Waters 996 Operator's Manual

Waters 996

PD A Detector

Operator’s Guide

34 Maple Street

Milford, MA 01757

053021TP, Revision 0

NOTICE

The information in this document is subject to change without notice and should not be construed as a commitment by Waters Corporation. Waters Corporation assumes no responsibility for any errors that may appear in this document. This manual is believed to be complete and accurate at the time of publication. In no event shall Waters Corporation be liable for incidental or consequential damages in connection with, or arising from, the use of this manual.

© 1997, 1993 WATERS CORPORAT ION. PRINTE D IN THE UNITED STATES OF AMERICA. ALL RIG H TS RESERVED. THIS BO OK OR PARTS THERE OF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.

Millennium, PowerStation, and Waters are registered trademarks and busLAC/E is a trademark of Waters Corporation.

All other trademarks are the sole property of their respective owners.

The quality management system of Waters’ chromatography applications software design and manufacturing facility, Milford, Massachusetts, complies with the International Standard ISO 9001 Quality Managemen t and Quality Assurance Stand ards. Waters’ quality management system is periodically audited by the registering body to ensure compliance.

STOP

Attention:

instrument. When you use the instrument, follow generally accepted procedures for quality control and

methods development. If you observe a change in the retention of a particular compound, in the resolution

between two compounds, or in peak shape, immediately take steps to determine the reason for the changes. Until you determine the cause of a change, do not rely upon the results of the separations.

This is a highly sensitive instrument. Read this user's manual before using the

STOP

Caution:

same type and rating.

Attention:

responsible for compliance could void the user’s authority to operate the equipment.

Attention:

A digital device, pursuant to Part 15 of the FCC rules. These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment. This equipment generates, uses, and can radiate radio frequency energy, and, if not installed and used in accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a residential area is likely to cause harmful interference, in which case you must correct the interference at your own expense.

Shielded cables must be used with this unit to ensure compliance with Class A FCC limits.

Note:

Level II category pertains to equipment that receives its electrical power from a local level, such as an electrical wall outlet.

For continued protection against fire hazard, replace fuses with those of the

Changes or modifications to this unit not expressly approved by the party

This equipment has been tested and found to comply with the limits for a Class

The Installation Category (Overvoltage Category) for t his instrument is Level II. The

Canadian Emissions Notice

This digital apparatus does not exceed the Class A limits for radio noise emissions from digital apparatus set forth in the Radio Interference Regulations of the Canadian Departme nt of Communications.

Le présent appareil numérique n’émet pas de bruits radioélectriques dépassant les limites applicables aux appareils numériques de la classe A prescrites dans les règlements sur le brouillage radioélectrique édictés par le Ministère des Communications du Canada.

Symbols Used on the Waters 996 Photodiode Array Detector

Direct current

Alternating current

Protective conductor terminal

Frame or chassis terminal

Caution, risk of electric shock (high voltage)

Caution or refer to manual

Caution, hot surface

Ultraviolet light

How to Use This Guide..................................................................... 10

Chapter 1

Installation ....................................................................................... 14

1.1 Installation Site Requirements .............................................. 14

1.2 Po wer Connections............................................................... 15

1.3 Millennium

1.3.1 Connecting the IEEE-488 Cable................................ 16

1.3.2 Setting the IEEE- 48 8 Addre ss............... ..... ..... .... ..... . 18

1.4 Non-IEEE-488 Communication Connections........................ 19

1.4.1 Connecting Analog Output Cables ............................ 19

1.4.2 Connecting Event Cables .......................................... 20

1.5 Fluidic Connections............................................................... 22

Workstation Connections................................. 16

1.6 Startup/Shutdown ................................................................. 24

Chapter 2

Diagnostics and Calibration ............................................................. 27

2.1 Startup Diagnostics............................................................... 27

2.2 User-Initiated Diagnostics..................................................... 30

2.3 PDA Calibration..................................................................... 31

Table of Content s 5

Chapter 3

Maintenance .................................................................................... 33

3.1 Flow Cell Maintenance.......................................................... 33

3.1.1 Flushing the Flow Cell ............................................... 33

3.1.2 Removing the Flow Cell........... .... .............................. 34

3.1.3 Disassembling and Cleanin g the Flow Cell. ..... .... ..... . 36

3.1.4 Installing the Flow Cell Assembly .............................. 38

3.2 Replacing the Lamp.............................................................. 39

3.2.1 Checking Lamp Usage .. ..... ..... .... ..... ......................... 39

3.2.2 Removing the Lamp....... ..... ..... .... ..... ..... ..... ..... .......... 41

3.3 Replacing the Fuses............................................................. 43

Chapter 4

Principles of the 996 PDA Detector Optics ...................................... 44

4.1 996 Detector Optics .............................................................. 44

4.2 Resolving Spectral Data ....................................................... 46

4.3 Measuring Light at the Photodiode ....................................... 47

4.4 Computing Absorbance Data Points..................................... 50

4.4.1 Calculatin g Absor ban ce........... .... ..... ..... .................... 50

4.4.2 Resolution.............................................. .................... 52

4.4.3 Filterin g Data ................. ..... ....................................... 53

Chapter 5

Spectral Contrast Theory ................................................................. 54

5.1 Comparing Absorbance Spectra........................................... 54

5.2 Representing Spectra as Vectors.......................................... 55

5.2.1 Vectors Derived from Two Wavelengths..................... 56

5.2.2 Vectors Derived from Multiple Wavelengths .............. 56

6 Table of Contents

5.3 Spectral Contrast Angles ...................................................... 57

5.4 Nonidealities ......................................................................... 60

5.4.1 Detector Noise............... ..... ..... .... ..... ..... ..... ..... .......... 60

5.4.2 Photom etric Error ..... .................................. ............... 61

5.4.3 Solvent Changes ...... ..... ............................................ 61

5.4.4 Thresho ld Angl e ... ..... ..... ..... ..... .... ..... ......................... 61

Appendix A

Detector S pecif icatio ns.. ..... ..... ...... ..... ..... ...... .. ...... ..... ..... ...... ..... 63

Appendix B

Spare Par ts ..... ...... ..... ..... ...... ..... ... ..... ..... ...... ..... ..... ...... ..... ..... ... 64

Appendix C

Warranty Informati on ..... ..... ..... ...... ..... ... ..... ..... ...... ..... ..... ...... ..... 65

C.1 Limited Express Warranty..................................................... 65

C.2 Shipments, Damages, Claims, and Returns......................... 69

Appendix D

Mobile P ha se A b so rb a nc e..... .. ... ... .. ................................... ........ 70

Index ................................................................................................ 74

Table of Contents 7

List of Figures

1-1 Waters 996 PDA Detector Dimensions..........................................15

1-2 Detector Rear Panel......................................................................16

1-3 Example of IEEE-488 Cable Connections.....................................17

1-4 Locating the IEEE-488 Address Switches.....................................18

1-5 Analog Output Terminals ...............................................................20

1-6 Event Input/Output Terminal Strip..................................................22

1-7 Compression Screw Assembly......................................................24

1-8 996 Detector Indicator Lights.........................................................25

2-1 996 PDA Detector Indicator Lights ................................................28

3-1 Flow Cell Access Door...................................................................35

3-2 Removing the Flow Cell Assembly ................................................35

3-3 Flow Cell and Fluidic Connections Assemblies.............................36

3-4 Disassembling the Flow Cell..........................................................37

3-5 Lamp Access Door........................................................................40

3-6 Lamp Usage Indicator....................................................................40

3-7 Lamp Power Cord and Mounting Screws.......................................42

3-8 Fuse Block.....................................................................................43

4-1 Optics Assembly Light Path...........................................................45

4-2 Benzene Spectrum at 1.2 nm Resolution......................................47

4-3 Photodiodes Discharged by Light..................................................48

4-4 Absorbance as a Function of Concentration..................................51

5-1 Comparing Spectra of Two Compounds........................................55

5-2 Plotting Vectors for Two Spectra....................................................56

5-3 Spectra with a Large Spectral Contrast Angle...............................58

5-4 Spectra with a Small Spectral Contrast Angle...............................59

5-5 Absorbance Spectra of a Compound at Two Concentrations........ 60

5-6 Effects of pH and Solvent Concentration on the Absorbance

Spectrum of p-Aminobenzoic Acid.................................................62

Table of Contents 8

List of Tables

1-1 Site Requirements ........................................................................14

1-2 Event In (Inject Start) Terminal Specifications on TTL

or Switch Closure.................................................................... 21

1-3 Event Out Terminal Specifications on Contact Closure............ 21

2-1 996 Detector Troubleshooting .......................................................28

4-1 Optics Assembly Components................................................. 45

A-1 996 Detector Specifications..................................................... 63

B-1 Spare Parts............................................................................. 64

C-1 Warranty Periods..................................................................... 68

D-1 Mobile Phase Absorbance Measured Against Air or Water...... 70

Table of Contents 9

How to Use This Guide

Purpose of This Guide

The

Waters 996 PDA Detector Operator’s Guide

maintaining, and troubleshooting the Waters optics and the principles of Spectral Contrast used in the Millennium analyzing the data from the PDA detector. Also included is information on vector analysis, mobile phase absorbance, specifications, and the warranty.

Audience

This guide is intended for individuals who need to install, operate, maintain, and troubleshoot the Waters 996 PDA Detector. It is also intended for users who need to understand the Spectral Contrast principles underlying the processi ng of PDA detector data by Millennium

soft wa r e .

Structure of This Guide

The

Waters 996 PDA Detector Operator’s Guide

Each chapter and appendix page is marked with a tab and a footer to help you quickly access information.

The following table describes the mater ial covered in each chapter and appendix of this guide.

describes the procedures for installing,

996 PDA Detector . It also describes detector

is divided into chapters and appendixes.

software for

Chapter/Appendix Description

Chapter 1, Installation De scr ibes how to install and set up the 996 detector. Chapter 2, Diagnostics

and Calibration Chapter 3,

Maintenance Chapter 4, Princi pl e s of

the 996 PDA Detector Optics

Chapter 5, Spectral Contrast Theory

10 How to Use This Guide

Describes how to troubleshoot the 996 detector.

Describes how to replace the flow cell, the lamp, and the fuse.

Explains the principles involved in resolving spectral data, measuring light at the photodiode, verifying wavelengths, and computing absorbance data.

Describes the calculations used for Spectral Contrast.

Chapter/Appendix Description

Appendix A, Detector

Provides the specifications of the Waters 996 PDA detector.

Specifications Appendix B, Spare

Provides a list of recommended and optional spare par ts.

Parts Appendix C, Warranty

Includes warranty and service information.

Information Appendix D, Mobile

Phase Absorbance

Provides a table of absorbances at several wavelengths for common mobile phases.

Installation

The Waters® 996 Photodiode Array (PDA) Detector operates in any standard laboratory environment. The detector requires electrical power, sample and waste fluidic lines, and the Millennium® communication with chart recorders, data integrators, and other instruments that are not compatible with Millennium software control.

1.1 Instal lation Site Requirements

Install the Waters 996 PDA Detector (Figure 1-1) at a site that meets the specifications listed in Table 1-1

Table 1-1 Site Requirements

Ambient temperature 4 to 40° C (39 to 104° F) Relativ e hum idi ty 20 to 80 percent,

Workstation. Optional connections on the detector rear panel allow

Factor Specification

noncondensing

Bench space Width: 11.5 in. (29 cm)

Bench support Capable of supporting

Clearance At least 4 in. (10 cm) on

14 Installation

Depth: 24 in. (61 cm) Height: 8.125 in. (22 cm)

31.5 pounds (14.3 kg)

the back and left sides for ventilation

11.5 in. (29 c m )

WATERS 996

Photodiode Array

8.5 in.

(22 cm)

24 in.

(61 cm)

Figure 1-1 Waters 996 PDA Detector Dimensions

1.2 Power Connections

Ensure that power connections for the 996 PDA Detector are made according to the procedures that follow.

LAMP

Sample Inlet

Sam ple Out let

Drain Line

Operating Voltage

The 996 PDA Detector has a universal input power supply that requires no voltage adjustment. The electrical power requirements for the Waters 996 PDA Detector are:

Voltage range:

•

Frequency range:

•

95 to 240 Vac (±10%)

50 to 60 Hz (±3 Hz)

Fuses

The Waters 996 PDA Detector is shipped with fuses rated for North American operation. If you operate the Waters 996 PDA Detector in another location, install the IEC-rated fuses (supplied in the Waters 996 Detector Startup Kit) in the fuse holder in the rear of the detector (refer to Section 3.3,

Replacing the Fuses).

Power Connections 15

Connecting the Power Cord

Connect one end of the 996 detector power cord to the rear panel power receptacle (Figure 1-2

) and the other end to a power outlet.

Power Cord

Receptacle

TP01452

Figure 1-2 Detector Rear Panel

1.3 Millennium32 Workstation Connections

The 996 detector requires signal connections to the Millennium32 Workstation over the IEEE-488 bus. All detector control and data acquisition communications take place over the IEEE-488 bus.

If an inject start signal is not available over the IEEE-488 bus, you must provide a

Note:

signal at the Event In terminals on the 996 detector rear panel (see Section 1.4.2,

Connecting Event Cables).

1.3.1 Connecting the IEEE-488 Cable

If the 996 detector is to be rack-mounted or stacked on other instruments, use the

Note:

right-angle adaptor included in the Star t up Kit when you make the IEEE-488 connection.

16 Installation

To connect the 996 detector to a Millennium32 Workstation:

1. Con nect one end of the IEEE-488 cable to the IEEE-488 receptacle on the rear panel of the 996 detector. Connect the other end of the cable (stackable connector for daisy-chaining additional instruments) to the IEEE-488 connector on any of the other instruments in your chromatographic system (Figure 1-3

Waters

IEEE-488

Cable

2690

IEEE-488 Cabl e

Wate r s 996

Detector

TP01544

IEEE-488

Connector

Millennium

busLAC/E Card

(on Workstati on)

Wate rs

Separations Module

Figure 1-3 Example of IEEE-488 Cable Connections

The order in which you connect IEEE-488 devices to the busLAC/E card on

Note:

the workstation is not important. For example, you can connect the 2690 separations module before or after the 996 detector.

2. Use another IEEE-488 cable to connect to the stackable connector on the first instrument and the IEEE-488 connector on another instrument.

3. Repeat step 2 for each IEEE-488 i nst rument in your chromatographic system, up to a maximum of 14 IEEE-488 instruments.

Keep in mind cable-length limitations when you set up your system. For a list

Note:

of IEEE-488 interface guidelines, refer to the Millennium

System

Installation/Configuration Guide, Section 2.3.1, Connecting IEEE-488 Devices.

4. Ensure that all IEEE-488 cable screws are fastened finger-tight.

Millennium32 Workstation Connections 17

1.3.2 Setting the IEEE-488 Address

To set the IEEE-488 address for the 996 detector:

1. Use a small screwdriver (or similar device) to set the DIP switches on the detector rear panel (Figure 1-4

) to the IEEE-488 address of the 996 detector. The address

must be a number from 2 to 29 and must be unique within your network. Refer to the

Millennium32 System Installation/Configuration Guide,

Section 2.3.1,

Connecting IEEE-488 Devices, for the correct IEEE-488 DIP switch settings.

IEEE-488 Cable

Connection

12345

IEEE-488

Address

Switches

– OPEN –

IEE 488 ADDRESS

2. To instruct the 996 detector to accept the new IEEE-488 address, power off, then power on the detector (see Section 1.6,

18 Installation

TP01457

Figure 1-4 Locating the IEEE-488 Address Switches

Startup/S hutdown).

1.4 Non-IEEE-488 Communication Connections

Non-IEEE-488 communication connections on the 996 detector include:

Analog Ou t pu ts

STOP

• absorbance-unit analog output channels to integrators, chart recorders, or other components.

Event Inputs and Outputs

• signals to and from other instruments.

You make all non-IEEE-488 communication cable connections to the 996 detector at the rear panel (see Figure 1-2

Attention:

disturbances that may affect the performance of this instrument, do not use cables longer than 9.8 feet (3 meters) when you make connections to the screw-type barrier terminal strips. In addition, ensure that you always connect the shield of each cable to chassis ground at one instrument only.

To me et the regulator y requirem ents of immunity from external electrical

– The 996 detector provides two, unattenuated, 1 volt-per

– The 996 detector sends and receives contact closure

1.4.1 Connecting Analog Output Cables

The values of the analog output signals generated by the 996 detector are specified by parameter valu es set from the Millen niu m “Waters 996 Detector Properties” topic in the

Required Materials

• One small, flat-blade screwdriver

Workstation. For details, refer to the

Millennium32 Online Help

Find tab.

• One electrical insulation stripping tool

• Analog signal cables (from Startup Kit)

Procedure

To connect the Waters 996 detector to a device that receives analog output signals from the 996 detector

1. Pull off the Analog output term inal str ip from the 996 detector rear panel (Figure 1-5

). This step simplifies the following steps.

Non-IEEE-488 Communication Connections 19

Removable An a l og O utput

Term inal Strip

Analog Ou t 1 –

IEE 488 ADDRESS

123456

Analog Out 2

–

Figure 1-5 Analog Output Terminals

2. Insert the bare wires at one end of an analog signal cable into the positive (+) and negative (–) terminals of Analog Out 1 (see Fi gure 1-5 screws to secure the + and – wires.

3. Connect the other end of the analog signal cable to the appropriate analog input terminal on the external device, being sure to maintain negative-to-negative and positive-to-positive continuity.

4. Reinstall the Analog Output strip.

1.4.2 Connecting Event Cables

The 996 detector has four terminal strip connections for contact closure signals:

TP01456

). Tighten the two

• Two input (inject start) signal terminals

• Two output (programmable event table) signal terminals

If an inject start signal is not avail a ble over the IEEE-488 bus, you must provide a signal at an Event In terminal on the 996 detector rear panel. Manual injectors such as the Rheodyne 7725i provide a cable that connects the injector to an Event In terminal on the 996 detector rear panel.

20 Installation

The values of the event output signals generated by the 996 detector are specified by parameter values set from the Millennium 996 Detector Properties” topic in the

Workstation. For details, refer to the “Waters

Millenn iu m32 Online Help

Find tab.

Electrical Specifications

Before you connect an external device to an event input or output terminal, refer to the electrical specifications in Table 1-2

Table 1-2 Event In (Inject Start) Ter m in al Specifications on TTL or Switch Closure

Parameter Specification

Low trigger <1.8 V High trigger >3.0 V Protected to ±30 V Minimum pulse width 30 msec Maximum current 5 mA

Table 1-3 Event Out Ter minal Spec ifications on Contact Closure

Parameter Specification

and Table 1-3.

Maximum power 10 W Maximum current 0.5 A at 20 V Maximum voltage 24 V RMS

Attention:

STOP

electrical connections as outlined in this section.

Required Materials

• Small flat-blade screwdriver

• One electrical insulation stripping tool

• Event signal cables (in Startup Kit)

To avoid damage to the 996 detector electronics, be sure you make the proper

Non-IEEE-488 Communication Connections 21

Making Event Input/Output Connections

To connect the 996 detector to an external event input or output device:

1. Pull off the Event Input/Output terminal strip from the rear panel (Figure 1-6 simplifies the following steps.

Removable Event

Input/Output

Terminal Strip

Event Out 2

Event Out 1

Event

–

In 2

1234567890

–

Event

In 1

IEE 488 ADDRESS

TP01455

Figure 1-6 Event Input/Output Terminal Strip

). This

2. Insert the bare wires at one end of the event signal cable into the positive (+) and negative (–) slots of the appropriate event input or output terminal (see

Figure 1-6

). Tighten the two screws to secure the +and – wires.

3. Connect the other end of the event signal cable to the appropriate event input or event output terminal on the external device.

4. Reinstall the Event Input/Output strip.

1.5 Fluidic Connections

Caution:

handing solvents. Refer to the Material Safety Data Sheets for the solvents in use.

22 Installation

To avoid chemical hazards, always observe safe laboratory practices when

Required Materials

• 5/16-inch open-end wrench

• 0.009-inch (0.23 mm) I.D. stainless steel tubing (in Startup Kit)

• Stainless steel tubing cutter or scribing file

• Pliers, plastic-covered, or with cloth

• Compression screw assembles, three

Procedure

To make fluidic connections to the 996 detector:

1. Measure the lengths of tubing needed to connect:

• The column outlet to the 996 detector inlet.

Be sure that you keep the length of this tubing as short as possible to

Note:

prevent band broadening.

• The 996 detector outlet to a waste collection bottle.

Ensure the length of this tubing is at least 1 to 2 feet (30 to 60 cm) to

Note:

prevent air bubbles from forming in the flow cell.

2. Cut the two lengths of tubing as follows:

• Use a Waters 1/16-inch stainless steel tubing cutter or a file with a cutting edge to scribe the circumference of the tubing at the desired break point.

• Grasp the tubing on both sides of the scribed mark with cloth- or plastic-covered pliers (to prevent marring the surface), then gently work the tubing back and forth until it separates.

• File the tubing ends smooth and straight to minimize dead volume and band broadening.

3. Assemble a compression fitting (as shown in Figure 1-7 column outlet line and at one end of the detector outlet line.

) at both ends of the

Fluidic Connec tions 23

+ 53 hidden pages

Waters 996 Operator's Manual

Specifications and Main Features

Frequently Asked Questions

User Manual

Table of Contents

List of Figures

List of Tables

How to Use This Guide

Installation

1.1 Instal lation Site Requirements

1.2 Power Connections

1.3 Millennium32 Workstation Connections

1.3.2 Setting the IEEE-488 Address

1.4 Non-IEEE-488 Communication Connections

1.4.1 Connecting Analog Output Cables

1.4.2 Connecting Event Cables

1.5 Fluidic Connections