SCIEX P/ACE MDQ Plus System Maintenance Manual
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SCIEX P/ACE MDQ Plus System Maintenance Manual

Capillary electrophoresis system
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P/ACE™ MDQ Plus Capillary
Electrophoresis System
System Maintenance Guide
RUO-IDV-05-4904-A
January 2019

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  • Page 1 P/ACE™ MDQ Plus Capillary Electrophoresis System System Maintenance Guide RUO-IDV-05-4904-A January 2019...
  • Page 2 This document is provided to customers who have purchased SCIEX equipment to use in the operation of such SCIEX equipment. This document is copyright protected and any reproduction of this document or any part of this document is strictly prohibited, except as SCIEX may authorize in writing.

  • Page 3: Table Of Contents

    Contents Chapter 1 Installation Procedures ........5 Install the UV or PDA Detector .
  • Page 4 SCIEX Support ........

  • Page 5: Chapter 1 Installation Procedures

    Prior to using the system, refer to the safety content in the System Overview Guide for detailed information on the safe use and operation of the system. This chapter contains the following installation procedures for the SCIEX P/ACE™ MDQ Plus capillary electrophoresis system. Begin by installing the detector, and then continue to remaining items.

  • Page 6: Install The Uv Or Pda Detector

    Install the UV or PDA Detector Required Materials • UV or PDA detector • Fiber optic cable (UV cable is 2-ended and the PDA cable is 3-ended) Verify the UV source optic module is installed. Verify that power is off and no capillary cartridge is installed. Lift the cartridge cover door and find the detector mounting location as shown in Figure 1.1 (for a UV detector) and...
  • Page 7 Figure 1.1 P/ACE™ MDQ Plus Instrument–Front View With UV Detector Installed 1. Outer Door Cover 4. Clamp Bar 2. UV Detector 5. Capillary Cartridge 3. Two-ended Fiber Optic Cable 6. Interface Block System Maintenance Guide P/ACE™ MDQ Plus Capillary Electrophoresis System RUO-IDV-05-4904-A | B54955AB 7 / 88...
  • Page 8 Figure 1.2 P/ACE™ MDQ Plus Instrument–Front View With PDA Detector Installed 1. Outer Door Cover 4. Clamp Bar 2. PDA Detector 5. Capillary Cartridge 3. Three-ended Fiber Optic Cable 6. Interface Block P/ACE™ MDQ Plus Capillary Electrophoresis System System Maintenance Guide 8 / 88 RUO-IDV-05-4904-A | B54955AB...

  • Page 9: Install Wavelength Filters For The Uv Detector

    Install Wavelength Filters for the UV Detector The P/ACE™ MDQ Plus system includes 200 nm, 214 nm, 230 nm, and 254 nm filters by default. The following section is for reference, if a different filter is required for your application. Required Materials •...
  • Page 10 Figure 1.3 Remove the Filter Wheel Access Cover 1. Open cover at the indicated point 2. Access Cover 3. Thumbscrews (2), one on each side Put on latex gloves and use small forceps to remove the filters from the filter wheel. Touching the optical surfaces of the filters without gloves leaves skin oil that can cause improper filter performance.
  • Page 11 Figure 1.4 Install a New Filter 11 When all filters have been inspected or replaced, reinstall the filter wheel access cover. 12 Place the optics source assembly in the mounting location. Align the two upper guide pins and tighten the two thumbscrews. 13 Reinstall the capillary cartridge in the interface block.
  • Page 12 20 Click the green arrow to move the selected detector icon from Available modules in the left pane to Configured modules in the right pane. 21 Double-click the icon in the Configured modules list to open the P/ACE MDQ plus System Instrument Configuration dialog (Figure 1.6).
  • Page 13 22 In the Filter area of the dialog, type the wavelength for each of the installed filters and then click OK to save the settings and close the dialog. Figure 1.7 Filter Area Detail System Maintenance Guide P/ACE™ MDQ Plus Capillary Electrophoresis System RUO-IDV-05-4904-A | B54955AB 13 / 88...

  • Page 14: Calibrate The Pda Detector

    Calibrate the PDA Detector The PDA detector must be calibrated following installation, after you change the lamp or other optical components, and when required as part of maintenance. After the OPCAL cartridge is installed, calibrate the PDA detector as follows: From the main menu, open an instrument configured with a PDA detector.

  • Page 15: Install The Lif Detector

    Click the rainbow above the lamp. The PDA Detector Parameters dialog appears. Figure 1.9 PDA Detector Parameters Dialog Click Calibrate to start the calibration sequence. The calibration requires approximately two minutes. Do not try to access the instrument during the calibration process. Install the LIF Detector WARNING It is not safe to operate the solid-state laser module on any system other...

  • Page 16: Installation Requirements

    Before you install the LIF detector module, install the filters and filter housing. The filters installed in channel 1 have a 488 nm rejection filter and a 520 nm emission filter. To use channel 2 requires a change to the detector. Contact SCIEX for assistance setting up and using channel 2.
  • Page 17 Install a Filter in the Filter Housing Hold the filter housing in the same orientation as shown in Figure 1.10. Figure 1.10 Install Filters in the Filter Housing 1. Notch Filter 2. Emissions (Band Pass) Filter (Bottom View) 3. Emissions (Band Pass) Filter (Top View) Hold the filter with the arrow on the outside edge of the filter pointing up.

  • Page 18: Install The Lif Detector In The P/Ace™ Mdq Plus Instrument

    Install the Filter Housing in the Detector Insert the filter housing into the opening in the right side of the detector (Figure 1.11). Make sure the keyed corner of the filter housing lines up with the keyed corner of the detector module. The housing is designed so it only fits in the opening in the orientation shown in the following figure.

  • Page 19: Connect The Probe To The Clamp Bar

    Connect the LIF detector to the P/ACE™ MDQ Plus instrument. Keep the two female connectors on the LIF detector level with the two male connectors found on the instrument. Lock the detector module in position by fastening the captive screws until finger-tight. Connect the Probe to the Clamp Bar IMPORTANT Be sure to install the LIF cartridge in the instrument and secure the clamp bar before connecting the probe.

  • Page 20: Calibrate The Lif Detector

    Click Configure in the instrument Configuration window. The 32 Karat Software Configuration dialog appears. Double-click the detector icon in the right pane of the P/ACE MDQ plus Configuration dialog. Click LIF Calibration Wizard in the P/ACE MDQ plus System Instrument Configuration dialog.
  • Page 21 Figure 1.13 Calibration Wizard–Step 1 Enter the target RFU value and capillary dimensions. Refer to the appropriate application guide for a recommended target RFU value. For example, the target value for an N-CHO- coated capillary is 7, not 35 as shown in Figure 1.14.
  • Page 22 Fill the reagents into the buffer vials and put the buffer vials in the following positions of the inlet buffer tray: • A1: DDI H • B1: LIF Performance Test Mixture Click Next to perform the automatic calibration (Figure 1.15). NOTE The wizard displays buffer in position A1.

  • Page 23: Troubleshoot The Lif Detector Calibration

    If the CCF is still less than 1 but not less than 0.1, there is probably no problem with the system. Run a standard and verify acceptable performance of the system. If the system performance is not acceptable or the CCF is less than 0.1, contact a SCIEX service representative.

  • Page 24: Install The Capillary Cartridge

    If there is no droplet seen, contact a SCIEX service representative. If flow through the capillary is verified, the detection system is the only possible cause when no step is detected. Verify the correct filters are installed in the LIF detector. Verify the laser supplied with the P/ACE™...
  • Page 25 • Cartridge components and tools shown in Figure 1.17 Figure 1.17 Capillary Cartridge Rebuild Components 1. Cartridge Assembly 6. Aperture O-ring 2. Coolant Tubing 7. O-ring Installation Tool 3. Capillary 8. Cleaving Stone 4. Capillary Rebuild Kit 9. Capillary Length Template 5.

  • Page 26: Remove The Capillary

    IMPORTANT If you plan to use the capillary again, prepare the capillary for storage as recommended in the appropriate application guide. IMPORTANT SCIEX recommends you do not install a pre-cut capillary. Doing so can cause the capillary window to break.
  • Page 27 Remove the aperture by pushing from the front of the cartridge and pulling out from the back (Figure 1.19). Figure 1.19 Removing Aperture from Cartridge (Rear View) Remove the o-ring from inside the aperture. To do this, gently tap the aperture on the work surface or use a small gripping tool.

  • Page 28: Replace The Coolant Tubing And Seals

    Figure 1.20 Removing the Capillary 1. Outlet Side Removal NOTE The following procedure is only necessary to replace damaged tubing, to replace an o-ring, or to use a different tubing length. Replace the Coolant Tubing and Seals Loosen and remove the tubing nut from the outlet side of the cartridge. Remove the coolant tubing from the outlet side of the cartridge (Figure 1.21).
  • Page 29 Figure 1.21 Coolant Tubing, Tubing Nut, Ferrule, and O-Ring 1. O-ring 2. Ferrule 3. Tubing Nut Remove the o-ring from the outlet side of the cartridge using a small gripping tool if necessary. Table 1.1 to match a pre-formed coolant tubing length and loop shape with a capillary length.
  • Page 30 Table 1.1 Capillary Length Length to Total Length Pre-Formed Coolant Tubing Description Detector 20 cm 30.2 cm Pre-formed coolant tubing (no loop) for a total capillary length of 30.2 cm 30 cm 40.2 cm Pre-formed coolant tubing for a total capillary length of 40.2 cm 40 cm 50.2 cm Pre-formed coolant tubing for a total capillary length of 50.2 cm...
  • Page 31 Slide one tubing nut, ferrule (tapered edge toward the middle of the tube length), and o-ring over each end of the new coolant tubing (Figure 1.22). NOTE Always install a new ferrule. Used ferrules can cause leaks. Figure 1.22 Proper Alignment of O-ring, Ferrule, and Tubing Nut 1.

  • Page 32: Replace The Capillary

    WARNING Do not over tighten. Over tightening the tubing nut can cause the cartridge to break. If leaking occurs, check the cartridge housing for cracks. If there is no visible damage to the cartridge, verify that the o-ring is centered on the tubing and replace any worn or damaged tubing or ferrule.
  • Page 33 Do not cut the capillary to size at this time. NOTE For consistent measurements, place tape on the edge of a workbench and mark the measurements on the tape. This allows you to line up the capillary with the marks on the tape when preparing a new capillary. Figure 1.24 Capillary Position Markers 1.

  • Page 34: For Uv And Pda Detectors: Install The Aperture Clip

    Pull the capillary through the cartridge from the inlet side until the capillary window appears in the center of the cartridge window (Figure 1.26). Figure 1.26 Centering Capillary Window in Cartridge Window 1. Capillary Window Install the Aperture Clip IMPORTANT Remove the previous o-ring before you re-install the clip. Select the proper aperture for the application as recommended in the appropriate application guide.
  • Page 35 Figure 1.27 Aligning Groove of Aperture Clip with Capillary Window 1. Aperture 2. Capillary 3. Aperture Clip Go to Trim the Capillary Ends on page System Maintenance Guide P/ACE™ MDQ Plus Capillary Electrophoresis System RUO-IDV-05-4904-A | B54955AB 35 / 88...

  • Page 36: For An Lif Detector: Install The Lif Probe Guide And Probe Retainer

    Install the LIF Probe Guide and Probe Retainer IMPORTANT For LIF detection, use a standard cartridge and in place of the aperture clip, use a probe guide and probe retainer. Carefully install the metal probe guide from the back of the cartridge. Insert the probe retainer so the lock pins snap into the dimples on the probe guide (Figure 1.28).

  • Page 37: Install The Seal-Retainer Clips

    Install the Seal-Retainer Clips Carefully insert a capillary seal-retainer clip over each capillary end and press to snap into position (Figure 1.29). NOTE Make sure both the front and back edges of each clip snap into position. Figure 1.29 Insert the Capillary Seal Retainer Clips 1.

  • Page 38: Trim The Capillary Ends

    Trim the Capillary Ends IMPORTANT Only trim the capillary when the sample, buffer trays, and sequence are ready and the instrument is ready to run. After you cut the capillary, store the cartridge so the protruding capillary ends are in a solution as recommended in the appropriate application guide.
  • Page 39 CAUTION Be sure to wear safety glasses when cleaving and breaking the capillary. Capillary ends are sharp; handle them with care. While holding the capillary against the template, score the inlet end of the capillary at the template cross-mark. Hold the cleaving stone at a 30 °C angle when scoring. NOTE Make sure to cut the measurement mark off the capillary.
  • Page 40 Figure 1.31 Capillary Cuts 1. Acceptable Cut 2. Unacceptable Cuts Smooth the capillary ends by burnishing them with the rough side of the cleaving stone. Repeat inspection and burnishing until the ends are smooth. 10 Wipe the ends of the capillary with methanol and then with deionized water to remove any traces of ink and debris.

  • Page 41: For Uv And Pda Detectors: Install The O-Ring

    Install the O-ring From the front side of the cartridge, place the retainer o-ring in the aperture clip hole. NOTE Use an o-ring only with 100 x 200 and 100 x 800 apertures. Do not use an o- ring with the LIF probe guide. WARNING The o-ring must lie flat on top of the capillary window.

  • Page 42: Install The Capillary Cartridge Into The P/Ace™ Mdq Plus Instrument

    Install the Capillary Cartridge into the P/ACE™ MDQ Plus Instrument Required Materials • A fully assembled capillary cartridge, which includes an installed capillary Lift the outer door and the cartridge cover door at the front of the P/ACE™ MDQ Plus instrument (Figure 1.33 Figure...
  • Page 43 Figure 1.34 Interface Block 1. Outer Door or Sample Cover Door 4. Clamp Bar 2. Detector 5. Capillary Cartridge 3. Two-ended Fiber Optic Cable 6. Interface Block Loosen the knobs and lift the clamp bar. Position the capillary cartridge over the cartridge interface block and carefully lower the cartridge into position.

  • Page 44: Install The Universal Vials And Caps

    Lower the clamp bar and tighten the knobs. Lower the cartridge cover door. NOTE To ensure correct performance, replace the capillary cartridge aperture after three months of use. Install the Universal Vials and Caps Fill the Universal Vials NOTE When you fill universal vials, remember: •...

  • Page 45: Fill The Micro Vials

    Figure 1.35 Universal Vials 1. Universal Vial Cap 2. Maximum Fill Level 3. Universal Vials Put the cap on the vial and press it into position. Fill the Micro Vials Required Materials • Micro vials • Universal vials • Universal vial caps NOTE Very small samples can evaporate into the space inside the micro vial holder.
  • Page 46 Using care as not to introduce any bubbles, fill the micro vial with at least 100 µL of sample (Figure 1.36). Figure 1.36 Micro Vials 1. Universal Cap 3. Universal Vial 2. Micro Vial 4. Micro Vial inside Universal Vial Place the micro vial in the universal vial.

  • Page 47: Chapter 2 Maintenance Procedures

    CHAPTER 2 Maintenance Procedures This chapter contains the following maintenance procedures for the SCIEX P/ACE™ MDQ Plus capillary electrophoresis system: • Clean the Ejecting Levers, Electrodes, and Interface Block on page 48 • Refill the Capillary Coolant on page 53 •...

  • Page 48: Clean The Ejecting Levers, Electrodes, And Interface Block

    Clean the Ejecting Levers, Electrodes, and Interface Block SCIEX recomends cleaning the ejecting levers and electrodes once a week and when switching applications. The interface block should be cleaned as needed. Required Materials • Lint-free wipes • Mirror • Pen light •...

  • Page 49: Remove The Ejecting Levers

    10 Inspect the interface block using a mirror and pen light. If needed, repeat the cleaning process until the interface block is clean. 11 Reinstall the capillary cartridge in the interface block. 12 Lower the clamp bar and tighten the two thumbscrews. 13 Close the cartridge cover door.

  • Page 50: Remove The Electrodes

    Figure 2.1 Interface Block, Electrodes, and Ejecting Levers 1. Interface Block 2. Electrodes 3. Ejecting Levers Remove the Electrodes If needed, follow the instructions in Remove the Ejecting Levers on page Align the electrode tool flush with the bottom of the interface block. Push straight forward under the interface block to capture the electrode in the tool.
  • Page 51 Make sure the nub at the end of the electrode tool handle fits into the notch on the interface block as shown in the circled area of Figure 2.2. Figure 2.2 Remove Electrodes from Interface Block 1. Electrode Tool 2. Electrode 3.

  • Page 52: Replace The Electrodes In The Interface Block

    Replace the Electrodes in the Interface Block Place an electrode in the tool with the electrode key facing you (Figure 2.3). Figure 2.3 Install an Electrode into the Interface Block 1. Electrode Tool 2. Electrode Key Align the tool below the notch and parallel to the bottom of the interface block. Press upward to snap the electrode into the interface block.

  • Page 53: Refill The Capillary Coolant

    Refill the Capillary Coolant Required Materials • Capillary cartridge coolant (PN 359976) • 150 mL syringe • Coolant Fill Adapter (PN 144647) Open the sample cover (Figure 2.4). Figure 2.4 Capillary Coolant Installation 1. Outer Door or Sample Cover (open) 5.

  • Page 54: Clean The Fiber Optic Cable (Uv And Pda Detectors)

    Fill the coolant supply until the coolant level in the sight-tube is approximately 75% full. Remove the syringe and close the sample cover. The coolant syringe comes with a plunger. When adding coolant, do not use the plunger. Gravity supplies enough force to pull the coolant into the system. Using the plunger can introduce too much pressure to the coolant system and can cause damage.
  • Page 55 Figure 2.5 Location of Fiber Optic Cable–UV Dectector 1. Outer Door (open position) 4. Clamp Bar and Cable Connection 2. UV Detector 5. Capillary Cartridge 3. Two-ended Fiber Optic Cable 6. Interface Block Turn the system power off. Loosen the two thumbscrews on the insertion bar and lift the insertion bar. Disconnect the fiber optic cable at the detector.

  • Page 56: Replace The Deuterium Lamp

    Disconnect the fiber optic cable at the clamp arm. Moisten the lens tissue with alcohol. Clean the surface of each fiber optic cable connector and allow each connector to dry before reconnecting. Reconnect the fiber optic cable at the clamp bar. 10 Reconnect the fiber optic cable at the detector.
  • Page 57 Figure 2.6 How to Change the Lamp 1. Outer Door or Sample Cover (open) 5. Sample Trays 2. Inner Door or Cartridge Cover (open) 6. Buffer Trays 3. Indicators 7. Fluid Bubble Indicator 4. Fluid Fill Port 8. Convenience 9. Switch System Maintenance Guide P/ACE™...
  • Page 58 Figure 2.7 Capillary Cartridge Door 1. Outer Door (open position) 4. Clamp Bar and Cable Connection 2. UV Detector 5. Capillary Cartridge 3. Two-ended Fiber Optic Cable 6. Interface Block Turn the system power off and allow enough time for the lamp to cool. Loosen the two thumbscrews on the clamp bar and lift the bar.
  • Page 59 Loosen the two thumbscrews and remove the optics source assembly (Figure 2.8). Figure 2.8 How to Remove the Optics Assembly 1. Access Door Catch 2. Access Door 3. Thumbscrews (one on each side) Pull the assembly forward and place it on a clean work surface. Open the deuterium lamp access cover on the back of the optics source assembly.
  • Page 60 Figure 2.9 Lamp Power Plug 1. Power Plug Using a 7/64 inch hex wrench, remove the two hex screws securing the deuterium lamp. Remove the lamp from the lamp housing. CAUTION Do not touch the envelope of the lamp as oils from your skin can damage the lamp.
  • Page 61 15 Lower the clamp bar and tighten the two thumbscrews. 16 Close the cartridge cover door. 17 Turn on the power. 18 Start the 32 Karat software. 19 Click Control > Diagnostics > View. The appropriate diagnostics dialog for your detector opens (Figure 2.10 Figure...

  • Page 62: Procedures For Instrument Care

    Figure 2.11 UV Detector Installed–Diagnostics Window 20 Click Control > Diagnostics > Set Lamp Hours. The Lamp Hours of Use dialog appears. (Figure 2.12). Figure 2.12 Lamp Hours of Use Dialog 21 In Lamp Hours, type 22 Click OK. Procedures for Instrument Care •...

  • Page 63: Replace The Fuses

    Make sure to replace the fuses with the correct type and rating for continued protection against risk of fire and/or improper instrument operation. CAUTION If the fuses continue to blow after being replaced, contact SCIEX service for additional assistance Required Materials • #2 flat screwdriver •...
  • Page 64 Using the flat-tip screwdriver, remove the fuse block (Figure 2.13). Figure 2.13 Replacing Fuses 1. Fuse Block 2. Air Conditioning Vents 3. External Connections Panel Replace the fuses. Reinstall the fuse block and reconnect the power cable. P/ACE™ MDQ Plus Capillary Electrophoresis System System Maintenance Guide 64 / 88 RUO-IDV-05-4904-A | B54955AB...

  • Page 65: Procedures For Lif Detector Care And Maintenance

    The 488 nm laser module emits laser light that can cause serious damage to the eyes. Servicing of the laser should be done only by a SCIEX Field Service Employee. Inspect the LIF The LIF detector system is designed to prevent laser light exposure outside the laser box, fiber cable, and detector.
  • Page 66 P/ACE™ MDQ Plus Capillary Electrophoresis System System Maintenance Guide 66 / 88 RUO-IDV-05-4904-A | B54955AB...

  • Page 67: Chapter 3 Connect The System To A Mass Spectrometer

    CHAPTER 3 Connect the System to a Mass Spectrometer This chapter contains the following procedures for interfacing the SCIEX P/ACE™ MDQ Plus capillary electrophoresis system with a mass spectrometer: • Install the Capillary in the Standard Cartridge on page 68 •...

  • Page 68: Install The Capillary In The Standard Cartridge

    Install the Capillary in the Standard Cartridge Required Materials • 100 cm capillary with a window 15 cm from the end • External detector adapter kit (PN A61216) • Standard cartridge (PN 144738) or CE/MS cartridge (PN 149044) • Capillary length template •...
  • Page 69 If it the aperture o-ring is present, remove it. Install the aperture by pressing it in squarely from the back side of the cartridge. Install the double seal and clip over the capillary and onto the cartridge. Trim the capillary at the bottom of the cartridge using the capillary length template and cleaving stone.
  • Page 70 15 Lower the clamp bar and tighten the screws. 16 Insert the capillary adapter body into the slot in the right side of the instrument. Secure the capillary adapter with the locking ring. 17 Slide the door closed, then close the cartridge cover door and the outer door. 18 Adjust the end of the capillary coming from the coolant tubing into the MS adapter so that the shortest length of capillary is exposed.

  • Page 71: Install The Capillary In The Eda-Only Cartridge

    Install the Capillary in the EDA-Only Cartridge Figure 3.3 EDA-only Cartridge Required Materials • 100 cm capillary • EDA cartridge (PN 144829), includes cartridge and external detector adapter • Capillary length template • Cleaving stone If a cartridge is already present in the P/ACE™ MDQ Plus instrument, remove it. Prepare a 100 cm capillary.
  • Page 72 Trim the capillary at the bottom of the cartridge using the capillary length template and cleaving stone. NOTE Do not trim the long end of the capillary now. Feed the long end of the capillary through the coolant tubing until the capillary emerges from the external detector adapter.

  • Page 73: Install The Ground Connection

    Install the Ground Connection A ground connection is provided on the P/ACE™ MDQ Plus instrument for additional safety. Required Materials • Grounding cable (PN 144999) Connect the grounding cable from the chassis of the mass spectrometer to the banana jack on the right side of the P/ACE™...
  • Page 74 P/ACE™ MDQ Plus Capillary Electrophoresis System System Maintenance Guide 74 / 88 RUO-IDV-05-4904-A | B54955AB...

  • Page 75: Chapter A Evaluating Uv Filters

    Evaluating UV Filters Overview By default, the SCIEX P/ACE™ MDQ Plus Capillary Electrophoresis System is configured with a UV detector. Absorbance wavelength is determined by the filters installed on the system. Seven positions are available for different wavelength filters. When a method is run, the instrument selects the specified filter and positions it in the optical path.
  • Page 76 The default filters for the UV detector are 200 nm, 214 nm, 230 nm, and 254 nm. Using the tool shown above, these filters can be scanned using a Beckman Coulter DU 800 or DU 700 Series spectrophotometer. The filters used for UV detection all have a center wavelength ( ) of ± 2 nm. To find , use the λ...
  • Page 77 Figure A.2 Calculating Center Wavelength using Full Width at Half Height The following equation shows the calculation of the center wavelength, using the values from the plot in Figure A.2: ) 220.8 nm 2 206.4 nm λ ----------------------------------------------------------- 213.2 nm 206.4 nm 220.8 nm System Maintenance Guide...
  • Page 78 Figure A.3 Calculated Result of Center Wavelength Because filter characteristics can change with time, heat, humidity, exposure to UV light, and handling it is important to periodically check or replace these filters. For most applications a small change is not significant. Pass/fail criteria for filter characteristics will vary depending on the application and sample.

  • Page 79: Chapter B Performance Tests For The Lif Detector

    A calibration can be performed to correct for these influences by using a SCIEX-supplied test mix. For this example, the environment is LIF calibration using the 488 nm laser with fluorescein.

  • Page 80: Applying The Ccf

    Applying the CCF In the 32 Karat software, calibration is available for systems with LIF detection. To access the LIF Calibration wizard, open the P/ACE MDQ plus System Instrument Configuration dialog. (Right-click the instrument, click Configuration, select the appropriate instrument type, and double-click the LIF detector icon in the right pane.)

  • Page 81: Calibration Method

    Calibration Method Automatic calibration can seem a bit mysterious because the method parameters are not shown and data is not displayed. However, the method is simple (Figure B.2). The capillary is first rinsed with a nonfluorescent solution. Next, a pressure separation is performed without voltage.

  • Page 82: Calibration Results

    Calibration Results Figure B.3 Representative Results from Automatic Calibration Using Method in Figure B.2 Non-fluorescent Solutions Typically, the non-fluorescent solution used in the calibration is the separation buffer. Always be careful that this solution is safe for use in the type of capillary being used. For example, borate buffer should never be used for calibration of coated capillaries such as those used with IEF, carbohydrate, and single-stranded DNA separations.

  • Page 83: Lif Calibration For Other Capillaries

    LIF Calibration for Other Capillaries For detector performance tests, a 75 µm i.d., 60 cm total length bare fused-silica capillary is used. Many applications require capillaries with different dimensions. Capillary dimensions can affect the test mix calibration in two ways: •...

  • Page 84: Standard System Performance Test

    Establishing a new target value is simple: • Use manual calibration to set the CCF to 1.0. • Run the pressure separation method described above six times. • Calculate the average response of the step change for the six runs. This value is the new target. If an application requires a coated capillary, do not use a borate solution.

  • Page 85: Calibration Results

    Calibration Results Figure B.6 Representative Results from System Performance Test Using Method in Figure B.5 If the suitability option has been selected in the instrument configuration, a pass/fail criteria can be programmed into the method. Suitability reports can only be generated as part of a sequence. System Maintenance Guide P/ACE™...

  • Page 86: Troubleshooting

    Troubleshooting Many variables can affect the quality of an LIF method. To simplify troubleshooting, it is necessary to isolate and verify the performance of each of these variables. Follow the steps in Figure B.7 to troubleshoot an LIF detector. Figure B.7 LIF Performance Troubleshooting Flow Chart P/ACE™...

  • Page 87: Contact Us

    SCIEXUniversity SCIEX Support SCIEX and its representatives maintain a staff of fully-trained service and technical specialists located throughout the world. They can answer questions about the system or any technical issues that might arise. For more information, visit the SCIEX website at sciex.com...

  • Page 88: Documentation

    Documentation for the hardware products can be found on the Customer Reference DVD that comes with the system or component. For the latest versions of the documentation, visit the SCIEX website at sciex.com. NOTE To request a free, printed version of this document, contact sciex.com/contact-us.

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