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Cary 100/300 Certification Manual

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1. vel Ss 235 257 313 amp 350 nm otdometric mode Abs Speetral bandwidth 1 0nm Signal averaging time 3 0 sec Lamp UV Vis Specifications Wavelength Absorbance Tolerance 235 nm 0 7480 Abs 0 010 Abs 257 nm 0 8650 Abs 40 010 Abs 313 nm 0 2920 Abs 0 010 Abs 350 nm 0 6400 Abs 0 010 Abs What if my instrument fails to meet the specification Q Repeat the test with a new solution then compare the results to determine if the error was introduced by the solution Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 6 4 Publication date 10 02 Cary 100 300 certification manual Australian TGA tests These tests available from the configure page TGA tests of the Cary WinUV Validate software are required by the Australian Therapeutic Goods Administration TGA good laboratory practices recommendations Many of these tests are duplicated in either the standard instrument performance tests or the other pharmacopoeia tests 7 1 Wavelength accuracy 7 1 1 Publication date 10 02 Poor wavelength accuracy can produce low quantitative results as the absorbance measurement may have been made on the side of the peak rather than at the peak Wavelength accuracy is determined by scanning a known wavelength peak and calculating the difference between the theoreticahwavelength peak and the wave
2. Recertification Kit description t Kit serial Comments references Name amp t date part number number signature Publication date 10 02 11 3 Cary 100 300 certification manual Certification Kit log sheet Page Number Recertification Kit description t Kit serial Comments references Name amp t date part number number signature 11 4 Publication date 10 02
3. To avoid injury ALWAYS ensure that safety practices governing the use handling and disposal of such materials are strictly observed Appropriate safety clothing and glasses should also be worn Instrument parameters Wavelength 241 15 287 45 x 361 50 526 Refer to tHe Sol cali SBW Photometric mode S Scan range NAN 231 15 to 546 30 Spectral bandwidth 0 10 nm Signal aver nd OBS 0 10 sec Data meng 0 10 nm Lamp AY UV Vis i A wa elength nm Tolerance 241 1 1 nm 287 1 1 nm 361 5 1 nm 536 3 3 nm What if my instrument fails to meet the specification m m Check that you have set the tolerances to values that are within the required specifications for the test Recalibrate the wavelength drive mechanism by turning the instrument off and on again Check the deuterium D and visible lamp alignments and perform the wavelength accuracy test again If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 7 3 Cary 100 300 certification manual 7 4 7 2 Resolution power The monochromator of the instrument separates light into a spectrum and sends a small band of that spectrum through to the sample If the spectral bandwidth is adjustable the smaller it is the better the resolution of the instrument The limiting resolution of an instrument is that sett
4. 9 4 Remote stray light Instrument parameters Wavelength 350 nm Photometric mode O T Spectral bandwidth 1 5 nm Signal averaging time 10s Lamp UV Vis Specifications Filter Error 2 T 20 0 10 T 10 0 20 T 10 0 What if my instrument fails to meet the specification QO Check the filter transmission values and tolerances have been correctly entered Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Stray light is the amount of light reachi detector that is of a wavelength other than that selected The stray light le es the minimum transmittance that the instrument is able to meastiye Tha detector in the accessory is located in the integrating sphere This test uses the blankingplate and UG11 filter Instrument paramet Wavelength 2 350 nm Photometric 140 OT Spectral i 1 5 nm Signal time 10s a AY UV Vis ification Stray light at 350 nm lt 0 01 T What if my instrument fails to meet the specification Q Ensure that the filters were inserted in the correct order as specified by the Cary WinUV software Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 9 1 3 4 9 2 Publication date 10 02 Cary 100 300 certification man
5. If you are using the multicell holder to automate the testing procedure then Select the Setup button to display the Setup dialog then select the Analyze tab and choose multicell accessory To start the Validate run 1 Select the Start button or select Start in the Commands menu to commence the testing procedure If you have chosen to perform an automated run with the Multicell Holder as per the previous step then the Cell Loading Guide will appear Place the specified solutions and or filters exactly as shown in this dialog and press OK Alternatively follow the instructions on the prompts that appear and select the OK button to continue with the performa ts Select the Cancel button at any time stop the Validate run For manual runs without the use tOxntated accessories the Validate application will automaticatly atra e tests so those requiring your filters or solutions into the sample your intervention wil performed later in the run The system will advise you when user intervepti The result ach test are displayed in the Report area immediately after the test is coryple After Za of the Validate run the Cary system automatically generates a report Phe report file is stored the Cary WinUV directory e Report file will be saved with the following format DATE TIME RVO For example 16 Apr 97 3 57 48 PM RVO The date and time will appear in the format set in the Windows Regional Settings Prope
6. To avoid injury ALWAYS ensurecthat safety practices governing the use handling and disposal materials are strictly observed Appropriate safety clotking and glasses should also be worn Warning 5 2 Instrument ze m 241 15 287 15 361 50 536 30 nm Refer to the Solution Test Certificate calibrated with 0 1 nm SBW Photometric mode Abs Scan range 231 15 to 546 30 nm Spectral bandwidth 0 10 nm Signal averaging time Data interval 10 sec 10 nm Lamp UV Vis Specifications Wavelength nm Tolerance 241 1 1 nm 287 1 1 nm 361 5 1 nm 536 3 3 nm Publication date 10 02 Cary 100 300 certification manual What if my instrument fails to meet the specification Check that you have set the tolerances to values that are within the specifications of the instrument Q Recalibrate the wavelength drive mechanism by turning the instrument off and on again C Check the deuterium D and visible lamp alignments and perform the wavelength accuracy test again Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 5 2 Control of absorbance 5 2 1 Publication date 10 02 K Cr 0 method The photometric accuracy of a spectrophotometer can be determined by measuring the absorbance of solutions of high purity compounds This method is limited because it requi
7. and 100 T reference disc in ection is required Please refer to the Cary WinUV online help for Instrument parameters Photometric mode lt R Scan range 360 to 830 nm Spectral ban widiy 2A 2 00 nm Signal se 2 00 s Data interya 10 nm Lamp BAY UV Vis ations 36Oto 830 nm 41 5 R What if my instrument fails to meet the specification Q Recheck the alignment of the diffuse reflectance accessory as described in the Cary WinUV online help Q Check the alignment of the instrument lamps then perform the test again Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 9 1 3 9 1 3 1 9 1 3 2 Publication date 10 02 Cary 100 300 certification manual Fabric diffuse reflectance accessory tests There are four tests performed on DRA when it is being used for fabric measurements They are wavelength accuracy photometric linearity stray light and signal to noise ratio These tests are all included in the Cary WinUV Validate application software by choosing the Fabric tests option These tests require the use of the following test equipment Q a holmium oxide filter part number 01 180207 90 Q aUG 11 and GG 420 filter combination part number 01 106543 00 linearity test filters part number 99 100990 00 Wavelength accuracy This test uses the holmium oxide filter By
8. 656 1 nm URA mode Trans e 2 to 2 484 to 488 654 to 658 nm Spectral Bandwidth 0 2 nm Signal averaging time 0 033 sec Data interval 0 02 nm Lamp UV Beam mode Single Specifications Zero order 0 7 nm 486 0 nm 0 2 nm 656 1 nm 0 2 nm 6 1 Cary 100 300 certification manual 6 2 6 1 2 What if my instrument fails to meet the specification m m Check that you have set the tolerances to values which are within the specifications of the Cary instrument Recalibrate the wavelength drive mechanism by turning the instrument off and on again Check the deuterium D lamp alignment and perform the wavelength accuracy test again If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Holmium oxide method This method scans the wavelength peaks of a holmium oxide filter between 269 4 and 647 5 nm This method requires the use of a holmium oxide filter part number 01 180207 90 Instrument parameters Wavelengths Photometric mode Scan range 269 4 to 646 8 nm Spectral bandwidth 2 00 nm Signal averaging ti 0 10 sec Data interv GY 0 10 nm Lamp S UV Vis ANALI Tolerance 3 nm 3 nm 3 nm 3 nm 1 nm 1 nm 1 nm What if my instrument fails to meet the specification m m Check that you have set the tolerances to values that are within th
9. Performance specifications for these tests are included in the Cary Guaranteed Specifications brochure part number 87 101604 00 8 1 Resolution power The monochromator of the instrument separates light into a spectrum and sends a small band of that spectrum through to the sample If the spectral bandwidth is adjustable the smaller it is the better the resolution of the instrument The limiting resolution of an instrument is that setting where a smaller setting of the Spectral Bandwidth SBW makes no difference This is important for high resolution work wh ery narrow SBW is needed The SBW is defined as the bandwidth at half ey elength peak height The following method is for maximum resolution 8 1 1 Maximum resolution m This test measures th m often called the limiting resolution of the instrument by sop the narrowest spectral bandwidth SBW that the instrument ca nieve The SBW is defined as the bandwidth at half power bandwidth ww The ins ans the 656 1 nm deuterium D emission line and determines the Geena i e the width of the peak at half height ument parameters Wavelength 656 1 nm Photometric mode Trans Scan range 654 6 to 657 6 nm Spectral bandwidth 0 20 nm Signal averaging time 0 20 sec Data interval 0 02 nm Beam mode Single Lamp UV Specifications Minimum HPBW gt 0 16nm Maximum HPBW lt 0 24nm What if my instrument fai
10. number 99 100852 00 Alternatively the Toluene Hexane solution can be prepared by the operator as follows 0 020 v v solution of toluene in hexane UV grade Warning To avoid injury ALWAYS ensure that safety practices governing the use handling and disposal of such materials are strictly observed Appropriate safety clothing and glasses should also be worn Publication date 10 02 Cary 100 300 certification manual Instrument parameters Photometric mode Abs Scan range 265 to 275 nm Spectral bandwidth 1 6 nm Signal averaging time 0 5 sec Data interval 0 10 nm Lamp UV Specifications Ratio gt 1 5 What if my instrument fails to meet the specification Q Make sure that you prepared the Toluene Hexane solution correctly Make up a fresh batch if necessary and repeat the measurement a If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 5 4 Stray light 5 4 1 Publication date 10 02 de r that is not of the selected ee stray light and light of the Stray light is defined as any light reaching wavelength The detector cannot distinguis selected wavelength and will add th reading than expected Stray lj Stray light is monitored by m solution is totally absorbing A es deviation from the Beer Lambert Law a test solution at a wavelength where the t that is detected is due to stray
11. a thermometer with an absolute accuracy of 0 1 C be used 2 2 3 Calibrated linear measuring device A calibrated vernier or other similar measuri ice capable of measuring up to 100 mm with an accuracy of 0 1 mm is r est the positional accuracy of the Varian sample transport accessory A X o Publication date 10 02 Cary 100 300 certification manual Instrument tests 3 1 Publication date 10 02 Note Note Varian establishes the performance factors and specifications for each instrument There is a precise procedure for determining whether each factor meets the specification The following sections of this document detail the tests that characterize instrument performance along with the Varian specifications and instrument parameter information Specifications are subject to change without notice Please refer to the latest issue of Cary UV Vis NIR Spectrophotometers Guaranteed Specifications part number 87 101604 00 for more information The tests outlined in the following sections require the use of the Cary WinUV Validate application software The tests may also be performed manually using the parameters stated for each test however no guidance is supplied in this manual for performing the tests manually es Some of the tests required by regulatory i e not representative of the capabilities of the Varian Cary UV Vi otometer Instrument specifications u in many cases exceed those specifi ry
12. averaging time 0 5 sec Data interval 0 10 nm Lamp UV Specification Ratio gt 1 5 What if my instrument fails to meet the specification Make sure that you prepared the toluene hexane solution correctly Make up a fresh batch if necessary and repeat the measurement Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 Cary 100 300 certification manual 7 3 Baseline flatness All double or dual beam spectrophotometers function by comparing a sample in one light path to a reference in the other If a wavelength scan is performed with the same material in both light paths the response would be the natural baseline of the instrument This baseline may not be a flat line To compensate for this all instruments have some form of baseline correction Baseline Flatness is a measure of how well the instrument compensates for the natural baseline of the instrument and the reference material The following method is recommended by the TGA to test for baseline flatness 7 3 1 Baseline flatness method In this test an instrument baseline is collected using air as the reference material followed by a corrected baseline From the corrected baseline scan the peak to peak signal levels over the wavelength range is monitored to determine the baseline flatness of the instrument gt Note A signal smoothing filter is
13. by e LONG dation osi varianinc com Australia is ISO9001 certified 1999 Varian Australia Pty Ltd A C N 004 559 540 All rights reserved Windows 95 98 2000 NT4 and XP are registered trademarks of Microsoft Corp Publication date 10 02 Cary 100 300 certification manual Publication date 10 02 Contents Introduction ssanie erae aeaea 1 1 Cary certification program 1 2 Available tests 1 2 Cary performance tests 1 3 Test kits and equipment sesssssesssssessssesssseesssssessseeessseessseeess 2 1 Test kits 2 1 Other test equipment 2 2 Instrument testsisersiinisc nieres sesio 3 1 Using the Cary WinUV Validate application software 3 1 Instrument performance tests Wavelength accuracy Wavelength reproducibility Baseline flatness Photometric noise European PharmacopociaNestS ccsssccecesseceeeesseeesesseeeeeees 5 1 C TESTS aera EE E E E 6 1 2 ngth accuracy 6 1 Photometric accuracy 6 3 Australian TGA ests essenin aseni ainas 7 1 Wavelength accuracy 7 1 Resolution power 7 4 Baseline flatness 7 5 Stray light 7 6 Photometric accuracy 7 8 Other instrument tests cecscesseessseecssescsseeessesesseeessseeseees 8 1 Resolution power 8 1 Stray light 8 2 Photometric accuracy 8 3 Photometric linearity 8 4 Photometric stability 8 5 Photometric noise 8 6 iii Cary 100 300 certification manual iv Qe LACCESSOFY LOSS veces dccteccsuvesauhedectetesevevnthedeeeseetsseestee
14. certification manual 7 8 7 5 7 5 1 Note Photometti The chief source of error in this measurement is fluorescence of the sample It is also important that the light beam is kept away from the edges of the cuvette that can act as a non attenuating light guide For this reason you must use masked microcells for this test With the cuvette in the cell holder hold the cell up to the light and look through the aperture You should not be able to see the cuvette walls Mask the cell holder if necessary to reduce the aperture width to 8 mm Instrument parameters Wavelength 370 nm Photometric mode oTrans Spectral bandwidth 2 00 nm Signal averaging time 10 sec Lamp UV Vis Specifications At 370 nm 50mg L NaNO At 370 nm 0 25 g L K Cr 0 What if my instrument fails to meet the sp ion Q Ensure that the sample compartment iy tly lt sealed Repeat the test with the room lights turned off If the in int Flasses the test this indicates that there is a light leak in thes 0 et somewhere Q As the test solutions ma gratl over time the accuracy of the test depends lt 1 0 T lt 0 05 T heavily on the correct han management and combination of materials Make up fresh sol ti ensuring cell cleanliness Mask the cell holder to reduce the aper to 8 mm and repeat test O If cones Fi cannot be obtained contact your local Varian Service office a wic call may be required A
15. light The following method i mended by the European Pharmacopoeia for this test KCI test Q Note Note Note This method eos the absorbance of potassium chloride KCI at a wavelength n is totally absorbing WinUV Pharma software version 2 5 measures this at a wavelength of 200 n To perform this test a solution of potassium chloride KCI and a water blank solution must be used These solutions are available as part of the solutions test kit part number 99 100852 00 Alternatively potassium chloride KCI solution can be prepared by the operator as follows KCl 12 g L KCI in water Warning To avoid injury ALWAYS ensure that safety practices governing the use handling and disposal of such materials are strictly observed Appropriate safety clothing and glasses should also be worn When performing this test it is critical that great care is taken during sample preparation and measurement Suprasil cuvettes must be used They should never be allowed to contact skin and must be cleaned with alcohol and dried with cotton wool never laboratory tissue Spectroscopy grade reagents must be used 5 5 Cary 100 300 certification manual gt Note The major source of error in this measurement is fluorescence of the sample It is also important that the light beam is kept away from the edges of the cuvette that can act as a non attenuating light guide For this reason do not use unmasked microcells for this test
16. scanning the filter this test monitors how close the indicated wavelength is to the actual wavelength The wavelength accuracy is determined by the Cary instrument itself not by the accessory This test uses the detector in the accessory to perform the test rather than the instrument s detector which would normally be used to perform the test Instrument parameters Wavelength 333 3 360 9 nm S Photometric mode Avsoreance Scan range ED to 38 Signal averaging time D Data interval 0 im AN Specification a accuraG f orrect results still cannot be obtained contact your local Varian Service fice as a service call may be required Linearity test This test determines how accurately the instrument measures increasing transmittance using the addition of filters technique To do this the Cary system measures the absorbance of two filters individually and then checks the sum of the result versus a measurement of the two filters together Poor photometric linearity will produce different results for these measurements This method requires the use of the three mesh filters included in the accessory test kit part number 99 100990 00 Enter the filter transmission values and test kit number on the linearity test setup page The results of the readings of each of the filters are compared to the known value and a percentage error in the results is given 9 3 Cary 100 300 certification manual
17. 10 Abs 350 nm 0 6400 Abs 0 010 Abs What if my instrument fails to meet the specification Q Repeat the test with a new solution then compare the results to determine if the error was introduced by the solution Q Check that the expected absorbance values have been correctly entered into the table on the Control of Absorbances tab LJ Ensure that the cuvette is clean Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required The monochromator of the instrument separa t into a spectrum and sends a small band of that spectrum through to thesar the spectral bandwidth is Bandwidth SBW makes no di This is important for high res SBW is defined as the bandwidt alf the wavelength peak height The following methodi mended by the European Pharmacopoeia for this test NN This methgt es the ratio of the absorbance of toluene hexane solution at its ima wavelengths approximately 268 nm and 266 nm respectively od a wavelength scan is performed from 275 to 264 nm then the ratio is calculated from the peak at 268 nm and the trough at 266 nm ratio does not meet the specified tolerance for the instrument then the SBW is reduced by 0 1 nm starting with a 1 6 nm SBW until this criterion is achieved The method requires the use of a Toluene Hexane solution The Toluene Hexane solution is available as part of the Solutions Test Kit part
18. 100477 00 UV VIS Filter set 99 100852 00 Solution Test Kit 01 180207 90 99 100477 00 99 100560 00 99 100561 00 99 100852 99 100477 00 UV Vis Filter set 99 100560 00 Double Aperture kit 99 100561 00 Linearity Neutral Density Filters 99 100811 00 Diffuse Reflectance Standards 99 100842 00 Calibrated Specular Standards 99 100843 00 Calibrated Color Standards 99 100852 00 Solution Test Kit 99 100991 00 Kit UV Vis Transmittance Minimum equipment required for testing the Fabric accessory 01 106543 00 Red filter disc UG11 GG420 01 180207 90 Holmium Oxide Filter 99 100990 00 Filter screen kit 3 10 20 T 2 1 Cary 100 300 certification manual 2 2 2 2 Other test equipment 2 2 1 NIST 930E Filters These filters are required for measuring the photometric accuracy of the instrument according to the USP and TGA regulatory authorities The filters can only be purchased directly from the National Institute of Standards and Technology NIST Standard Reference Materials Contact details for NIST can be found in section 10 4 of this manual 2 2 2 Calibrated Precision Thermometer A calibrated precision thermometer is required to accurately determine the temperature accuracy for the Varian thermostatted accessories The absolute accuracy of this thermometer is to be determined by the intended use of the accessory but it is suggested that
19. Cary 100 300 Certification For Cary 100 3 UV Vis Spec ometers VY g NOTICE This document contains references to Varian Please note that Varian Inc is now part of Agilent Technologies For more information go to 85 101747 00 www agilent com chem October 2002 ig Agilent Technologies Cary 100 300 certification manual ii Varian offices Varian has offices in most countries The major offices for optical spectroscopy products are listed below Varian Australia Pty Ltd Manufacturing site 679 Springvale Road Mulgrave Victoria 3170 Australia International telephone 61 3 9560 7133 International fax 61 3 9560 7950 Varian Instruments 2700 Mitchell Dr Walnut Creek CA 94598 USA Phone 1 800 926 3000 International telephone 1 925 939 2400 International fax 1 925 945 2102 Varian BV Herculesweg 8 4338 PL Middelburg Netherlands International telephone 31 118 67 1000 International Fax 31 118 62 3193 Internet The Varian Internet home page can ba fotind at http www varianinc com Varian Australia Pty Lt associated software owner of copyright on this document and any the written permission of Varian Australia Pty Ltd er the documentation or the software is copied r converted to electronic or other machine readable form in First p evlay 1999 in Australia Revised in October 2002 Comments about 3 hould be directed to Varian Australia at the address above or
20. MS which is approximately one fifth of the peak to peak value over a two minute period To perform this test at 3 0 to 5 0 Absorbance yo will be prompted to ensure there is approximately 1 6 Abs of rear beam attenuation You can achieve the appropriate rear beam attenuation by either one of the methods to 1 7 Abs Q a UV Vis Filter Set part er 0477 00 for up to 3 0 Abs Q a solution that absorbs thecorrg ct Abs 1 5 Abs at the required wavelength the Rear Beam Attenuator RBA A part number 00 100411 00 for up Instrument parameters Wavelength os 500 nm Photometric Abs Abscissa m Min Spectral idt 2 00 nm Scan oa 1 00 min Pel ets time 1 00 sec Vis Specifications Cary 100 at 500 nm 0 Abs lt 0 000085 Abs 1 Abs lt 0 000200 Abs 2 Abs lt 0 000300 Abs 3 Abs lt 0 000400 Abs with 1 6 Abs rear beam attenuation Publication date 10 02 Cary 100 300 certification manual Cary 300 at 500 nm 0 Abs lt 0 000060 Abs 1 Abs lt 0 000200 Abs 2 Abs lt 0 000300 Abs 3 Abs lt 0 000400 Abs with 1 6 Abs rear beam attenuation 4 Abs lt 0 003000 Abs with 1 6 Abs rear beam attenuation 5 Abs lt 0 008000 Abs with 1 6 Abs rear beam attenuation What if my instrument fails to meet the specification Q Check the visible lamp alignment then perform the test again Q Replace the visible lamp if necessary Q If co
21. Neutral Density Filters are approximate values only when measured in the visible region Instrument parameters QO Wavelength 50071 Awe Photometric mode Spectral bandwidth 2 00 Signal averaging time N00 sec Lamp is WS Specifications BO D Not available oS ary 100 300 series instruments Q es in the Cary WinUV software for each absorbance are as 0 0010 Abs 0 0018 Abs 0 0081 Abs What if my instrument fails to meet the specification Q Recalibrate the wavelength drive mechanism by turning the instrument off and on again Q Check the result a second time paying special care with the positioning of the neutral density filters Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 8 4 Publication date 10 02 Cary 100 300 certification manual 8 5 Photometric stability Photometric stability provides an indication of how stable your photometric results will be over time by measuring the photometric stability over an extended period The following method is recommended to test for photometric stability 8 5 1 Photometric stability test method The test measures the change in photometric signal at 500 nm over a one hour time period Instrument parameters Wavelength 500 nm Photometric mode Abs Abscissa mode min Spectral Bandwidth 2 0 nm Measuremen
22. O must be prepared by the A R in distilled water and diluted to give solutions Ntsations of 1 065 w v 0 751Abs 0 710 w v 0 500Abs To avoid injury ALWAYS ensure that safety practices governing the use handling and disposal of such materials are strictly observed Appropriate safety clothing and glasses should also be worn Instrument parameters Wavelength 302 nm Photometric mode Abs Spectral bandwidth 2 00 nm Signal averaging time 3 00 sec Lamp UV Vis 7 10 Publication date 10 02 Publication date 10 02 Cary 100 300 certification manual Specifications KNO method 14 2 w v KNO 0 250 Abs 0 355 w v 0 010 Abs 0 500 Abs 0 710 w v 0 010 Abs 0 010 Abs 0 751 Abs 1 065 w v What if my instrument fails to meet the specification Q Repeat the test with a new solution then compare the results to determine if the error was introduced by the solution Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required amp A X o Cary 100 300 certification manual A This page is intentionally A X o 7 12 Publication date 10 02 Cary 100 300 certification manual 8 Other instrument tests These tests are provided to check additional performance parameters that are not routinely tested as part of the pharmacopoeia tests or the Varian instrument performance tests
23. S kdtometric accuracy is determined by measuring a known absorbance value and calculating the difference between the theoretical absorbance value and the absorbance value as reported by the instrument There are three methods recommended by the TGA to test for photometric accuracy Q NIST filters method Q KCr 0 method Q KNO method NIST filters The photometric accuracy of a spectrophotometer can be determined by measuring the absorbance of calibrated NIST 930E standard reference materials issued by one of the national standards organizations This is a set of filters with various optical densities of know absorbance values NIST provides a certificate with the filters showing the absorbance value at specific wavelengths along with their tolerances They suggest the filters are returned to them annually for recertification This method is limited because it requires accuracy determinations to be performed at a specific temperature wavelength and SBW Refer to the NIST Certificate supplied with the NIST 930E standard reference materials Publication date 10 02 Cary 100 300 certification manual gt Note To perform this test a set of NIST 930E Filters or filters calibrated against a reference set of filters must be used This filter set can be obtained from the National Institute of Standards and Technology NIST See section 10 4 of this manual for ordering details gt Note Due to the design of the Cary 100 300 series ins
24. UT ENGLAND FREE CALL 1 800 252 284 Telephone 61 2 9484 0033 Telephone 44 0 181 501 5550 Facsimile 61 2 9484 0055 Facsimile 44 0 181 501 1118 Email sales starna com au Email sales optiglass co uk Web site www starna com au www optiglass co uk Starna Cells Inc Stana GmbH 5950 Traffic Way chilerstrasse 10A ATASCADERO 64311 Pfungstadt CA 93423 GERMANY USA Telephone 1 805 Facsimile Telephone 06157 7953 Email info star Facsimile 06157 85564 Web site w CI T Wien lauberg S Telephone 03335 4850 Facsimile 03335 4851 Email starna austria netway at Publication date 10 02 Cary 100 300 certification manual 10 4 Ordering NIST reference materials Purchase orders in English for all NIST SRMs RMs should be directed to National Institute of Standards and Technology Standard Reference Materials Program 100 Bureau Drive Stop 2322 Gaithersburg MD 20899 2322 USA Telephone 1 301 975 6776 Facsimile 1 301 948 3730 Email SRMINFO nist gov Web site http www nist gov Each purchase order should give the number of units catalogue number and name of each reference material requested Example 1 each SRM 930E Glass Filter Set Transmittance The following information must be included with each order a billing address a shipping address name of customer telephone number N fax number purchase order number For US customers also includ OoOoooo Q a customer identifica
25. With the cuvette in the cell holder hold the cell up to the light and look through the aperture You should not be able to see the cuvette walls Mask the cell holder if necessary to reduce the aperture width to 8 mm Instrument parameters Wavelength 200 nm Photometric mode Trans Spectral bandwidth 1 00 nm Signal averaging time 10 00 sec Lamp UV Specifications At 200nm 12 g L KCI lt 1 000 T gt 2 0 Abs Pharmacopoeia method Q Preparing your own potassi C1 solution is not recommended as the quality of the Salt ca considerably the test is sensitiv ary contamination of the KCI and the test solutions may d grade over time For best results tified potassium chloride KCI solution such as the one contain standards kit part number 99 100852 00 is un t or cold in the laboratory you should wait until the te returns to normal room temperature 22 25 C or that printed on ficate supplied with the kit before repeating the test Q The abs potassium chloride KCI is influenced by temperature If it sy are that the sample compartment is tightly sealed Repeat the test with the oom lights switched off If the instrument passes the test this indicates that there is a light leak in the instrument cover somewhere Q As the test solutions may degrade over time the accuracy of the test depends heavily on the correct handling management and combination of materials Ma
26. applied to the corrected baseline for the Varian specification For a description of the smoothing operation refer to the Cary WinUV online help Instrument parameters O X Photometric mode Aby Scan range OXo 850 nm Spectral Bandwidth 4 0m Signal averaging time NO18 sec Data interval AN 1 00 nm Lamp QWY UV Vis eatin Specificatio NX Baseline Ss 200 a t if my instrument fails to meet the specification 40 001 Abs Q Ensure that there are no samples or other obstructions in the sample compartment Q Check the deuterium D and visible lamp alignments then perform the test again Q Check the cleanliness of the sample compartment windows and clean with a soft cloth dampened with distilled water if necessary Q Check for possible sources of contamination in the air Changes in the baseline could be caused by the absorption of vapors in the air Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 7 5 Cary 100 300 certification manual 7 6 7 4 Stray light Stray light is defined as any light that gets to the detector that is not of the selected wavelength The detector cannot distinguish between stray light and light of the selected wavelength and will add the two together to give a higher photometric reading than expected Stray light also causes deviation from
27. asuring the standard 0 T sample position with no sample and 100 T calibrate position corrections are required Only 24 of the reflectance values indicated by asterisks provided in the calibration data booklet are certified on an NIST master Instrument parameters Photometric mode Absolute R Scan Range 250 850 nm Spectral bandwidth 2 00 nm Signal averaging time 2 00 s Data interval 10 nm Lamp UV Vis 9 5 Cary 100 300 certification manual 9 6 Specifications lt 300 nm 3 0 R 300 to 900 nm 1 5 R What if my instrument fails to meet the specification m m Recheck the alignment of the VW absolute specular reflectance accessory as described in the Cary WinUV online help then perform the test again Check the alignment of the instrument lamps then perform the test again If correct results still cannot be obtained contact your local Varian Service office as a service call may be required amp A X o Publication date 10 02 10 10 1 General care 10 2 Cleaning the liquid standards Publication date 10 02 Cary 100 300 certification manual Care and handling To maintain optimum optical and reflectance properties of the optical standards the following care and cleaning instructions must be followed Although the materials used for these standards are quite durable care should be taken to prevent contaminants such as finger oil
28. authorities The tests outlined in the follo tion require the use of the Cary WinUV Validate application software The tests may also be performed manually using the parameters stated for Cate application software about the Cary WinUV Validate application software is pplication s online help system To set up the tests follow these 2 Programs gt Cary WinUV gt Validate Alternatively you can double click on the Validate icon in the Cary WinUV folder on the desktop 3 If you are running a GLP system you may be prompted to enter a password before accessing the Validate application To set up the test parameters 1 Select the Tests menu or the Tests button to display the Configure page and then select the type of instrument performance tests that you require The various tests that are available will appear as tabs across the top of this dialog 2 If your access privileges are restricted you may find that fields and buttons in the application software are not accessible See your administrator to gain access Cary 100 300 certification manual gt Note 3 2 Click on each tab and select the tests that you require to be performed by selecting the appropriate checkbox Change any test parameters as required If you wish to save the tests you have set up for future use select Save Method As in the File menu to display the Save As dialog box and save the method MVO To set up automated runs 1 2
29. d data calibrated for wavelength ded for this purpose part number 99 100811 00 Prior to AN of the standard a O T sample reflectance port open Ceri eference disc in sample port correction is required Please refer to the AUY online help for more details Photometric mode R Scan range 250 to 850 nm Spectral bandwidth 2 00 nm Signal averaging time 2 00 s Data interval 10 nm Lamp UV Vis Cary 100 300 certification manual 9 2 Specifications lt 300 nm 3 0 R 300 to 900 nm 1 5 R What if my instrument fails to meet the specification Q Recheck the alignment of the diffuse reflectance accessory as described in the Cary WinUV online help Q Check the alignment of the instrument lamps then perform the test again Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 9 1 2 Color diffuse reflectance accessory test Note Note When the DRA is being used for color measurement the operation of the DRA is tested by measuring the diffuse reflectance of Red Yellow Green and Blue color standards at incremental wavelengths or wavenumbers This data is then compared with the calibration data that is supplied with the Color Standards This method requires the use of a color reflectange standards kit with its associated Prior to measurement of the standard aOoaMSample reflectance port open
30. ding It determines the precision of the measurement and the detection limit of the instrument Poor signal to noise performance makes it very difficult to determine the actual absorbance value as it fluctuates It introduces errors into both quantitative and qualitative spectroscopy The following method is used at Varian s factory to test for photometric noise 4 4 1 Photometric noise method Note This test measures the photometric noise specified as Absorbance Root Mean Square RMS which is approximately one fifth of the peak to peak value over a two minute period Instrument parameters Wavelength 500 nm Photometric mode Abs Abscissa mode Min Measurement period 2 00 min S Spectral Bandwidth 2 00 nn _ Signal averaging time ENR Data interval Ase Lamp vis Specifications Cary 100 At 0 Abs RO lt 0 000085 Abs Cary 300 Z lt 0 000060 Abs y instrument fails to meet the specification heck the alignment of the Visible lamp and perform the test again Replace the Visible lamp if necessary Q Prior exposure of the detector to large amounts of light may cause high noise levels Allow some time before retesting Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Photometric noise can be affected by a decrease in the signal received by the detector so the electronic noise is grea
31. e specifications of the instrument Recalibrate the wavelength drive mechanism by turning the instrument off and on again Perform the wavelength accuracy test again If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 Cary 100 300 certification manual 6 2 Photometric accuracy 6 2 1 Publication date 10 02 Note Photometric accuracy is determined by measuring a known absorbance value and calculating the difference between the expected absorbance value and the absorbance value as reported by the instrument Varian provides a kit of solutions 99 100852 00 containing certified standard solutions suitable for performing the photometric accuracy tests If you are preparing your own solutions the expected absorbance and wavelength values will be advised in the relevant pharmacopoeia documentation When using the Cary WinUV Validate software application ensure the correct wavelength and absorbance values are entered by selecting Tests then the Photometric accuracy or Control of absorbances page DO NOT use the default values There are two methods recommended by the United States of America Pharmacopoeia to test for photometric accuracy Q NIST filters method a KCr 0 method NIST filters Note Note The photometric accuracy of a spectrophoto n be determined by measuring the absorbance of calibrated NIST 930E Sta d refgrenc
32. e Materials issued by one of the national standards organizations Dyi a set of filters with various optical densities of known absorbance yal I bxOvides a certificate with the filters showing the absorbance value a avelengths along with their tolerances They suggest that the filters a ed to them annually for recertification This method is limited because jt requires accuracy determinations to be performed at a specific temperature w gth and SBW Refer to the NIST Certificate supplied with the NIST 930E eference Materials To perform th Cg of NIST 930E Filters or filters calibrated against a set of reference filt be used NIST filters can be obtained from the National Institute oes s and Technology NIST See section 10 4 of this manual for a ordering de g design of the Cary 100 300 series instruments the maximum SBW is 4 0 Instrument parameters Wavelength nm 440 00 465 00 546 10 590 00 635 00 SBW nm 2 2 2 7 4 0 4 0 4 0 Photometric mode Abs Abs Abs Abs Abs Signal averaging 3 00 sec 3 00 sec 3 00 sec 3 00 sec 3 00 sec time SAT Specifications at 1 0 Abs 40 003 Abs at 0 5 Abs 0 002 Abs What if my instrument fails to meet the specification Q The absorbance of the filters is influenced by the temperature and the age of the filters If it is unusually hot or cold in the laboratory then you should wait until the temperature returns to nor
33. erial is required to have high resistance to deep UV radiation the material should also be prepared as follows Flush the material with gt 18 mohm distilled deionised water for 24 hours Vacuum bake the material at 75 C for 12 hours at approximately 1 Torr then purge the vacuum oven with clean dry air or nitrogen 10 7 Re calibrating the diffuse reflectance sta gt Note 10 4 Labsphere recommends that t a be recalibrated at least once per year sooner if the standards are visibly soiled or damaged Recalibrations will be completed within two weeks of receipt of the standards Call Labsphere for pricing infofthation prices depend on each type of standard to be recalibrated For recalibration aysphere standards please contact your nearest Labsphere distributor O KS 7 Telephone 1 603 927 4266 Facsimile 1 603 927 4694 Be ready to give Labsphere the serial number of the standards to be recalibrated Publication date 10 02 Cary 100 300 certification manual 11 Certification kit log sheet The following certification kit log sheet can be used to record the recertification history of the certification kits used with this package if appropriate sf A X o Publication date 10 02 11 1 Cary 100 300 certification manual amp This page is intentio left blank A X o 11 2 Publication date 10 02 Cary 100 300 certification manual Certification Kit log sheet Page Number
34. fication Manual outlines the tests and results expected from the instrument including wavelength accuracy wavelength reproducibility baseline flatness photometric noise stray light photometric linearity and photometric accuracy These tests are automated using the Cary WinUV Validate application software to improve the measurement calculation and reporting of these results The instruments published guaranteed specifications are used throughout this document An instrument will age and its performance may change over time The absolute amount of the change is of less importance than the rate of change It is more important to track the instrument s performance during its life to establish confidence in analytical results As an instrument ages performance may be improved by additional instrument service such as cleaning of the optics but it may not have any effect on improving the overall analysis Ultimately the instruments operator will need to determine what level of instrument performance is required for his her analysis This will then determine what out of specification parameters if any require additional service t t the specification Regulatory authorities demand tha Laboratory Practices GLP A fange af performance tests for UV Visible spectrophotometers are specified e British European Pharmacopoeia BP EP and the US Pharmacopoeia USP tMese tests are included in this manual Also included in this re a series of
35. filter part number 01 180207 90 Instrument parameters Wavelength 637 5 536 Photometric mode Scan range 269 4 to 647 5 nm Spectral bandwidth bs 00 nm Signal averaging time 0 10 sec Data interval S5 0 10 nm Lamp UV Vis sana Wav Tolerance S Z 7 3 nm 3 nm 7 3 nm 360 9 1 nm 287 5 1 nm 279 4 1 nm What if my instrument fails to meet the specification m m Check that you have set the tolerances to values that are within the specifications of the instrument Recalibrate the wavelength drive mechanism by turning the instrument off and on again Check the deuterium D and visible lamp alignments and perform the wavelength accuracy test again If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 7 1 3 Cary 100 300 certification manual Holmium perchlorate method This method scans the wavelength peaks of a holmium perchlorate solution between 231 1 and 546 3 nm This method requires the use of a holmium perchlorate solution and a blank solution These solutions are available as part of the Solutions Test Kit part number 99 100852 00 Alternatively the operator can prepare the holmium perchlorate solution as follows Publication date 10 02 4 w v solution of holmium oxide 99 5 in 1 4 M perchloric acid Warning
36. h 656 1 nm Photometric X Trans Scan range 654 6 to 657 6 nm es 0 20 nm Sign ee g time 0 2 sec KAAI 0 020 nm ode Single UV Specifications Peak separation lt 0 08 nm Standard deviation lt 0 02 What if my instrument fails to meet the specification Q Ensure that the instrument has been warmed up for at least one hour Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 Cary 100 300 certification manual 4 3 Baseline flatness All double or dual beam spectrophotometers function by comparing a sample in one light path to a reference in the other If a wavelength scan is performed with the same material in both light paths the response would be the natural baseline of the instrument This baseline may not be a flat line To compensate for this all instruments have some form of baseline correction Baseline Flatness is a measure of how well the instrument compensates for the natural baseline of the instrument and the reference material The following method is used at Varian s factory to test for baseline flatness 4 3 1 Baseline flatness method In this test an instrument baseline using air as the reference material is first collected followed by a corrected baseline Then from the corrected baseline scan the peak to peak signal levels over the wavelength range are monito
37. ing light guide For this reason you must use masked microcells for this test With the cuvette in the cell holder hold the cell up to the light and look through the aperture You should not be able to see the cuvette walls Mask the cell holder if necessary to reduce the aperture width to 8 mm Instrument parameters Wavelength 200 nm Photometric mode oTrans Spectral bandwidth 1 00 nm Signal averaging time 10 sec Lamp UV Publication date 10 02 Cary 100 300 certification manual 7 4 2 Publication date 10 02 Specification At 200nm 12 g L KCI lt 1 000 T gt 2 0 Abs What if my instrument fails to meet the specification Q Manually scan down to 198 nm to reflect the limits of the current European Pharmacopoeia method Q Preparing your own potassium chloride KCI solution is not recommended as the quality of the salt can vary considerably the test is sensitive to any contamination of the KCl and the test solutions may degrade over time For best results use a certified potassium chloride KCI solution such as the one contained in the standards kit part number 99 100852 00 Q The absorbance of potassium chloride KCI is influenced by temperature If it is unusually hot or cold in the laboratory you should wait until the temperature returns to normal or that printed on the certificate supplied with the kit before repeating the test Q Ensure that the sample compar
38. ing where a smaller setting of the Spectral Bandwidth SBW makes no difference to the resolution This is important for high resolution work where a very narrow SBW is needed The SBW is defined as the bandwidth at half the wavelength peak height The following method is recommended by the TGA to test for resolution power 7 2 1 Toluene hexane method Note This method measures the ratio of the absorbance of toluene hexane solution at its maxima and minima wavelengths approximately 268 nm and 266 nm respectively For this method a wavelength scan is performed from 275 to 265 nm then the peak trough ratio is calculated from the peak at 268 nm and the trough at 266 nm If this ratio does not meet the specified toleranc fer the instrument then the SBW is reduced by 0 1 nm starting with a 1 6 nm ntil this criterion is achieved This method requires the use of a Toluene D ne solution The Toluene Hexane solution is avai o t of the Solutions Test Kit part number 99 100852 00 Alternatively the Toluene Hexane s liition can be prepared by the operator as follows 0 020 v v solution luene in hexane UV grade Warning To avoid oe AYS ensure that safety practices governing the use haz dNng and disposal of such materials are strictly observed j afety clothing and glasses should also be worn ment parameters Photometric mode Abs Scan range 265 to 275 nm Spectral bandwidth 1 6 nm Signal
39. ke up fresh solutions ensuring cell cleanliness Mask the cell holder to reduce the aperture width to 8 mm and repeat test Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 5 6 Publication date 10 02 6 1 6 1 1 Publication date 10 02 Cary 100 300 certification manual US Pharmacopoeia tests These tests available from the configure page USP tests of the Cary WinUV Validate software are required by the United States of America Pharmacopoeia Many of these tests are duplicated in either the standard instrument performance tests or the other pharmacopoeia tests Wavelength accuracy Poor wavelength accuracy can produce low quantitative results as the absorbance measurement may have been made on the side of the peak rather than at the peak Wavelength accuracy is determined by scanning a known wavelength peak and calculating the difference between the theoretical wavelength peak and the wavelength peak as reported by the instrument There are two methods recommended by the SS of America Pharmacopoeia to test for wavelength aco Q Deuterium D emission line meth Q Holmium oxide method Deuterium D emission i od m D emission lines at 486 0 and 656 1 nm in the Vis region as well g so order line This method does not require the use of additional equip Instrument a ers Wave QS 0 486 0
40. le Specifications Zero order 0 7 nm 486 0 nm 0 2 nm 656 1 nm 0 2 nm Publication date 10 02 ion lines at 486 0 and 656 1 nm in the e This method does not require the use of Cary 100 300 certification manual 4 2 What if my instrument fails to meet the specification Q Check that you have set the tolerances to values that are within the specifications of the instrument Q Recalibrate the wavelength drive mechanism by turning the instrument off and on again Q Check the deuterium D lamp alignment and perform the wavelength accuracy test again Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Wavelength reproducibility Poor wavelength reproducibility introduces errors in analytical results due to wavelength shifts This test monitors the ability of the instrument to correctly return to the set wavelength repeatedly The following method is used at Varian s factory to test for wavelength reproducibility Wavelength reproducibility method This method scans the deuterium D emis gain of the detector until it detects an a level the scan is repeated ten times deviation of the data and use This dispersion is then compaked with the specified tolerance ine peak at 656 1 nm increasing the ate p ak Then maintaining that gain are will then calculate the standard Instrument parameters Wavelengt
41. length peak as reported by the instrume There are three methods recommended b Ayo test for wavelength accuracy Q Deuterium D emission line met Q Holmium oxide method d Q Holmium perchlorate met Deuterium D emissi method This method scan Cs rium D emission lines at 486 0 and 656 1 nm in the Vis region as a zero order line This method does not require the use of additional K Instru eters AKA 0 486 0 656 1 nm ae tric mode O oTrans Seay range 2 to 2 484 to 488 654 to 658 nm Spectral Bandwidth 0 2 nm Signal averaging time 0 033 sec Data interval 0 02 nm Lamp UV Beam mode Single Specifications Zero order 0 7 nm 486 0 nm 0 2 nm 656 1 nm 0 2 nm 7 1 Cary 100 300 certification manual 7 2 7 1 2 What if my instrument fails to meet the specification m m Holmium Check that you have set the tolerances to values that are within the specifications of the Cary instrument Recalibrate the wavelength drive mechanism by turning the instrument off and on again Check the deuterium D lamp alignment and perform the wavelength accuracy test again If correct results still cannot be obtained contact your local Varian Service office as a service call may be required oxide method This method scans the wavelength peaks of a holmium oxide filter between 269 4 and 647 5 nm This method requires the use of a holmium oxide
42. ls to meet the specification Q If correct results cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 8 1 Cary 100 300 certification manual 8 2 8 2 Stray light Stray light is defined as any light that gets to the detector that is not of the selected wavelength The detector cannot distinguish between stray light and light of the selected wavelength and will add the two together to give a higher photometric reading than expected Stray light also causes deviation from the Beer Lambert Law Stray light is monitored by measuring a test solution at a wavelength where the solution is totally absorbing Any light that is detected is due to stray light There is one non pharmocopoeia method to test for stray light This is the following ASTM test method 8 2 1 Nal at 220 nm test Note Note This method measures the absorbance of a Sodium Iodide Nal solution at a wavelength of 220nm This method requires the use of a Sodium Iodide Nal solution This solution is available as part of the solutions test kit part number 99 100852 00 Alternatively the Sodium Iodide Nal solution can be prepared by the operator as follows Nal 10 g L Nal in water Warning To avoid injury ALWAYS ensure er X practices governing the use handling and disposal c ials are strictly observed Appropriate safety clothing and glasses should also be worn When perfor
43. mal room temperature 22 25 C or that printed on the certificate supplied with the filters before repeating the test 6 3 Cary 100 300 certification manual Q If you have had the filter set for several years you may need to return it to the issuing body for recalibration Q Check the result a second time paying special care with the positioning of the NIST filter Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 6 2 2 K Cr 0 method The photometric accuracy of a spectrophotometer can be determined by measuring the absorbance of solutions of high purity compounds This method is limited because it requires accuracy determinations to be performed at a specific temperature and wavelength gt Note This method requires the use of a potassium dichromate K Cr 0 solution and a blank solution The potassium dichromate K Cr 0 and blank solutions are available in the Solutions Test Kit part number 99 100852 00 Alternatively the potassium dichromate K Cr 0 solution can be prepared by the operator as follows Standard 60 06 mg L in 0 01N sulphuric acid Blank 0 01N sulphuric aci Both solutions should be placed in clean silies es Nee Warning To avoid injury ALWAYS ensure that safety practices governing the use handling and di of such materials are strictly observed Appropriate Ke ing and glasses should also be worn oer
44. ming thi critical that great care is taken during sample preparation and ement Suprasil cuvettes must be used They should never be allowed to co and must be cleaned with alcohol and dried with cotton wool never lahoxratory tissue Spectroscopy grade reagents must also be used iefsOurce of error in this measurement is fluorescence of the sample It is M t that the light beam is kept away from the edges of the cuvette that xet as a non attenuating light guide For this reason you must use masked acells for this test With the cuvette in the cell holder hold the cell up to the light and look through the aperture You should not be able to see the cuvette walls Mask the cell holder if necessary to reduce the aperture width to 8 mm Instrument parameters Wavelength 220 nm Photometric mode oTrans Spectral Bandwidth 2 0 nm Signal averaging time 10 0 sec Specifications Cary 100 At 220 nm 10 g L Nal ASTM method lt 0 0200 T Cary 300 At 220 nm 10 g L Nal ASTM method lt 0 0005 T Publication date 10 02 Cary 100 300 certification manual What if my instrument fails to meet the specification Q Ensure that the sample compartment is tightly sealed Repeat the test with the room lights switched off If the instrument passes the test this indicates that there is a light leak in the instrament cover somewhere Q As the test solutions may degrade over time the accuracy of the test depends heavily on the co
45. nd is run using Cary WinUV software Throughout this document except w itly stated otherwise all references to Cary system and Cary in eh Cary 3 Cary 100 and Cary 300 systems Also other than in Binder 1 Section 3 a reference to Cary 100 will be t include Cary 1 and Cary 300 to include Cary 3 The various module hin each of the Cary 1 3 100 and 300 series e g Cary 1E Cary 3G Bio Cary 300 Conc reflect the software applications and accessori ith the instrument uly qualified Varian trained personnel to perform instrument testing ully documented instrument test results to fulfill your auditing requirements e Instrument certification services that can be added to your existing Varian Support Agreement e Standards used in instrument certification that are either primary standards or are traceable back to recognized Standards Institutes such as the U S National Institute for Standard and Technology NIST Contact your local Varian office for full details Publication date 10 02 1 1 Cary 100 300 certification manual 1 2 1 1 Cary certification program 1 2 Available tests Note Good Laboratory Practices quality control methods and other regulatory requirements mandate proof of performance from your Cary system The following procedures are designed to help you meet those needs A spectrophotometer must meet several criteria to demonstrate that it is operating correctly The Cary 100 300 Certi
46. oxide z 7 E 3 Holmium perchlorate 7 FA Wavelength reproducibility amp Resolution power l l 1 Maximum resolution v lt S 2 Toluene hexane v v Baseline flatness v v Photometric noise v v 3 NaNO K Cr 0 370 nm OOo T 0 v Photometric linearity Photometric accuracy pis 1 Double aperture Q v 2 NIST filters z AN 3 K Cr 0 method v O WY 4 KNO method A v Photometric stability SAZ vx Accessory tests Stray light 1 Nal 230m Po 2 KCI 200 nm i ew v VW absolute specular reflectance v accessory ue Diffuse reflectance accessory AS v general N Diffuse reflectance o v ZT Diffuse reflectance detessory v fabric sued for these tests which are performed at the factory before shipping 7 XT A certificateNs Publication date 10 02 1 3 Cary 100 300 certification manual A This page is intentionally A X o 1 4 Publication date 10 02 Cary 100 300 certification manual 2 1 Test kits Publication date 10 02 Test kits and equipment Kits are available to assist in the testing of your instrument and accessories They are as follows 99 100878 00 Cary Basic Recertification kit Minimum equipment required for testing to US BP EP and TGA regulatory requirements 01 180207 90 Holmium Oxide Filter 99
47. rator as follows Standard 60 06 mg L in 0 01N sulphuric acid Blank 0 01N sulphuric acid Both solutions should be placed in clean silica cuvettes Warning To avoid injury ALWAYS ensure that safety practices governing the use handling and disposal of such materials are strictly observed Appropriate safety clothing and glasses should also be worn Publication date 10 02 7 9 Cary 100 300 certification manual Instrument parameters Wavelength 235 257 313 amp 350 nm Photometric mode Abs Spectral bandwidth 2 00 nm Signal averaging time 3 00 sec Lamp UV Vis Specifications Wavelength Absorbance Tolerance 235 nm 0 7480 Abs 40 010 Abs 257 nm 0 8650 Abs 0 010 Abs 313 nm 0 2920 Abs 40 010 Abs 350 nm 0 6400 Abs 0 010 Abs What if my instrument fails to meet the specification Q Repeat the test with a new solution then compare the results to determine if the error was introduced by the solution Q If correct results still cannot be obtained ct your local Varian Service office as a service call may be required 7 5 3 KNO method The photometric accuracy of photometer can be determined by measuring the absorbance of solutions o rity compounds This method is limited because it requires accura amp y determinations to be performed at a specific temperature and wavele gt Note To perform t lution of potassium nitrate KN
48. red to determine the baseline flatness of the instrument gt Note A signal smoothing filter is applied to the corrected baseline for the Varian specification For a description of the smoothing operation refer to the Cary WinUV online help Instrument parameters Photometric mode Abs S Scan range Q 8 Spectral bandwidth Ro G Signal averaging time NO 1b sec Data interval nm Lamp ES UV Vis Specification Baseline Fla ae to Cam fails to meet the specification 40 001 Abs nsure that there are no samples or other obstructions in the sample compartment Q Check the alignment of the deuterium and Visible lamps and perform the test again Q Check the cleanliness of the sample compartment windows and clean with a soft cloth dampened with distilled water if necessary Q Check for possible sources of contamination in the air Changes in the baseline could be caused by the absorption of vapors in the air Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 4 3 Cary 100 300 certification manual 4 4 4 4 Photometric noise Photometric noise is a measure of the signal to noise ratio of the instrument The higher the noise the harder it is to distinguish between small changes in the signal and the noise of the system The noise level indicates the stability of a rea
49. res accuracy determinations to be performed at a specific temperature and wavelength The following method is recommended by the European Pharmacopoeia for this test This method measures the absorbance of a and a blank solution The certified potassi solutions are available in the Solutio Alternatively the potassium di Cr 0 operator as follows dichromate K Cr 0 solution ighromate K Cr 0 and blank iN part number 99 100852 00 solution can be prepared by the Standard 60 06 mg L in sulphuric acid Blank 0 01N s uric acid Both solutions shoul d in clean silica cuvettes with a path length of 10 mm TV yoidinjury ALWAYS ensure that safety practices governing the pb narMing and disposal of such materials are strictly observed propriate safety clothing and glasses should also be worn If you are using a potassium dichromate K Cr 0 certified solution enter the expected absorbance values into the table on the Control of Absorbance page Instrument parameters Wavelength 235 257 313 amp 350 nm Photometric mode Abs Spectral bandwidth 2 00 nm Signal averaging time 3 00 sec Lamps UV Vis 5 3 Cary 100 300 certification manual 5 4 5 3 Resolution power 5 3 1 Toluene hexane Y Note Specifications Wavelength Absorbance Tolerance 235 nm 0 7480 Abs 0 010 Abs 257 nm 0 8650 Abs 0 010 Abs 313 nm 0 2920 Abs 40 0
50. rrect handling management and combination of materials Make up fresh solutions ensuring cell cleanliness Mask the cell holder to reduce the aperture width to 8 mm and repeat test Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 8 3 Photometric accuracy 8 3 1 Publication date 10 02 Double aperture Note Photometric accuracy is determined by measuring a known absorbance value and calculating the difference between the expected absorbance value and the absorbance value as reported by the instrument The following method is recommended for testing photometric accuracy The Double aperture method was first used een A at the National Physical Laboratory in England and is used by the ajo naWdnal standards laboratories to measure the absolute accuracy of theif reference spectrophotometers This test takes several measurements with the A sample beam interrupted with several apertures and then mathematically cd cutates the photometric accuracy The advantage of the Double Ape od of determining photometric accuracy is that there is no limitation Pary standa wavelength or SBW as with the other methods This method is aprNnary standard and therefore does not require traceability Varian certifie e Double Aperture method for photometric accuracy To perform 11 Q fitted in the G NS te arameters 590 nm
51. rrect results still cannot be obtained contact your local Varian Service office as a service call may be required gt Note Photometric noise can be affected by ade in the signal received by the detector so the electronic noise j Publication date 10 02 8 7 Cary 100 300 certification manual 8 8 A This page is intentionally A X o Publication date 10 02 9 1 9 1 1 Publication date 10 02 Cary 100 300 certification manual Accessory tests The Cary 100 300 series instruments can be used with a number of accessories to extend the capabilities of the instrument For example the Cary WinUV Validate application software includes tests specifically designed for Fabric applications using the diffuse reflectance accessory In the case of the reflectance accessories such as the specular and diffuse reflectance accessories specific test methods are recommended for monitoring their performance These methods are outlined below Diffuse reflectance accessory tests The following methods are recommended to test the diffuse reflectance accessory DRA These tests vary depending upon the purpose for which the DRA is to be used There are three sets of tests U General QO Color O X Q Fabric General diffuse reflectance ac ry test Note Note This method requir of a diffuse reflectance standard with its associated calibration cegtifi separate certified reflectance standard an
52. rties dialog which is accessible from the Windows Control Panel You can save your data at the end of the Validate run To do this select Save Data As from the File menu to display the Save As dialog and save the current method data and report in a batch file BVO Publication date 10 02 Cary 100 300 certification manual 4 Instrument performance tests The following tests are listed in the Cary WinUV Validate application software as Instrument performance tests On installation of the instrument it is recommended that these tests are repeated and the results compared to those achieved at the factory 4 1 Wavelength accuracy Poor wavelength accuracy can produce low quantitative results as the absorbance measurement may have been made on the side of the peak rather than at the peak Wavelength accuracy is determined by scanning a known wavelength peak and calculating the difference between this peak and the wavelength peak reported by the instrument The following method is used at Varian s fa est for wavelength accuracy 4 1 1 Deuterium D emission line J tiss t This method scans the deuteria D Vis region as well as the zero f additional equipment Instrument paramet Wavelength C2 0 486 0 656 1 nm Photometric Trans Scan range 2 to 2 484 to 488 Zi 654 to 658 nm SPE ndwidth 0 2 nm Sign veraging time 0 033 sec Data interval 0 02 nm Lamp UV Beam mode Sing
53. s from contacting the material surface Always wear clean gloves when handling these materials The liquid standards must not be allowed to freeze and it is recommended that they be stored within the temperature range of 5 to 50 Celsius Dirty cells are the greatest single sou error in spectrophotometry Avoid handling the cells by the polised X In the event of the faces becoming contaminated clean by soakin eutral detergent solution according to the manufacturer s instructions ringe horoughly with de ionized water and dry with ethanol DO NOT use acetdne Solution spillage sho be allowed to dry on the cells and should be wiped off with a OH r lint free cloth 10 1 Cary 100 300 certification manual 10 3 Re calibrating the liquid standards 10 2 Starna the recommended supplier of liquid standards recommend that their liquid standards sets be recalibrated every two years However most accreditation authorities require that standards be recalibrated at least once per year The calibration of the liquid standards does not usually change significantly within this time interval unless the cell becomes cracked marked or scratched and will then require repolishing to restore its accuracy For recalibration of the Starna liquid standards please contact your nearest Starna distributor Starna Pty Ltd Optiglass Ltd PO Box 113 52 54 Fowler Road Thornleigh Hainault Industrial Estate HAINAULT NSW 2120 AUSTRALIA ESSEX 1G6 3
54. t period 120 min Signal averaging time 1 00 sec Data interval 2 00 sec Lamp Vis A Specification SS Stability What if my instrument fails tome t the specification aperature variation in the laboratory does not d the test exceed d m aan ontaminants are present in the air that may absorb at 500 nm an open beaker of solvent f correct results still cannot be obtained contact your local Varian Service office as a service call may be required Publication date 10 02 8 5 Cary 100 300 certification manual 8 6 8 6 Photometric noise Photometric noise is a measure of the signal to noise ratio of the instrument The higher the noise the harder it is to distinguish between small changes in the signal and the noise of the system The noise level indicates the stability of a reading It determines the precision of the measurement and the detection limit of the instrument Poor signal to noise performance makes it very difficult to determine the actual absorption value as it fluctuates It introduces errors into both quantitative and qualitative spectroscopy This method provides an extension of the photometric noise test offered in the Instrument performance tests and allows measurements at up to 3 0 Abs for the Cary 100 instrument and up to 5 0 Abs for the Cary 300 instrument 8 6 1 Photometric noise method Note This test measures the photometric noise specified as Absorbance Root Mean Square R
55. ter in comparison Low light throughput can be due to a faulty or misaligned source mirror so replacing or realigning the source mirror may improve the noise performance Prolonged exposure to UV light will slowly degrade the optical surfaces It is recommended that you tum off the deuterium lamp when it is not in use The lamps can be automatically turned off at the end of each run by checking the Auto lamps off checkbox on the Options tab of the Setup dialog Publication date 10 02 Cary 100 300 certification manual 5 European Pharmacopoeia tests These tests available from the configure page EP BP tests of the Cary WinUV Validate software are required by the European and British Pharmacopoeia Many of these tests are duplicated in either the standard instrument performance tests or the other pharmacopoeia tests 5 1 Control of wavelengths Poor wavelength accuracy can produce low quantitative results as the absorbance measurement may have been made on the side of the peak rather than at the peak Wavelength accuracy is determined by scanning a known wavelength peak and calculating the difference between this peak and the wavelength peak reported by the instrument There are two methods recommended by the n Pharmacopoeia to test for wavelength accuracy Q Deuterium D emission line 29 Q Holmium perchlorate meth d method m D emission lines at 486 0 and 656 1 nm in the o order line This method does not require
56. teses 9 1 VW absolute specular reflectance accessory test 9 2 Diffuse reflectance accessory tests 10 Care and handling ee eceseeeeeessceeeeseeeeeeeseees 10 1 General care 10 2 Cleaning the liquid standards 10 3 Re calibrating the liquid standards 10 4 Ordering NIST reference materials 10 5 Re calibrating the NIST Filter standards 10 6 Cleaning the diffuse reflectance standards 10 7 Re calibrating the diffuse reflectance standards 11 Certification kit log sheet eee seeeeeeseeeeeeeeee A X o Publication date 10 02 Cary 100 300 certification manual 1 Introduction The Cary 100 300 Certification Manual can be used in conjunction with the Cary 100 300 Validation Binder 2 to qualify your Cary instrument and accessories Varian can provide an Installation and Operational Qualification service and provide ongoing operational testing throughout the life of the instrument This manual assists you to run operational checks on the instrument and the accessories You can run these tests to verify that the instrument is performing to the specifications required Certification of instrument analytical methods is not included This validation package is suitable for use with Cary 100 300 series instruments running Cary WinUV Version 2 0 or Cary WinUV Pharma Version 2 5 software A Cary 1 3 series instrument is regarded as bei e same as a Cary 100 300 series instrument when it has Version 8 0 or hig are installed a
57. tests that were originally performed at manufacture RO designed to check that all components of the spectrophotorfgtet ate Working correctly and within the specifications set by Varian These tests aya aum set of criteria for a general purpose UV Vis spectrophi ed Perfor tests should be performed routinely or whenever the instrument has d or serviced or when malfunctioning is suspected The frequency of esting should be determined based on the type of application and the frequency of use of the instrument The table on the opposite page summarizes the tests available in the Cary WinUV software listed by the pharmacopoeia requirements e g EP USP and TGA The Cary WinUV Validate application software includes the Australian Code of GMP for Therapeutic Goods TGA Guidelines for Laboratory Instrumentation TGA tests On 1 January 1999 the following change to the Manufacturing Principles of the Therapeutic Goods Act 1989 took effect Part 2 of the Australian Code of GMP for Medicinal Products was replaced with Annex 1 of the EU Guide to GMP The Cary WinUV software still reflects the tests which were recommended by the TGA prior to 1 January 1999 Publication date 10 02 Cary 100 300 certification manual 1 3 Cary performance tests Instrument Test USP BP EP TGA Instrument Other Tests Tests Tests performance tests tests Wavelength accuracy tests l l 1 D emission line v v v v 2 Holmium
58. the Beer Lambert Law Stray light is monitored by measuring a test solution at a wavelength where the solution is totally absorbing Any light that is detected is due to stray light There are two methods recommended by the TGA to test for stray light Q KA 12 g L KC in water test a NaNO K Cr 0 for 370 nm test 7 4 1 KCI test Note Note Note Note This method measures the absorbance of potassium chloride KCI at a wavelength where the solution is totally absorbing The European Pharmacopoeia recommend a wavelength of 198 nm The Cary WinUV software version 2 5 measures 94 a wavelength of 200 nm To perform this test a solution of potassium ehtoride KCI and a water blank solution must be used These solutions ar dvailgble As part of the solutions test kit part number 99 100852 00 Alternatively potassium chloride KCI solution can be prepared by the operator as fol gt KCI 12 g L KCI in water Warning forming this test it is critical that great care is taken during sample preparation and measurement Suprasil cuvettes must be used They should never be allowed to contact skin and must be cleaned with alcohol and dried with cotton wool never laboratory tissue Spectroscopy grade reagents must be used The chief source of error in this measurement is fluorescence of the sample It is also important that the light beam is kept away from the edges of the cuvette that can act as a non attenuat
59. the use of 5 1 1 Deuterium D emission This method scans th Vis region as well he additional eq e us Instrume ers Wav CBs 0 486 0 656 1 nm o j mode O oTrans ges 2 to 2 484 to 488 654 to 658 nm Spectral Bandwidth 0 2 nm Signal averaging time 0 033 sec Data interval 0 02 nm Lamp UV Beam mode Single Specifications Zero order 0 7 nm 486 0 nm 0 2 nm 656 1 nm 0 2 nm Publication date 10 02 5 1 Cary 100 300 certification manual What if my instrument fails to meet the specification Q Check that you have set the tolerances to values that are within the specifications of the Cary instrument Q Recalibrate the wavelength drive mechanism by turning the instrument off and on again Q Check the deuterium D lamp alignment and perform the wavelength accuracy test again Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 5 1 2 Holmium perchlorate method This method scans the wavelength peaks of a holmium perchlorate solution between 231 15 and 546 30 nm The method requires the use of a holmium perchlorate solution and a blank solution These solutions are available as part of the Solutions Test Kit part number 99 100852 00 Alternatively the operator can prepare the holmium perchlorate solution as follows 4 w v solution of holmium oxide 99 5 ik 1 4 M perchloric acid X
60. tion i e a social securi mber SSN for consumer customers tax identificatio ex TIN for commercial customers or agency code S government customers 10 5 Re calibrating the er standards National Institute of Standards and Technology Calibrations Program 100 Bureau Drive Stop 2330 Gaithersburg MD 20899 2330 c ion of the NIST filter standards please contact your nearest NIST er Or contact USA Telephone 1 301 975 2002 Facsimile 1 301 869 3548 Email CALIBRATIONS nist gov Web site http www nist gov Publication date 10 02 10 3 Cary 100 300 certification manual 10 6 Cleaning the diffuse reflectance standards gt Note If the surface of the standards material should ever become contaminated it may be blown clean with a jet of clean dry air or nitrogen Do not use Freon If this is not sufficient then the material surface can be sanded under running water with 220 240 grit water proof emery cloth until the surface is totally hydrophobic the water beads and runs off the material immediately If the material is grossly contaminated or scratched then the material surface can be sanded with 220 320 grit wet sand paper under a stream of running water The materials surface should then be blown dry with clean dry air or nitrogen or alternatively allow the material to air dry For low reflectance material i e lt 10 sand it dry rather than under running water If the standard mat
61. tment is tj sealed Repeat the test with the room lights turned off If the instrument p the test this indicates that there is a light leak in the instrument QY somewhere Q As the test solutions may degrad ON ime the accuracy of the test depends heavily on the correct havidtivfg wandgement and combination of materials Q If correct results st not be obtained contact your local Varian Service office as a servi ay be required N NaNO K Cr 70 nm Note Note measures the absorbance of either sodium nitrite NaNO or potassium te K Cr 0 at a wavelength of 370nm To perform this test a solution of either sodium nitrite NaNO or potassium dichromate K Cr 0 must be prepared by the operator The sodium nitrite NaNO solution is prepared as follows 50 g L NaNO in water The potassium dichromate K Cr 0 solution is prepared as follows 0 25 g L K Cr 0 in aqueous 0 05 M KOH Warning To avoid injury ALWAYS ensure that safety practices governing the use handling and disposal of such materials are strictly observed Appropriate safety clothing and glasses should also be worn When performing this test it is critical that great care is taken during sample preparation and measurement Suprasil cuvettes must be used They should never be allowed to contact skin and must be cleaned with alcohol and dried with cotton wool never laboratory tissue Spectroscopy grade reagents must also be used 7 7 Cary 100 300
62. toxetric mode Trans Spextral bandwidth 3 00 nm Signal averaging time 3 00 sec Replicates 3 minimum Lamp Vis Specifications at 0 3 Abs 0 0006 Abs at 1 0 Abs 0 0012 Abs What if my instrument fails to meet the specification Q Ensure that the double aperture kit is fitted and aligned correctly as described in the operation manual Also refer to the Cary WinUV online help for more details Q If correct results still cannot be obtained contact your local Varian Service office as a service call may be required 8 3 Cary 100 300 certification manual 8 4 Photometric linearity This test determines how accurately the instrument measures absorbance with increasing concentration Poor photometric linearity will produce incorrect results The following method is recommended for testing photometric linearity 8 4 1 Photometric linearity The linearity of the instrument is tested using the addition of filters technique at absorbance levels of 1 0 2 0 and 3 0 Abs i e the sum of individual filters readings is subtracted from the reading of all filters combined This is the error at the combined absorbance gt Note This test can be performed on the Cary 100 300 series instruments however there are no guaranteed specifications provided gt Note The photometric value of each of the filters within the Neutral Density Filter Kit part number 99 100561 00 will vary with wavelength The marked values on these
63. truments the maximum SBW is 4 0 nm Instrument parameters Wavelength nm 440 00 465 00 546 10 590 00 635 00 Photometric mode Abs Abs Abs Abs Abs SBW nm 2 2 2 7 4 0 4 0 4 0 Signal averaging 3 00 sec 3 00 sec 3 00 sec 3 00 sec 3 00 sec time SAT Specification at 1 0 Abs 40 005 Abs What if my instrument fails to meet the specification Q The absorbance of the filters is influence he temperature and the age of the filters If it is unusually hot or cold boratory then you should wait until the temperature returns to nor that printed on the certificate supplied with the filters before pep a e test Q If you have had the filter or ral years you may need to return it to the issuing body for recalibration Q Check the result a se nd time paying special care with the positioning of the NIST filter Q If correct res 5 nnot be obtained contact your local Varian Service office Ci may be required 7 5 2 K Cr 0 metho epared by the operator This method is limited because it requires accuracy determinations to be performed at a specific temperature and wavelength gt Note This method requires the use of a potassium dichromate K Cr 0 solution and a blank solution The potassium dichromate K Cr 0 and blank solutions are available in the Solutions Test Kit part number 99 100852 00 Alternatively the potassium dichromate K Cr 0 solution can be prepared by the ope
64. ual Signal to noise ratio VW abso Note Note Note The noise level is an indication of the stability of a reading It determines the precision of the measurement and the detection limits of the instrument Poor signal to noise performance causes fluctuations in the instrument readings and makes it very difficult to determine the real T value No additional filters are required to perform this test Instrument parameters Wavelengths 280 350 380 nm Photometric mode Absorbance Spectral bandwidth 1 0 nm Signal averaging time 1 0s Data interval 1 0s Measurement period 1 minute Lamp UV Vis Specification Signal to noise ratio at gt 50 1 280 350 and 380 nm What if my instrument fails to mee peeification Q Check the alignment of th inst lamps and perform the test again Q Check the alignment of th ory mirrors and perform the test again J Ensure that the filt Cary WinUV sof ere inserted in the correct order as specified by the annot be obtained contact your local Varian Service Q If correct res j office NS call may be required operation of the VW absolute specular reflectance accessory is tested by meas ring the specular reflectance of a certified reflectance standard at 24 wavelengths This method requires the use of a specular reflectance standard with its associated calibration certificate part number 99 100842 00 Prior to me

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