient Start Time Figure 51 Timing for Stator Wash Step From the chromatographic point of view it is Important to understand the optional cleaning step in detail It is assumed that the valve stator between ports 1 and 6 example standard Chemi
26. an Time Solvent 2 Needle Gap is a parameter from Rinse Inj Atom The variable in this macro is Needle Gap Valve Clean The function of this parameter is to lift the needle in the injection port to allow rinsing around the needle tip The pressure of the spring loaded balls in the DLW Syringe Holder Assembly is released by moving approx 3 mm up default This leaves a gap between the needle tip and the valve bottom of approx 1 mm to enable a flush at this contact point Fig 46 The Injection Unit is moved to the DLW Wash Wash2 Station position Wash2 Syr Eject Speed The needle is rinsed inside and out with Wash Post Clean Time Solvent 2 Solvent 2 Fig 47 The Injection Unit is moved back to the Injection Wash1 Valve The Inlet Port and engraving to waste Port Inject to is flushed with Wash Solvent 1 to prepare the Needle Gap Valve Clean valve for the next injection Valve Clean Time Solvent 1 Fig 48 PAL is a registered trademark of CTC Analytics AG Switzerland 45 syste The Injection Unit is moved back to the DLW Wash Station position Wash1 to flush the syringe needle inside and out with Wash Solvent lee This is a preparation step for next injection Especially important for biofluid samples Cycle end for LC Inj DLW Standard macro An optional cleaning step is attached to the DLW Standard injection cycle Stator Wash or Valve Toggle A Repeat End loop
27. and ready for use All the points described below are not applicable for new systems For all field upgrades make sure the instructions given below are followed For field upgrades only The injection unit the Z axis must be prepared in order to install the DLW option The upper needle guide must be removed and the lower needle guide must be lowered to adapt to a needle length of 73 mm The standard setting for the needle guide is 50 5 0 5 mm Detailed descriptions for these maneuvers are given in the PAL Service Manual A brief outline is provided below 1 Remove the Injection Unit from the Y axis 2 Open the top cover to disconnect the long tension cord 3 Pull out the slider to access the Anti Twist Device of the upper needle guide Open the Torx screw of the device 4 Pull out the slider to access the Mechanical stop for Needle Guide Carefully open the Torx screw of the stop 5 Pull out the lower needle guide assembly until the Upper Needle Guide can be removed The new Upper Needle Guide designed for use in combination with the PAL DLW Option has a longer rod This rod length is critical if the new version of the Upper Needle Guide Is used and in cases where the PAL System Is retrofitted to standard liquid injection using a Wash Station other than the PAL DLW Option The longer rod would touch an electronic component from the Z axis PCB Therefore the system must be adjusted to avoid this problem with
28. ar Volume Holding Loop SL volume Airgap Volume The Front air segment is aspirated Filling Speed Pullup Delay Fig 40 The Injection Unit moves to the DLW Wash Station position Wash1 The needle is inserted dipped for 1 second to wash the outer needle surface No plunger movement at this step Fig 41 The Injection Unit moves to the specified Inject to Injection Valve Front Volume The Front and Airgap Volume are ejected Airgap Volume Injection Speed Fig 42 The PAL System waits for the signal from the data Wait for DS system The Injection Valve is switched to position Inject to Active Pre Inject Delay Fig 43 The system pauses according to the delay time Pre Inject Delay The loop is filled with the sample volume as SL volume specified in the Sample List Injection Speed The Injection Valve is switched to position Post Inject Delay Standby the loop contents are injected Timer 1 Fig 44 Timer 1 Delay Stator Wash is started and a start signal to the HPLC system is sent The plunger of the DLW Syringe is pushed down Syr Eject Speed to dispense the Rear Sample and Air Segment to Waste The Holding Loop is still filled with Wash Solvent 1 Fig 45 The DLW Actuator Solenoid is activated to deliver Wash2 Wash Solvent 2 into the Holding Loop to clean Inject to the Injection Valve from Port 1 to Port Waste Needle Gap Valve Clean Valve Cle
29. arts The syringe and Holding Loop are preloaded with Wash Solvent 1 at the start The sample is picked up and remains separated from Wash Solvent 1 by an air gap After loading the loop and injection Wash Solvent 1 is pushed into the system followed directly by Wash Solvent 2 to flush the critical valve paths The DLW Syringe Assembly is then moved to the wash station for further cleaning steps and to prepare the Syringe and Holding Loop for the next cycle Further details are given in section G Operation point 5 Cycle Step by Step PAL is a registered trademark of CTC Analytics AG Switzerland syste 1 1 Specifications Part Number e DLW Pumps Flow rate range Wetted parts e Transfer Tubing Kit e Solvent Reservoir e DLW Wash Station Wetted parts e DLW Syringe Assembly o Syringe Wetted parts o Syringe Needle o DLW Actuator Solenoid Wetted parts o DLW Manifold adapter assembly Wetted parts o Holding Loop o DLW Flow Diverter Wetted part Addendum to PAL System User Manual DLW Option PAL DLW Option Two pcs solenoid diaphragm pump self priming 100 mL min at atmospheric pressure Pump Itself no connections to HPLC system Pumps Ryton PPS and Kalrez FFPM Two pcs 1 8 PFA tubes including connection fittings 1000 mL borosilicate glass including Glass Filter 40 um pore size Wash Station with two solvent ports and one Waste port T
30. bubbles are observed Repeat the steps to prime with Solvent 1 by activating Wash 1 39 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual A SYSTEM DLW Option 1 3 Other Priming functions or Troubleshooting Another priming function is available in section Utilities Injector Select Clean Injector and follow the dialog with similar logic as described above In case any problems are observed follow the instructions given in Section H Troubleshooting Dedicated tests to verify the DLW functionality are described 2 Location of Solvent and Waste Bottles The DLW Option contains self priming membrane pumps The solvent bottles can be placed either in the Fast Wash Station Holder or on the lab bench The Waste bottle must be placed gt 30 cm below the injection valve Make sure that the waste liquid can flow into the waste bottle without restriction The waste tubing must be placed above the level of the liquid Ideally the tube should be fixed at the neck of the waste bottle Use good lab practice to avoid contaminating the wash solvents and bottles Avoid the growth of biological contaminants in pure water by either replacing it regularly or adding a small amount of organic solvent such as methanol or acetonitrile Certain buffer solutions can decompose at room temperature when exposed to light Filtering the wash solvents before filling the bottles especially if
31. cal samples The first wash Is always an aqueous solution followed by organic solvents Check the connections from the Holding Loop Is the pilot distance appropriate Is the tube end damaged Is the tube end cut absolutely square Are the right nuts and ferrules used Verify the functionality of the DLW Actuator Is solvent flowing when active Use command Utilities Wash Station for tests Clean the frits in an ultrasonic bath with an appropriate solvent If flow is not ensured after cleaning replace frits Replace wash solvent and clean reservoir bottle and tubing carefully 68 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 2 Special Considerations for Troubleshooting 2 1 Tubing Connections Special attention must be paid to all tubing connections Common mistakes in routine practice are Nut and ferrule not connected tightly and leak Pilot distance incorrect Tubes not cut square Wrong type of ferrule or nut used Air bubbles trapped Male Nut internal Ferrule Tubing Pilot Figure 70 Definition of Terms for Tubing Connections Disregarding these basic rules could lead to e Dead volume e Peak deformation or peak splitting e Carry over effects e Decreased resolution of the tubing between column and detector 2 1 1 Nut and Ferrule Tightening 2 1 1 2 Stepwise Tightening The first important rule when
32. cals However the following solvents are not recommended for use with PEEK DMSO THF methylene chloride dichloromethane nitric acid sulfuric acid For more details see the compatibility tables provided by the manufacturer of PEEK materials or components A green LED is activated when current is applied from the actuator control PCB to the actuator solenoid This activation does not mean that the solenoid will open or close In cases of defects such as a defective or clogged Actuator Solenoid would the LED lighten up but the solvent path is not opened 4 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Closed Solvent NC a ii i ii ty eet PET z224 I 3 DLW Flow c Diverter ee E TI So vent 2 Figure 35 DLW Manifold and Actuator Solenoid Syringe amp Se Holding Loop Figure 36 Inserting the DLW Flow Diverter PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 4 Installing Cycle Composer Macros or ICC Cycles used for the DLW Option The PAL DLW Option can only be operated in combination with PAL control software such as Cycle Composer or the Cycle Editor for ICC interpretation in another data handling system software For software control two macros or two cycles are provided The first macro is used for Standard
33. can be activated with the Count If Stator Wash is activated the following steps will be executed The Injection Unit is moved to the Injection Valve From the last step above the Holding Loop is filled with Wash Solvent 1 Timer 1 is awaited to switch the valve Toggle into Active position fill loop The DLW Actuator Solenoid is activated to deliver Wash Solvent 2 to the Holding Loop and into the valve system The first solvent flush arriving at the valve Is Wash Solvent 1 followed by Wash Solvent 2 The Wash Solvent is changed to Wash Solvent ge The Injection Valve is switched back to the Standby position Table 4 LC Inj DLW Standard Macro Addendum to PAL System User Manual DLW Option Wash Post Clean Time Solvent 1 Stator Wash Fig 49 Fig 50 Stator Wash count 1 Cleaning step active Stator Wash count 0 Cleaning step disabled Inject to Delay Stator Wash Active Inject to Wash2 Stator Wash Time Solvent 2 Inject to Wash1 Stator Wash Time Solvent 1 Inject to Standby PAL is a registered trademark of CTC Analytics AG Switzerland Fe 55 Fig 54 Fig 55 Fig 56 Fig 57 46 Addendum to PAL System User Manual DLW Option system 4 4 Fast DLW Injection Cycle The Fast injection cycle differs from the Standard cycle in three points e The needle is not dipped in the Wash station
34. d trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Fast Steps 11 12 Valve Clean with Wash Solvent 2 Figure 66 Fast Steps 13 14 Valve Clean with Wash Solvent 1 Figure 67 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Fast Step 15 Active Syringe Needle Wash with Wash Solvent 1 a E Column Figure 68 Fast Cycle End Figure 69 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual A SYSTEM DLW Option H Troubleshooting The PAL System is always one of several components that make up a complete chromatography system The following Troubleshooting Guide is limited to the DLW Option only Troubleshooting for the PAL System is covered in the corresponding PAL User Manual 1 Troubleshooting based on observed Symptoms or Error Messages Symptom or Error Message Possible Cause Recommended Action No or very low Clogged Syringe Needle or Remove syringe and aspirate dispense liquid manually detector signal is Holding Loop Clean syringe and or needle Holding Loop observed Bent needle Inspect and or change syringe No sample liquid injected Check and or adjust Needle Penetration into sample vial see Section F point 6 2 Sample volume too low Increase sample volume The valve n
35. de of the Injection Unit centers on top of the Wash Station assembly Fine tune the Z Position until the bottom of the lower needle guide is flush with the top surface 3 Press F1 to verify the position and F4 for Home 55 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual a SYSTEM DLW Option 4 Injection Valve Reference Points For an LC Valve the reference position is the valve needle guide fitting mounted on the top valve port see Fig 31 Figure 31 LC Valve Reference Point The lower needle guide of the Injection Unit is centered on the valve needle guide fitting Adjust the Z Position so that the bottom of the lower needle guide just touches the surface of the valve needle guide fitting Then reduce this value by 2 0 mm See Figure 32 oe 2mm gap Figure 32 Positioning the Z Axis Needle Guide on the Valve Needle Guide To adjust the Valve Needle penetration value see point 5 below Defining the Valve Needle Penetration Value 56 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual A SYSTEM DLW Option 5 Defining the Valve Penetration Value Before performing the following steps make sure that Object positions X Y and Z for LC Vivi are properly defined and that the complete DLW syringe assembly is inserted into the injection unit Complete the following step
36. ded to use precut tubing which is cut smooth clean and passivated Polymer tubes such as PEEK Teflon PFA etc are soft and entice the user to cut them with any handy tool Commercially available tubing cutters from 71 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option many manufacturers are available If the blade does not provide a clean right angle cut use a different technique or tool The best and most commonly used method for cutting tubing of any material is to use a cutter with an adjustable blade Carry out the initial turns readjust the blade a little turn once more and adjust again until approximately half of the tubing wall is cut Hold the tube on both sides of the cut with flat nose pliers and twist the tube until it breaks For polymer tubing the same can be done by replacing the flat nose pliers with tweezers with a flat tip Tube end cutting 90 Correct X Angled Wrong 7 Rough or pressed edges Wrong Figure 72 Illustration of various tubing cuts 72 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option 2 1 4 Wrong Nuts or Ferrules used Do not use a nut or ferrule from a vendor other than specified for the product Figure 73 below illustrates the resulting dead volume that occurs when an incorrect type Is used for connecting The second illustrat
37. e the crucial DLW parts are attached to the injection unit at any available position 2 9 2 Electrical Connections for 2 or 4 Injection Valves Connect the valve drives as shown below in Figure 27 Note that the DLW option can only be used in combination with PAL xt systems Therefore connect the first cable from the PAL Interface MODBUS to the serial valve drive connector A Each drive is connected with daisy chain cabling The last serial drive connector B remains open or unused Electrical current and interface logic is provided by the interface MODBUS 30 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual wy SYSTEM DLW Option TERMINAL SER1 LAN MODBUS wasn INTERFACE 1 INTERFACE 2 AUX 1 AUX 2 D CC M mo _ e Figure 27 Electrical Connections to 4 Injection Valves Daisy Chaining with PAL x System gt ow gt ow B notused A OT PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option F PAL Firmware and PAL Firmware Object Installation for the DLW Option 1 PAL Firmware The operation of the DLW option is only possible with the PAL xt System which requires the APR Control xt board and PAL Firmware version 4 1 X The DLW option is NOT compatible with a PAL System operating with PAL Firmware version 3 0 X or lower The DLW
38. e second waste line can be connected to the lower waste port in front of the DLW Wash Station If only one connector is used apply a dummy plug to the second port on the Wash Station 2 Place the Waste Tube from the DLW Wash Station into the waste container The waste container must be positioned below the injection valve Make sure that the waste liquid flows into the waste container without restriction 27 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Wash Station Figure 22 Solvent Line Plumbing on Cheminert Valve Types Wash Station Figure 23 Solvent Line plumbing on Valco W type Valves PAL is a registered trademark of CTC Analytics AG Switzerland note note Addendum to PAL System User Manual oY SYSTEM DLW Option Wash Station Waste Pos 2 Figure 24 Solvent Line plumbing on Rheodyne Valve PD Series The Agilent specific Rheodyne Valve with Agilent PNo 5067 412 is connected in the same manner to the HPLC system as shown in the corresponding HTS or HTC PAL User Manual for the Rheodyne type PD7991 For details see the specific valve description 2 8 Electrical Connections for the Single Injection Valve Setup Always switch the PAL power supply OFF before connecting or disconnecting the PAL DLW cable or any other PAL accessory cables The electrical connection of the DLW option Is identical to t
39. eedle guide and or Check valve needle guide and seal see Section F needle seal are not properly point 6 2 5 installed The valve ports are not plumbed Check plumbing connections see Section F point 6 2 correctly to the pump and or detection system Wrong valve type specified Check valve type by selecting the path Menu setup Objects Injectors LCVIv1 valve Sample is backing up Syringe OD too small Check the syringe needle for correct Gauge on the valve needle Gauge 22 OD 0 72 mm The needle penetration depth for Adjust the Injection Valve Needle Penetration the injection valve is not set see Section F point 6 2 6 correctly The Syringe Plunger speed is too Reduce Inject Speed in method high resulting in excessive pressure in inlet Syringe does not fill DLW Actuator Solenoid not PAL Firmware level lt 4 0 or PAL Firmware Object for properly functioning DLW Option not loaded onto the system Solvent path blocked Prime the system or inspect it to identify the reason for clogging Air bubbles trapped in the system caused by loose connections or tubing not cut square Syringe plunger tip worn out Replace syringe plunger Check if glass barrel is scratched damaged If in doubt replace the entire syringe at once DLW Actuator Control PCB If possible verify the DLW Actuator Holder Assy on defect another PAL System If defect is definite replace PCB DLW Flow Diverter Check the Flow Diverter s the Flow Dive
40. eliver approximately 10 mL This short test will give an indication of whether the system Is performing as expected If none or very little flow is observed perform a systematic check using a process of elimination Consult the table in the Troubleshooting section for more details 2 4 Verify the Functionality of the DLW Actuator Solenoid The DLW Actuator Is electrically connected to the circuit of the heated syringe Increasing the heated syringe temperature does activate the DLW Actuator The green LED is the indication that current is applied but please note that this is not proof that the DLW Actuator opens the seat to allow a solvent flow To check the functionality the following two tests are described e Test Electrical activation of the DLW Actuator 1 Select the DLW syringe by following path and change the standby temperature to 65 C Path from local terminal Menu Utilities Syringe Standby Temp select 65 C 2 Expected result The green LED on the Syringe Holder assembly is turned ON Please do not reset the temperature if the second test Is activated as well In case this result is not observed verify the electrical connection from the DLW Actuator to the PCB PAL is a registered trademark of CTC Analytics AG Switzerland 715 Addendum to PAL System User Manual A SYSTEM DLW Option e Test 2 Verify the seal tightness of the DLW Actuator Close the DLW Actuator by setting the standby tempera
41. enced at using automated systems to run existing analytical procedures The Appendices provide information on PAL DLW macros for multiple injection valve combinations and on spare parts PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option E PAL DLW Option Installation 1 General System Overview p Figure 1 PAL DLW Option General System Overview DLW Dynamic Load amp Wash The DLW Option represents a new concept in wash stations combining an injection cycle with wash steps The combination of both injection and wash steps usually separate procedures minimizes cycle time and carry over The two characteristic features of the DLW Option are e The sample solution never comes in contact with the syringe itself it is held sandwiched in a so called Holding Loop e Wash solvents are pumped from the rear to the front of the DLW system to intensely purge all critical surfaces which have come in contact with the sample The DLW option consists of two self priming micro pumps mounted on a dedicated bracket the wetted parts are Ryton PPS and Kalrez FFPM The IN ports are connected to the wash solvent bottles and the OUT ports are connected to the DLW manifold which is part of the dedicated DLW Syringe Holder assembly A Holding Loop separates the syringe and the DLW actuator to avoid the sample coming in contact with these p
42. er part of a standard PAL configuration e g Combi PAL HTS PAL or an optional addition e g Cooled Stack MALDI Tool Dilutor The term Module is intentionally used to differentiate from Object which is reserved for the PAL Firmware Object 87 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 2 Naming Convention This section recommends the standard naming convention for the PAL System Following this convention will allow the PAL setup to be pre configured for certain applications This will simplify software backups and application development and will improve technical support and training Tray Type Tray Description VT200 VT98 VI78 V154 V121 V132 10 V132 20 M196 DW96 M1384 Vial Tray 200 positions 10 x 20 For 7 mm micro vials 1 mL Vial Trays 98 positions 7 x 14 For 12 mm vials 2 mL Vial Tray 98 positions 6 x 13 For 7 mm micro vials 1 mL opposite side of 98 positions Tray Vial Tray 54 positions 6 x 9 For 12 mm vials 2 mL Vial Tray 21 positions 7 x 14 For 12 mm vials 2 mL opposite side of 32 positions Tray Vial Tray 32 positions 4 x 8 For 23 mm headspace vials 10 mL Vial Tray 32 positions 4 x 8 For 23 mm headspace vials 20 mL Standard 96 position shallow microplate Deep well 96 position microplate High density 384 position shallow microplate Table 11 Naming Conventi
43. es are given below to ensure uninterrupted operation If you use the system extensively for example throughout nights and weekends or If you use corrosive solvents you may need to perform the maintenance procedure more frequently The points listed below are dedicated to the DLW Option Maintenance for the PAL system Is listed in the specific PAL User Manual Maintenance Step Prime the entire DLW system by using the Utilities functions Wash Stations or Syringe Clean the outside of the instrument Use only a soft lint free cloth dampened with mild soap and water Clean Instrument syringe and surfaces Replace the DLW Syringe plunger Replace the DLW Syringe Needle Observe the correct placement of the Teflon seal Replace the Valve Needle Seal Check all connections for tightness Replace the DLW Holding Loop Check the spring loaded Needle Holder Assembly for functionality The spring compression in particular must be checked Wash and Waste Solvent reservoirs Check flow restriction of frits Quality of solvents biological growth Check the functionality of the DLW Actuator Solenoid Table 10 Maintenance Check List Interval Daily or as needed Required before routine running Weekly or as needed Weekly or as needed The Syringe Plunger for a gas tight syringe polymer tip has to be replaced on a regular basis The interval is highly dependent on the application
44. escription PAL DLWLoop Kit PAL DLW Holding Loop with Needle Adapter mounted Needle Kit for PAL DLW Option Gauge 22 PST 3 length 51 mm PAL DIC al Package of 3 pcs incl Needle Retaining Nut MZ 30 21 Tension Cord 280mm 1 each MZ 30 23 Tension Cord 90 95 mm 1 each MV 30 13 Needle Seal Gauge 22 for Valco Valve pkg of 3 Kit SealCap 10 caps and 10 seals for 100 VW solvent bottle PAL BC TestL CTC Test Label for Barcode Reader pkg of 3 PAL LubKit PAL Lubrication Kit incl brush BN 4851 Screws for mounting Injection Unit M4x8 pkg of 2 PAL PM DocLC PM Document HPLC Technique Revision F Table 13 PM Kit for PAL xt System including PAL DLW Option Note To reorder Needle Seals please use the following information PNo PAL NdlSeal Description Needle Seal Gauge 22 for Valco Valve pkg of 10 Needle Seals for valves made out of PAEK material Gauge 19 and Rheodyne type valves are available 92 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option This page intentionally left blank 93 PAL is a registered trademark of CTC Analytics AG Switzerland
45. hat of the PAL HTS xfand the HTC xt models The same board is used for both models The schematic in Figure 25 shows an installation using the valve drive connected to the AUX interface If a serial or multiposition valve drive Is used see schematics for 2 or 4 injection valve combinations Figures 26 and 27 o o INTERFACE 1 INTERFACE 2 a AUX 1 Tes pee gt eam gt DO o 8 8 Figure 25 Electrical Connection DLW Option on a PAL xf 29 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oY SYSTEM DLW Option 2 9 DLW Option Installation in Combination with 2 or 4 Injection Valves 2 9 1 General System Overview Valve 2 Waste line CH Valve 3 Figure 26 DLW Option in Combination with 4 Injection Valves The DLW Option can be combined with PAL Systems using a setup of 2 or 4 injection valves See Figure 26 The concept of the DLW option is that the crucial parts of the wash station are attached to the injection unit The sample is taken up into the Holding Loop the syringe is actually the prepared reservoir for the following wash step after the loop filling and injection steps Following this concept allows for more than one injection valve in the PAL configuration No special setups or additional modules are required to inject into and clean one or more valves connected to the PAL System As described abov
46. ion Figure 74 shows the various forms of the ferrules from different manufacturers Be aware that some of them are very similar in shape but not similar enough to exchange without risk m Tubing seats correctly at the bottom Tubing doesn t reach the bottom introducing dead volume Tubing reaches the bottom before ferrule seats Figure 73 Dead Volume created due to wrong Ferrule Type Pee eS Valco Rheodyne Parker Swagelok Figure 74 Ferrule Types from various Vendors 73 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 2 1 5 Avoiding Trapped Air Bubbles It is common practice to make a connection only if the ports are wetted The liquid helps to prevent trapped air bubbles which are often very difficult to remove or work out of a system 2 2 Solvent Delivery Pumps are not priming It has been observed that the solvent delivery pumps of the Pump Module can fail to prime at initial operation The pump has to be wetted at the solvent inlet port or preferably prime the tubing from the Solvent reservoir to the pump and make the connection Dry running the pump for a short time will not harm tt 2 3 Solvent Delivery Pump is not functioning The pump flow rate depends greatly on the viscosity of the selected wash solvent and the given backpressure of the entire system The valve bore size standard Cheminert valve 0 2
47. is deficiency Please observe the market to find a technical suitable solution 70 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 2 1 2 Pilot Distance Incorrect When making a new tube connection it is critical to follow these guidelines 1 Slip the nut and ferrule over the tube The end of the tube should stick out approximately 50 mm 2 Inches 2 Press the tube end firmly into its female counterpart for the connection 3 Move the nut and ferrule together into the female counterpart for connection At this stage Is it important to maintain pressure on the tube to avoid the tube slipping backwards out of position 4 Tighten the nut as described above ae Figure 71 Pressing Tube into Place Failure to follow this procedure will lead to an incorrect pilot distance which will result in actual dead volume not delay volume 2 1 3 Tube Not Cut Square All tubes must be cut absolutely square Any deviation from square causes a dead volume which yields carry over and other chromatographic effects This applies to stainless steel PEEK or Polymer tubing It is important to use the correct tool for each material when cutting a tube Stainless steel tubes are often cut using pliers Often an egg shaped profile will result which will no longer seal and will cause dead volume Dedicated pliers for HPLC tubing are available nevertheless it is recommen
48. jection Loop O O O O Total delay volume 205 uL Loop content volume The second valve toggle back to starting conditions should be done before the system equilibration time has started The Loop contents should ideally be a solvent of a low elution power when switched back If isocratic chromatography is applied it is possible that the remaining contaminants are washed into the system and can build up higher background noise for the column and or detector over a longer period of time SH PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual wa SYSTEM DLW Option 5 2 2 Additional Cleaning step Stator Wash or Valve Toggle Step by Step Figures 52 57 Additional Cleaning Step Stator Wash Step by Step Stator Wash End of Standard Injection Cycle Start for additional Cleaning Step Valve Toggle Figure 52 Stator Wash Steps 1 2 Valve Switched to Load Position toggle Valve clean with Wash Solvent 1 Figure 53 PAL is a registered trademark of CTC Analytics AG Switzerland 58 Addendum to PAL System User Manual A SYSTEM DLW Option Stator Wash Step 3 Valve Clean with Wash Solvent 2 Figure 54 Stator Wash Step 4 Wash Solvent 2 dispensed by Wash Solvent 1 Solenoid Actuator Valve Figure 55 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Opti
49. lder and select the DLW Option folder as the Method folder If you wish to add the DLW Option macros to an existing Method folder just copy the macros pma and the methods pme files from the DLW Option folder on the CD ROM to the appropriate folder If the PAL System Is integrated with a data system software that controls the PAL using the Cycle Editor for PAL ICC interpretation e g Analyst ChemStation Empower EZChrom MassLynx Xcalibur an ICC Cycle is used and not the Cycle Composer Macro the cycle extension Is cyx Follow the instructions as described above The folder for the cycle Is usually located within the data system application folder 43 PAL is a registered trademark of CTC Analytics AG Switzerland note note Addendum to PAL System User Manual A SYSTEM DLW Option A Cycle Composer macro can be converted to a Cycle file format extension cyx using the Cycle Editor software Conversion is possible starting with Cycle Editor Version 1 4 0 4 The macros provided are written for standard injection valve drives which are controlled and activated through the AUX interface 4 2 General Considerations The duration of the wash steps has to be established for each configuration and application The viscosity and surface tension of the individual wash solvents and the backpressure of the system are factors to consider Be aware that a higher backpressure build
50. lectronic waste WEEE Table 1 Commonly Used Safety Symbols PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option B Table of Contents A Safety Information 2 B Table of Contents 4 C List of Figures and Tables 7 D How to Use this Manual 9 E PAL DLW Option Installation 10 1 General System Overview 10 1 1 Specifications 11 1 2 Hardware Requirements 12 1 3 Software Requirements 12 2 Installation of the DLW Option 13 2 1 Unpacking the Components 13 2 2 Preparation of the Injection Unit for the DLW Option 14 2 3 Mounting the Pump Holder 17 2 4 Installing the DLW Wash Station 19 2 5 Installing the DLW Syringe Holder Assembly 21 2 6 Installing the Valve Needle Guide Assembly J 2 7 Injection Valve Plumbing and Connection to the DLW Wash Station 27 2 8 Electrical Connections for the Single Injection Valve Setup 29 2 9 DLW Option Installation in Combination with 2 or 4 Injection Valves 30 2 9 2 Electrical Connections for 2 or 4 Injection Valves 30 F PAL Firmware and PAL Firmware Object Installation f or the DLW Option 32 1 PAL Firmware 32 2 Loading the PAL DLW Object 32 3 Wash Station Reference Points 34 3 1 Wash1 Wash2 Reference Points 34 4 Injection Valve Reference Points 36 5 Defining the Valve Penetration Value 37 G Operating Instructions 39 1 Priming the Solvent Lines 39 1 1 Priming with Utilities Function Wash Station
51. les are processed The elements of a Method are a Cycle a Syringe and a Parameter List Methods have names with up to eight characters and can be edited copied and deleted Method Parameters Method Parameters are associated with the Cycle operations User assigned Parameter values define how a processing operation is performed A zero Parameter value will disable a Cycle operation Cycle Parameters are application specific Tray Holder A Tray Holder can hold one or more trays Each Tray Holder has a reference position X Y Z coordinates that defines its location 85 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Tray A Tray holds multiple samples Trays are defined by designating the Tray Type see below and the Tray Holder Tray names are used to identify the sample source within a PAL Job Tray Type A Tray Type defines the pattern and sampling sequence of sample locations within a Tray Stack A Stack is a particular type of Tray Holder that is designed to hold micro plates A six drawer Stack holds 12 standard micro plates two in each drawer A three drawer Stack holds six deep well micro plates two in each drawer PAL Object Manager Software to load a PAL Object List to an instrument if a Module hardware module has been added to the PAL System In a special mode Object Manager can also be used to create and maintain Object Lists
52. low path such as an injection valve will contribute to the system backpressure For example rather short tubing with an ID 0 13 mm 0 005 can build up high back pressure when standard wash pump flow rates are used The wash solvent pumps operate in the range of mL min Such a flow rate in combination with a high system backpressure caused by small ID tubing will slow down the pump and thus the flow rate The above described points are only critical if an HPLC setup dedicated for nano flow is selected 84 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option L Appendices 1 Definition of Terms Job Queue A Job Queue is a list of sample processing Jobs Jobs are executed in the order displayed on the JOB QUEUE menu screens New Jobs may be added to the queue while samples are being processed Job A Job contains the information needed by the PAL System to process multiple samples using the same processing steps The elements of a Job are a Method and a Tray that define the location of the samples to be processed Jobs are automatically numbered from 01 to 99 and then restart with number 01 when they are added to the Job Queue Cycle A Cycle consists of the specific operations necessary to process one sample The Cycle operations are repeated for each sample within a Job Cycles are designed for specific applications Method A Method defines how the samp
53. m of the lower needle guide lightly touching the surface of the Wash Station assembly Figure 29 DLW Wash Station Wash1 Wash2 Reference Points 1 Press HOME on the handheld controller and select the sequence Menu Setup Objects Wash Stations Wash1 2 Enter Wash1 and adjust the X Y Z positions until the lower needle guide of the Injection Unit centers on top of the Wash station assembly Fine tune the Z position until the bottom of the lower needle guide Is flush with the top surface 3 Repeat steps 1 3 for Wash2 using the following path Menu Setup Objects Wash Stations Wash2 54 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual A SYSTEM DLW Option 3 2 Waste Reference Point Injectors Waste The Waste position is defined as an Injector by the PAL software It is classitied in the Object category Injectors The reference position for the Waste Port a hole slightly larger than the needle guide has been placed in the middle positions of Wash1 and Wash2 see Fig 30 The lower needle guide should be centered on this hole with the bottom of the lower needle guide lightly touching the surface of the Wash Station assembly Figure 30 Waste Reference Position 1 Using the hand held controller select the sequence Menu Setup Objects Injectors Waste 1 2 Enter Waste and adjust the X Y Z positions until the lower needle gui
54. nert Valve is contaminated and cannot be cleaned during the standard injection process The valve toggle brings the engraving back between the two ports Flushing the valve with both wash solvents eliminates any remaining sample material located between Stator ports 1 and 6 56 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option What points need to be considered when using the Stator Wash or Valve Toggle option Observe the rules if biofluid samples are injected The first sample contact should always be with an aqueous solution to avoid protein precipitation After washing with an organic solvent higher elution power the system must be flushed again with Wash Solvent 1 The first toggle near the end of the chromatographic cycle provides the advantage that the sample loop is already flushed out first using the mobile phase with a solvent of high elution power assuming gradient application The second valve toggle time follows immediately after the second solvent flush is completed A second switching time cannot be programmed The waiting period before the second valve toggle should be long enough so that the entire system is flushed out by both wash solvents Consider the entire delay volume to determine the second valve toggle time The DLW internal volumes are Manifold 90 uL Holding Loop 108 uL Syringe Needle Gauge 22 6 7 uL Installed In
55. on Stator Wash Step 5 Second Valve Clean with Wash Solvent 1 Figure 56 Stator Wash Step 6 Valve switched back to Inject Position toggle Figure 57 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual my SYSTEM DLW Option 5 3 Fast Injection Cycle Step by Step Figures 58 69 The Fast Injection Cycle Step by Step Fast Cycle Start Figure 58 Fast Step 1 Aspirate Rear Air Segment Figure 59 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Fast Step 2 Get Sample Aspirate Rear Inject and Front Volume Holding Loop Figure 60 Fast Steps 3 4 Aspirate Front Air Segment Figure 61 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option Fast Steps 5 6 Dispense Font Air Segment and Front Sample Volume to Waste Figure 62 Fast Steps 7 8 Valve Is switched to LOAD position Loop Is filled with Inject Volume Figure 63 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oY SYSTEM DLW Option Fast Step 9 Valve is switched to INJECT position start chromatographic process Figure 64 Fast Step 10 Rear Sample Volume and Air Segment are dispensed to Waste Figure 65 PAL is a registere
56. ons 88 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oY SYSTEM DLW Option 3 DLW Option Spare Parts and PM Kit 3 1 Spare Parts Order Information Illustration DLW Pump Module PAL DLWPump PAL DLW Pump Kit single module indl electrical connections 24V PAL DilAspKit Aspiration Tube Kit consisting of 1 pc Tube PFA 1 pc PEEK solvent filter 10 um 1 pc Plug with feed through hole Cbl R4LX 1300 Cable for PAL xfWash Station round connector length 130 cm PAL DLWPCB PAL DLW Pump Control Module PCB Kit PAL TubeDLW PAL DLW Wash Station Tubing Kit Consisting of 2 tubing wire supports and connections PAL is a registered trademark of CTC Analytics AG Switzerland 89 Addendum to PAL System User Manual a SYSTEM DLW Option Illustration DLW Wash Station PAL DLWWash PAL DLW Wash Station Block complete assembly incl centering plate PAL DLW Insert PAL DLW Wash Station Insert Kit consisting of 2 pcs Inserts 1 pc Waste Connector 5 pcs Dummy Plugs Tube Waste for Wash Station length 2 m 90 PAL is a registered trademark of CTC Analytics AG Switzerland syste Addendum to PAL System User Manual DLW Option Illustration DLW Syringe SyrC DLW100 R PLG DLW 100 PAL DLWPlg PAL DLWNdI CTC Syringe for PAL DLW Option 100 uL gastight 87 7 mm Scale Length 60 mm Thread 4 28 UNF Removable needle not
57. option can be controlled with either a Cycle Composer macro or with any data system that controls the PAL using Cycle Composer macros or PAL ICC cycles e g Analyst ChemStation Empower EZChrom MassLynx Xcalibur 2 Loading the PAL DLW Object If the version of the PAL Object Manager List installed on the computer Is not PAL xt Object Lists Rev B then copy the provided folder Wash Station Option see Fig 28 trom the CD ROM to the Object Lists folder which has been installed with the Object Manager This software Is usually installed in the following path C Program Files PAL Object Manager Object Lists Using PAL xt Object List Rev B or higher the DLW Option object will be integrated into the list in the Wash Stations folder The following Object List is then available PAL DLW DLW Option Firmware Object List 1 Open the Object Manager and verify the revision version number of the Object Lists If PAL xf Object Lists Revision level B or higher is available continue as described below If a PAL Object Lists version Is used e g Rev K or L copy the provided DLW Object List into the Object Manager folder as described above 2 Select the PAL DLW Object list and click Send selected Object Lists to PAL 3 Close the Object Manager 32 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual a SYSTEM DLW Option The DLW Objec
58. own in Figure 4 below 7 Tighten the mechanical stop for the needle guide securely III YN K T wn on 4 T O A tf Figure 6 Adjusting the Needle Guide to Needle Length for the DLW Option 1 F 8 Insert the syringe slider into the Z Tube and attach the long tension cord to the top cover 9 Reinstall the Injection Unit on the Y Axis note Figure 6 illustrates the position of the mechanical stop for the needle guide shown outside the unit for illustrative purposes When assembled this part is mounted to the rod This illustration emphasizes the fact that the stop must first be opened and then fixed again 10 Adjusting the Upper Needle Guide is illustrated in Figure 7 This position was found to be ideal for changing the syringe needle without bending it at the Upper Needle Guide PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual wy SYSTEM DLW Option Figure 7 Adjusting the Upper Needle Guide 2 3 Mounting the Pump Holder The pump holder should always be mounted on the far left of the X axis The solvent lines are connected to the holder Moving across the pump holder with the Z axis could kink the stabilizing wire To install the holder we recommend dismounting the drive from the PAL X axis 1 Locate the pump holder and the
59. p Stator Wash Step by Step Fast Injection Cycle Step by Step Definitions of Terms for Tubing Connections Pressing Tube into Place Illustration of various Tubing Cuts Dead Volume created due to wrong Ferrule Type Ferrule Types trom various Vendors PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oY SYSTEM DLW Option List of Figures and Tables cont d Table 1 Commonly used Safety Symbols Table 2 DLW Object List Table 3 The DLW Macro Definitions Table 4 Macro LC In DLW Standard Table 5 Macro LC In DLW Fast Table 6 Troubleshooting Table 7 Carry over Test UV Detection Table 8 Repeatability Test UV Detection Table 9 Test with Benzophenone and UV detection Table 10 Maintenance Check List Table 11 Naming Convention Table 12 DLW Spare Parts Table 13 PM Kit for PAL xt System including PAL DLW Option PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual A SYSTEM DLW Option D How to Use this Manual The PAL System must be installed and properly set up before the DLW Option instructions can be implemented This manual is divided into the following sections e Installing the DLW Option Operating the DLW Option Troubleshooting Replacing Parts Maintenance Limitations Appendices The DLW Installation Instructions are intended for frequent PAL users or new users who are already experi
60. re done at the factory 7 Insert the cable provided into the middle socket between the other two sockets on the PCB For details see Figure 10 Pump 1 To PAL Figure 10 Electrical Connections at the Pump Holder Bracket PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option 8 The other end of the cable is connected to the Wash Station connector on the Control x board as shown below in Figure 11 SER1 LAN MODBUS suzzr L msa INTERFACE 1 INTERFACE 2 d AUX 1 AUX 2 ER a D STATION Le dap im com gt 2 LO CEB lt a b dm e Figure 11 Electrical Connection from the DLW Pump Holder to the PAL xt Board 2 4 Installing the DLW Wash Station The DLW Wash Station can be connected anywhere along the X axis however it is strongly recommended to install it as close as possible to the Injection Valve The travel path of the Injection Unit from the valve to the Wash Station should be as short as possible to keep the cycle time as brief as possible Figure 12 shows two possible configurations for installing DLW modules favoring a short connection from the injection valve to the other HPLC system components on the one hand and always having the DLW Wash Station close to the injection valve on the other It is important that the DLW Pump Module with the connected DLW tubing from the kit always be attached on the far
61. rrule will bite into the polymer tube and block the flow When installing a tube with a narrow bend typically a loop it is advisable to fix one connection first without inserting the second nut into the female counterpart After tightening open the connection Start preparing the second connection without inserting the first nut Open the second connection again and check the pilot distance The last step is to insert the complete tubing with the two connections and make a final seal as described above This stepwise installation of a bended tube is mandatory for PEEKsil tubing fused silica tubing sheathed with PEEK polymer 2 1 1 2 Re use of Installed Nuts Ferrules The second rule to be observed is Do not re use an installed nut ferrule for any other connection Any tube with an Installed nut ferrule used for one purpose should not be installed for any other application Cut a portion of the tube end until straight tubing Is reached and remove the ferrule This rule also applies to loops Never re use a loop with another valve or a valve after exchanging the stator If a finger tight nut has been installed replace it as well If a stainless steel nut has been used ensure that the correct type is used see details below Newly installed finger tight ferrules can hold a backpressure of up to 200 bar This remark Is valid for the well known finger tight nuts which are used since a long time Recent developments try to overcome th
62. rter inserted Inserted in full up position Is the solvent flowing freely in the tube Fitting replaceable needle Check the fixation and seal of the replaceable syringe needle Is the retaining nut tight Is the PTFE seal installed Is the needle entry free of debris not clogged Is the correct needle diameter used Gauge 22 67 PAL is a registered trademark of CTC Analytics AG Switzerland syste Addendum to PAL System User Manual DLW Option iu Ye A Possible Cause Sample Dirty syringe and or Holding peaks responses are Loop Recommended Action Increase Pst Cln Slv1 Slv2 values in method Use PreClnSlv1 Slv2 and PreClnSpl not reproducible Syringe pressure differences Increase Pullup Delay value Vacuum created in sample vial Method Parameters Excessive carryover between samples Loose instable or wrong connection Inappropriate wash solvent s Waste tubing ID at injection valve is too small causing waste liquid to be pulled back by capillary action into the rotor groove Air gaps not formed Liquids of different type diffusing into each other Dirty needle Holding Loop and or valve injection port Excessive carryover between samples Inappropriate wash solvent s Waste tubing ID at injection valve is too small causing waste liquid to be pulled back by capillary action into rotor groove Damaged or grooved valve rotor Valve needle seal leaks
63. s to define the Valve Needle Penetration Depth 1 Using the hand held controller select the sequence Menu Setup Objects Injectors LC Vlv1 2 Press the F1 button Check Pos The Injection Unit moves to the previously defined LC VIv1 position 3 Highlight the item Needle Penetr with the cursor bars by pressing ENTER 4 Slowly rotate the outer knob to adjust the needle penetration depth The syringe moves down stepwise into the injection port 5 When the syringe needle tip enters the valve needle guide slow down the Z movement again Always observe the syringe needle during this adjustment step 6 Move down stepwise until the bottom of the cross bar of the DLW Needle Holder Assembly ts flush with the lower line of the DLW Needle Adapter Block See Figure 33 below 7 Press ENTER to save the needle penetration depth value 8 Verity the defined Needle Penetration Depth value by repeating Steps 3 to 6 Figure 33 Teaching the Needle Penetration Depth Value 57 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual oY SYSTEM DLW Option If a click sound occurs this means the ball spring loaded Needle Holder Is completely compressed STOP the Z axis downwards movement immediately Rotate the outer knob stepwise in the opposite direction until the gap in the spring loaded Needle Holder is approximately 1 mm or the bottom of the
64. s up if the valve bore size or the installed loop internal diameter are lower standard valve bore 0 25 mm The standard loop internal diameter ID for loops provided by CTC Analytics with a volume of 5 10 and 20 uL is 0 25 mm The loop with 2 uL content volume has an ID of 0 125 mm Keep the internal diameters of the tubing in line with the valve dimensions loop ID and flow rate For details see the poster Tips amp Hints for HPLC Technique 44 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual DLW Option system 4 3 Standard DLW Injection Cycle Macro Name LC Inj DLW Standard Macro Description For this step the needle tip is lifted releasing the sealing pressure to enable rinsing around the tip sealing point Remark The Atom Rinse Inj is new available starting with FW 4 1X The DLW Actuator Solenoid is activated the Wash Solvent pump the Needle Gap and the Rinse Time are selectable Macro Variable The PAL System waits for the Sync Signal Ready Remark before the injection cycle is started Sync Signal setting Start Fig 37 The Injection Valve is brought to a defined Inject to position Standby Standby The Rear air segment is pulled into the Airgap Volume Holding Loop Filling Speed Fig 38 The combined volumes of Rear Sample List Front Volume Fig 39 and Front Volume are aspirated into the Re
65. salt buffers are used is mandatory to avoid clogging of the solvent paths 40 PAL is a registered trademark of CTC Analytics AG Switzerland note note Addendum to PAL System User Manual A SYSTEM DLW Option 3 Functionality of the DLW Option 3 1 DLW Pumps From the control point of view the DLW pumps respond in the same manner as the Fast Wash Station The pumps are activated by Power out Signals The electric current setting for the DLW is different this means that the corresponding PAL Firmware Objects have to be loaded for each DLW wash station type See Section F point 2 Loading PAL DLW Objects for further details The wetted parts in the pump are made of the following materials e Membrane Kalrez FFPM e Body Valves Ryton PPS The pumps are self priming with a suction lift of up to 3 m water column 3 2 DLW Actuator Solenoid The DLW Actuator Solenoid separates and completely shuts off the lines in the direction of the syringe sample loading and the wash solvent After opening the DLW Actuator Solenoid for the wash solvent lines the desired wash solvent can be pumped into the system by activating the corresponding DLW pump The schematic in Figures 35 and 36 illustrates this function The wetted parts in the DLW Actuator Solenoid are made of the following materials e Solenoid body PEEK e Seal material FFKM Simriz PEEK exhibits excellent resistance to most commonly used chemi
66. siderations Changes or modifications to this unit not expressly approved by the party responsible for compliance could nullify the user s authority to operate the equipment The user should be aware that using the equipment in a manner not specified by the manufacturer may compromise its safety features A Electrical Hazards Every analytical instrument has specific hazards so please read and comply with the precautions as described in the corresponding PAL User Manual Only use fuses of the type and current rating specified Do not use repaired fuses and do not short circuit the fuse holder A i F F in AN Fi r r M Other Hazards To avoid injury while operating the PAL System keep your hands away from the syringe For detailed Safety Information see the additional warnings in the corresponding PAL User Manual PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Commonly Used Symbols A Caution Risk of Needle Stick Puncture FR pone Fuse Electrical Power ON Used with Main PAL Power Supply Electrical Power OFF Used with Main PAL Power Supply Electrical Power ON for only part of the System Used with Optional Device s Electrical Power OFF for only part of the System ihono AN Caution Risk of Electrical shock high voltage Disposal Do not dispose in municipal waste X Follow local waste regulations to reduce electrical and e
67. ssing Is recommended Important Is to observe the sequence of steps to ensure that Solvent 1 will be the last solvent in the DLW System Good practice Is to have the wash solvent with lower chromatographic strength elution power defined as Wash Solvent 1 and Wash Solvent 2 logically consistent as the solvent with a stronger elution power 1 1 Priming with Utilities Function Wash Station Prime first the PAL DLW System with the Utilities function Wash Station Select Menu Utilities Wash Station select Wash 2 Function Key F3 Move to Wash Function Key F2 Activate Valve Observe the green LED as an indicator that the DLW Actuator is activated Prime with Solvent 2 until no further air bubbles are observed Repeat the steps to prime with Solvent 1 by activating Wash 1 1 2 Priming with Utlitities Function Syringe As second priming step for the PAL DLW System is the Utilities function Syringe recommended This function allows controlling the plunger movement It is important that the priming through Wash Station is completed the solvent lines are free of air bubbles Select Menu Utilities Syringe Function Key F2 Cln Syr select Wash 2 select the desired Clean Count recommended a minimum of 3 counts Observe the green LED as an indicator that the DLW Actuator is activated and the smooth plunger movement Prime with Solvent 2 until no further air
68. t list contains the following PAL Firmware Objects which are loaded with a single command as described above Object Class Object Name Motor Drive MPlgMed Heater HDLW Syringe 100ulIDLW Wash Station Wash1 Wash 2 Injector Waste PWR Event Pwr Outl Pwr Out2 Table 2 DLW Object List File Help C Program Files PAL O0bject Manager PAL xt Object Lists Rev A Wash Stations Object List Folder Object List LHS Upgrade PAL Wash ct2 Multi Position Valve Drives PAL WashAct4 Output Expansion Box PAL Washactive Solvent Reservoirs PAL WashF ast SPME Fiber Conditioning Stations PAL WashLarge SPME Upgrade PAL WashStd Stack Coolers Stacks Syringes Tray Coolers Tray Holders Tray Types Valve Drives Wash Stations NT Send Selected Objects to PAL Connected on COM1 Figure 28 PAL Object Manager choosing Object List Folder Wash Stations For more details on the PAL Object Manger software see the Addendum to the PAL User Manual PAL Object Manager Software 55 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oY SYSTEM DLW Option 3 Wash Station Reference Points 3 1 Wash1 Wash2 Reference Points For the DLW Wash Station Option the reference positions for Wash1 and Wash2 are the two holes on top of the Wash Station assembly See Figure 29 The lower needle guide should be centered on these holes with the botto
69. tem that controls the PAL using PAL ICC e g Analyst ChemStation Empower EZChrom MassLynx Xcalibur For details see Section F point 1 PAL Firmware and Firmware Object Installation for PAL DLW Option and Section K Limitations PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option 2 Installation of the DLW Option 2 1 Unpacking the Components The PAL DLW option is shipped in one box Check for the following items DLW Pump Module DLW Syringe Needle DLW Syringe DLW Plunger Holder DLW Flow Diverter installed in the DLW Syringe Assembly DLW Syringe Holder Assembly CD ROM Wash Station cable to PAL System DLW Tubing Kit DLW Holding Loop installed in the DLW Syringe Assembly Needle Guide Length Tool DLW Wash Station incl Waste Tube 2 pcs Solvent Bottle Transfer Line including Glass Filter Solvent Inlet 40 um pore size 2 pcs 1 liter Solvent reservoir bottles At OY Og SS SS a Se N Figure 2 PAL Active Wash Station Components Figure 1 illustrates how the various DLW components as shown in Figure 2 are mounted on the PAL System PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual A SYSTEM DLW Option 2 2 Preparation of the Injection Unit for the DLW Option New PAL xt Systems equipped with a PAL DLW Option are already configured
70. the DLW Wash Station Module can be individually exchanged Wash Station Block Wash station Inserts Waste Tube Waste Tube Valve to Wash station Dummy plugs The inserts and individual parts of the Wash Station Block are not yet available This will change with future updates 8l PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 3 DLW Syringe The following parts keywords from the DLW Syringe assembly can be individually exchanged DLW Syringe 100 uL DLW Syringe Plunger for syringe 100 pL DLW Syringe Plunger Holder DLW Syringe Retaining Nut DLW Syringe Replaceable Needles 4 DLW Syringe Holder Assembly The following parts keywords from the DLW Syringe Holder assembly can be individually exchanged DLW Syringe Holder Assembly complete assembly DLW Needle Adapter Block DLW Needle Holder Assembly Holding Loop DLW Manifold Entire unit assembled consisting of DLW Solvent Selector DLW Syringe Adapter with Perfluor O ring installed DLW Flow Diverter DLW Actuator Solenoid DLW Actuator Control PCB incl cable DLW Syringe Holder Fixation Screw 82 PAL is a registered trademark of CTC Analytics AG Switzerland syste Addendum to PAL System User Manual DLW Option J Maintenance Regularly performing maintenance helps ensure the accuracy and precision of the PAL System Suggested intervals for maintenance procedur
71. the longer needle guide length See Figure 3 Upper needle guide blocked Original PCB Figure 3 New Upper Needle Guide touching electronic Components PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Starting with shipments March 2010 onwards for the PAL Z axis with PNo MZ 01 00F a modified PCB was installed PNo APR Head Revision KM If this board Is installed the new type of Upper Needle Guide with the longer rod can be installed without any restrictions See Figure 4 Repositioned for free movement of upper needle guide Figure 4 New Upper Needle Guide installed in Models with the modified PCB If the older version of the Z axis PCB Is installed Revision KL We advise you to store the original Upper Needle Guide assembly for future use Replace the current Upper Needle Guide with the new version containing the longer rod AN DLW Figure 5 Preparing the Injection Unit for the DLW Option PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 6 The needle length has to be re adjusted to 73 0 mm A tool is provided with the DLW option the DLW Needle Length Guide Tool Insert the tool in one of the guide bars as sh
72. throughput quality of sample solution particles etc and washing solvent It is advisable to check the tightness of the plunger on a regular basis to gain experience with the application Only as needed e g if bent Check the tightness of the needle in the needle seal See section F point 6 2 5 Weekly or as needed Recommended to be changed once a year as a preventative maintenance step Weekly or as needed Weekly or as needed Weekly or as needed Recommended to be changed once a year as a preventative maintenance step 85 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option K Limitations 1 PAL System The DLW Option can be operated only with a PAL xfsystem which requires PAL Firmware level 4 1 X and the APR Control xfboard An existing PAL System can be upgraded to this level For details contact your CTC Analytics representative 2 Sample Volume The current version of the DLW Option can handle sample volumes from 2 to 100 pL 3 Wetted Parts and Material Some components are not compatible with certain solvents or chemicals that are used in the given applications The material for wetted parts of the individual component Is described in the section Installation Specification and in the specific Point for this component 4 Tubing Internal Diameter ID and System Backpressure Any tubing and component in the f
73. tightening a nut and ferrule Is to proceed stepwise 1 Tighten the nut by hand as much as possible 2 Continue to tighten using a wrench until resistance Is felt Add one or two quarter turns to reach the sealing point 3 Open the connection and remove the assembly to inspect the pilot distance see Fig 70 above 4 Install the assembly again and tighten the nut manually at first Use the wrench to reach the same sealing or resistance point as in the step before and add one extra quarter turn to obtain the final seal 5 Check the seal when liquid is pumped through the system In case a leak IS observed tighten once more by another quarter turn The rule is to always go from leaking to properly sealed and never try to release an over tightened seal 69 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option Do not over tighten the nut ferrule If a connection proves to be leaky use another quarter turn to tighten Step by step tightening is the correct approach Over tightening even once will damage the seat and the next connection can only be sealed by force If the seat is damaged there is a considerable chance that the ferrule will stick and cannot be removed without force or special tools such as a drill If different materials are used such as polymer tubing and nut together with a stainless steel ferrule then over tightening is always a danger The sst fe
74. ture to 35 C Use the following path from local Terminal Menu Utilities Syringe Standby Temp select 35 C Expected result The green LED on the Syringe Holder assembly is turned OFF Place a beaker or other adequate vessel under the DLW Syringe Needle Select the Power Out signal and activate both DLW Pumps Path Menu F3 Setup Objects Events PwrOut1 for Pump or PwrOut2 for Pump2 Activate both pumps by changing the status to the Active State Active State OFF Pump Is activated start Active State ON Pump is stopped Expected result The DLW Actuator has been closed step 1 Activating the DLW Pumps delivers solvent up to the point of the DLW Actuator No solvent should pass the DLW Actuator which verifies the tightness of the seal In case a solvent flow Is observed during the test remove the DLW Actuator and check if particles are breaking the seal If nothing special is observed replace the device The PNo Is listed in the Appendix 76 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 3 Standard Chromatographic Tests In this section standard chromatographic tests for carry over and repeatability are described These tests should provide basic information for support technicians and users The tests described are examples only and should be adjusted accordingly if the configuration of the HPLC
75. two supplied Torx screws 2 Attach the holder to the X axis as shown below in Figure 8 Figure 8 Attaching the Pump Holder Bracket to the X Axis 3 The Solvent line assembly is not preinstalled at the factory The correct mounting Is shown in Figure 9 below Select the end of the of the guide wire with the longer tubing ends and connect the guide wire with the screw M4 x 8 and the corresponding serrated lock washer to the back side of the module Note the guide groove which orients the wire PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual a SYSTEM DLW Option Always install the DLW Tubing from the kit to the pump module first The other end connected to the DLW Syringe Assembly should be connected while the assembly is not yet installed onto the injection unit are I ae Figure 9 Installing the Solvent Tube Assembly to the Pump Holder 4 Connect the solvent tubing of the DLW Tubing kit to the pump module using the pre mounted nut Maintain this order Solvent 1 connected to the left and Solvent 2 to the right pump module when viewed from the front of the PAL system This avoids any confusion when refilling or changing solvents 5 Connect the solvent lines from the solvent reservoir bottle to the Inlet connectors of the pump module 6 The electrical connections from the pumps to the PCB sockets mounted on the bracket a
76. um to PAL System User Manual oY SYSTEM DLW Option Wash Solvent 1 follows to prepare the valve for Wash1 the next injection Inject to Needle Gap Valve Clean Valve Clean Time Solvent 1 Fig 67 The Injection Unit is moved back to the DLW Wash1 Wash station position Wash1 to flush the Post Clean Time Solvent 1 syringe needle inside and out with Wash Solvent 1 This is a preparation step for the next injection Especially important for biofluid samples Fig 68 Cycle end for LC Inj DLW Fast macro Fig 69 Table 5 Macro LC Inj DLW Fast 48 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual a SYSTEM DLW Option 5 PAL DLW Cycle Step by Step 5 1 Standard Cycle Figures 37 to 50 Standard Injection Cycle Step by Step Standard Cycle Start Figure 37 Standard Step 1 Aspirate Rear Air Segment Figure 38 PAL is a registered trademark of CTC Analytics AG Switzerland 49 Addendum to PAL System User Manual oY SYSTEM DLW Option Standard Step 2 Get Sample Aspirate Rear Inject and Front Volume Figure 39 Standard Steps 3 4 Aspirate Front Air Segment Figure 40 Eo PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual oY SYSTEM DLW Option Standard Steps 5 6 Passive needle clean outside dip in wash position 1 Figure 41 Standard Steps 7
77. without any restriction The waste tube must always be placed above the liquid level in the waste container 20 PAL is a registered trademark of CTC Analytics AG Switzerland note Addendum to PAL System User Manual a SYSTEM DLW Option 2 5 Installing the DLW Syringe Holder Assembly Before a syringe Is inserted the position Change Syr should first be verified The position for Change Syringe should not occur above an object for example where the needle could collide with a vial Path Menu Utilitites Syringe function key F3 Change Pos Move the PAL Injection Unit to a location where no collision can occur with the Z axis by selecting the X and Y axes appropriately The value for the Z axis Is given as a default and a change of this position is not necessary in standard operation This precaution helps avoid needle damage during routine operation Nevertheless the description below on how to install the DLW Syringe Assembly recommends switching off the unit and inserting the assembly without using the PAL command Change Syringe The DLW Syringe Holder differs from standard liquid syringe holders because the DLW manifold is attached to the syringe holder The DLW Actuator Solenoid and the Holding Loop connecting the syringe inlet bottom and Syringe Needle are attached to the DLW manifold All of these parts are preinstalled at the factory See Figure 14 below DLW Syringe Holder Assembly
78. wo additional horizontal Waste inlet ports in front to be combined with Valve waste port Wash Station block PVDF Inserts sst 1 4404 Connections at bottom of inserts PEEK Waste Tube Polyethylene PE Gastight Syringe 100 uL with Valflon tip Glass PTFE and Valflon Gauge 22 OD 0 72 mm ID 0 41 mm Length 51 mm PTFE Seal 2 2 way flipper solenoid valve Body PEEK Seal material FFKM Simriz Solvent selector module and Syringe Block sst 1 4435 Perfluor O ring FEP Tubing OD 1 16 ID 0 50 mm length 55 cm Loop Volume 108 uL Tube dimensions Gauge 22 OD 0 72 mm ID 0 41 mm Length 20 5 mm Stainless chromium steel o DLW Needle Holder Assembly Wetted part sst 1 4435 PAL is a registered trademark of CTC Analytics AG Switzerland Addendum to PAL System User Manual A SYSTEM DLW Option 1 2 Hardware Requirements The PAL DLW option can be used with HTS xtor HTC xt PAL instruments in combination with one or several injection valves of the following types VICI DCxxWK conical rotor VICI Cheminert or Rheodyne ultra high pressure valves The PAL Injection Unit needs to be modified in order to install the DLW Option Details are described in Section E point 2 2 Preparation of Injection Unit for DLW Option 1 3 Software Requirements The PAL DLW Option can only be operated with PAL x Firmware version 4 1 X or higher It can be controlled by the Cycle Composer or any data sys

Addendum to the PAL User Manual PAL DLW Option Installation

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