Agilent Technologies 8453 User manual

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Agilent 8453 UV-visible
Spectroscopy System
Operator’s Manual

Agilent 8453 UV-visible Spectroscopy System Operator’s Manual
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2003-2007
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Manual Part Number
G1115-90030
Edition
04/07
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book.

Agilent 8453 UV-visible Spectroscopy System Operator’s Manual 3
In This Guide…
To be able to use your new Agilent 8453 UV-visible spectroscopy system
quickly, this book gives you step-wise procedures and examples for basic
operations and tasks.
This book shall not replace the detailed manuals available for installation:
Installing Your UV-visible Spectroscopy System and operation of your
software Understanding Your UV-visible Spectroscopy System nor your
Agilent 8453 Service Manual.
1 Introduction to Your System
In this chapter you will find an introduction to your Agilent 8453
spectrophotmeter and the concept of your Agilent ChemStation software.
2 Installation and Start Up
In this chapter you will find a summary of system installation and start-up of
a measurement session.
3 Good Measurement Practices
Good measurement practices are discussed in this chapter.
4 Using your Agilent 8453 UV-visible Spectroscopy System
Stepwise examples for basic measurements and related tasks are given in this
chapter.

4 Agilent 8453 UV-visible Spectroscopy System Operator’s Manual

Agilent 8453 UV-visible Spectroscopy System Operator’s Manual 5
Contents
1 Introduction to Your System 9
Agilent 8453 Spectrophotometer — Overview 10
Optical System Overview 10
Spectrophotometer Description 14
General Purpose Agilent ChemStation Software for UV-visible Spectroscopy —
Overview 18
User Interface Elements 19
Software Structure 23
Standard Mode Tasks 25
Standard Mode Data Processing 28
2 Installation and Start Up 35
Installation Summary for Your Agilent 8453 General Purpose UV-visible
System 36
General 36
Spectrophotometer 36
PC 37
Starting a Measurement Session 38

6 Agilent 8453 UV-visible Spectroscopy System Operator’s Manual
Contents
3 Good Measurement Practices 39
General Considerations 40
Spectrophotometer Design 40
Making Measurements 40
Sample Cell Material 41
Optical Specifications of Cells 42
Apertured Cells 43
Flow Cells 44
Handling and Maintaining Cells 45
Solvents 47
Sample Preparation 48
Photosensitive Samples 49
Stirring and Temperature Control 50
Checklist for Best Results 50
Inserting a Cell 53
4 Using your Agilent 8453 UV-visible Spectroscopy System 55
Starting Your First Measurement Session 56
Starting Your UV-visible Software 58
Measuring Caffeine Absorbance at 273 nm 59
Saving Your Parameters as a Method 62
Retrieving and Printing a Method 64
Saving and Retrieving Data 67
Saving your Samples 67
Saving a Selected Spectrum 69
Retrieving Spectra 71
Deleting Current Spectra 72
Print Preview of Reports 73
Finding the Caffeine Absorbance Maximum 76

Contents
Agilent 8453 UV-visible Spectroscopy System Operator’s Manual 7
Entering your Cell’s Path Length 80
Controlling your Sipper System 81
Using your Multicell Transport 83
Quantitative Analysis using a Calibration with Standards 86
Setup 87
Calibration 89
Analysis 91
How Can I Be Sure That My Agilent 8453 Works Properly? 93
Agilent 8453 Self test 93
How Can I Get a Deeper Understanding of UV-visible Spectroscopy? 96
When Do I Have to Measure a Blank? 98
Index 99

8 Agilent 8453 UV-visible Spectroscopy System Operator’s Manual
Contents

9
Agilent 8453 UV-visible Spectroscopy System
Operator’s Manual
Agilent Technologies
1
Introduction to Your System
Agilent 8453 Spectrophotometer — Overview 10
General Purpose Agilent ChemStation Software for UV-visible
Spectroscopy — Overview 18
Operation of the system is much easier if you understand the implementation
models. The mind-models of data acquisition, data evaluation and data
handling will help you to run the system successfully.
Your spectroscopy system is based on an Agilent 8453 spectrophotometer and
general purpose Agilent ChemStation software for UV-visible spectroscopy
running on a PC with the supported Microsoft operating system(s). These two
components are linked together by a network connection. This type of link is
very flexible: it can be used for a direct connection between the
spectrophotometer and the PC as well as the integration in a enterprise
network with network access from the PC to the spectrophotometer.
The tasks are split between these devices such that the spectrophotometer
acquires and provides absorbance data which are handled by the PC’s
application software. All of the data display, evaluation and longterm storage
is done under software control on the PC.

10 Agilent 8453 UV-visible Spectroscopy System Operator’s Manual
1 Introduction to Your System
Agilent 8453 Spectrophotometer — Overview
Agilent 8453 Spectrophotometer — Overview
This section gives an overview of the optical system and explains the
spectrophotometer front and back panels. It also explains the layout and
construction of the spectrophotometer including the electronic and
mechanical assemblies inside the spectrophotometer.
Optical System Overview
Optical System
The optical system of the spectrophotometer is shown in Figure 1. Its
radiation source is a combination of a deuterium-discharge lamp for the
ultraviolet (UV) wavelength range and a tungsten lamp for the visible and
short wave near-infrared (SWNIR) wavelength range. The image of the
filament of the tungsten lamp is focused on the discharge aperture of the
deuterium lamp by means of a special rear-access lamp design which allows
both light sources to be optically combined and share a common axis to the
source lens. The source lens forms a single, collimated beam of light. The beam
passes through the shutter/stray-light correction filter area then through the
sample to the spectrograph lens and slit. In the spectrograph light is dispersed
onto the diode array by a holographic grating. This allows simultaneous access
to all wavelength information. The result is a fundamental increase in the rate
at which spectra can be acquired.