SCIEX PA 800 Plus Operating manual

PA 800 Plus Pharmaceutical Analysis
System
System Maintenance Guide
RUO-IDV-05-5519-A
January 2018

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© 2018 AB Sciex
PA 800 Plus Pharmaceutical Analysis System System Maintenance Guide
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AB Sciex LLC
500 Old Connecticut Path
Framingham, Massachusetts 01701
USA

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Contents
CHAPTER 1: Installation Procedures, 5
Overview, 5
Introduction, 6
Install UV or PDA Detectors, 7
Calibrate the PDA Detector, 10
Install UV Detector Wavelength Filters, 11
Integrated 488 nm Solid-State Laser, 17
Installation Requirements, 17
Install the Solid-State Laser Module, 17
Install the LIF Detector Filters, 17
Install the Filter Housing into the Detector, 19
Install the LIF Detector into the PA 800 Plus System, 20
Connect the Probe to the Clamp Bar, 20
LIF Detector Calibration, 21
Troubleshooting Procedures, 24
Capillary Cartridge, 25
Rebuild a Capillary Cartridge, 25
Install a Capillary Cartridge into the PA 800 Plus System, 44
Universal Vials and Caps, 46
Fill the Universal Vials, 46
Fill the Micro Vials, 47
CHAPTER 2: Maintenance Procedures, 49
Maintenance Procedures, 49
Opening Levers, Electrodes, and the Interface Block, 50
Clean Electrodes, Opening Levers, and the Interface Block, 53
Coolant, 54
Fiber Optics (UV and PDA Detectors), 56
Lamp, 58
Instrument Care, 65

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Contents
LIF Care and Maintenance, 65
Inspect the LIF, 65
How to Clean the LIF, 65
How to Store the LIF, 65
Replace the Quad-Rings, 66
Replace the Fuses, 67
CHAPTER 3: Connect the System to a Mass Spectrometer, 69
CE-MS/MS Mode Cartridge Procedures, 69
Install the Capillary in the Standard Cartridge, 70
Ground Connection, 72
APPENDIX A: LIF Test Mix Calibration and Performance Tests, 73
Relative Fluorescent Units (RFU), 73
Calibration Correction Factor (CCF), 73
Applying the CCF, 74
Automatic Calibration, 74
Calibration Method, 75
Non-fluorescent Solutions, 76
LIF Calibration for Different Dimension Capillaries, 76
LIF Calibration for Different Fluorescent Solutions, 77
Standard System Performance Test, 77
Troubleshooting, 79
APPENDIX B: Evaluating PA 800 Plus System UV Filters, 81
Overview, 81
Revision History, 85

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CHAPTER 1
Installation Procedures
CAUTION
Prior to using the system, refer to the PA 800 Plus Pharmaceutical Analysis System
Overview Guide for detailed information on the safe use and operation of the
system.
Overview
The PA 800 Plus Methods Development Guide is a self-paced introduction to the operation of the SCIEX
PA 800 Plus Pharmaceutical Analysis System, which includes the 32 Karat software. It contains a
brief introduction to the technique of capillary electrophoresis (CE), an overview of the PA 800 Plus
System hardware, tutorials for the most frequently used software features, and appendices of useful
information.
The Methods Development Guide has been designed to assist users who are new to CE or to 32 Karat
software. It contains step-by-step exercises that allow you to create methods, to generate and
analyze data, and to report results. It does not cover every feature in the system or every aspect of
capillary electrophoresis.
The on-instrument activities contain specific instructions for users with ultraviolet (UV),
photodiode array (PDA), and laser-induced fluorescent (LIF) detector systems. The activities are
progressive. Each builds on the previous one. To get the most from this guide, work through all the
activities in order.
The activities that teach data reprocessing, calibration, and reporting, are all based on data collected
with an ultraviolet detector system. The steps in these procedures are very similar regardless of the
detector type used, so these exercises are important for users of all detector types. Data files for the
post-run exercises are installed with the 32 Karat software.
Additional information can be found in the 32 Karat Software Online Help. This can be accessed at any
time while using the 32 Karat software. You can access Help through the Help menu located in the
menu bar, by pressing the F1 key while working within the software, or any time a Help button is
displayed in an application window.

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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
• Install the Solid-State Laser Module
• LIF Detector Calibration
• Capillary Cartridge
WARNING
Disconnect the power before any instrument disassembly. Failure to do so can
cause electrical shock or other injury.
WARNING
Maintenance or repair procedures not specifically described in this guide present
a risk of electrical shock or injury. Refer additional servicing to SCIEX.
WARNING
Do not attempt to defeat any of the instrument interlocks or safety mechanisms.

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Installation Procedures
Install UV or PDA Detectors
NOTE The following procedure covers installation of the UV and PDA detectors. For the capillary cartridge
installation procedure, refer to Capillary Cartridge.
For information on how to install the LIF Detector, refer to Install the LIF Detector into the PA 800
Plus System 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)
• UV source optic module installed
1 Verify that power is off and no capillary cartridge is installed.
2 Lift the cartridge cover door and find the detector mounting location as shown in Figure 1.1 and
Figure 1.2.
3 Remove the detector from the package and remove the protective connector caps.
4 Position the detector on the mounting location. Carefully slide the detector back to make
contact with the back-plane receptacle.
5 Tighten the thumb screw until tight.
6 Unpack the fiber optic cable and remove the protective covers from each end (two covers for
the UV cable and three covers for the PDA cable). Make sure to keep the protective covers for
storage when not in use. Connect the cable to the clamp arm first, then to the detector.

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Installation Procedures
Figure 1.1 Installing the UV Detector
1. Outer Door Cover
2. UV Detector
3. Two-ended Fiber Optic Cable
4. Clamp Bar
5. Capillary Cartridge
6. Interface Block

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Installation Procedures
Figure 1.2 PDA Detector Installation
1. Outer Door Cover
2. PDA Detector
3. Three-ended Fiber Optic Cable
4. Clamp Bar
5. Capillary Cartridge
6. Interface Block

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Installation Procedures
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 after the OpCal cartridge is installed in its place:
1 From the main menu, open an instrument configured with PDA detection.
2 From the Control menu, select Direct Control. The Direct Control window opens.
Figure 1.3 PDA Detector - Direct Control

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Installation Procedures
3 Select the rainbow above the lamp. The PDA Detector Parameters dialog opens.
Figure 1.4 PDA Detector Parameters
4 Select Calibrate to start the calibration sequence. The calibration requires approximately two
minutes. Do not try to access the instrument during the calibration process.
Install UV Detector Wavelength Filters
When you install, the PA 800 Plus system includes 200 nm, 214 nm, 254 nm, and 280 nm filters. The
following section is for reference.
Before you start, you need:
• Filters
• Powder-free gloves (nitrile or neoprene recommended)
1 Using Direct Control, move the trays to the load position.

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Installation Procedures
2 Lift the outer door and then the cartridge cover door. The cartridge cover door is to the left of
the UV Detector as shown in Figure 1.5.
Figure 1.5 Installing UV Detector Wavelength Filters
3 Loosen the two thumb-screws and lift the clamp bar.
1. Outer Door Cover
2. Three-ended Fiber Optic Cable
3. UV Detector
4. Clamp Bar
5. Capillary Cartridge
6. Interface Block

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Installation Procedures
4 Remove the capillary cartridge from the interface block.
5 Turn the system power off.
6 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.
7 Remove the filter wheel access cover and rotate the filter wheel until the correct filter is visible
(Figure 1.6).
Figure 1.6 How to Remove the Filter Wheel Access Cover
8 Put on powder-free gloves (nitrile or neoprene recommended). Use the small forceps to remove
the desired filters from the filter wheel. Touching the optical surfaces of the filters without
gloves leaves skin oil that can cause improper filter performance.
1. Open cover at the indicated point
2. Access Cover
3. Thumb screws (2), one on each side

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Installation Procedures
9 Inspect each wavelength filter for a burned spot. Replace any filters that appear damaged.
UV filters are affected by heat and time. It is good practice to periodically check filter
performance with a spectrometer or to replace the filters.
10 Insert the new filter with the reflective side facing inward (Figure 1.7).
Figure 1.7 How to 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 thumb-screws.
13 Reinstall the capillary cartridge in the interface block.
14 Lower the insertion bar and tighten the two thumb screws.
15 Close the cartridge cover door.

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Installation Procedures
16 Turn the system power on and start the 32 Karat software.
17 Enable System Administration mode by selecting Tools > Enterprise Login on the enterprise
screen.
18 Right-click on an instrument icon and select Configure > Instrument. System Administration
mode must be enabled, and the user must have instrument administration privileges to
configure the instrument.
19 Select Configure on the instrument Configuration dialog.
The PA 800 Plus System Configuration dialog opens (Figure 1.8).
Figure 1.8 Configuration Dialog
20 Drag selected detector icons from the left pane to the right pane.

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Installation Procedures
Figure 1.9 System Configuration Dialog
21 In the Filter dialog, enter the position and type for each of the installed filters.

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Installation Procedures
Integrated 488 nm Solid-State Laser
WARNING
It is not safe to operate the solid-state laser module on any CE system other than
the PA 800 Enhanced Protein Characterization System or the PA 800 Plus
Pharmaceutical Analysis System. The 488 nm solid-state laser draws more current
than the interconnect module, previously used with the 488 nm argon ion laser,
and damages PA 800 systems that are not a PA 800 Enhanced Protein
Characterization System upgrade or a PA 800 Plus Pharmaceutical Analysis
System. The PA 800 Enhanced Protein Characterization System upgrade and the
PA 800 Plus Pharmaceutical Analysis System include a higher-rated wiring
harness to safely operate with the increased current.
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.
Before installing the solid-state laser, close the 32 Karat software instrument application.
WARNING
To prevent a possible shock hazard, power down the instrument before continuing
with component installation.
Installation Requirements
A SCIEX service engineer must complete the initial installation of the 488 nm solid-state laser. This
procedure includes proper laser configuration, installation of the updated wiring harness, setup of
the LIF detector (if necessary), and verification of operation including system safety features.
Install the Solid-State Laser Module
WARNING
To prevent a possible shock hazard, power down the instrument before continuing
with component installation.
If currently installed, remove the UV or PDA detector module from the PA 800 Plus system. Insert
the solid-state laser module and secure by finger tightening the thumb screws at the front of the
module.
Install the LIF Detector Filters
Before you install the LIF Detector Module, install the filters and filter housing.

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Installation Procedures
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. Contact SCIEX for assistance setting up and using channel 2.
The Filter Housing sits in the right side of the LIF Detector and has two filter holders, one for each
detector channel. Refer to Figure 1.10 for a complete diagram of the Filter Housing. You can use one
or more filters in each filter holder, depending on the application and the width of the filter. Typical
operation is with two filters: one rejection filter and one emission filter. The rejection filter prevents
scattered laser light from passing into the detector. The emission filter is a band-pass filter that
allows only the fluorescent signal at a selected wavelength to pass into the detector.
Depending on the application and laser, you can use any standard 0.5 inch diameter filters in the
range of 350 nm to 750 nm. The total thickness of the filters cannot exceed 0.350 inches. To install
filters into channel 1 of the filter holder. Refer to Figure 1.10.
Install a Filter in the Filter Housing
1 Hold the filter housing in the same orientation as shown in Figure 1.10.
Figure 1.10 Install Filters in the Filter Housing
2 Hold the filter with the arrow on the outside edge of the filter pointing up.
NOTE Make sure to handle filters by the edges.
3 Carefully put the filter into the filter holder.
1. Notch Filter
2. Emissions (Band Pass) Filter (Bottom View)
3. Emissions (Band Pass) Filter (Top View)

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Installation Procedures
4 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. The more reflective side has a wide-edge rim that extends beyond the
filter surface. Install the filter with the more reflective side toward the light source (the laser). In LIF
detection, laser light comes from the center of the filter housing. While holding the housing in the
correct orientation, install a filter with the arrow — or reflective side with the wider rim — pointing up
(toward the filter housing).
NOTE If you install a filter incorrectly, performance degrades even though the filter appears operational.
Install the Filter Housing into the Detector
Insert the filter housing into the opening in the right side of the detector. Refer to Figure 1.11. Make
sure the keyed corner of the filter housing lines up with the keyed corner of the Detector Module.
To ensure correct installation, the housing only fits into the opening in the orientation shown in
Figure 1.11. After you change the filters, allow the system to stabilize for 15 minutes before use.
Figure 1.11 Install the Filter Housing

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Installation Procedures
Install the LIF Detector into the PA 800 Plus System
WARNING
To prevent a possible shock hazard, power down the PA 800 Plus instrument
before continuing with component installation.
Before installing the LIF Detector, remove the UV source optics module by loosening the knobs at
the sides of the interface block. Remove the module using the handle.
Connect the LIF Detector into the PA 800 Plus system. 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.
To connect the probe to the LIF Cartridge, align the two pins on the left side of the probe and probe
body to the two pin holes and the probe hole in the clamp bar. Squeeze the probe to open its jaws
and carefully attach the probe to the clamp bar. Refer to Figure 1.12.
When not in use, install the probe in the probe holder on the 488 nm laser module. This provides
mechanical protection for the probe and shields it from dust and other particles.
Figure 1.12 Connect the Probe to the LIF Cartridge