Agilent Technologies 8453 Manual de usuario

Agilent 8453 UV-visible

Spectroscopy System

Operator’s Manual

Agilent 8453 UV-visible Spectroscopy System Operator’s Manual

Notices

2003-2007

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G1115-90030

Edition

04/07

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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

Agilent 8453 UV-visible Spectroscopy System

Operator’s Manual

Agilent Technologies

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

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.

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