Beckman Coulter PA 800 plus User Manual
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Beckman Coulter PA 800 plus User Manual

Beckman Coulter PA 800 plus User Manual

Pharmaceutical analysis system system maintenance
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A51964AC
February 2011
Beckman Coulter, Inc.
250 S. Kraemer Blvd.
Brea, CA 92821
User's Guide
PA 800 plus
Pharmaceutical Analysis
System
System Maintenance

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Summary of Contents for Beckman Coulter PA 800 plus

  • Page 1 User’s Guide PA 800 plus Pharmaceutical Analysis System System Maintenance A51964AC February 2011 Beckman Coulter, Inc. 250 S. Kraemer Blvd. Brea, CA 92821...
  • Page 2 Beckman Coulter Ireland, Inc. Mervue Business Park, Mervue Galway, Ireland 353 91 774068 Beckman Coulter do Brasil Com e Imp de Prod de Lab Ltda Estr dos Romeiros, 220 - Galpao G3 - Km 38.5 06501-001 - Sao Paulo - SP - Brasil CNPJ: 42.160.812/0001-44...

  • Page 3: Revision History

    Revision History Second Revision, A51964AC, February 2011 32 Karat Software version 9.1 patch PA 800 plus Software version 1.1 patch PA 800 plus Firmware version 9.2 Numerous syntax and grammatical edits First Revision, A51964AB, December 2009 Revised corporate address. Initial Issue, A51964AA, April 2009 32 Karat Software version 9.1...
  • Page 4 Revision History A51964AC...

  • Page 5: Safety Notices

    Safety Notices Symbols and Labels Introduction The following is a description of symbols and labels used on the Beckman Coulter PA 800 plus Pharmaceutical Analysis System or shown in this manual. WARNING If the equipment is used in a manner not specified by Beckman Coulter, Inc., the protection provided by the instrument may be impaired.
  • Page 6 THIS PRODUCT CONFORMS TO APPLICABLE REQUIREMENTS OF 21 CFR 1040 AT THE DATE OF MANUFACTURE. MANUFACTURED: 726024-C A016350L.EPS Sharp Object Label A label reading “CAUTION SHARP OBJECTS” is found on the PA 800 plus. CAUTION SHARP OBJECTS A16558-AA A016351L.EPS A51964AC...
  • Page 7 For Beckman Coulter products bearing this label, please contact your dealer or local Beckman Coulter office for details on the take back program that facilitates the proper collection, treatment, recovery, recycling, and safe disposal of this device.
  • Page 8 Safety Notices Symbols and Labels Restriction of Hazardous Substances (RoHS) Labels These labels and materials declaration table (the Table of Hazardous Substance's Name and Concentration) are to meet People's Republic of China Electronic Industry Standard SJ/T11364-2006 "Marking for Control of Pollution Caused by Electronic Information Products" requirements. RoHS Caution Label This logo indicates that this electronic information product contains certain toxic or hazardous elements, and can be used safely during its environmental protection use period.
  • Page 9 Safety Notices Symbols and Labels Alerts for Warning, Caution, Important, and Note WARNING WARNING indicates a potentially hazardous situation which, if not avoided, could result in death or serious injury. The warning can be used to indicate the possibility of erroneous data that could result in an incorrect diagnosis (does not apply to all products).
  • Page 10 Safety Notices Symbols and Labels A51964AC...

  • Page 11: Table Of Contents

    Contents Revision History, iii Safety Notices, v Symbols and Labels, v Installation Procedures, 1-1 CHAPTER 1: Introduction, 1-1 Install UV or PDA Detectors, 1-2 Calibrate the PDA Detector, 1-5 Install UV Detector Wavelength Filters, 1-6 Integrated 488 nm Solid-State Laser, 1-12 Installation Requirements, 1-12 Install the Solid-State Laser...
  • Page 12 LIF, 2-17 How to Store the LIF, 2-17 Replace the Quad-Rings, 2-18 Replace the Fuses, 2-19 Interfacing PA 800 plus with Mass Spectrometry, 3-1 CHAPTER 3: CE-MS/MS Mode Cartridge Procedures, 3-1 How to Install the Capillary in the Standard Cartridge, 3-2...

  • Page 13: Installation Procedures

    CHAPTER 1 Installation Procedures Introduction The installation procedures covered are listed as follows. Start with the type of detector, then continue to remaining items. Install UV or PDA Detectors • Calibrate the PDA Detector • Install UV Detector Wavelength Filters •...

  • Page 14: Install Uv Or Pda Detectors

    Capillary Cartridge. For information on how to install the LIF Detector, see Install the LIF Detector into the PA 800 plus procedure. Before you start, you need: • UV or PDA Detector • Fiber optic cable (UV cable is 2 ended and the PDA cable is 3 ended) •...
  • Page 15 Installation Procedures Install UV or PDA Detectors Figure 1.1 Installing the UV Detector 1. Outer Door Cover 4. Clamp Bar 2. UV Detector 5. Capillary Cartridge 3. Two-ended Fiber Optic Cable 6. Interface Block A51964AC...
  • Page 16 Installation Procedures Install UV or PDA Detectors Figure 1.2 PDA Detector Installation 1. Outer Door Cover 4. Clamp Bar 2. PDA Detector 5. Capillary Cartridge 3. Three-ended Fiber Optic Cable 6. Interface Block A51964AC...

  • Page 17: Calibrate The Pda Detector

    Installation Procedures 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. Calibrate the PDA detector as follows: From the main menu, open an instrument configured with PDA detection.

  • Page 18: Install Uv Detector Wavelength Filters

    Do not try to access the instrument during the calibration process. Install UV Detector Wavelength Filters When you install, the PA 800 plus includes 200 nm, 214 nm, 254 nm, and 280 nm filters. The following section is for reference.
  • Page 19 Installation Procedures Install UV Detector Wavelength Filters Figure 1.5 Installing UV Detector Wavelength Filters 1. Outer Door Cover 4. Clamp Bar 2. Three-ended Fiber Optic Cable 5. Capillary Cartridge 3. UV Detector 6. Interface Block Loosen the two thumb-screws and lift the clamp bar. Remove the capillary cartridge from the interface block.
  • Page 20 Installation Procedures Install UV Detector Wavelength Filters Loosen the two thumb-screws and remove the optics source assembly. Put the assembly on a clean work surface. The optics source assembly is located right behind the interface block and has a horizontal bar handle as shown in Figure 1.6.
  • Page 21 Installation Procedures Install UV Detector Wavelength Filters Figure 1.7 How to Install a New Filter When all filters have been inspected or replaced, reinstall the filter wheel access cover. Place the optics source assembly in the mounting location. Align the two upper guide pins and tighten the two thumb-screws.
  • Page 22 Installation Procedures Install UV Detector Wavelength Filters Right-click on an instrument icon and select . System Administration Configure > Instrument mode must be enabled, and the user must have instrument administration privileges to configure the instrument. Select on the instrument Configuration dialog box. Configure The 32 Karat System Configuration dialog box displays.
  • Page 23 Installation Procedures Install UV Detector Wavelength Filters Figure 1.9 System Configuration Dialog Box In the Filter dialog box, enter the position and type for each of the installed filters. A51964AC 1-11...

  • Page 24: Integrated 488 Nm Solid-State Laser

    The 488 nm solid-state laser is part of the interconnect module in the PA 800 Enhanced release and in the new PA 800 plus Pharmaceutical Analysis System. The solid-state laser is compatible only with these two instruments. However, the new laser module does allow for connection of an external laser.

  • Page 25: Install The Lif Detector Filters

    Before you install the LIF Detector Module, install the filters and filter housing. The PA 800 plus system ships with a single color LIF Detector. The filters installed for channel 1 have a 488 nm rejection filter and a 520 nm emission filter. The use of channel 2 requires a change to the detector.

  • Page 26: Install The Filter Housing Into The Detector

    Installation Procedures Integrated 488 nm Solid-State Laser When all filters are in the filter holder, insert the spring and install the filter holder in the appropriate location (channel 1 or 2) in the filter housing. NOTE If there is no arrow on the edge of the filter, look at both sides of the filter and find which side has the more reflective surface.

  • Page 27: Install The Lif Detector Into The Pa 800 Plus

    Remove the module using the handle. Connect the LIF Detector into the PA 800 plus. 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.
  • Page 28 Installation Procedures Integrated 488 nm Solid-State Laser Figure 1.12 Connect the Probe to the LIF Cartridge 1-16 A51964AC...

  • Page 29: Lif Detector Calibration

    Installation Procedures Integrated 488 nm Solid-State Laser LIF Detector Calibration LIF detector calibration allows for normalization of the signal generated by an analyte relative to a known concentration of sodium fluorescein. Perform calibration after installation of a new capillary or after changing a detector or cartridge. Items required: •...
  • Page 30 Installation Procedures Integrated 488 nm Solid-State Laser Next, enter the target RFU value and capillary dimensions. See 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 31 Installation Procedures Integrated 488 nm Solid-State Laser Figure 1.15 Calibration Wizard Screen - Step 3 Select once the calibration completes. See Figure 1.16. Accept Figure 1.16 Calibration Wizard Screen - Step 4 A51964AC 1-19...

  • Page 32: Troubleshooting Procedures

    Verify the correct filters are installed in the LIF detector. Verify the laser supplied with the PA 800 plus is connected and the LASER ON light is lit. Manually set the CCF to 1.0. Run the calibration test with a new capillary and chemistries. If there is no step change detected, contact a Beckman Coulter service representative.

  • Page 33: Capillary Cartridge

    Capillary Cartridge You must install the appropriate capillary for the application to run properly. To determine the capillary needed, consult the specific application documentation for your PA 800 plus instrument. IMPORTANT The Capillary Cartridge rebuild procedures are also in video form. They are accessible from the PA 800 release CD, from the www.ceLeader.com web site, and from within the PA 800...
  • Page 34 Installation Procedures Capillary Cartridge Before rebuilding the cartridge, make sure you have the following items. See Figure 1.17: • Magnifying Lens • Safety Glasses • Forceps • Ruler • Felt-Tip Marker • Cartridge Assembly • Coolant Tubing Kit • Capillary •...
  • Page 35 Installation Procedures Capillary Cartridge 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. Aperture Plug (UV/PDA) 100 x 200, 100 x 800 A51964AC 1-23...
  • Page 36 If you plan to use the capillary again, prepare the capillary for storage as recommended in the appropriate application guide. IMPORTANT Beckman Coulter recommends you do not install a pre-cut capillary. Doing so can cause the capillary window to break. Carefully pull the seal-retainer clips from the capillary cartridge. See Figure 1.18.
  • Page 37 Installation Procedures Capillary Cartridge Figure 1.19 Removing Aperture from Cartridge (Back 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. Be careful not to damage the aperture. Grasp the capillary at the cartridge outlet and pull the capillary firmly to remove it.
  • Page 38 Installation Procedures Capillary Cartridge IMPORTANT To prevent breaking the capillary, do not pull it out of the cartridge from the inlet side. Remove the capillary from the outlet side of the cartridge only. 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.
  • Page 39 Installation Procedures Capillary Cartridge Remove the coolant tubing from the inlet side of the cartridge. See Figure 1.21. NOTE The tubing nut, ferrule, and o-ring are still attached to the coolant tubing. Figure 1.21 Coolant Tubing, Tubing Nut, Ferrule, and O-Ring 1.
  • Page 40 Installation Procedures Capillary Cartridge Table 1.1 Capillary Length Capillary Length Total Capillary Pre-Formed Coolant Tubing Description to Detector Length 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...
  • Page 41 Installation Procedures Capillary Cartridge NOTE Use the pre-formed tubes supplied in the coolant tubing kit to prevent kinks and restrictions. 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. See Figure 1.22.
  • Page 42 Installation Procedures Capillary Cartridge On the inlet side, push the tubing nut, ferrule, and o-ring in and tighten the tubing nut finger tight. 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 43 Installation Procedures Capillary Cartridge Measure the required length from the middle of the capillary window to the inlet end. Using a felt-tip pen, mark the capillary just outside the measured length. This is the long side of the capillary farthest from the window. See Figure 1.24.
  • Page 44 Installation Procedures Capillary Cartridge Carefully push the capillary in through the cartridge outlet until the capillary appears at the cartridge inlet. Pull the capillary through the cartridge from the inlet side until the capillary window appears in the center of the cartridge window. See Figure 1.26.
  • Page 45 Installation Procedures Capillary Cartridge From the back side of the cartridge, carefully align the center groove of the aperture clip with the capillary window and press into place. See Figure 1.27. Figure 1.27 Aligning Groove of Aperture Clip with Capillary Window 1.
  • Page 46 Installation Procedures Capillary Cartridge Figure 1.28 Install the LIF Probe Guide and Probe Retainer 1. Probe Retainer 2. Probe Guide 1-34 A51964AC...
  • Page 47 Installation Procedures Capillary Cartridge Install the Seal-Retainer Clips Carefully insert a capillary seal-retainer clip over each capillary end and press to snap into position. See 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 48 Installation Procedures Capillary Cartridge Trim the Capillary Ends IMPORTANT Only trim the capillary when the wet chemistry 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 49 Installation Procedures Capillary Cartridge CAUTION Be sure to wear safety glasses when cleaving and breaking the capillary. Capillary ends are sharp; handle them with care. IMPORTANT Do not saw back and forth with the cleaving stone. While you hold the capillary against the template, score the inlet end of the capillary at the template cross-mark.
  • Page 50 Installation Procedures Capillary Cartridge Figure 1.31 Capillary Cuts 1. Acceptable Cut 2. Unacceptable Cuts To smooth the capillary ends, burnish them with the rough side of the cleaving stone. Repeat inspection and burnishing until the ends are smooth. Install the O-Ring From the front side of the cartridge, place the retainer o-ring in the aperture clip hole.

  • Page 51: Install A Capillary Cartridge Into The Pa 800 Plus

    Before starting the cartridge installation, you must have a fully assembled capillary cartridge, which includes an installed capillary. Capillary Cartridge Installation Lift the outer door and the cartridge cover door at the front of the PA 800 plus. See Figure 1.33 Figure 1.34.
  • Page 52 Installation Procedures Capillary Cartridge Figure 1.33 Cartridge Cover Door 1. Outer Door or Sample Cover Door 5. Sample Trays 2. Inner Door or Cartridge cover Door 6. Buffer Trays 3. Indicators 7. Fluid Bubble Indicator 4. Fluid Fill Port 8. Power Switch 1-40 A51964AC...
  • Page 53 Installation Procedures Capillary Cartridge 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. A51964AC 1-41...

  • Page 54: Universal Vials And Caps

    Installation Procedures Universal Vials and Caps Position the capillary cartridge over the cartridge interface block and carefully lower the cartridge into position. 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. See Rebuild a Capillary Cartridge.

  • Page 55: Fill The Micro Vials

    Installation Procedures Universal Vials and Caps 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 Items required: • Micro vials •...
  • Page 56 Installation Procedures Universal Vials and Caps 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. Put the cap on the universal vial. Put the capped vial in the tray.

  • Page 57: Maintenance Procedures

    CHAPTER 2 Maintenance Procedures Maintenance Procedures This section describes maintenance of the following hardware for the PA 800 plus: • Opening Levers, Electrodes, and the Interface Block Coolant • Fiber Optics (UV and PDA Detectors) • Lamp • Instrument Care •...

  • Page 58: Opening Levers, Electrodes, And The Interface Block

    Maintenance Procedures Maintenance Procedures Opening Levers, Electrodes, and the Interface Block Remove and Replace Opening Levers to move the trays to the load position. Direct Control Lift the cartridge cover door. Loosen the two thumb screws and lift the clamp bar. Remove the capillary cartridge from the interface block.
  • Page 59 Maintenance Procedures Maintenance Procedures Figure 2.1 Interface Block, Electrodes, and Opening Levers 1. Interface Block 2. Electrodes 3. Opening Levers CAUTION Do not place fingers directly below the electrodes when installing opening levers. Electrode ends are sharp; handle them with care. Install an opening lever.
  • Page 60 Maintenance Procedures Maintenance Procedures Remove and Replace Electrodes Repeat steps 1-6 of Remove and Replace Opening Levers. Remove the electrodes one at a time as shown in Figure 2.2. Figure 2.2 Remove Electrodes from Interface Block 1. Electrode Tool 2. Electrode 3.

  • Page 61: Clean Electrodes, Opening Levers, And The Interface Block

    Maintenance Procedures Maintenance Procedures Figure 2.3 Install an Electrode into the Interface Block 1. Electrode Tool 2. Electrode Key a. Place an electrode in the tool with the electrode key facing you. See Figure 2.3. b. Align the tool below the notch and parallel to the bottom of the interface block. c.

  • Page 62: Coolant

    Maintenance Procedures Coolant Sonicate these four parts for five minutes. Retrieve the parts and dry them thoroughly with a Kim Wipe. Perform the steps under Install an electrode into the interface block. CAUTION Do not place fingers directly below the electrodes when installing opening levers. Electrode ends are sharp;...
  • Page 63 Maintenance Procedures Coolant Figure 2.4 Capillary Coolant Installation 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. Power Switch Attach the coolant fill adapter to the coolant fill port.

  • Page 64: Fiber Optics (Uv And Pda Detectors)

    Maintenance Procedures Fiber Optics (UV and PDA Detectors) Fiber Optics (UV and PDA Detectors) Items required: • Non-abrasive lens tissue • Analytical methanol, ethanol, or isopropanol CAUTION Do not touch the fiber optic surface with your fingers as oils from your skin can degrade performance.

  • Page 65: Detectors),

    Maintenance Procedures Fiber Optics (UV and PDA Detectors) Figure 2.5 Location of Fiber Optic Cable 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.

  • Page 66: Lamp

    • 7/64” hex head wrench WARNING If the PA 800 plus has been on for any length of time, the deuterium lamp will be very hot. To prevent burning your fingers, allow the lamp to cool before replacing. to move the trays to the load position.
  • Page 67 Maintenance Procedures Lamp 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.
  • Page 68 Maintenance Procedures Lamp 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 thumb-screws on the clamp bar and lift the bar.
  • Page 69 Maintenance Procedures Lamp Loosen the two thumb screws and remove the optics source assembly. See Figure 2.8. To remove, pull the assembly forward. Put the assembly on a clean work surface. Figure 2.8 How to Remove the Optics Assembly 1. Access Door Catch 2.
  • Page 70 Maintenance Procedures Lamp Figure 2.9 How to Disconnect the Lamp Power Plug 1. Power Plug Using a 7/64” hex head 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. CAUTION Make sure an orange O-ring is installed on the lamp flange before you install the lamp.
  • Page 71 Maintenance Procedures Lamp Reinstall the capillary cartridge in the interface block. Lower the clamp bar and tighten the two thumb screws. Close the cartridge cover door. Turn on the power. Start the 32 Karat software. Select from the Instrument screen control menu. The PDA (PDA detector installed) Diagnostics or UV (UV detector installed) diagnostic dialog box displays.
  • Page 72 Maintenance Procedures Lamp Figure 2.11 UV Diagnostics Screen Select . The Lamp Hours of Use dialog box displays. See Control > Diagnostics > Set Lamp Hours Figure 2.12. Figure 2.12 Lamp Hours of Use Display , enter a value of ‘ ’.

  • Page 73: Instrument Care

    Instrument Care LIF Care and Maintenance WARNING The Beckman Coulter 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 Beckman Coulter Field Service Representative.

  • Page 74: Replace The Quad-Rings

    Maintenance Procedures Instrument Care Replace the Quad-Rings If coolant is leaking between the interface block and cartridge, the quad-rings might require replacement. The following procedure describes how to change the quad-rings. You must have new quad-rings to do the replacement. How to Replace the Quad-Rings to move the trays to the load position.

  • Page 75: 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 Beckman Coulter service for additional assistance Items required: •...
  • Page 76 Maintenance Procedures Instrument Care Figure 2.13 Replacing Fuses 1. Fuse Block 2. Air Conditioning Vents 3. External Connections Panel Replace the fuses: Table 2.1 Fuse Type and Rating Line Voltage 100 - 120 VAC 200 - 240 VAC Fuse Type and Rating 8.0 A Slow Blow;...

  • Page 77: Chapter 3: Interfacing Pa 800 Plus With Mass Spectrometry

    In CE-MS mode, the coolant tubing and capillary extend outside the PA 800 plus to the EDA block. The EDA block is a device that allows the capillary to exit the PA 800 plus instrument and supplies a return path for the capillary coolant. The EDA block can be used with either the standard cartridge or the EDA cartridge.

  • Page 78: How To Install The Capillary In The Standard Cartridge

    Interfacing PA 800 plus with Mass Spectrometry CE-MS/MS Mode Cartridge Procedures How to Install the Capillary in the Standard Cartridge Figure 3.1 Standard Cartridge 901027.AI Prepare a 1 meter capillary with a window at 15 cm from one end. Remove the clip and double seal from the bottom of the cartridge on the aperture side.

  • Page 79: How To Install The Capillary In The Eda-Only Cartridge

    Interfacing PA 800 plus with Mass Spectrometry CE-MS/MS Mode Cartridge Procedures Put the nut, ferrule, and o-ring on the coolant tubing. Feed the long end of the capillary through the coolant tubing until the capillary emerges from the CE/MS-MS coolant adapter.
  • Page 80 Interfacing PA 800 plus with Mass Spectrometry CE-MS/MS Mode Cartridge Procedures Remove the clip and double seal from the bottom of the cartridge on the aperture side. Remove the nut and seal from the CE/MS-MS coolant adapter. Hold the capillary at one end. Put this end of the capillary into the bottom of the cartridge and push the capillary through the cartridge.

  • Page 81: Ground Connection

    Ground Connection A ground connection is provided on the PA 800 plus for additional safety. A grounding cable runs from the chassis of the MS system to the banana jack on the right side of the PA 800 plus. A51964AC...
  • Page 82 Interfacing PA 800 plus with Mass Spectrometry CE-MS/MS Mode Cartridge Procedures A51964AC...

  • Page 83: Appendix A: Lif Test Mix Calibration And Performance Tests

    APPENDIX A LIF Test Mix Calibration and Performance Tests LIF detection differs from absorbance detection in some important ways. An absorbance detector measures a small intensity difference in a high-intensity light source. This is an effective technique because the percentage of light does not change as the source light intensity changes. This means peak response for a sample remains constant as the lamp ages or when a lamp is changed.

  • Page 84: Applying The Ccf

    LIF Test Mix Calibration and Performance Tests Applying the CCF Applying the CCF In 32 Karat instrument configuration, LIF calibration is available for systems with LIF detection. To access the LIF calibration wizard, open the LIF instrument configuration. (Right-click on instrument, select configuration, select appropriate instrument type, and double-click LIF detector icon in right pane).

  • Page 85: Calibration Method

    LIF Test Mix Calibration and Performance Tests 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. The capillary is first rinsed with a non- fluorescent solution.

  • Page 86: Non-Fluorescent Solutions

    LIF Test Mix Calibration and Performance Tests Non-fluorescent Solutions 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 ssDNA separations.

  • Page 87: Lif Calibration For Different Fluorescent Solutions

    LIF Test Mix Calibration and Performance Tests LIF Calibration for Different Fluorescent Solutions LIF Calibration for Different Fluorescent Solutions A given application can require use of a special fluorescent marker. Depending on the marker, the emission wavelength can be significantly different than fluorescein. In this case, it is likely that different filters are required.

  • Page 88: Troubleshooting

    LIF Test Mix Calibration and Performance Tests Troubleshooting 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. Troubleshooting Many variables can affect the quality of an LIF method.
  • Page 89 LIF Test Mix Calibration and Performance Tests Troubleshooting A51964AC...
  • Page 90 LIF Test Mix Calibration and Performance Tests Troubleshooting A51964AC...

  • Page 91: Appendix B: Evaluating Pa 800 Plus Uv Filters

    1 cm x 1cm square by some varying degree of height. To use a spectrophotometer to measure the UV filters for the PA 800 plus, it is necessary to position an individual filter at a height such that the optical beam of the spectrophotometer passes through the center of the filter. A special filter holder tool is available for Beckman Coulter model DU 700 series and model DU 800 spectrophotometers.
  • Page 92 Figure B.1 Filter Holder Tool (Part Number A32981) The default filters shipped with PA 800 plus systems with UV detection are 200 nm, 214 nm, 254 nm, and 280 nm. Using the tools shown above, a set of these filters was scanned using a DU 800 spectrophotometer.
  • Page 93 Evaluating PA 800 plus UV Filters Overview Figure B.3 254 nm filter, Lambda max 253 nm Figure B.4 214 nm filter, Lambda max 211 nm A51964AC...
  • Page 94 Evaluating PA 800 plus UV Filters Overview Figure B.5 200 nm filter, Lambda max 200 nm All of these filters meet the expected accuracy of +/- 2 nm for a new filter except for the 214 nm filter. For most applications, this small change is not significant. Pass/Fail criteria should be established as part of method validation depending on the application and sample.
  • Page 95 Evaluating PA 800 plus UV Filters Overview Figure B.6 Diagnostic Screen The PDA detector uses an array of 256 photodiode elements. The diagnostic screen shows the output of the array correlating detector response vs. diode. For calibration, the system correlates known mercury emission wavelengths to diode numbers.
  • Page 96 Evaluating PA 800 plus UV Filters Overview Figure B.7 Spreadsheet for Out-of-Calibration Detector A51964AC...
  • Page 98 © 2009-2011 Beckman Coulter, Inc. All Rights Reserved...

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