Waters 2475 - C Solvent Considerations

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C-1

C Solvent Considerations

Contents:

Topic Page

Introduction C-2

Solvent miscibility C-3

Buffered solvents C-6

Head height C-6

Solvent viscosity C-6

Mobile phase solvent degassing C-7

Wavelength selection C-9

Main Page

Copyright Notice2
Trademarks2
Customer Comments3
Safety Considerations3
Safety Advisories4
Operating this Instrument4
Audience and Purpose4
Intended Use of the Instrument5
Calibrating5
ISM Classification5
ISM Classification: ISM Group 1 Class B5
EC Authorized Representative6
Table of Contents7
Theory of Operation16
Fluorescence Theory16
Fluorescence Detection17
Overview17
Excitation Sources17
Types of Light Sources17
Excitation Wavelength Selection17
Exciting the Sample18
Flow Cell18
Measuring Fluorescence18
Quantitation18
Emission Wavelength Selection18
Photomultiplier Tube18
Scanning19
Multichannel Operation19
Fluorescence Data19
References20
Detector Description21
Features21
Principles of Operation23
Detector Optics23
Optics Assembly Light Path24
Photomultiplier (PMT) Calibration26
PMT Sensitivity26
Filtering Noise26
Electronics27
Wavelength Verification and Test28
Operational Modes28
Single-Channel Mode28
Multichannel Mode29
Spectrum Scanning30
Lamp Energy and Performance30
Auto-Optimize Gain31
Method Optimization32
Example of Recommended Method Development Approach33
Ensuring Gain Optimization for each Peak of Interest34
Startup Diagnostic Tests35
Mobile-Phase Solvent Degassing35
Wavelength Selection36
Setting up the Detector37
Before You Begin38
Installing the Detector39
Plumbing the Detector39
Connecting Columns39
Assembling Fittings40
Making Tubing Connections41
Making Signal Connections41
Component Connection Overview42
Connecting the Ethernet Cable43
Choosing Signal Connections46
Making I/O Signal Connections47
Signal Connections48
Connecting an Alliance Separations Module50
Connecting RS-232 Devices54
Connecting Ethernet Devices56
Connecting Other Devices59
Required Materials59
Connecting Cables59
Connecting a Data System Using a Bus SAT/IN Module60
Connecting a 746 Data Module63
Connecting a Chart Recorder64
Connecting a 600-Series Pump65
Connecting a 717Plus Autosampler69
Connecting to the Electricity Source72
Using the Detector73
Starting the Detector74
Initializing the Detector74
Startup Failure75
Idle Mode76
Using the Operator Interface76
Using the Display76
Fluorescence and Message Icons77
Using the Keypad81
Navigating the User Interface88
Navigating to and from the Home Screen88
Preparing to Start a Run90
Setting up a Run90
Accessing Primary and Secondary Functions91
Operating the Trace and Scale Functions95
Configuring the Detector97
Configuring Event Inputs and Contact Closures97
Setting Pulse Periods98
Setting the Display Contrast99
Displaying System Information100
Using Online Help100
Operating the Detector100
Two Operating Modes101
Standalone Operation101
Remote Control Operation Via Ethernet Connection Using 2475 Instrument Control Software106
Verifying the Detector106
Manual Wavelength Calibration107
Normalizing Emission Units108
Operating the Detector in Single-Channel Mode109
Operating the Detector in Multichannel Mode110
Setting Gain and EUFS112
Programming Methods and Events115
Storing Methods115
Programming Timed Events116
Programming Threshold Events119
Storing a Method121
Retrieving a Method122
Viewing Events Within a Method122
Resetting a Method123
Clearing Events123
Scanning Spectra124
Types of Scanning124
Before You Begin124
Scanning New Spectra129
Parameters Used for Sample and Zero-Scans130
Programming a Zero-Scan131
Running a Sample Scan132
Scanning Using a Static Flow Cell136
Managing Results137
Storing a Spectrum137
Getting Information about a Stored Spectrum138
Reviewing a Stored Spectrum138
Creating a Difference Spectrum (Subtracting a Spectrum)139
Replaying a Spectrum139
Conserving Lamp Life139
Manually Extinguishing the Lamp140
Manually Lighting the Lamp141
Using a Timed Event Method to Program the Lamp142
Shutting down the Detector142
Maintenance Procedures144
Contacting Waters Technical Service144
Maintenance Considerations144
Safety and Handling144
Spare Parts145
Routine Maintenance145
Removing the Front-Left-Hand Panel Cover146
Inspecting, Cleaning, and Replacing the Flow Cell147
Flushing and Passivating the Flow Cell147
Removing the Flow Cell Assembly148
Replacing the Flow Cell149
Replacing the Lamp150
When to Replace the Lamp150
Removing the Lamp151
Installing the New Lamp154
Recording the New Lamp's Serial Number155
Replacing the Fuses156
Cleaning the Instruments Exterior157
Error Messages, Diagnostic Tests, and Troubleshooting159
Startup Error Messages160
Operational Error Messages161
User-Selected Diagnostic Tests and Settings166
Overview of Diagnostic Tests and Settings166
Sample and Reference Energy Diagnostic Tests169
Raman Signal-To-Noise Test Diagnostic Test170
Input and Output Diagnostic Tests and Settings170
Change-Lamp Function173
Testing the Keypad174
Testing the Display175
Other Diagnostic Tests and Settings175
Generating Test Peaks175
Overriding the Optical Filter Setting176
Reducing PMT Sensitivity177
Troubleshooting178
Introduction178
Information Needed When You Contact Waters178
Power Surges179
Hardware Troubleshooting179
A Safety Advisories184
Warning Symbols184
Task-Specific Hazard Warnings184
Specific Warnings185
Caution Symbol187
Warnings that Apply to All Waters Instruments188
Electrical and Handling Symbols194
Electrical Symbols194
Handling Symbols195
B Specifications197
C Solvent Considerations202
Introduction202
Clean Solvents202
Solvent Quality202
Preparation Checklist202
Water202
Buffers203
Tetrahydrofuran (THF203
Solvent Miscibility203
How to Use Miscibility Numbers205
Buffered Solvents206
Solvent Viscosity206
Mobile Phase Solvent Degassing207
Gas Solubility207
Solvent Degassing Methods208
Solvent Degassing Considerations209
UV Cutoffs for Common Solvents210
Index215

Waters 2475 - C Solvent Considerations

Table of Contents

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