Agilent Technologies Cary 8454 Operator's Manual
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Agilent Technologies Cary 8454 Operator's Manual

Uv-visible spectroscopy system
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Summary of Contents for Agilent Technologies Cary 8454

  • Page 1 Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual ...

  • Page 2: Safety Notices

    1987) or any equivalent agency regulation or contract clause. Use, duplication or disclo- sure of Software is subject to Agilent Tech- nologies’ standard commercial license terms, and non-DOD Departments and Agencies of the U.S. Government will Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 3 In This Guide… This book is an introduction to your Agilent Cary 8454 UV-Visible spectroscopy system. It includes step-wise procedures and examples for basic operations and tasks. For more detailed information see the following manuals: • Agilent Cary 8454 UV-Visible Spectroscopy System Installation Guide •...
  • Page 4 Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 5: Table Of Contents

    Introduction to Your System Agilent Cary 8454 UV-Visible Spectrophotometer — Overview Optical System Overview Spectrophotometer Description General Purpose Agilent ChemStation Software for UV-Visible Spectroscopy — Overview User Interface Elements Software Structure Standard Mode Tasks Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 6 Inserting a Cell Using your UV-Visible Spectroscopy System Starting Your First Measurement Session Starting Your UV-Visible Software Measuring Caffeine Absorbance at 273 nm Saving Your Parameters as a Method Retrieving and Printing a Method Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 7 How Can I Be Sure That My Spectrophotometer Works Properly? Spectrophotometer Self test What Maintenance Does My Spectrophotometer Require? Cleaning Spare Parts How Can I Get a Deeper Understanding of UV-Visible Spectroscopy? When Do I Have to Measure a Blank? Index Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 8 Contents Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 9: Safety Information

    Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual Safety Information Safety Information 10 Warning Symbols 10 Precautions 12 Solvent Information 13 Other Precautions 15 Environmental Conditions and Power Requirements 17 Regulatory Information 18 Agilent Technologies...

  • Page 10: Safety Information

    This symbol is placed on the product where it is necessary for you to refer to the manual in order to understand a hazard. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 11 Indicates a protective ground terminal. Eye damage may result from directly viewing light produced by deuterium lamps used in detectors and spectrophotometers. Always turn off the deuterium lamp before opening the lamp door on the instrument. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 12: Precautions

    Do not operate the instrument in the presence of flammable gases or fumes. Operation of any electrical instrument in such an environment constitutes a definite safety hazard. Do not install substitute parts or make any unauthorized modification to the instrument. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 13: Solvent Information

     2CHCl3 + O2 2COCl2 + 2HCl This reaction, in which stainless steel probably acts as a catalyst, occurs quickly with dried chloroform if the drying process removes the stabilizing alcohol. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 14 For example, a 1-% solution of acetic acid in methanol will attack steel. • Solutions containing strong complexing agents (for example, EDTA, ethylene diamine tetra-acetic acid). • Mixtures of carbon tetrachloride with 2-propanol or THF. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 15: Other Precautions

    Shock Hazard WARNING Connecting the Agilent Cary 8454 UV-Vis to a power source that is not equipped with a protective earth contact creates s shock hazard for the operator and can damage the instrument. Always operate your instrument from a power outlet which has a ground connection.

  • Page 16: Allowing Hot Instrument To Cool

    The Lithium battery can be found on the Spectrometer Processor Main Board (SPM). Danger of explosion if battery is incorrectly replaced. Replace only with the same or equivalent type recommended by the equipment manufacturer. Do not dispose of lithium batteries in domestic waste. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 17: Environmental Conditions And Power Requirements

    Avoid air circulation of varying temperatures (like from refrigerators or air conditioners) and exposure to direct sun light to ensure the instrument operates within performance specifications. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 18: Pollution Level

    Group 1 ISM equipment: group 1 contains all Industrial, Scientific and Medical (ISM) equipment in which there is intentionally generated and/or used conductively coupled radio- frequency energy which is necessary for the internal functioning of the equipment itself. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 19 4 Make sure that all peripheral devices are also certified. 5 Make sure that appropriate cables are used to connect the device to peripheral equipment. 6 Consult your equipment dealer, Agilent Technologies, or an experienced technician for assistance. Changes or modifications not expressly approved by Agilent Technologies could void the user’s authority to operate the equipment.
  • Page 20 이 기기는 업무용 (A 급 ) 전자파적합기기로서 판 매자 또는 사용자는 이 점을 주 의하시기 바라 며 , 가정외의 지역에서 사용하는 것을 목적으 로 합니다 . Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 21: Introduction To Your System

    Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual Introduction to Your System Agilent Cary 8454 UV-Visible Spectrophotometer — Overview 22 General Purpose Agilent ChemStation Software for UV-Visible Spectroscopy — Overview 30 This chapter introduces the principles of data acquisition, data evaluation and data handling to help you to run your system successfully.

  • Page 22: Agilent Cary 8454 Uv-Visible Spectrophotometer - Overview

    This allows simultaneous access to all wavelength information. The result is a fundamental increase in the rate at which spectra can be acquired. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 23 If the measurement rate is very fast, you can force the shutter to remain open (using Agilent ChemStation software) or it stays open automatically (handheld controller software). Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 24 The spectrograph lens is the first of the parts which are collectively known as the spectrograph. It is mounted on the housing of the spectrograph. The spectrograph lens refocuses the collimated light beam after it has passed through the sample. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 25 The photodiode array is the heart of the spectrograph. It is a series of 1024 individual photodiodes and control circuits etched onto a semiconductor chip. With a wavelength range from 190 nm to 1100 nm the sampling interval is about 0.9 nm. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 26: Spectrophotometer Description

    The front view of the spectrophotometer is shown in Figure 2. Notice that the sample compartment is open. Unlike conventional spectrophotometers the Agilent Cary 8454 UV-Visible spectrophotometer does not suffer from ambient false light. The open sample area makes it easier to access for cuvette handling and to connect tubing to a flow cell or thermostatable cell holder.
  • Page 27 Additional information such as concentration and so on, have to be entered in the operating software. • Stop—the spectrophotometer and/or software aborts any ongoing activity and returns to a to ready state. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 28 The UV-Visible ChemStation software currently supports LAN communication only. • The MIO board slot is reserved for a LAN interface board. • The accessory board slot is reserved for future use. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 29 For exchanging lamps both doors, the plastic door and the sheet-metal door must be opened. Safety light switches are automatically turning off the lamps when the inner sheet metal door is opened. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 30: General Purpose Agilent Chemstation Software For Uv-Visible Spectroscopy - Overview

    Agilent ChemStation software and the data analysis concept behind. It explains how data are processed and what the advantages of this processing are from a practical point of view. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 31: User Interface Elements

    This user interface consists of a number of elements as described in the following sections. Method name Tool bar Menu bar Analysis panel Data files Navigation bar Modes Instrument panel Tabular results Spectra Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 32 The analysis panel gives you a graphical visualization of the current working context. In addition, it provides access to the setup dialog of your actual task by means of the Setup button. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 33 In addition you can select a sampling system from the list of available systems. Its parameters can be adjusted by means of the Setup button. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 34 Views are usually handled automatically by the operation you performed, but you may also use the view menu’s commands to select the view you prefer. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 35: Software Structure

    PC. In addition a data analysis session, which has the suffix (Offline), can be launched. The offline session allows recalculations based on stored data, and is useful in the development of analytical methods. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 36 A set of parameters and data can be saved as a method. This allows you to repeat your analysis task under defined conditions simply by loading a method and running your samples. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 37: Standard Mode Tasks

    A task switch within the Standard mode is much faster than an entire mode switch. An additional benefit of these tasks is that the definition of a method is done within a single dialog. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 38 Spectrum/Peaks In the Spectrum/Peaks task, you can view absorbance minima and maxima. The focus here is more on the wavelength scale, but the relevant absorbance readings are also provided. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 39 The calibration can be optimized for your concentration range of interest by changing the wavelength used for calibration. A new calibration and a new analysis is automatically performed, based upon your current standard data. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 40: Standard Mode Data Processing

    The raw data container held the originally acquired data as specified with your acquisition parameters and stamped with, for example, acquisition date and time and the acquisition operator’s name. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 41 This data is stored in the used wavelength container. In the Fixed Wavelength task, for example, a tabular view of this data is available with the Sample/Results Table window. An additional step is the evaluation of the accessed data. Results Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 42 These steps are always performed in the above order identically for all spectra in the raw data container. The results are placed in the results container. The previous content of the processed spectra, used wavelength and results container is replaced. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 43 Agilent ChemStation memory. This means that results now become a function of a measured set of standards and of the standard concentrations specified. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 44 Also, by simply removing this standard from your standard data set, you can eliminate outliers in your calibration. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 45 This optimization can be done quickly, just by setting new method parameters. You get new answers almost instantaneously with consistent and reliable results. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 46 Introduction to Your System General Purpose Agilent ChemStation Software for UV-Visible Spectroscopy — Overview Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 47: Installation And Start Up

    Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual Installation and Start Up Installation Summary for Your General Purpose UV-Visible System 48 Starting a Measurement Session 50 This chapter does not replace the information available with the Agilent Cary 8454 UV-Visible Spectroscopy System Installation Guide. It is intended as a reminder of the key steps of the installation and system startup.

  • Page 48: Installation Summary For Your General Purpose Uv-Visible System

    Check that your spectrophotometer is either connected to your PC directly using a crossover LAN cable, or to your LAN using a direct connection. The Agilent Cary 8454 UV-Visible spectrophotometer does not have a CAN interface. CAUTION Do not connect the LAN adapter of your PC to the CAN interface of the Agilent 8454...
  • Page 49 If you connected your spectrophotometer directly to your PC be sure that the Agilent Bootp Service is installed and configured. If you connect to your spectrophotometer using a LAN make sure that your network administrator assigns the IP address to your spectrophotometer. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 50: Starting A Measurement Session

    After this alignment you are ready to measure absorbance data and spectra. It takes about 15 minutes for the lamps to reach stable state conditions. For best results, do NOTE not perform measurements before this period of time has elapsed. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 51: Good Measurement Practices

    Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual Good Measurement Practices General Considerations 52 Inserting a Cell 65 This chapter describes • making measurements • selecting material, optical specification and type of cell • handling and maintaining cells • checklist for good results •...

  • Page 52: General Considerations

    Chemically, the only difference between the blank and the sample should be the presence of the analyte(s). For measurements with liquid samples, the blank should be a sample cell filled with the solvent you plan to use. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 53: Sample Cell Material

    350 to 1100 nm, you can use good quality glass cells. Disposable plastic sample cells, for measurements in the range 400–1100 nm, are also available. The quality of these cells varies and they are generally not recommended. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 54: Optical Specifications Of Cells

    Light Beam out Light Beam In Light Beam in Non-parallel Non-parallel Cell Walls Cell Walls Light Beam Out Light Beam out Figure 4 Shift of the spectrophotometer light beam due to non-parallel cell walls Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 55: Apertured Cells

    Quartz cells with black Figure 5 Recommended cells * Quartz cells with black apertures smaller than 2 mm, when used with a multicell CAUTION transport, can lead to measurements with poor reproducibility. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 56: Flow Cells

    Also, the cell can be rinsed thoroughly with the solution to be measured. The design of the flow cell should minimize entrapment of bubbles and flow channeling to provide the most reliable results. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 57: Handling And Maintaining Cells

    Floating particles in the cell will deflect the light beam and result in very poor quality of the measured spectrum. Floating particles will deflect and scatter the light beam Figure 7 Floating particles in a cell Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 58 If glass pasteur pipettes are used, make sure that the optical windows of the cell are CAUTION not touched or scratched by the pipette. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 59: Solvents

    Cyclohexane Ethyl ether 245–260 nm Chloroform Ethyl acetate Methyl formate 265–275 nm Carbon tetrachloride Dimethyl sulfoxide Dimethyl formamide Acetic acid 280–290 nm Benzene Toluene m-Xylene Above 300 nm Pyridine Acetone Carbon disulfide Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 60: Sample Preparation

    Samples which contain colloidal dispersions, dust or other particulate matter should be filtered, centrifuged or allowed to settle. If not, the overall attenuation-of-transmittance spectrum due to light scattering and/or reflection will hide the spectral information from the analyte. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 61: Photosensitive Samples

    Tungsten lamp is sufficient for a good signal to noise ratio over the wavelength range 400–1100 nm. When using cells with small apertures, you should check the signal to noise ratio by making sample measurements under the conditions of your application. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 62: Stirring And Temperature Control

    Measurements: Solution in cell is free of floating particles Solution in cell, and cell walls are free of bubbles Solution in cell is mixed homogeneously Blank measured on same solvent as sample Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 63 Time between blank and sample measurement should be short Figure 9 Example of a blank on water showing a good baseline Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 64 Example of a blank on water with bubbles causing a poor baseline If your blank or spectra shows artifacts similar to those in Figure 10, see “Solvents" on NOTE page 59 to optimize the measurement procedure. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 65: Inserting A Cell

    The cell holder is designed for cells with 15 mm center height. If applicable, these cells should not be removed between the respective reference (blank) measurement and the sample measurement. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 66 Good Measurement Practices Inserting a Cell Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 67: Using Your Uv-Visible Spectroscopy System

    Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual Using your UV-Visible Spectroscopy System Starting Your First Measurement Session 68 Starting Your UV-Visible Software 70 Measuring Caffeine Absorbance at 273 nm 71 Saving Your Parameters as a Method 74 Retrieving and Printing a Method 76...

  • Page 68: Starting Your First Measurement Session

    Starting Your First Measurement Session Starting Your First Measurement Session 1 Make sure that your spectrophotometer has been installed correctly. For details of installation see your Agilent Cary 8454 UV-Visible Spectroscopy System Installation Guide. 2 Switch on your PC, monitor and printer.
  • Page 69 Instrument Panel’s Sample button, or by pressing the spectrophotometer’s Sample button. For details on how to mount your cell, see “Inserting a Cell" on page 65. NOTE Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 70: Starting Your Uv-Visible Software

    4 Enter your name to log on to your Agilent ChemStation session. If you protected your manager level by password, you must enter the correct password. The system will then come up with the last-used mode and method. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 71: Measuring Caffeine Absorbance At 273 Nm

    1 Make sure that you are in Standard mode. The mode is indicated on the toolbar of your Agilent ChemStation session. 2 Select the Fixed Wavelength task in the analysis panel’s selection box. 3 Click Setup in the analysis panel to open the parameter dialog. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 72 Put the cell in the cell holder and make sure the transparent windows face towards the font and the back of the spectrophotometer. Push the lever down to secure your cell in the cell holder. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 73 9 The view shows you the spectrum of your caffeine sample with a vertical line indicating your wavelength of interest. Below the spectrum is the Sample/Result Table which shows the absorbance reading at 273 nm. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 74: Saving Your Parameters As A Method

    This allows you to load such a method on your system or even transfer such a method to your colleague with a Agilent ChemStation system. 1 Choose Save Method As… from the File menu or click the icon. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 75 This displays the Method Options & Information dialog box. In the Method Information section you can enter a descriptive text which becomes part of your method. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 76: Retrieving And Printing A Method

    1 Choose Load Method… from the File menu or click the icon in the toolbar. If your current method has been modified, a dialog box will ask you whether you want to NOTE save or ignore these changes. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 77 3 If you want to load this method, click OK. Whenever you change a parameter of the current method, an indication is displayed in the NOTE toolbar’s modification field. This triggers the reminder dialog mentioned above. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 78 4 To print a method, choose Print, Method from the File menu. To print your method report, your printer must be properly configured and online. If your NOTE printer is currently not online, you can view the print preview on the screen. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 79: Saving And Retrieving Data

    In addition a single spectrum can be selected for storage. Saving your Samples 1 Choose Save, Samples As… from the File menu or click the toolbar icon. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 80 Usually the extension .std is used for standards only. If the file name exists, a message box is displayed allowing you to abort or continue with the operation. 3 Click OK to start the operation. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 81: Saving A Selected Spectrum

    Table window. you may get the warning, Select/activate a window!, on the message line. 3 Select a data files in the File name selector box of the Save Spectra As… dialog or enter a valid file name into the File name edit box. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 82 Using your UV-Visible Spectroscopy System Saving and Retrieving Data 4 Press OK to start the operation. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 83: Retrieving Spectra

    Saving and Retrieving Data Retrieving Spectra 1 Choose Load, Samples… from the File menu. 2 Select the data file in the File name selector box of the Load Spectra to SAMPLES dialog box. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 84: Deleting Current Spectra

    1 Choose Clear, Samples from the Edit menu or click the toolbar icon. Clear, Standards and Clear, Math. Results can be used to delete the current standards and NOTE the current mathematical results spectra from the Agilent ChemStation memory. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 85: Print Preview Of Reports

    Printing a results report requires you to have set all parameters properly and have data NOTE available for evaluation. If you do not have data, you may get the message No results present! on the message line. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 86 In addition, you may use a different size for your preview display. Three sizes are available with the size selection box. Depending on your display resolution select the one which best fits your needs. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 87 The Print button sends the report to the printer, which is displayed in the lower left corner of the print preview window. • The Close button closes your print preview window and discards the report. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 88: Finding The Caffeine Absorbance Maximum

    Agilent ChemStation session. 2 Select the Spectrum/Peaks task in the analysis panel’s selection box. 3 Use the Setup button of the analysis panel to open the parameter dialog. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 89 Put the cell in the cell holder and make sure the transparent windows face towards the front and the back of the spectrophotometer. Push the lever down to secure your cell in the cell holder. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 90 Close the cell holder’s level. 8 Press the Sample button on the front of your spectrophotometer or click Sample on the Instrument Panel to start the measurement. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 91 9 The view shows you the spectrum of your caffeine sample. Two peaks were found and are annotated with the wavelength. Below the spectrum graph the Sample/Result Table shows the wavelength of the peaks found and the measured absorbance values. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 92: Entering Your Cell's Path Length

    1 Click Setup on the Instrument Panel. 2 Type the path length in cm in the Setup Manual dialog box. 3 Click OK to set the specified path length. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 93: Controlling Your Sipper System

    OK. 3 Click Setup again, and access the Sipper Parameter dialog by clicking Parameter. The parameters required can be determined using the Flow Test task of your Verification and Diagnostics mode. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 94 Every measurement you start by clicking one of the Instrument Panel measurement buttons NOTE or by pressing the spectrophotometer buttons uses the sipper for sample introduction. Sipper introduction is also used by an automated sequence. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 95: Using Your Multicell Transport

    Transport manual. 1 Select Multicell (8-cell) in the Instrument Panel selection box. 2 Click Setup on the Instrument Panel. Type the path lengths of all cells used in cm and click OK. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 96 Instrument Panel or choose Multicell Transport Position from the Spectrophotometer menu to access the Multicell transport Control dialog. In the Multicell Transport Control, press one of the position buttons to move the multicell transport. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 97 In an automated sequence, the multicell transport can be used for automatic sample NOTE introduction. A maximum of 8 samples can be introduced. If you specify more than 8 measurements this warning is displayed: Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 98: Quantitative Analysis Using A Calibration With Standards

    Agilent ChemStation. For the practical experiment we use the IQ caffeine sample as standard and a 1:1 dilution with distilled water as sample. For the calibration we use absorbance data at 273 nm. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 99: Setup

    3 A new task panel is displayed and the Quantification Parameters dialog is opened automatically. If you are already in the Quantification task, use the Setup button on the analysis panel to NOTE open the parameter dialog. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 100 Prompt for sample information. Select Absorbance as Data type and set Display spectrum From 190 nm To 340 nm. 5 Click OK to set your parameters. Now you are ready to run your measurements. NOTE Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 101: Calibration

    Close the cell holder’s level. 4 Press the Standard button on the front of your spectrophotometer, or click Standard on the Instrument Panel to start the measurement. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 102 NOTE to the samples or standards view. At this point you can also save your method for future use. See “Saving Your Parameters as a Method" on page 74 for more information. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 103: Analysis

    Sample on the Instrument Panel to start the measurement. 3 Enter your sample’s information in the Sample Information dialog box and click OK. The view switches to the samples and your quantitative results will be displayed with the Sample/Result table. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 104 Using your UV-Visible Spectroscopy System Quantitative Analysis using a Calibration with Standards To save your data for future use or for documentation purposes, see “Saving and Retrieving NOTE Data" on page 79 for more information. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 105: How Can I Be Sure That My Spectrophotometer Works Properly

    Standard kits are available for operational qualification and performance verification (OQ/PV). OQ/PV kits for the Agilent Cary 8454 UV-Visible spectrophotometer are available as spare parts. A quick check without the need of standards can be performed by the Verification and Diagnostics mode’s self test.
  • Page 106 Self-test results can be stored with the spectrophotometer. Storing the self-test results NOTE with the spectrophotometer allows you to monitor the performance over time. Graphical representations of the self-test histories can be generated. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 107: What Maintenance Does My Spectrophotometer Require

    Do not use organic solvents or abrasive cleaning products. Spare Parts For spare parts and consumables ordering information, refer to the Agilent Technologies website at www.agilent.com. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 108: How Can I Get A Deeper Understanding Of Uv-Visible Spectroscopy

    This covers Basic Principles through to details of Derivative Spectroscopy. In addition, solutions to selected topics in UV-Visible analysis are described. There you can find help on topics such as Enhancing Sensitivity or Determining Purity. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 109 How Can I Get a Deeper Understanding of UV-Visible Spectroscopy? More specific examples, including data, are available in the Examples section of the Standard Mode help. These can be used to run your Agilent ChemStation software for the special purpose described. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 110: When Do I Have To Measure A Blank

    To ensure the highest accuracy of sample measurement, the blank measurement should be taken immediately prior to the actual sample measurement. Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...

  • Page 111: Index

    35 coefficients, 43 session online, 69 curve, 39 workstation, 44 carbon disulfide, 59 ambient light, 26 carbon tetrachloride, 59 analysis, 103 CE compliance, 18 analysis panel, 32, 105 cell changer, 95 Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 112 27 making measurements, 52 door to access lamps, 29 installation, 48 manager level, 36, 70 dust, 60 installation qualification, 71 maximum, 37 measure standard, 101 enhancing sensitivity, 108 standard information, 102 Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 113 36 passivating new cells, 57 rapid absorbance changes, 62 thermal denaturation, 36 path length ratio, 37 verification and diagnostics, 36, 105 setup, 92 raw data, 40 mouse cursor, 33 PC, 50 Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 114 65 sheet-metal used wavelength, 41 standard push button, 27 door, 29 useful wavelength range of solvents, 59 standard single-cell cell holder, 65 shine-through aperture, 23 user definable equation, 39 shutter, 22, 23 Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 115 86 Warning Symbols, 10 warning symbols, 10 water, 59 wavelength, 41 wavelength reproducibility, 45 wedge shaped cells, 54 weight, 39 width of slit, 24 window, 34 sample/results table, 41 Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 116 Index Agilent Cary 8454 UV-Visible Spectroscopy System Operator’s Manual...
  • Page 118 In This Book To be able to use your new Agilent Cary 8454 UV-Visible spectroscopy system quickly, this book gives you step-wise procedures and examples for basic operations and tasks.  Agilent Technologies 2002, 2003-2008, 2011, 2013, 2014, 2016...

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