Certified UV/Vis Reference Materials - Dialab · Certified UV/Vis Reference Materials Guidelines //...
Transcript of Certified UV/Vis Reference Materials - Dialab · Certified UV/Vis Reference Materials Guidelines //...
CertifiedUV/Vis Reference Materials Guidelines
// easy handlinG// Reliable measuRement Results // inteRnational compaRability
High Precision in Spectro-Optics
// Glass filteRs
// liquid filteRs
// ceRtified RefeRence plates
accredited accordance todin en iso 17025
tRaceable
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contents
1. IntroductIon page 4–10
1.1 hellma analytics calibration laboratory page 4
1.2 certified test equipment page 6
1.3 Glass filter applications page 6
1.4 liquid filter applications page 6
1.5 dakks calibration certificate page 8
1.6 expanded absorbance range – from 0 to < 3.1 for uV-Vis reference materials page 11
2. Glass fIlters page 12–21
2.1 holmium oxide glass filter for checking wavelength accuracy page 12
2.2 didymium glass filter for checking wavelength accuracy page 13
2.3 didymium glass filter for checking photometric accuracy page 14
2.4 neutral density glass filters for checking photometric accuracy page 15
2.5 Glass filter sets page 16
2.6 General usage guidelines for glass filters page 17
2.7 calibration with glass filters page 18–21
3. lIquId fIlters page 22–35
3.1 holmium oxide liquid filter for checking wavelength accuracy page 22
3.2 potassium dichromate liquid filter for checking photometric accuracy page 23
3.3 liquid filters for checking for stray light page 24
3.4 liquid filters for checking spectral resolution page 25 3.5 liquid filter set compliant with the european pharmacopoeia page 26
3.6 liquid filter set compliant with the united states pharmacopeia page 27
3.7 General usage guidelines for liquid filters page 27 3.8 calibration with liquid filters page 28–35
4. certIfIed reference plates page 36–41
4.1 Reference plate for checking photometric accuracy page 36
4.2 Reference plate for checking photometric and wavelength accuracy page 37
4.3 General usage guidelines for reference plates page 38
4.4 calibration with reference plates page 39–41
5. recertIfIcatIon page 42–43
6. faqs page 44–45
7. Glossary page 46
8. recommendatIons for further readInG page 47
9. product overvIew page 48–50
accRedited to
din en iso 17025
although checking measuring equipment to ensure thatresults are accurate has long been common practice foranalytical balances, it still tends to take something ofa backseat where spectrophotometers are concerned.Spectrophotometers are important instruments that playa major role in health care, the life sciences, environmentalanalysisandprocessessuchasproductioncontrolandensu-ring product quality. Over the last two years, many labora-torieshavebecomeconsiderablymoreawareof theneed tocheck their spectrophotometers, making it all the more im-portant to know that these precision tools are also subjectto mandatory checks under DIN eN ISO 9001. The standardclearly stipulates that measuring equipment be calibratedor verified, either at regular intervals or before use, usingmeasurementstandards thatcanbe tracedback to interna-tional or national standards. For an overview of the measu-rementstandardsforUV-Visspectrophotometers,pleasere-fertoourreferencematerialsinthisbrochure.anincreasingnumber of laboratories are turning to this easy method forensuringhighstandardsofwork–notonlytosatisfyrequire-ments in time for theirnextaudit,butalso tobesafe in theknowledge that theyare takingaccuratemeasurementsandthus basing their actions and responses on correct results.We are delighted that our products are helping to achie-ve this. Details of our product range, as well as usage gui-delines,helpful tips,and recommendations,areall includedin this Handbook. We VeRy much hope that you enjoy ReadinG it!
1.1 hellma analytics calibration laboratory: accredited to din en iso 17025
Our lab is a DakkS calibration laboratory and is accredited toDIN eN ISO 17025, a comprehensive quality management sy-stem that acts as a seamless continuation of other systemssuchas ISO9000.Byachieving thisaccreditation,wehavede-monstratedproofofexpertiseinthecalibrationactivitiesthatweperform and are authorized to issue internationally recognizedDakkS calibration certificates. accreditation is the key to highqualitymeasurements, internationalcomparability,andtrust inboththeworkofthecalibrationlaboratoryandthetransparencyofresults.
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1. IntroductIon
dear Readers,
din en iso 17025
accReditationstands for trust – and trust is
built on reliability(in latin accredere means giving credibility
to something)
accRedited to
din en iso 17025
birgit Kehl,HeadofHellmaanalyticscalibrationlaboratory
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1.2 certified test equipment
Qualityassuranceandqualitycontrolregulations,suchasISO9000,gLp,gMp,andpharmacopoeias,requirecompaniestoverify the consistently excellent performance of any spectro-photometerinuse.Thetwomostimportantfactorsforobtai-ningprecisespectrophotometerdataarethephotometricac-curacy(absorbanceaccuracy)andwavelengthaccuracyofthespectrophotometer,whichshouldbetestedonaregularbasis.
IntheHellmaanalyticscalibrationlaboratory,whichisacc-redited toDINeN ISO17025,wemanufacturecertified re-ferencematerialsbasedon the regulatorycodes issuedbyNIST(NationalInstituteofStandardsandTechnology),aSTM(americanSocietyforTestingandMaterials)andpharmaco-poeias (ph.eur.,DaB,USp).all certifiedmeasurement re-sultscanbetracedbacktoNISTstandardreferencemateri-als(SRMs).(photometricaccuracy:SRM®930eandSRM®1930,wavelengthaccuracy:SRM®2034).
choose between glass filters and liquid filters for your reference materials:
1.3 Glass filter applications
glass filters are certified reference materials made of glassmanufactured specifically for calibration. They are, above all,extremelyrobust.allglassfilterscertifiedbyHellmaanalyticsare traceable to NIST primary standards. Certified glass fil-tersaresuitableforcheckingthefollowingparametersofyourspectrophotometer:
» Wavelength accuracy » photometric accuracy (absorbance)
1.4 liquid filter applications
Liquidfiltersarecertifiedliquidreferencematerialsthataremanufacturedincompliancewithpharmacopoeiasand/orNISTstandardsandfilledintoquartzglasscuvettesundercontrolledconditions.Thecuvettesarethenpermanentlysealedtobecomeairtight.Liquidfiltershavethedistinctad-vantageofequatingtorealmeasuringconditions.CertifiedHellmaanalyticsliquidfiltersaresuitableforcheckingthefollowingparametersofyourspectrophotometer:
» Wavelength accuracy » photometric accuracy (absorbance) » stray light levels » spectral resolution
You should regularly check your UV/Vis spectrophotome-ter forallof these factors,especiallyphotometricandwa-velength accuracy, while observing the relevant require-mentsinyourdevicehandbook.Thankstotheireaseofuseand long service life, certified Hellma analytics referencematerialsprovideanexcellentaidforallroutinechecks.
hellma analytics’ dakks-certified refe-rence materials comply with the provi-sions stipulated by quality management systems and pharmacopoeias, meeting the highest quality requirements and ensuring the international comparability of measurement results.
1. IntroductIon
hellma analytics RefeRence mateRials in accoRdance With the most impoRtant set of Rules and ReGulations
mateRial checKinG of RanGe ph.euR. dab usp 851 astm
Glass filteRs
holmium oxide glass Wavelengthaccuracy UV/Vis
didymium glass filter Wavelengthaccuracy UV/Vis
didymium glass filter photometricaccuracy UV
neutral density glass photometricaccuracy Vis
liquid filteRs
holmium oxide solution Wavelengthaccuracy UV/Vis
potassium dichromate solution photometricaccuracy UV/Vis
toluene in hexane Spectralresolution UV
potassium chloride solution Straylight UV
sodium iodide solution Straylight UV
sodium nitrite solution Straylight UV
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FO-Labor-062, Rev.:5– 17.12.14
Kalibriergegenstand: Kalibrierstandard-Satz, bestehend aus drei Neutralglas-filtern NG11, NG5 und NG4.
Calibration Object: Set of calibration filters, consisting of three neutral density glass filters NG11, NG5 and NG4.
Kalibrierverfahren: Messung der optischen Dichte. Diese Kalibrierstandards wurden gegen Luft als Referenz gemessen.
Calibration Method:Measurement of optical density. These calibration standards were measured using air as reference.
Messtechnische Bedingungen bei der Kalibrierung: Conditions of Calibration: Die in diesem Kalibrierschein angegebenen Werte wurden mit dem verwendeten Spektralphotometer und den nach-folgenden Einstellungen ermittelt:
The following settings were used on the spectrometer employed to obtain the data quoted on this calibration certificate:
UV/VISModus der Ordinatenskala: Optische Dichte (Abs) Spaltbreite: 1,00 nm Spaltmodus: Fix Integrationszeit: 3,0 s
UV/VISOrdinate mode: Optical density (Abs)
mn 00.1 :tilS xiF :edoM tilS
Integration time: 3.0 s
Für die Kalibrierung dieses Kalibriergegenstandes wurde ein UV/VIS/NIR-Spektralphotometer Varian Cary-5000 mit der Seriennummer UV1101M202 eingesetzt.
This calibration object was calibrated on a UV/VIS/NIR spectrophotometer Varian Cary-5000 with serial number UV1101M202.
Dieses Gerät wird regelmäßig auf die Einhaltung seiner Spezifikationen überprüft. Datum der letzten technischen Überprüfung: 27. November 2014
This instrument is regularly checked for the compliance with its specifications. Most recently technical check: 27 November 2014
Für die regelmäßige Überprüfung der photometrischen Richtigkeit werden die Bezugsnormale des NIST SRM 930e Filter Nr. 2115, gültig bis März 2015 und SRM 1930 Filter Nr. 202, gültig bis März 2016 eingesetzt.
A set of NIST SRM 930e Filter No. 2115, valid until March 2015 and SRM 1930 Filter No. 202, valid until March 2016 standard reference materials is used to regularly check the photometric accuracy of the spectrophotometer.
Zur regelmäßigen Überprüfung der Wellenlängenrichtigkeit wurde das intrinsische Bezugsnormal
Hellma UV5 S.Nr. 0861 / 87450-PTB-14, gültig bis Dezember 2015 eingesetzt.
The intrinsic standard reference material Hellma UV5 serial no. 0861 / 87450-PTB-14, valid until
December 2015 is used to regularly check the wavelength accuracy.
Zusätzlich werden die Emissionslinien von Deuterium, Quecksilber und Argon zur Überprüfung der Wellen-längenrichtigkeit verwendet.
In addition, the emission lines of deuterium, mercury and argon are used to check the wavelength accuracy.
:snoitidnoC latnemnorivnE :negnugnidebsgnubegmU
Die Messungen wurden bei einer Umgebungstemperatur von 22°C ± 2°C und einer relativen Luftfeuchtigkeit von 30% bis 65% durchgeführt.
Measurements were performed at an ambient temperature of 22°C ± 2°C and a relative humidity of 30% to 65%.
1.5 dakks calibration certificate
after careful production, reference materials are certifiedintheHellmaanalyticscalibrationlaboratory(accreditedtoDIN eN ISO 17025) using a high-performance UV-Vis/NIRspectrophotometer. Reference materials are only conside-
redtobecertifiediftheyhavebeenissuedwithaDakkSca-libration certificate and bear a calibration mark. Using themeasurement valuesdocumentedandcertified in thecali-brationcertificate,userscancheckandcalibratetheirspec-trophotometersaccordingly.
1 description of calibrated item
2 measurement conditions
3 type of device used to carry out
measurements
4 type, serial number and validity of
calibrated nist/ptb reference
standards used to regularly check
reference photometers; details of
additional checking methods
5 ambient conditions during
measurement
1 name of accreditation body
2 name and address of issuing
laboratory
3 description of calibrated item
4 clear details of measured item
5 customer
6 date of measurement
7 internal document reference,
including date of issue and last
modified date
8 date of issue of calibration certificate
9 calibration certificate serial numbers
10 dakks registration number
11 year and month of calibration
12 authorized persons
the dakks calibration certificate from the national accreditation body for the federal Republic of Germany may only be issued by accredited partners. the hellma analytics calibration laboratory is the only such laboratory in Germany that is accredited to certify uV-Vis re-ference materials.
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1. IntroductIon
accuRate measuRement
Results GuaRanteed.
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21112SeitePage 2 / 3
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FO-Labor-062, Rev.:5– 17.12.14
Kalibriergegenstand: Kalibrierstandard-Satz, bestehend aus drei Neutralglas-filtern NG11, NG5 und NG4.
Calibration Object: Set of calibration filters, consisting of three neutral density glass filters NG11, NG5 and NG4.
Kalibrierverfahren: Messung der optischen Dichte. Diese Kalibrierstandards wurden gegen Luft als Referenz gemessen.
Calibration Method:Measurement of optical density. These calibration standards were measured using air as reference.
Messtechnische Bedingungen bei der Kalibrierung: Conditions of Calibration: Die in diesem Kalibrierschein angegebenen Werte wurden mit dem verwendeten Spektralphotometer und den nach-folgenden Einstellungen ermittelt:
The following settings were used on the spectrometer employed to obtain the data quoted on this calibration certificate:
UV/VISModus der Ordinatenskala: Optische Dichte (Abs) Spaltbreite: 1,00 nm Spaltmodus: Fix Integrationszeit: 3,0 s
UV/VISOrdinate mode: Optical density (Abs)
mn 00.1 :tilS xiF :edoM tilS
Integration time: 3.0 s
Für die Kalibrierung dieses Kalibriergegenstandes wurde ein UV/VIS/NIR-Spektralphotometer Varian Cary-5000 mit der Seriennummer UV1101M202 eingesetzt.
This calibration object was calibrated on a UV/VIS/NIR spectrophotometer Varian Cary-5000 with serial number UV1101M202.
Dieses Gerät wird regelmäßig auf die Einhaltung seiner Spezifikationen überprüft. Datum der letzten technischen Überprüfung: 27. November 2014
This instrument is regularly checked for the compliance with its specifications. Most recently technical check: 27 November 2014
Für die regelmäßige Überprüfung der photometrischen Richtigkeit werden die Bezugsnormale des NIST SRM 930e Filter Nr. 2115, gültig bis März 2015 und SRM 1930 Filter Nr. 202, gültig bis März 2016 eingesetzt.
A set of NIST SRM 930e Filter No. 2115, valid until March 2015 and SRM 1930 Filter No. 202, valid until March 2016 standard reference materials is used to regularly check the photometric accuracy of the spectrophotometer.
Zur regelmäßigen Überprüfung der Wellenlängenrichtigkeit wurde das intrinsische Bezugsnormal
Hellma UV5 S.Nr. 0861 / 87450-PTB-14, gültig bis Dezember 2015 eingesetzt.
The intrinsic standard reference material Hellma UV5 serial no. 0861 / 87450-PTB-14, valid until
December 2015 is used to regularly check the wavelength accuracy.
Zusätzlich werden die Emissionslinien von Deuterium, Quecksilber und Argon zur Überprüfung der Wellen-längenrichtigkeit verwendet.
In addition, the emission lines of deuterium, mercury and argon are used to check the wavelength accuracy.
:snoitidnoC latnemnorivnE :negnugnidebsgnubegmU
Die Messungen wurden bei einer Umgebungstemperatur von 22°C ± 2°C und einer relativen Luftfeuchtigkeit von 30% bis 65% durchgeführt.
Measurements were performed at an ambient temperature of 22°C ± 2°C and a relative humidity of 30% to 65%.
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1. IntroductIon
1 measurement value and smallest attributed measurement uncer-tainty that can be specified. this value only refers to hellma analytics measurements and applies solely to the company’s specific measu-rement conditions. in justified cases, calibration certificates may also show measurement results that do not fall within the calibration laboratory’s scope of accreditation. these must be clearly labeled as such on the calibration certificate.
2 notes on determining expanded measurement uncertainty.
3 notes on initial measurements of filters used to determine optical den-sity. initial measurements are not taken for filters used to determine wavelength accuracy.
4 calibration certificates shall not contain recommended recertification in-tervals (in accordance with dakks-dKd-5). exceptions may be made if re-quested by the customer or required by legislation.
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21112SeitePage 2 / 3
D-K- 18752-01-00
2014-12
FO-Labor-062, Rev.:5– 17.12.14
Kalibriergegenstand: Kalibrierstandard-Satz, bestehend aus drei Neutralglas-filtern NG11, NG5 und NG4.
Calibration Object: Set of calibration filters, consisting of three neutral density glass filters NG11, NG5 and NG4.
Kalibrierverfahren: Messung der optischen Dichte. Diese Kalibrierstandards wurden gegen Luft als Referenz gemessen.
Calibration Method:Measurement of optical density. These calibration standards were measured using air as reference.
Messtechnische Bedingungen bei der Kalibrierung: Conditions of Calibration: Die in diesem Kalibrierschein angegebenen Werte wurden mit dem verwendeten Spektralphotometer und den nach-folgenden Einstellungen ermittelt:
The following settings were used on the spectrometer employed to obtain the data quoted on this calibration certificate:
UV/VISModus der Ordinatenskala: Optische Dichte (Abs) Spaltbreite: 1,00 nm Spaltmodus: Fix Integrationszeit: 3,0 s
UV/VISOrdinate mode: Optical density (Abs)
mn 00.1 :tilS xiF :edoM tilS
Integration time: 3.0 s
Für die Kalibrierung dieses Kalibriergegenstandes wurde ein UV/VIS/NIR-Spektralphotometer Varian Cary-5000 mit der Seriennummer UV1101M202 eingesetzt.
This calibration object was calibrated on a UV/VIS/NIR spectrophotometer Varian Cary-5000 with serial number UV1101M202.
Dieses Gerät wird regelmäßig auf die Einhaltung seiner Spezifikationen überprüft. Datum der letzten technischen Überprüfung: 27. November 2014
This instrument is regularly checked for the compliance with its specifications. Most recently technical check: 27 November 2014
Für die regelmäßige Überprüfung der photometrischen Richtigkeit werden die Bezugsnormale des NIST SRM 930e Filter Nr. 2115, gültig bis März 2015 und SRM 1930 Filter Nr. 202, gültig bis März 2016 eingesetzt.
A set of NIST SRM 930e Filter No. 2115, valid until March 2015 and SRM 1930 Filter No. 202, valid until March 2016 standard reference materials is used to regularly check the photometric accuracy of the spectrophotometer.
Zur regelmäßigen Überprüfung der Wellenlängenrichtigkeit wurde das intrinsische Bezugsnormal
Hellma UV5 S.Nr. 0861 / 87450-PTB-14, gültig bis Dezember 2015 eingesetzt.
The intrinsic standard reference material Hellma UV5 serial no. 0861 / 87450-PTB-14, valid until
December 2015 is used to regularly check the wavelength accuracy.
Zusätzlich werden die Emissionslinien von Deuterium, Quecksilber und Argon zur Überprüfung der Wellen-längenrichtigkeit verwendet.
In addition, the emission lines of deuterium, mercury and argon are used to check the wavelength accuracy.
:snoitidnoC latnemnorivnE :negnugnidebsgnubegmU
Die Messungen wurden bei einer Umgebungstemperatur von 22°C ± 2°C und einer relativen Luftfeuchtigkeit von 30% bis 65% durchgeführt.
Measurements were performed at an ambient temperature of 22°C ± 2°C and a relative humidity of 30% to 65%.
1.6 extended absorption range – from 0 to < 3.1 for uV/Vis reference materials
Underthepreviousaccreditation,thecompanywascertifiedto manufacture UV-Vis reference materials for wavelengthaccuracyandopticaldensityuptoanabsorbanceof2.05a.However, pharmaceutical applications, biotechnology andspectrophotometer manufacturers all increasingly requirereferencematerialswithacertifiedabsorbanceupto3a.
SinceNIST(NationalInstituteofStandardsandTechnology)is no longer producing the relevant SRM 2930 filter set,Hellmaanalyticshasmanufacturedanequivalentreferencematerial,whichhasbeenofficiallycertifiedbyNIST,and isavailableastheir666S300filterset.
aRticle no. 666S300
applicaton Filtersetfortestingthephotometricaccuracy
contentF390:glassfilter(0.04abs)F301:Neutraldensitiyglassfilter(2.5abs)F303:Neutraldensitiyglassfilter(3.0abs)
standaRd ceRtification Wavelengths: 440,465,546.1,590,635nmslit width:1nm
extended abs.range till < 3,1! neW
PREVIOUS ABSORBANCE RANGE: < to 2,05
190 300 400 500 600 700 800 8900
0,5
1
1,5
2
2,5
3
3,5Abs.
ERWEITERTE DAKKS-AKKREDITIERUNG FÜRZERTIFIZIERTE UV/VIS- REFERENZMATERIALIEN
nm
NEW ABSORBANCE RANGE: < to 3,1
extended dakks accreditation for certified uV-Vis reference materials
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230 430 530 6303300
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Abso
rptio
n
Wavelength (nm)
2
279.3360.8 453.4
536.4637.5
270 470 570 8703700
1
Abso
rptio
nWavelength (nm)
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329
2
3
770670
472
512
681
875
2.1 holmium oxide glass filter for checking wavelength accuracy
applicationTo measure wavelength accuracy, the filter absorbs thelightbeamof thespectrophotometer toagreaterextentatcertainwavelengthscreatingabsorbancepeaks.Ideally,anyreferencematerialsusedtodeterminewavelengthaccuracyshouldhavenarrow,well-definedpeaksatavarietyofwa-velengthsintheUVandvisiblerange.
pRoduct descRiptionHolmiumoxideglassfilter666-F1hasarangeofnarrow,well-definedpeaksintheUVandvisiblerange,makingholmiumanexcellentchoiceforcheckingthewavelengthscaleofspectro-photometers. In comparison to filters that use holmium oxi-de solutions, the holmium oxide glass filter has a somewhatweakerspectrumwithfewerpeaks.InthelowUVrangeinpar-ticular, theabsorbancebehaviorof theglassmatrix issuper-imposedontheholmiumpeaks.Themainadvantageofusingaglassfilteroveraliquidfilteristhatitismorerobust.
notesThepositionsofholmiumoxidepeaksmayvaryslightlydepen-dingontheglassbatchused.ThisiswhyHellmaanalyticscerti-fieseachholmiumoxideglassfilterindividually.
2.2 didymium glass filter for checking wavelength accuracy
applicationpleasesee2.1formoredetailsoftheapplication.
pRoduct descRiptionDidymiumglassfilter666-F7WismadefrommaterialspeciallymanufacturedbySchottag.Likeholmiumoxideglass,didymi-umglasshasavarietyofcharacteristicpeaksintheultravioletand visible rangeand is therefore typicallyused for checkingwavelengthaccuracy. Itspeaksarenotasnarrowasthoseofholmium oxide glass filters, however. The filter’s absorbancebehavior in theultraviolet rangealsomakes itsuitableasanabsorbancefilterforcheckingphotometricaccuracy(see2.3).
notesThe positions of didymium glass peaks may vary slight-ly depending on the glass batch used. This is why Hellmaanalyticscertifieseachdidymiumglassfilterindividually.
aRticle no. 666F1-339
application TestingthewavelengthaccuracyintheUVandVisrange(279nmto638nm)ataspectralbandwidthupto2nm.
content Holmiumoxideglassfilterwithmetalframe
standaRd ceRtification Wavelength accuracy at: 279;361;453;536;638nmslit width: 1nm
possible ceRtification additional possible wavelengths: 287;418;445;460nmslit width: allupto2nm
aRticle no. 666F7W-323or666F7-323
application TestingthewavelengthaccuracyintheUVandVisrange(379nmto875nm)ataspectralbandwidthupto2nm.
content Didymiumglassfilterwithmetalframe
standaRd ceRtification Wavelength accuracy at:379;472;512;681;875nm slit width:1nm
possible ceRtification additional possible wavelengths: 291;302;430;446;482;626nmslit width:allrecommendedupto2nm
Typicalspectrumforaholmiumoxideglassfilter Typicalspectrumforadidymiumglassfilter
2. Glass fIlters WaVelenGth accuRacy
Gla
ss f
ilte
Rs
Gla
ss f
ilte
Rs
1514
2.3 didymium glass filter for checking photometric accuracy
applicationTo measure photometric accuracy (absorbance), the filterreduces the light beam from the spectrophotometer. an ab-sorbancevalue(abs)canbededucedfromthelightextinctioncausedbythefilter.pRoduct descRiptionDidymiumglassfilter666-F7aismadefrommaterialspeciallymanufactured by Schott ag. The didymium glass filter’s ab-sorbancebehavior in theultravioletrangealsomakes itsui-table for use as an absorbance filter. Didymium glass filtersarethereforesuitableforcheckingwavelengthaccuracyintheUV-VisrangeaswellascheckingphotometricaccuracyintheUVrange.
absorbance behavior in the UV range can be checked at270nm,280nm,297nm,320nm,and340nm.Filtersarerou-tinelysetatathicknessthatproducesanominalopticaldensi-tyof0.5absat340nm.Thisresultsinincreasinglylargerab-sorbancestheshorterthewavelengthsbecome.
notesThese absorbance values vary greatly depending on theglass batch used, however, and can only be compared forfiltersderivedfromthesameglassmeltingprocess.Thisiswhyalldidymiumglassfiltersarecertifiedindividually.
Abso
rptio
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Wavelength (nm)
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ca. 0.9 A ca. 0.8 A
ca. 0.6 A
ca. 0.5 A
aRticle no. 666F7a-323or666F7-323
application TestingthephotometricaccuracyintheUVrange(270nmto340nm)
content Didymiumglassfilterwithmetalframe
standaRd ceRtification photmetric accuracy: approx.0.5to1abs.at wavelengths: 270;280;297;320;340nmslit width: 1nm
possible ceRtification additional possible wavelengths:from270to290nmand310to320nm slit width:allpossibleupto5nm
Selectedspectrumforadidymiumglassfilterbetween270nmand370nm
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Abso
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Wavelength (nm)
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0.6
0.8
1.0
1.2
1.4
1.6
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666-F2
666-F203
666-F3
666-F4
666-F202
666-F3666-F201
2.4 neutral density glass filter for checking photometric accuracy
applicationTo measure photometric accuracy (absorbance), the filterreduces the light beam from the spectrophotometer. an ab-sorbancevalue(abs)canbededucedfromthelightextinctioncausedbythefilter.
pRoduct descRiptionHellmaanalyticsneutraldensityglassfiltersaremadefromfilter materials produced by Schott ag, which were selec-ted on account of their homogeneity and stability. Thankstoarelativelyconstanttransmittancewithinthewavelengthrangeof405nmto800nm, theyhavebeenused tocheckphotometricaccuracyandlinearityinthevisiblewavelengthrange(>405nm)fordecades.
notesIfyouhaveseveralneutraldensityglassfilterswithdifferentnominal absorbances, you can check the linearity of yourabsorbancescalebyplottingtheabsorbancevaluesmeasu-redforeachwavelengthagainstthemeasurementvaluesontheDakkScalibrationcertificateinadiagram.
Typicalspectrumforneutraldensityglassfilters,measuredwithaslitwidthof1nm
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2. Glass fIlters photometRic accuRacy
aRticle no. 666F2-39,666F3-38,666F4-37,666F201-39,666F202-36,666F203-36
application TestingthephotometricaccuracyintheVisrange(405nmto890nm)
contentf2: neutraldensityglassfilterNg11(0.25abs),f3:neutraldensityglassfilterNg5(0.5abs),f4: neutraldensityglassfilterNg4(1.0abs),f201:neutraldensityglassfilterNg11(0.3abs)f202: neutraldensityglassfilterNg3(1.5abs),f203:neutraldensityglassfilterNg3(2.0abs)
standaRd ceRtification Wavelengths:440;465;546.1;590;635nmslit width: 1nm
possible ceRtificationall wavelengths possible from 405 to 890 nm.alsopossibleabove890nm,withHellmaanalyticscalibrationcertificateslit width: allpossibleupto5nm
Gla
ss f
ilte
Rs
Gla
ss f
ilte
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2.5 Glass filter sets
Specificallycreatedtomeetcustomerrequirements,Hellmaanalyticsglassfiltersetsconsistofexistingindividualfilterssuitableforstandardorcustomvalidationprocedures.
Toensurethat filterscanbeeasily identified, thesetnum-ber isengravedoneachfilter frame.Theabsorbance/peakpositionvaluesmeasuredforeachfiltercanbefoundontheDakkScalibrationcertificateprovided.
2.6 General usage guidelines for glass filters
glass filtersaremadeofglassdopedwithmetal ions/rareearthmetals,whichareassembledstress-freeinblackan-odizedprecisionframesmadeofaluminum.Theyaredesi-gnedtofitintoallspectrophotometersequippedwithahol-derforstandardcuvetteswitha10-mmopticalpathlength.To ensure that filters can be easily identified, each filterframe is engraved with the filter type and serial number.Detailsoftheabsorbanceandpeakpositionvaluesmeasu-redforeachfiltercanbe foundontherespectivecalibrati-oncertificate.pleaseensurethatyoudonottouchtheglasssurfacesofthefilter.Dirt,dust,anddamagecansignificant-ly impair the accuracy of measurement results. anodizedaluminumholdersshouldnotcome intocontactwithacidsoralkalis.
stoRaGeafter use, we strongly recommend storing the filters atroom temperature, in their packaging, and in a dry, dust-freearea.
otheR factoRs that may influence measuRementsDirt(e.g.fingerprints)andduston,ordamage(scratches,cor-rosion)to,polishedopticalsurfacescansignificantlyimpairtheaccuracy of measurement results. always store the filters intheiroriginalpackagingandprotect theopticalwindows fromcontamination.Onlyhandlethefiltersbytheirframes.
cleaninGDirtoftenaccumulatesonopticalsurfacesasaresultofre-gularuse.This isbest removedusinga lint-freeclothandalcoholorHellmanexIIIcleaningsolution.
influence of tempeRatuRe on measuRementsTemperature has a very small influence on certified measu-rementvalues,andtemperaturesbetween20°Cund24°Cfallwithin themeasurementuncertaintystatedonthecalibrationcertificate. Measurements should therefore be taken in thisrange tokeepanypotential temperature influenceon there-sultstoaminimum.
aRticle no. 666S000 666S001
applicationCompleteglassFilterSetfortestingthephotometricaccuracyundthewavelengthaccuracyofthespectrophotometer
glassFilterSetfortestingthewavelengthaccuracyandthephotometricaccuracyofthespectrophotometer
content
F1:holmiumoxideglassfilterF2:neutraldensityglassfilterNg11(0.25abs)F3:neutraldensityglassfilterNg5(0.5abs)F4:neutraldensityglassfilterNg4(1.0abs)F0:filterframewithoutglass(referencefilter)
F3:neutraldensityglassfilterNg5(0.5abs)F4:neutraldensityglassfilterNg4(1.0abs)F7:didymiumglassfilter(0.5-1.0abs)
standaRd ceRtification
f1: holmium oxide glass filter:Wavelength accuracy at:279;361;453;536;638nmslit width: 1nm
f2, f3, f4: neutral density glass filter:wavelengths: 440;465;546.1;590;635nmslit width: 1nm
f3, f4: neutral density glass filter:wavelengths: 440;465;546.1;590;635nmslit width:1nm
f7: didymium glass filter:wavelength accuracy at: 329;472;512;681;875nmphotometric accuracy:from0.5to1absslit width: 1nm
possible ceRtification
Wavelength accuracy; possible wavelengths: 287;418;445;460nmslit width:upto2nmrecommended
photometric accuracy:wavelengths: allpossiblefrom405to890nmalsopossibleabove890nm,withHellmaanalyticscalibrationcertificateslit width: allpossibleupto5nm
f3, f4: neutral density glass filter:wavelength accuracy: allpossiblefrom405to890nm.alsopossibleabove890nm,withHellmaanalyticscalibrationcertificateslit width: allpossibleupto5nm
didymium glass filter:wavelength accuracy:possible wavelengths: 291;302;430;446;482;626nmslit width: upto2nmrecommendedphotometric accuracy:
possible wavelengths:from270to290nmandfrom310to320nmslit width: allpossibleupto5nm
for further sets, please see our product overview on pages 48 to 50.
hellmanex iii cleaning concentrate// the cleaning concentrate significantly lowers surface
tension and ensures optimum surface wetting even when used on complex shapes or on very large surfaces
// it is extremely easy to rinse off, leaving no trace of the cleaner on the glass surface
// it uses special surfactants that cause no absorption above 288 nm, and does not interfere with subsequent measurements in the uV/Vis range// it is extremely compatible with materials, reduces corrosion of glass and surfaces, and preserves optical surface quality
aRticle no. pRoduct
9-307-011-4-507 HellmanexIII,1l(1.3kg)
9-307-011-5-507 HellmanexIII,10l(14kg)
9-307-011-6-507 HellmanexIII,25l(35kg)
oRdeR hotline+49 7631 [email protected]
2. Glass fIlters sets
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steps to taKe befoRe peRfoRminG calibRation With Glass filteRs
1. Warmupthespectrophotometeruntilthecorrectoperatingtemperaturehasbeenreachedandremainsconstant(e.g.foronehour),takingcaretoobservethedevicemanufacturer’sguidelines.
2. Makesurethatyouuseastablecuvetteholderfor10mmstandardcuvettestomeasurethefilters,asthisistheonlywaytoguaranteethebestpositioningofthefiltersinthelightpath.Checkthattheholderissecureandstableinthesamplecompartment.
3. Tobeginwith,carryoutabaselinecorrectionwithanemptysamplecompartment.
4. CheckthatthefilteriscorrectlypositionedinthelightpathbyfirstplacingemptyfilterholderF0inthecuvetteholder.TheF0markingmustbevisiblefromabove.ensurethatallfilterframesarealwayspositionedinthesameway,i.e.withserialnumbersfacingthelightsource.
5. Checkthatthedevice’sdisplayhasnotchanged.Inspectrophotometerswithverylargebeams,themeasurementbeammaytouchthefilterframe(beamclipping).Ifthisisthecase,youwillnoticeachangeinthedevice’sdisplay.
» Ifnecessary,adjusttheheightofthecuvetteholderuntilthelightbeamshinesthroughtheaperture unimpeded.Tohelp,youcanswitchthedevice’smeasurementbeamtovisiblei.e.byadjustingthemonochromatorto
500nm.Theremaybeotherwaysofdoingthisdependingonthedevice. » Ifthelightbeamtouchesthesidesoftheaperture,adjustthehorizontalpositionofthecuvetteholderuntil thelightbeamshinesthroughthecenteroftheaperture.Thefilterframeiscorrectlypositionedifthedisplayvalues,
fromthezeroadjustmentperformedinstep3(baselinecorrection),donotchange.Inrarecaseswherethezero readingcannotberetainedafterinsertingtheemptyfilterframeandcarryingouttheaboveproceduresitispermissible tore-zerotheinstrumentwiththeemptyfilterframeinplaceandthencontinuewiththefiltermeasurements.
6. Carryoutthefiltermeasurementinaclosedsamplecompartmentascarefullyasyouwouldcarryoutasamplemeasurement
(opensamplecompartmentsproduceincorrectresults).
7. pleasenotethat,ifyouareusingadiodearrayspectrophotometerwithastand-alonecuvetteholderconnectedviaafiber-opticcable,extraneouslightandvibrations(e.g.movementoffiber-opticcables)mayalsoimpairtheaccuracyofmeasurementresults.
steps foR checKinG WaVelenGth accuRacy With a holmium oxide Glass oR didymium Glass filteR
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithglassfilters’.
2. Runthescanprogramonyourspectrophotometer,observingtheguidelinesintheusermanual.Selectascanningrangethatcoversallofthepeakslistedonthefilter’scalibrationcertificate.
3. Setyourspectrophotometertothemeasurementparametersthatappearonthecalibrationcertificateprovided.Selecttheslowestscanningspeedandasmalldatainterval.
4. Ifpossible,carryoutabaselinecorrection.
5. Measurementsaretakenusinganairblankwhichmeans,whichmeansthatthereferencecuvetteholderremainsemptyindoublebeamphotometers,whileareferencemeasurementistakenusingtheemptycuvetteholderinsinglebeamphotometers.
6. Inserttheholmiumoxideglassordidymiumglassfilterintothecuvetteholder.ensurethatthefilterisinsertedintotheholderasfarasitwillgo,andthatthefilterIDcanbeseenfromabove.Thefiltersmustalwaysbepositionedinthecuvetteholderinthesameway,i.e.withtheserialnumberfacingthelightsource.
7. Startthemeasurement.
8. Calculatethepositionsofthepeaksatthewavelengthsstatedonthecalibrationcertificate.(Takeseveralmeasurementsandthenusethemeanofthemeasuredvaluestoavoiderrors).
9. Compareyourmeasurementvalueswiththecertifiedones.
measuRement paRameteRs foR checKinG WaVelenGth accuRacyensurethatyouhaveselectedthecorrectmeasurementpara-metersbeforeplottingtheabsorbancecurvetocalculatepeakpositions. Incorrect parameters may distort the absorbancecurveandthusshifttheactualpositionsofpeaks.pleaseusethesettingsstatedontheaccompanyingcalibrationcertificate.Itshouldbenotedthatchangingtheslitwidthofthespectro-photometercancausetheabsorptionmaximatoshiftslightly.Ignoreanyinfluencethatthespectralbandwidthfrom1nmto2nmhasonpeakpositions.peakheights,however,mayvarygreatlyfollowingchangestotheslitwidthduetotheirnarrownature. as a result, filters for checking wavelength accuracyareusuallyunsuitableforcheckingabsorbanceaccuracy.
»Generally speaking, filters can also be mea-sured using a slit width that differs from the in-formation provided on the calibration certificate. however, please note that large slit widths will prevent weaker peaks from being resolved. in ca-ses of doubt, it is therefore advisable to choose as small a slit width as possible. We recommend taking several measurements and then using the mean value to avoid errors during evaluation.«
thomas brenn,productManager
We look forward to hearing any comments,
criticism or suggestions you may have:
2. Glass fIlters2.7 calibration with glass filters
pRepaRations
Watch a video of theindividual steps here.
Watch a video of theindividual steps here.
2. Glass fIlters2.7 calibration with glass filters
WaVelenGth accuRacy
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Therearemanydifferentmakes,modelsand
designsofspectrophotometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
Therearemanydifferentmakes,modelsand
designsofspectrometer,thesestepsareonly
generalguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
2120
steps foR checKinG photometRic accuRacy With neutRal density Glass oR didymium Glass filteRs
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithglassfilters’.
2. Runthewavelengthselectionprogramonyourspectrophotometer,observingtheguidelinesintheusermanual.Selectthewa-velengthsprovidedonthecalibrationcertificate.
3. Setyourspectrophotometertothemeasurementparametersthatappearonthecalibrationcertificateprovided.
4. adjusttozero.
5. Measurementsaretakenusinganairblankwhichmeansthatthereferencecuvetteholderremainsemptyindoublebeamspec-trophotometers,whileareferencemeasurementistakenusingtheemptycuvetteholderinsinglebeamspectrophotometers.
6. Inserttheneutraldensityglassordidymiumglassfilterintothecuvetteholder.ensurethatthefilterisinsertedintotheholderasfarasitwillgo,andthatthefilterIDcanbeseenfromabove.Thefiltersmustalwaysbepositionedinthecuvetteholderinthesameway,i.e.withtheserialnumberfacingthelightsource.
7. Starttheprogramformeasuringtheabsorbancevaluesatthewavelengthsstatedonthecalibrationcertificate.(Takeseveralmeasurementsandthenusethemeanofthemeasuredvaluestoavoiderrors).
8. Compareyourmeasurementvalueswiththecertifiedones.
inteRpRetinG the measuRement Results of Glass filteRs foR checKinG photometRic and WaVelenGth accuRacy Themeasurementuncertaintiesthatappearonthecalibrationcertificate only refer to measurements conducted by Hellmaanalytics and apply solely to the measurement conditions atthe company (spectrophotometer used, environmental influ-encessuchas temperature,airhumidity,user influence,andreferencematerialsused).
The smallest possible measurement uncertainty can then bederivedbystatisticallycombiningthemeasurementuncertaintystatedonthecalibrationcertificateandalloftheuser’suncer-tainty contributions. These include the wavelength scale tole-rance of the spectrophotometer used and other influences onmeasurementaccuracy(environmentalfactorssuchastempe-rature,airhumidity,user influence,etc.).For further literatureoncorrectlycalculatingmeasurementuncertainty,pleaserefertochapter8ofthisusermanual.
see the faqs for more details.
2. Glass fIlters2.7 calibration with glass filters
photometRic accuRacy
2. Glass fIlters2.7 calibration with glass filters
inteRpRetinG measuRement Results
Watch a video of theindividual steps here.
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»in practice it is easier to simply add up uncer-tainty contributions than to combine their stati-stics. however, the method used to determine measurement uncertainty depends on the speci-fications of your quality system and your measu-rement accuracy requirements.«
carola steinger,ChemicalLaboratory Technician
Therearemanydifferentmakes,modelsand
designsofspectrophotometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
2322
3.1 holmium oxide liquid filter for checking wavelength accuracy
applicationTomeasurewavelengthaccuracy, the filterreduces the lightbeamofthespectrophotometertoagreaterextentatcertainwavelengths(peaks).Ideally,anystandardsusedtodeterminewavelengthaccuracyshouldhavenarrow,well-definedpeaksatavarietyofwavelengthsintheUVandvisiblerange.
pRoduct descRiptionTheholmiumoxideliquidfilterconsistsofasolutionofhol-miumoxidedissolvedinperchloricacid.Thisfilterisideallysuited tochecking thewavelengthaccuracyofspectropho-tometersintheUVandvisiblerange.Ithasaspectrumwitha variety of characteristic, very well-defined peaks in therangebetween240nmand650nm.
3.2 potassium dichromate liquid filter for checking photometric accuracy
applicationphotometricaccuracy (absorbance) ismeasuredbyshiningalight beam from the spectrophotometer through the insertedfilter.anabsorbancevalue(abs)canbededucedfromthelightextinctioncausedbythefilter.
pRoduct descRiptionpotassium dichromate in perchloric acid is very suitable forcheckingthephotometricaccuracyofspectrophotometers. Inthe UV range, the potassium dichromate spectrum has cha-racteristicmaximaat257nmand350nmandcharacteristicminimaat235nmand313nm.Thespectrumreachesapla-teauat430nm,whichisusedtodeterminephotometricaccu-racy in the visible range. Hellma analytics purchases the re-ferencematerial forthis filterdirectly fromNIST(SRM®935a
“potassiumDichromate”).Thefiltersolutionsaremanufactu-redinstrictcompliancewithNISTrequirementsandfilledun-der controlled conditions. The cuvettes are then immediatelyfusedtobecomeairtight.
notesas the filters are certified individually, measurement resultsare free fromsystematicerrorsmadewhenpreparingsoluti-onsandwithregardstotheopticalpathlengthofthecuvette.The measurement values of reference filter UV14 (perchloricacidmeasuredagainstanairblank)appearseparatelyontheDakkS calibration certificate. To check absorbance linearity,take measurements using potassium dichromate filters withdifferentconcentrations.plottheabsorbancevaluesmeasuredforeachfilterandwavelengthagainstthemeasurementvaluesthatappearontheDakkScalibrationcertificateinagraph.
230 430 530 6303300
0.5
Abso
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Wavelength (nm)
1
640.51
536.59
485.25
451.47
416.30
361,28
345.41
333.49
287.20278.12
241.15
aRticle no. 667005
application TestingthewavelengthaccuracyaccordingtotheeuropeanpharmacopoeiaintheUV/Visrange
content Holmiumoxideinperchloricacid
standaRd ceRtification wavelengths:241;287;361;536;640nmslit width: 1nm
possible ceRtifcation additional wavelengths: 250;278;333;345;386;416;451;468;485nmslit width:allupto2nm,abovepeaksbecomeindistinct
aRticle no. 667020,667040,667060,667080,6670100,667600,667014(referencefilter)
application TestingthephotometricaccuracyintheUVrange(235to350nm)undVisrange(measurementwavelength430nm)ataspectralbandwidthof2nmorless
content
UV20,20mgpotassiumdichromateinHCL04(0.25abs)UV40,40mgpotassiumdichromateinHCL04(0.5abs)UV60,60mgpotassiumdichromateinHCL04(0.75abs),inaccordancewiththeeuropeanpharmacopoeiaUV80,80mgpotassiumdichromateinHCL04(1.0abs)UV0100,100mgpotassiumdichromateinHCL04(1.25abs)UV600,600mgpotassiumdichromateinHCL04(1.0abs)inaccordancewiththeeuropeanpharmacopoeiaUV14,perchloricacid(HCLO4),referencefilter
standaRd ceRtificationat uV20 - uV100 wavelengths: 235;257;313;350nmat uV600 wavelength: 430nmslit width:2nm
possible ceRtifcation wavelength: fixedslit width:allupto2nm
Typicalspectrumofholmiumoxidedissolvedinperchloricacid,measuredataslitwidthof1nm.
0200 300 400 500
257
350235
313
Abso
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Wavelength (nm)
0.5
0.9
3
2
1
0380 420 460 500
430 nm
Abs
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Wavelength (nm)
Typicalspectrumofa0.006%potassiumdichromatesolution
Typicalspectrumofa0.06%potassiumdichromatesolution
We would be interested to hear about your
experiences with reference materials.
Write to us at [email protected]
3. lIquId fIlters WaVelenGth accuRacy
3. lIquId fIlters photometRic accuRacy
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3.3 liquid filters for checking for stray light
applicationIn a spectrophotometer, stray light is light that passes bythesampleand fallsdirectlyon thedetector.Thiscan lead toincorrect measurement results. Stray light may be caused byscatteringordiffraction,bypooropticalalignment,theuseofin-correctordamagedcuvettes,incorrectlyfittedsamplingacces-soriesordamagedsealsaroundalight-tightsamplechamber.Straylightisproblematic,asitreducestherangeofmeasurab-leabsorbanceandimpairsthelinearitybetweenconcentrationand absorbance. Cut-off filters (filters with a strictly definedspectrum)arerequiredtocheckthedeviceforstraylight.
pRoduct descRiptionHellma analytics stray light filters do not allow light topassthroughthembelowacertainwavelength(cut-offwa-velength).anytransmittancevaluesdisplayedinthecut-offwavelengthrangethereforerepresentstraylight.
Due to their strictly defined spectrum, potassium chloridefilters, sodium iodide filters, and sodium nitrite filters areideally suited to qualifying the stray light level of spectro-photometersincompliancewithpharmacopoeias.Thestepsarethesameforallstraylightfilters.
3.4 liquid filters for checking spectral resolution
applicationRegularly checking the spectral resolution of spectrophoto-meters ensures, for example, that neighbouring peaks areresolvedandnotsuperimposedonthepeaksofborderingwa-velengths.Thisalsopreventsabsorbanceerrors.
pRoduct descRiptionThetolueneinhexaneliquidfilterhasaprominentpointinitsspectrum,whichisexcellentfordeterminingthespectralreso-lutionand/oractualslitwidthofspectrophotometersincomp-liancewiththeeuropeanpharmacopoeia.
notesaspectrophotometer’sspectralresolutionisverycloselycon-nected to the correct slit width setting and characterized byitsabilitytoresolve(recognize)twoverycloselyrelatedpeaks.Thesmallertheslitandcorrespondingspectralbandwidth,thehighertheresolution.
aRticle no. 667001,667010,667011,667012(referencefilter)
application TestingforstraylightintheUVrange(UV1accordingtotheeuropeanpharmacopoeia)(atwavelengthsfrom198nmto370nm,dependingonthefilterselected)
content
UV1,potassiumchlorideinpurewaterUV10,sodiumiodideinpurewaterUV11,sodiumiodideinpurewaterUV12,purewater(referencefilter)
standaRd ceRtification
uV1:cut-offat200nmuV10:cut-offat259nmuV11: cut-offat385nmslit width: 2nm
possible ceRtifcation wavelengths: fixedpossible slit widths: allupto5nm
aRticle no. 667006,667009
application Testingtheresolutionaccodingtotheeuropeanpharmacopoeia
content UV6,tolueneinn-hexaneUV9,n-hexane(referencefilter)
standaRd ceRtification
wavelength: scanfrom265to270nmslit width: 0.5;1.0;2.0nmwithHellmaanalyticsCalibrationCertificate(noDakkSCalibrationCertificate)
possible ceRtifcation
wavelength: fixedpossible slit widths: from0.5to3nm
200 250 300 350 400 450 500
KCl(667-UV1)
NaI(667-UV10)
NaNO2(667-UV11)
H2O(667-UV12)
Abso
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1
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Wavelength (nm)
0.5
1.5
2.5
Abso
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265 270269268267266
0.3
0.1
0.2
0.5
0.4
Wavelength (nm)
slit 0.25
slit 4.0
slit 3.0
slit 2.0
slit 1.0
slit 0.5
0.3
0.1
0.2
0.5
0.4
Typicalspectraoftheliquidfiltertoluene,measuredwithdifferentslitwidths
3. lIquId fIlters stRay liGht
3. lIquId fIlters spectRal Resolution liquid filteRs
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»as a rule of thumb, the slit width should be no more than 10% of the peak width at half maximum in order to be able to determine its absorbance with an accuracy of 99.5%. two peaks are deemed to be resolved separately if the minimum absorbance between them amounts to less than 80% of the maximum peak. impairments to the spectrophoto-meter’s spectral resolution will cause two different peaks to be shown as a combined peak, leading to inaccurate measurement results.«
timo Rapp,ChemicalLaboratoryTechnician
2726
3.5 liquid filter set compliant with the european pharmacopoeia
The complete liquid filter set 667003 was compiled on thebasis of european pharmacopoeia requirements and con-tainsallfiltersrequiredtocarryoutacompletespectropho-tometercheck.
checking photometric accuracy (uV60, uV600, uV14)
checking wavelength accuracy (uV5) checking for stray light (uV1 and uV12)
checking spectral resolution (uV6 and uV9)
all liquid filters consist of reference materials that arefilled into precision Hellma cuvettes made of quartz glass(SUpRaSIL®). These cuvettes are permanently sealed, andthecompletesetisdeliveredinahigh-qualitystoragebox.Toensureeasyidentification,eachfilterisengravedwithitsserialnumber.ThecalibrationvaluesmeasuredforeachfiltercanbefoundontheDakkSandHellmaanalyticscalibrationcertifica-tesprovided.
3.6 liquid filter set compliant with the united states pharmacopeia
3.7 General usage guidelines for liquid filters
Liquidfiltersbearamarkingononesideshowingthechemi-calformulaofthesubstancecontainedinthecuvette. Ifafil-terbreaks,pleaseobservethecodesofconductandsafetyin-structionsthatapplytothissubstance.Thisinformationcanbefoundinthesafetyinstructions.Up-to-datesafetyinstructionsforallsubstancesusedtomanufactureliquidfiltersareavai-lableatwww.hellma-analytics.com/download.
stoRaGeafter use, we strongly recommend storing the filters atroom temperature, in their packaging, and in a dry, dust-freearea.Liquidfiltersmustnotbeexposedtotemperatu-resbelow4°Corabove40°C.Thisalsoapplieswhentrans-portinganddeliveringliquidfiltersforrecertification.
otheR factoRs that may influence measuRementsDirt (e.g. fingerprints) and dust on, or damage (scratches,corrosion)to,polishedsurfacescansignificantly impairtheaccuracy of measurement results. always store the filtersin theiroriginalpackagingandprotect theopticalwindowsfromcontamination.Onlyhandlethefiltersbytheircapsormattsurfaces.
cleaninGDirtoftenaccumulatesonopticalsurfacesasaresultofregularuse.Thisisbestremovedusingalint-freeclothandalcoholorHellmanexIIIcleaningsolution.
influence of tempeRatuRe on measuRementsTemperaturehasaverysmall influenceoncertifiedmeasu-rementvalues,andtemperaturesofbetween20°Cund24°Cfall within the measurement uncertainty stated on the cali-brationcertificate.Measurementsshould thereforebe takeninthisrangetokeepanypotentialtemperatureinfluenceontheresultstoaminimum.
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for further information, please see page 17
aRticle no. 667003
application Completefiltersetfortestingthespectrophotometeraccordingtotheeuropeanpharmacopoeiaphotometricaccuracy,wavelengthaccuracy,straylightandresolution
content
UV1,potassiumchlorideinpurewaterUV12,purewater(referencefilter)UV5,holmiumoxideinHClO4UV6,tolueneinn-hexaneUV9,n-hexane(referencefilter)UV60,60mgpotassiumdichromateinHClO4UV600,600mgpotassiumdichromateinHClO4UV14,perchloricacid(referencefilter)
standaRd ceRtification
uV1/uV12: wavelength: 200nm(cut-off)uV5: wavelenght:241;287;361;536;640nm;slit width:1nmuV6/uV9: wavelength: scanfrom265to270nm; slit width: 0.5;1.0;2.0nmuV60: approx.0.75abs; wavelengths:235;257;313;350nm; slith width: 2nmuV600: approx.1.0abs; wavelength: 430nm; slit width: 2nm
possible ceRtification
uV1/uV12: wavelength: fixed; slit width:allupto5nmuV5: wavelenght: 250,278,333,345,386,416,451,468,485nm; slit width: allupto2nm;above,peaksbecomeindistinctuV6/uV9: wavelength: fixed;slit width:0.5to3nmuV60: wavelength: fixed; slit width:allupto2nmuV600: wavelength: fixed; slit width:allupto2nm
aRticle no. 667004
application FiltersetfortestingthespectrophotometeraccordingtoUSp851,photometricaccuracyandwavelengthaccuracy
content
F2:neutraldensityglassfilter(0.25abs)F3:neutraldensityglassfilter(0.5abs)F4:neutraldensityglassfilter(1.0abs)F0:filterframewithoutglass(referencefilter)UV60:60mgpotassiumdichromateinHClO4UV14:perchloricacid(HClO4,referencefilter)UV5:holmiumoxideinHClO4
standaRd ceRtification
f2, f3 and f4: wavelengths: 440,465,546.1,590,635; slit width:1nmuV60: wavelengths:235,257,313,350; slit width: 2nmuV5: wavelengths: 241,250,278,287,333,345,361,385,416,452,468,485,536,640nm;slit width: 1 nm
possible ceRtification
f2, f3, f4: wavelengths: allpossiblefrom405to890nmalsopossibleabove890nmwithHellmaanalyticsCalibrationCertificateslit width:allpossibleupto5nmuV60: wavelength:fixed; slit width: upto2nmuV5: wavelength: fixed, slit width: allupto2nm;above,peaksbecomeindistinct
always take great care when placing li-quid filters in the sample holder of your spectrophotometer. Wherever possible, only touch filters by their caps or matt sides. take care not to touch the polis-hed surfaces. the filters are fragile and should be handled with the utmost care.
3. lIquId fIlters sets
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steps to taKe befoRe peRfoRminG calibRation With liquid filteRs
1. Warmupthespectrophotometeruntilthecorrectoperatingtemperaturehasbeenreachedandremainsconstant(e.g.foronehour),takingcaretoobservethedevicemanufacturer’sguidelines.
2. Makesurethatyouuseastablecuvetteholderfor10mmstandardcuvettestomeasuretheliquidfilters,asthisistheonlywaytoguaranteethebestpositioningofthefiltersinthelightpath.Checkthattheholderissecureandstableinthesamplecompartment.
3. Thefiltersshouldalwaysbepositionedinthecuvetteholdersinthesameway,i.e.withtheHellmaletteringfacingthelightsource.Thelightbeammustpassthroughthepartofthefilterfilledwithliquid(solution).
4. Carryoutthefiltermeasurementinaclosedsamplecompartmentascarefullyasyouwouldcarryoutasamplemeasure-ment(opensamplecompartmentsproduceincorrectresults).
5. pleasenotethat,ifyouareusingadiodearrayspectrophotometerwithastand-alonecuvetteholderconnectedviaafiber-opticcable,extraneouslight,extraneouslightandvibrations(e.g.movementoffiber-opticcables)mayalsoimpairtheaccuracyofmeasurementresults.
measuRement paRameteRs foR checKinG WaVelenGth accuRacyensurethatyouhaveselectedthecorrectmeasurementpa-rameters before plotting the absorbance curve to calcula-te peak positions. Incorrect parameters may distort the ab-sorbancecurveandthusshift theactualpositionsofpeaks.pleaseusethesettingsstatedontheaccompanyingcalibra-tioncertificate.Itshouldbenotedthatchangingtheslitwidthof thespectrophotometercancause theabsorptionmaximatoshiftslightly. Ignoreany influencethat thespectralband-widthfrom1nmto2nmhasonpeakpositions.peakheights,however,mayvarygreatlyfollowingchangestotheslitwidthdue to their narrow nature. as a result, filters for checkingwavelengthaccuracyareusuallyunsuitableforcheckingab-sorbanceaccuracy.
share your experience of our
products by writing to:
see the faqs for more details.
steps foR checKinG WaVelenGth accuRacy With a holmium oxide liquid filteR
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithliquidfilters’.
2. Runthescanprogramonyourspectrophotometer,observingtheguidelinesintheusermanual.Selectascanningrangethatcoversallofthepeakslistedonthefilter’scalibrationcertificate.
3. Setyourspectrophotometertothemeasurementparametersthatappearonthecalibrationcertificateprovided.Selecttheslowestscanningspeedandasmalldatainterval.
4. Ifpossible,carryoutabaselinecorrection.
5. MeasurementsaretakenagainsttheperchloricacidblankUV14,whichmeansthatthereferencecuvetteholderremainsemptyindoublebeamphotometers,whileareferencemeasurementistakenusingtheemptycuvetteholderinsinglebeamphotometers.
6. Inserttheholmiumoxideliquidfilterintothecuvetteholder,observingthegeneralusageguidelinesforliquidfilters.Thefiltersshouldalwaysbepositionedinthecuvetteholdersinthesameway,i.e.withtheHellmaletteringfacingthelightsource.
7. Startthemeasurement.
8. Calculatethepositionsofthepeaksatthewavelengthsstatedonthecalibrationcertificate.(Takeseveralmeasurementsandthenusethemeanofthemeasuredvaluestoavoiderrors).
9. Compareyourmeasurementvalueswiththecertifiedones.
3. lIquId fIlters3.8 calibration with liquid filters
pRepaRations
3. lIquId fIlters3.8 calibration with liquid filters
WaVelenGth accuRacy
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»Generally speaking, filters can also be mea-sured using a slit width that differs from the in-formation provided on the calibration certificate. however, please note that large slit widths will prevent weaker peaks from being resolved. in cases of doubt, it is therefore advisable to choose as small a slit width as possible. We recommend taking several measurements and then using the mean value to avoid errors during evaluation.«
carola steinger,ChemicalLaboratory Technician
Therearemanydifferentmakes,modelsand
designsofspectrophotometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
Therearemanydifferentmakes,modelsand
designsofspectrometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
3130
steps foR checKinG photometRic accuRacy With a potassium dichRomate liquid filteR
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithliquidfilters’.
2. Runthewavelengthselectionprogramonyourspectrophotometer,observingtheguidelinesintheusermanual.Selectthewavelengthsprovidedonthecalibrationcertificate.
3. Setyourspectrophotometertothemeasurementparametersthatappearonthecalibrationcertificateprovided.
4. adjusttozero.
5. asageneralrule,measurementsaretakenusingareferencefilterfilledwithperchloricacidobservingthegeneralusageguidelinesforliquidfilters.Thefiltersshouldalwaysbepositionedinthecuvetteholdersinthesameway,i.e.withtheHellmaanalyticsletteringfacingthelightsource.
6. Measurementsinasinglebeamspectrophotometer:Carefullyinserttheperchloricacidreferencefilterprovidedintothecu-vetteholder.Startthemeasurement.Next,measurethecertifiedreferencematerial,whichcontainspotassiumdichromatedissolvedinperchloricacid.Thensubtractthereferencemeasurementvaluesfromthemeasurementvaluesofthecertifiedreferencematerial.Orsetzero/blanktheinstrumentwiththeperchloricacidreferencefilterUV14fitted,thenreplacewiththecertfiedreferencematerialandreadthevaluedirectlyfromthedisplay.
7. Measurementsinadoublebeamspectrophotometer:Carefullyinsertthecertifiedreferencematerial,whichcontainspotassiumdichromatedissolvedinperchloricacid,intothesampleholder,andtheperchloricacidreferencefilterintothereferencesampleholder.
8. Starttheprogramformeasuringtheabsorbancevaluesatthewavelengthsstatedonthecalibrationcertificate.(Takeseveralmeasurementsandthenusethemeanofthemeasuredvaluestoavoiderrors).
9. Compareyourmeasurementvalueswiththecertifiedones.
measuRement paRameteRs foR checKinG photometRic accuRacyasthedifferencebetweenthemaximaandminimaintheab-sorbancespectrumisrelativelylarge,thepotassiumdichro-mateliquidfiltersmayalsobemeasuredwithaslitwidththatdiffers from the one on the calibration certificate. However,pleasenote thatusing largeslitwidths (>2nm)mayresultinslightdeviationsfromthevaluesstatedonthecalibrationcertificate.Incasesofdoubt,itisthereforeadvisabletochoo-setheslitwidthquotedonthecalibrationcertificate.Were-commend takingseveralmeasurementsand thenusing themeanvaluetoavoiderrorsduringevaluation.
inteRpRetinG the measuRement Results of liquid filteRs foR checKinG photometRic and WaVelenGth accuRacy The measurement uncertainties that appear on the calibrati-oncertificateonlyrefertomeasurementsconductedbyHellmaanalyticsandapplysolelytothemeasurementconditionsatthecompany (spectrophotometer used, environmental influencessuchastemperature,airhumidity,userinfluence,andreferencematerialsused).
The smallest possible measurement uncertainty that canbeachievedbytheusercanthenbederivedbystatistical-ly combining the measurement uncertainty stated on thecalibration certificate with all the user’s uncertainty con-tributions, such as the wavelength scale tolerance of thespectrophotometerusedandother influencesonmeasu-rementaccuracy(environmentalfactorssuchastempera-ture, air humidity, user influence, etc.). For further lite-rature on correctly calculating measurement uncertainty,pleaserefertochapter8ofthisusermanual.
»in practice it is easier to simply add up uncer-tainty contributions than to combine their stati-stics. however, the method used to determine measurement uncertainty depends on the speci-fications of your quality system and your measu-rement accuracy requirements.«
birgit Kehl,HeadHellmaanalyticscalibrationlaboratory
share your experience of our
products by writing to:
see the faqs for more details.
3. lIquId fIlters3.8 calibration with liquid filters
photometRic accuRacy
3. lIquId fIlters3.8 calibration with liquid filters
inteRpRetinG measuRement Results
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Therearemanydifferentmakes,modelsand
designsofspectrophotometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
3332
measuRement paRameteRs foR checKinG stRay liGht leVelFor a realistic calculation of the stray light level, choose afilter with a cut-off wavelength as close above the requiredwavelength as possible. This enables the stray light test tobecarriedoutat thewavelengthatwhichthestray light fil-ter can fully absorb light. The remaining transmittancedisplayed by the device at the measurement wavelengthrepresents the stray light level. Since this value differs de-pending on the properties of the measuring system, filterscan only be certified with regard to their suitability for useas a stray light filter. Certification therefore demonstra-tes that filters have virtually full absorbance in the measu-ring range and steep peaks at high transmittance values.
You can check the lower absorbance range of your spectro-photometerusingreferencefilter667-UV12,whichisfilledwithultrapure water. The filter’s absorbance characteristics from200 nm to NIR are practically only determined by the reflec-tion losseson the twoair/glasssurfaces. Youcancheck yourdevice’sdisplayatverylowabsorbancevaluesagainstthecer-tifiedvaluesat198nm,200nm,300nmand400nm.Ifyourre-sultsdiffersignificantlyfromthecertifiedvalues,particularlyifthemeasuredvaluesaresmallerthan0.02a,youshouldcon-tactthedevicemanufacturer.
inteRpRetinG measuRement Results When checKinG foR stRay liGhtToestimatethesamplemeasurementerrorduetostraylight,comparethecalculatedstraylightleveltothesignalstrengthfromthesamplemeasurement.Forexample,astraylightva-lueof0.1%transmittanceandasamplewithanabsorbanceofaround1abswouldequatetoameasurementerrorduetostraylightofaround0.4%.Ifyouhavecalculatedastraylightlevel that is considerably higher than the level stated in thedevice specifications, check whether extraneous light couldhaveinterferedwiththisresult.Ifyoucanruleoutextraneouslight,pleasecontactaservicetechnician.
please note that the reference for the measuring system to be tested is not provided by the entire transmittance characteristic of the stray light filter, but solely by the transmittance value measured in the range of virtually full absorbance.
steps foR checKinG the stRay liGht leVel
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithliquidfilters’.
2. Runthescanprogramonyourspectrophotometer,observingtheguidelinesintheusermanual.Selectascanningrangethatcoversallofthevalueslistedonthefilter’scalibrationcertificate.
3. Setyourspectrophotometertothemeasurementparametersquotedonthecalibrationcertificateprovided
4. Setthespectrophotometertoawavelengthofapprox.20nmabovethecut-offwavelengthforthestraylightfilterused(forpotassi-umchloride(UV1),forexample,startat220nm)andscanuptothewavelengthforwhichyouwishtodeterminethestraylightlevel.
5. Ifpossible,carryoutabaselinecorrection.
6. asageneralrule,takethemeasurementsusingareferencefilterfilledwithwater(UV12)observingthegeneralusagegui-delinesforliquidfilters.Thefiltersshouldalwaysbepositionedinthecuvetteholdersinthesameway,i.e.withtheHellmaanalyticsletteringfacingthelightsource.
7. Measurementsinasinglebeamspectrophotometer:Carefullyinsertthereferencefilterprovidedintothecuvetteholder.Startthemeasurement.Next,measurethecertifiedreferencematerial.Thensubtractthereferencemeasurementvaluesfromthemeasurementvaluesofthecertifiedreferencematerial.
8. Measurementsinadoublebeamspectrophotometer:Carefullyinsertthecertifiedreferencematerialintothesampleholderandthereferencefilterintothereferencesampleholder.Startthemeasurement.
9. Scanuptothewavelengthforwhichyouwishtodeterminethestraylightlevel.
Thelightlevel(remainingtransmittancevalue)measuredbelowthecut-offwavelengthrepresentsstraylight.10.
3. lIquId fIlters3.8 calibration with liquid filters
stRay liGht leVel
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Therearemanydifferentmakes,modelsand
designsofspectrophotometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
3534
measuRement paRameteRs When checKinG spectRal ResolutionWhen measuring spectral resolution, the liquid filter ab-sorbs the light beam from the spectrophotometer to si-gnificantly different extents in a narrow wavelength range(5nm).Thefilterwillshowaclearmaximumandminimumwithinthenarrowrange.afterplacingtheliquidfilterinthespectrophotometer, run the scan program in the definedwavelengthrangeanddividethemaximumpeakmeasuredatλmax=269nmbytheminimumpeakmeasuredatλmin266nm.Theresultingratiorepresentstheabsorbanceratio,whichisdirectlylinkedtotheslitwidth.Iftheratioisconsi-derablylower(e.g.15%),pleasecontactthedevicemanufac-turer.pleasenote,however,thattheresultalsodependsonthe measurement conditions. Therefore, please make surethatyouselectasufficiently long integration time,particu-larlyifusingasmallslitwidth.
inteRpRetinG measuRement Results When checKinG spectRal ResolutionRegulatory codes or internal applications and measuringproceduresmayplacerequirementsontheratiosthatmustbe achieved. In addition, comparing calculated ratios withcertified valuesmayprovidean indicationof theactual slitwidthofthedeviceused.
steps foR checKinG spectRal Resolution
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithliquidfilters’.
2. Runthescanprogramonyourspectrophotometer,observingtheguidelinesintheusermanual.Selectascanningrangethatcoversbothoftherequiredpeaks.
3. Setyourspectrophotometertothemeasurementparametersstatedonthecalibrationcertificateprovided.
4. Ifpossible,carryoutabaselinecorrection.
5. Takethemeasurementusingareferencefilterfilledwithhexane–ifthespectrumiscorrectedtozeroat300nm,measu-rementscanalsobetakenusingair–observingthegeneralusageguidelinesforliquidfilters.Thefiltersshouldalwaysbepositionedinthecuvetteholdersinthesameway,i.e.withtheHellmaanalyticsletteringfacingthelightsource.
6. Measurementsinasinglebeamspectrophotometer:Carefullyinsertthehexanereferencefilterprovidedintothecuvetteholder.Startthemeasurement.Next,measurethecertifiedreferencematerial,whichcontainstolueneinhexane.Thensubtractthereferencemeasurementvaluesfromthemeasurementvaluesofthecertifiedreferencematerial.
7. Measurementsinadoublebeamspectrophotometer:Carefullyinsertthetolueneinhexaneliquidfilterintothesampleholderandthehexanereferencefilterintothereferencesampleholder.Startthemeasurement.
8. Measuretheactualminimumoftheabsorbancevaluesat266nmandtheactualmaximumat269nm.(Takeseveralmeasure-mentsandthenusethemeanofthemeasuredvaluestoavoiderrors).
9. Ifpossible,carryoutabaselinecorrectioneachtimeyouchangetheslitwidth.
Determinetheratiousingbothofthemeasuredvalues,asstatedonthecalibrationcertificateprovided.10.
please note that the filter set for de-termining spectral resolution does not fall within our scope of accredit-ation, and therefore cannot be issued with a dakks calibration certificate or calibration mark.
slit Width absoRbance Ratio
0,25 2,3
0,5 2,2
1,0 2,0
2,0 1,4
3,0 1,1
4,0 1,0
absoRbance Ratio of maximum/minimum peaK in Relation to slit Width
(seeStandardsandBestpracticeinabsorptionSpectrometry,editedbyC.Burgess&T.Frost)
share your experience of our
products by writing to:
3. lIquId fIlters3.8 calibration with liquid filters
spectRal Resolution
3. lIquId fIlters3.8 calibration with liquid filters
inteRpRetinG the measuRement Results
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Therearemanydifferentmakes,modelsand
designsofspectrophotometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
3736
4. certIfIed reference plates photometRic accuRacy
4. certIfIed reference plates photometRic and WaVelenGth accuRacy
4.1 Reference plates for checking photometric accuracy
applicationTheHellmaanalyticsreferenceplate666R013canbeusedtocheckthephotometricaccuracyofmicrotitreplatereaders.
pRoduct descRiptionThe reference plate dimensions are suitable for a 96-wellmicrotitre plate with a diameter of 6.6 mm per window (H14.5mmxD125mmxL85.5mm).eachofthefiveneutraldensityglassfilters(fieldsS1–S5)inthereferenceplatecanmeasuretheabsorbancevaluefor16windows.Theother16windowsdonotcontainglass(S0)andserveasreferences.
notesThereferenceplatehasfiveneutraldensityglassfilterswithdifferentnominalabsorbancevalues,allowingyoutocheckthe linearity of your absorbance scale by plotting the ab-sorbancevaluesmeasuredforeachwavelengthagainstthemeasurementvaluesontheDakkScalibrationcertificateinadiagram.
4.2 Reference plate for checking photometric and wavelength accuracy
applicationTheHellmaanalyticsreferenceplate666R113canbeusedtocheckthephotometricandwavelengthaccuracyofmicrotitreplatereaders.
pRoduct descRiptionThe reference plate dimensions are suitable for a 96-wellmicrotest plate with a diameter of 6.6 mm per window (H14.5mmxD125mmxL85.5mm).eachofthefourneutraldensityglassfiltersused(fieldsS1–S4)canmeasuretheab-sorbancevaluefor16windows.Holmiumoxideglass(S5)isusedtotestthewavelengthaccuracyin16windowswhileafurther 16 windows (S0) do not contain glass and serve asreferences.
notesThe reference plate has four neutral density glass filterswith different nominal absorbance values, allowing you tocheckthelinearityofyourabsorbancescalebyplottingtheabsorbance values measured for each wavelength againstthemeasurementvaluesontheDakkScalibrationcertifica-teinadiagram.
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aRticle no. 666R013
application Referenceplateformicroplatereadersfortestingthephotometricaccuracy
content
neutraldensityglassfilterNg11(0.25abs)neutraldensityglassfilterNg5(0.5abs)neutraldensityglassfilterNg4(1.0abs)neutraldensityglassfilterNg3(1.5abs)neutraldensityglassfilterNg3(2.5abs)framewithoutglass(referencefilter)
standaRd ceRtification photometric accuracy certified at wavelengths:405;450;490;650nmat 8 points in a row
possible ceRtificationwavelengths: allpossiblefrom405to890nmalsopossibleabove890nm,butonlywithaHellmaanalyticsCalibrationCertificateslit widths: allpossibleupto5nm
aRticle no. 666R113
application Referenceplateformicroplatereadersfortestingthewavelengthaccuracyandthephotometricaccuracy
content
neutraldensityglassfilterNg5(0.5abs)neutraldensityglassfilterNg4(1.0abs)neutraldensityglassfilterNg3(1.5abs)neutraldensityglassfilterNg3(2.0abs)holmiumoxideglassfilterframewithoutglass(referencefilter)
standaRd ceRtificationphotometric accuracy certified at 8 points in row at wavelengths:405;450;490;650nmwavelength accuracy certified at: 279;361;453;536;638nmslit width: 1nm
possible ceRtification
photometric accuracy:additionalwavelengths:allpossiblefrom405to890nmalsopossibleabove890nm,butonlywithaHellmaanalyticsCalibrationCertificateslit widths: allpossibleupto5nmwavelength accuracy:additional possible wavelengths: 287;418;445;460nmslit widths: allupto2nmrecommended
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4.3 General usage guidelines for reference plates
Referenceplatesaremadeofglassdopedwithmetal ionsorrareearthmetals,whichisannealedandassembledinblackanodized precision frames made of aluminum. They are de-signed to fit into all microtitre plate readers. To ensure easyidentification,each referenceplate isengravedwith the refe-renceplatetypeandserialnumber.Detailsoftheabsorbanceandpeakpositionvaluesmeasuredforeachfiltercanbefoundontherespectivecalibrationcertificate.pleaseensurethatyoudonottouchtheglasssurfacesofthefilter.Dirt,dust,andda-mage can significantly impair the accuracy of measurementresults.anodizedaluminumframesshouldnotcomeintocon-tactwithacidsoralkalis.
stoRaGeafter use, we recommend storing reference plates at roomtemperature,intheirpackaging,andinadry,dust-freearea.
otheR factoRs that may influence measuRementsDirt(e.g.fingerprints)andduston,ordamage(scratches,cor-rosion)to,polishedsurfacescansignificantly impairtheaccu-racyofmeasurementresults.alwaysstorereferenceplatesintheiroriginalpackagingandprotect theopticalwindows fromcontamination.Onlyhandlereferenceplatesbytheirframes.
cleaninGDirtoftenaccumulatesonopticalsurfacesasaresultofre-gularuse.This isbest removedusinga lint-freeclothandalcohol.
influence of tempeRatuRe on measuRementsTemperaturehasaverysmallinfluenceoncertifiedmeasure-ment values.Measurements takenat temperaturesbetween20°Cund24°Cfallwithinthemeasurementuncertaintystatedonthecalibrationcertificate.Measurementsshouldthereforebetakeninthisrangetokeepanypotentialtemperatureinflu-enceontheresultstoaminimum.
4. certIfIed reference plates for mIcrotItre plate readers
steps to taKe befoRe peRfoRminG calibRation With RefeRence plates
1. Warmupthemicrotitreplatereaderuntilthecorrectoperatingtemperaturehasbeenreachedandremainsconstant(e.g.foronehour),takingcaretoobservethedevicemanufacturer’sguidelines.
2. Tobeginwith,carryoutabaselinecorrectionwithanemptysamplecompartment.
3. Checkthatthereferenceplateiscorrectlypositionedinthelightpathbyfirstmeasuringthewindowswithoutglass(usuallyrows1and2).Thelabelshowingthereferenceplatetypemustbevisiblefromabove.
4. Checkthatthedevice’sdisplayhasnotchanged.Inmicrotitreplateswithverylargebeams,themeasurementbeammaytouchthewindowframe.Ifthisisthecase,youwillnoticeachangeinthedevice’sdisplay.
» Ifnecessary,adjustthepositionofthereferenceplateholderuntilthelightbeamshines throughtheemptywindowunimpeded. » Thereferenceplateiscorrectlypositionedifthedisplayvaluesfromthezeroadjustmentperformedinstep2
(baselinecorrection)donotchange.
5. Carryoutthefiltermeasurementinaclosedsamplecompartmentascarefullyasyouwouldcarryoutasamplemeasure-ment(opensamplecompartmentsproduceincorrectresults).
ideas, criticism, experiences?
We want to hear your thoughts!
4. certIfIed reference plates4.4 calibration with reference plates
pRepaRations Therearemanydifferentmakes,models
anddesignsofspectrometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
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inteRpRetinG measuRement Results With RefeRence plates foR checKinG photometRic and WaVelenGth accuRacy The measurement uncertainties stated on the calibrationcertificate only refer to measurements conducted by Hellmaanalytics and apply solely to the measurement conditions atthe company (spectrophotometer used, environmental influ-encessuchas temperature,airhumidity,user influence,andreferencematerialsused).
The smallest possible measurement uncertainty that canbeachievedbytheusercanthenbederivedbystatisticallycombiningthemeasurementuncertaintystatedonthecali-brationcertificatewithalltheuser’suncertaintycontributi-ons,suchasthewavelengthscaletoleranceofthemicrotestplate reader used and other influences on measurementaccuracy (environmental factors such as temperature, airhumidity,userinfluence,etc.).Forfurtherliteratureoncor-rectlycalculatingmeasurementuncertainty,pleaserefertochapter8ofthisusermanual.
measuRement paRameteRs foR checKinG photometRic accuRacygenerally speaking, reference plates can also be measuredusing a slit width that differs from the information providedonthecalibrationcertificate.However,pleasenotethatusinglargeslitwidthsmayresult inslightdeviations fromtheva-luesstatedonthecalibrationcertificate.Incasesofdoubt,itisthereforeadvisabletochooseassmallaslitwidthaspos-sible.Werecommendtakingseveralmeasurementsandthenusingthemeanvaluetoavoiderrorsduringevaluation.
measuRement paRameteRs foR checKinG WaVelenGth accuRacyensure that you have selected the correct measurementparameters before plotting the absorbance curve to cal-culate peak positions. Incorrect parameters may distorttheabsorbancecurveandthusshift theactualpositionsofpeaks.pleaseusethesettingsstatedontheaccompanyingcalibrationcertificate. Itshouldbenoted thatchanging theslit width of the microtitre plate reader can cause the ab-sorbance maxima to shift slightly. Ignore any influence
thatthespectralbandwidthfrom1nmto2nmhasonpeakpositions.peakheights,however,mayvarygreatlyfollowingchanges to the slit width due to their narrow nature. as aresult, filters forcheckingwavelengthaccuracyareusuallyunsuitableforcheckingabsorbanceaccuracy.
steps foR checKinG WaVelenGth accuRacy With RefeRence plates
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithreferenceplates’.
2. Runthescanprogramonyourmicrotitreplatereader,observingtheguidelinesintheusermanual.Selectascanningran-gethatcoversallofthepeakslistedonthefilter’scalibrationcertificate.
3. Setyourmicroplatereadertothemeasurementparametersquotedonthecalibrationcertificateprovided.Selecttheslowestscanningspeedandasmalldatainterval.
4. Ifpossible,carryoutabaselinecorrection.
5. placethereferenceplateintheplateholder.ensurethatthefilterIDisvisiblefromabove.Referenceplatesmustalwaysbepositionedintheplateholdersinthesameway.
6. Startthemeasurementforthepositionswhereholmiumoxideglassfiltersareinserted(usuallyS5).
7. Calculatethepositionsofthepeaksatthewavelengthsstatedonthecalibrationcertificate.
8. Takeseveralmeasurementsandthenusethemeanofthemeasuredvaluestoavoiderrors.
9. Compareyourmeasurementvalueswiththecertifiedones.
»in practice it is easier to simply add up uncer-tainty contributions than to combine their stati-stics. however, the method used to determine measurement uncertainty depends on the speci-fications of your quality system and your measu-rement accuracy requirements.«
see the faqs for more details.
birgit Kehl,HeadHellmaanalyticscalibrationlaboratory
4. certIfIed reference plates4.4 calibration with reference plates
photometRic accuRacy
4. certIfIed reference plates4.4 calibration with reference plates
WaVelenGth accuRacy
steps foR checKinG photometRic accuRacy With RefeRence plates
1. Firstofall,followthe‘stepstotakebeforeperformingcalibrationwithreferenceplates’.
2. Runthewavelengthselectionprogramonyourmicrotitreplatereader,observingtheguidelinesintheusermanual.Selectthewavelengthsstatedonthecalibrationcertificate.
3. Setyourmicrotestplatereadertothemeasurementparametersquotedonthecalibrationcertificateprovided.
4. adjusttozero.
5. placethereferenceplateintheplateholder.ensurethatthereferenceplateIDisvisiblefromabove.Referenceplatesmustalwaysbepositionedintheplateholdersinthesameway.
6. Starttheprogramformeasuringabsorbancevaluesatthewavelengthsstatedonthecalibrationcertificate–thepositionsmeasuredarethosewhereneutraldensityglassfiltersareinserted.
7. Takeseveralmeasurementsandthenusethemeanofthemeasuredvaluestoavoiderrors.
8. Compareyourmeasurementvalueswiththecertifiedones.
Therearemanydifferentmakes,modelsand
designsofspectrophotometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
Therearemanydifferentmakes,modelsand
designsofspectrometer,thesestepsare
onlygeneralguidelines.pleaserefertoyour
manufacturer'sinformationtogetmoredetails.
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5. recertIfIcatIon
Rec
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ReceRtification inteRVals foR RefeRence mateRials
as is the case for all measuring devices, the referencematerials used to verify spectrophotometers must alsobecheckedand recertifiedat regular intervals (see forex-ampleISO9001:2008“ControlofMonitoringandMeasuringequipment”).Thisallowsyoutoensurethatyouconsistent-lyfulfillyourin-housequalityrequirementsandguaranteeshighlevelsofaccuracyandreliabilityinyourmeasurements.
The length of intervals between the recertification of re-ference materials depends on how frequently materialsare used, the wear associated with this, accuracy require-ments, and the requirements of a company’s internal au-diting. Therefore, only clients themselves can determinethese. In general, a recertification interval of 12 months isrecommendedforcheckingandrecertifyingglassfiltersdu-ringthefirsttwoyearsofuse,withanintervalof24monthsthereafter. We recommend verifying and recertifying liquidfilterswithinamaximumof12months.IntervalsshouldbespecifiedindividuallyinaccordancewithyourQasystem.
RetuRninG youR RefeRence mateRials foR ReceRtification
30-yeaR WaRRanty
allofourreferencematerialscomewitha30-yearwarranty,providedthattheyareregularlyrecertified(atleasteverytwo years) at the Hellma analytics calibration laboratory.Certifiedreferencematerials(filters)sentforrecertificationarecarefullycleanedandrecertifiedbeforebeingsentbackwith a new DakkS calibration certificate and calibrationmark. Damaged filters and filters that deviate significantlyfrom nominal values are usually replaced in consultationwiththecustomer.
1. yeaR 2. yeaR 3. yeaR 4. yeaR 5. yeaR 6. yeaR 7. yeaR 8. yeaR
liquid filteRs
ReCeRTIFYINgeVeRY12 months
Glass filteRs
1. yeaR 2. yeaR 3. yeaR 4. yeaR 5. yeaR 6. yeaR 7. yeaR 8. yeaR
ReCeRTIFYINgeVeRY24 months
01.Completethereturnshipmentforminfull.Whenreturningseveralfiltersorfiltersets,pleaseuseoneformforeach.
04.FiltersarecleanedandrecertifiedintheHellmaanalyticsca-librationlaboratory.Ifnecessary,filterswillalsoberepairedorexchangedfollowingaconsultation.
05.you will receive your filter with a new dakks calibration certificate or a hellma analytics calibration certificate in the case of filters for verifying spectral resolution.
03.SendyourfiltertotheHellmaanalyticscalibrationlabora-toryviayourlocalHellmaoffice.Usetheaddresslabelprintedontherearofthereturnship-mentformtodoso.
02.encloseacopyofthecurrentcalibrationcertificate.
12 MONTHS12 MONTHS
12 MONTHS
24 MONTHS 24 MONTHS 24 MONTHS
COmPAny:
nAmeOfCOntACtPeRSOn:
StReet/StReetnumBeR:
POStAlCODe/CIty:
COuntRy:
emAIl:
PHOne:
tHISADDReSSIS:SHIPPInGADDReSS BIllInGADDReSS
ARtIClenumBeR/fIlteRtyPe*:
Set/SeRIAlnumBeR*:
yOuRORDeRnumBeR:
High Precision in Spectro-Optics
RetuRnfORm
PleASenOte!manyqualitysystemsstipulatearegular
inspectionofthereferencematerialsfor
anidealmeasuringcertainty.Itisrecom-
mendtorecertifyglassfilterservery24
monthsandliquidfiltersevery12months.
RecertificationintheaccreditedHellmaAnalyticsCalibrationlaboratory
I’dliketoreceivethefollowingrecertification
DAkkSCalibrationcertificate (according to the already existent calibration certificate; please enclose a copy of it)
IndividualrecertificationwithDAkkSCalibrationcertificate Please specify:
SlItWIDtH/S:
WAVelenGtH/S:
OtHeR:
Additionaloptions I’dliketoreceiveadocumentationofthemeasureddatauponreceipt(priortocleaning)
measurementreportoftheabsorptionvalues for neutral density glass filters (at 1 nm slit width and the following wavelengths 440, 465, 546.1, 590, 635) (charge 18.90 € – depending on the country and exchange rate) Individual documentation of the measureddatauponreceiptcombinedinameasurementreport (charges depending on the type of filter and extent of measurement) SlItWIDtH/S:
WAVelenGtH/S:
Documentation of the measureddatauponreceiptwithaDAkkSCalibrationcertificate for all measurement parameters, according to the recertification stated above (charges depending on the type of filter and extent of measurement)
thankyouverymuchfortrustingtheHellmaAnalyticsCalibrationlaboratory!
Pleasecompletethisformandencloseitwiththereturnshipment.
*You’ll find this information on your calibration certificate
Returnformavailablefor
download:
www.hellma.com/return
4544
6. faqs
6.1 Why do holmium oxide glass filters become cloudy? Will this interfere with the measurement?The glass material used for this filter is somewhat hygro-scopic,whichmeans that the filtersbecomecoatedwithakindofwaterfilm.Thefilmdoesnotinterferewithmeasure-mentsorchangethecharacteristicpeakpositionsofholmi-umoxide.Thefiltercanbeeasilywipeddownusingalcoholand a soft cloth. The filter should generally be stored in adryplace.6.2 how long can a calibration standard be used for in total?Depending on the conditions in which they are used andstored,aswellashowtheyaremaintained,filtersusuallylastformanyyears.Werecommendhavingfiltersregularlyrecer-tifiedsothatanysignsofdeteriorationcanberecognizedatanearlystage.6.3 how often should filters be recertified?Certifiedreferencematerialsshouldberecertifiedat regu-lar intervalstocheckthatthevaluesstatedonthecalibra-tion certificate are still valid. It is up to the user to decideon the regularityof these intervals,whichshould take intoaccount theuse,storageandusageconditionsof the filterinthelaboratory.Toestablishastatisticaldatabaseforde-termining recertification intervals, we recommend havingallreferencematerialsrecertifiedat leastevery12monthsduringtheirfirsttwoyearsofuse,andthenselectingasui-tablerecertificationintervalbasedonthevaluesmeasured.(pleaseseechapter9).6.4 What do the tolerances on the calibration certificate tell us and how can they be correctly interpreted?The measurement uncertainties that appear on calibrati-on certificates only refer to measurements conducted byHellmaanalyticsandapplysolelytothemeasurementcon-ditions at the company (spectrophotometer used, environ-mental influences such as temperature, air humidity, userinfluence,referencematerialsused,etc.).Consequently,themeasurementuncertaintiesoftheNISTreferencematerialsusedtoensuretraceabilityhavebeenmathematicallycom-binedwith themeasurementuncertaintystatisticscalcula-tedbyHellmaanalytics.Thevalueprovided is thereforeanexpandedmeasurementuncertainty(doublestandarddevia-tion,coveragefactork=2).Thismeansthattheactualvalueis95%certaintofallwithinthisrange.Tocorrectlycalcula-tethemeasurementuncertaintiesvalidfortheirmeasuringsystem, reference material users should follow the samesteps,mathematically/statistically combining themeasure-ment uncertainties provided with the measurement uncer-tainty statistics they have calculated themselves for a par-ticularspectrophotometerandrelevantconditions(seeISO/IeCguide98-3:2008‘guidetotheexpressionofUncertaintyinMeasurement’).
6.5 What is a baseline correction?Baseline corrections are carried out with an empty cuvetteholder tocompensate for the lamps.Since lampsemit lightat different strengths at various wavelengths, baseline cor-rections(alsoknownasautozero)arecarriedouttodetermi-neazerovalue.Baselinecorrectionsareusuallyperformedautomaticallywhenthespectrophotometerisstartedup,butcanalsobecarriedoutmanually.6.6 What is background correction?Backgroundcorrectioniscarriedouttoeliminateanyinflu-encesthatextendbeyondthesample’sproperties.Indoub-le beam photometers, background correction is performedbysimultaneouslymeasuringthecomparisoncuvetteinthereferencebeampath.Thiscomparisoncuvetteusuallycon-tainspuresolvent.Insinglebeamphotometers,backgroundcorrectioniscarriedoutbeforetheactualsamplemeasure-mentistakenbymeasuringthecomparisoncuvette.Theva-luesobtainedforthecomparisoncuvettearethendeductedfromthevaluesofthesamplemeasurement.6.7 Why does the calibration certificate for the filter set used to determine spectral resolution look different to other calibration certificates? Determiningspectralresolutiondoesnotfallwithinourscopeofaccreditation.The filterset fordeterminingspectral resolutionthereforecannotbeissuedwithaDakkScalibrationcertificateorcalibrationmark.Thatiswhythiscalibrationcertificatelooksdif-ferentfromothercalibrationcertificatesforfiltersets.6.8 Why does hellma analytics no longer offer potassium dichromate filters for checking photometric accuracy with sulfuric acid as a solvent, as described in the european pharmacopoeia?Inthepast,certifiedreferencematerialsforcheckingphoto-metricaccuracy intheUVrangecontainedasolutionofpo-tassiumdichromate insulfuricacidandweremanufacturedin strict compliance with european pharmacopoeia require-ments. Over a number of years, Hellma analytics noticeda continuous decrease in the absorbance values of the ‘po-tassium dichromate dissolved in sulfuric acid’ filter duringdaily calibrations. We do not have a sufficient explanationforwhythishappens,butweassumethatthecomparativelyhigh ionic strength of sulfuric acid causes mixed chromium(VI)complexestoform.Tocompensateforthisbehavior,overwhichwehavenocontrol,filterswouldneedtoberecertifiedmuchmoreregularly.anotherpossibilitywouldbepreparingnewsolutionseverytimethespectrophotometer ischecked.asasimplealternative,weofferaliquidfilterthatuses‘po-tassiumdichromatedissolvedinperchloricacid’.Thistypeofliquidfilterforcheckingphotometricaccuracyhasprovenit-selfasareliableandverystablestandardformanyyears.Nochangesinabsorbancepropertiescomparabletothoseofthe
sulfuricacidmodelareknownforthisfilter.Hellmaanalyticscuvettesarepermanentlysealed,eliminatingconcernsaboutthe toxicity of perchloric acid. Furthermore, the europeanpharmacopoeiastates that “suitablecertified referencema-terials”mayalsobeused,whichundoubtedlyapplies toourperchloricacidsolventmodel.Thismodelalsocontainsafor-mulationdescribedbyNIST.6.9 Why does the weight of potassium dichromate filters seem to change after every recertification?Due to measurement uncertainties, measurement valuesmay fall within a specific range. This leads to an apparentchange in weight from qualification to qualification, as theinitial weight is calculated directly from the measured ab-sorbance values. earlier versions of regulatory codes stipu-lated that filters for checking photometric accuracy had tocontain60.06mg/lpotassiumdichromate,andallowedato-lerance of 0.01 abs. More current versions of the europeanpharmacopoeiahavereplaced this verystrictprovision,nowacceptingweightsbetween57.0mg/land63.0mg/l.Thespe-cific absorbance calculated (see european pharmacopoeia,chapter2.2.25)isnowstatedwithamarginoftolerance.6.10 Why are these peaks measured for certifying holmium oxide glass and didymium glass filters?Measurement errors are low in medium to high transmit-tanceranges.asaresult,peaks in therange from0abs to1.0abs(correspondsto100%-Tto10%T)arepreferredforcertification.6.11 how do i calculate my measurement uncertainty?The measurement uncertainties stated on the calibration cer-tificate only refer to measurements conducted by Hellmaanalyticsandapplysolelytothemeasurementconditionsatthecompany (spectrophotometer used, environmental influencessuch as temperature, air humidity, user influence, referencematerialsused,etc.).Thesmallestpossiblemeasurementun-certaintythatcanbeachievedbytheusercanthenbederivedbystatistically combining the measurementuncertainty stated onthecalibrationcertificatewithalltheuser’suncertaintycontribu-tions,suchasthewavelengthscaletoleranceofthespectropho-tometer used and other influences on measurement accuracy(environmentalfactorssuchastemperature,airhumidity,userinfluence,etc.).
example of calculatinG standaRd measuRement unceRtainty foR a neutRal density Glass filteR (hiGhly simplified): the calibRation ceRtificate sta-tes the folloWinG measuRement Values and measu-Rement unceRtainties:
here, a wavelength of 440 nm produces the following parameters:target measurement value (xs) : 0,2542 absexpanded measurement uncertainty: +/- 0,0024 abs (coverage factor k=2)standard measurement uncertainty (xa): +/- 0,0012 abs
Next, you must calculate the measuring error specific to yourspectrophotometer(xb)–refertotheoperatinginstructionsformoredetails–anddefineavalueforthemeasuringerrorduetoenvironmentalinfluencesatyourcompany(xu)(suchastempe-ratureandairhumidity).
example of measuring error parameters:spectrophotometer (xb): +/- 0,01 absenvironmental influences (xu): +/- 0,001 abscalculating standard measurement uncertainty (mu):mu = √xa² + xb² + xu² = 0,0101expanded measurement uncertainty is calculated by multip-lying this value by coverage factor k.
asshownhere,inpracticeitisofteneasiertosimplyaddupun-certaintycontributionsthantocombinetheirstatistics.However,the method used to determine measurement uncertainty de-pends on the specifications of your quality system and yourmeasurement accuracy requirements. For further literatureoncorrectlycalculatingmeasurementuncertainty,pleaserefertotherecommendationsforfurtherreadinginchapter8ofthisusermanual.
seRial numbeR 3524
optische dichte (abs)optical density (abs)
440nm 465nm 546.1nm 590nm 635nm
measuRed Value 666-f2 0.2542
±0.00240.2254±0.0024
0.2254±0.0024
0.2415±0.0024
0.2416±0.0024
4746
7. Glossary 8. recommendatIons for further readInG
abbreviations:
a: absorbanceastm: americanSocietyforTestingandMaterialsbG: SpecifictermforSchottglassdab: Deutschesarzneibuch(germanpharmacopoeia)dakks: Deutscheakkreditierungsstelle(Nationalaccreditation bodyfortheFederalRepublicofgermany)daR: Deutscherakkreditierungsrat (germanaccreditationbody)dKd: DeutscherKalibrierdienst(germancalibrationbody)ph. eur.:europeanpharmacopoeiafaqs: FrequentlyaskedquestionsGlp: goodlaboratorypracticeGmp: goodmanufacturingpracticei: Intensityi0: Originalintensity0: Originalk: Coveragefactorformeasurementuncertaintyλmax: Maximumpeakatdefinedwavelengthλmin: MinimumpeakatdefinedwavelengthniR: Near-infrarednist: NationalInstituteofStandardsandTechnologyptb: physikalisch-TechnischeBundesanstalt (germany’snationalmetrologyinstitute)sRm®: StandardReferenceMaterial (registeredtrademarkofNIST)usp: UnitedStatespharmacopeiauV: UltravioletVis: Visible(visiblewavelengthrange)
absorbance (abs): Whenlightfallsonorpassesthroughasample,thequantityofabsorbed light is equal to the difference between the originalintensityI0andtheintensityIafterinteractionwiththesample.Thisisbecausepartoftheirradiatedlightistransferredtothemolecules,causingthebeamtohaveasmalleroutputwhenitexitsthesample.Theextenttowhichlightisabsorbedisdeter-minedbytheprinciplesoftheBeer-Lambertlaw.Theamountofabsorbedlightcanbeexpressedastransmittance(seedefiniti-on)orabsorbance.absorbanceisdefinedasabs=-logT.accordingtotherelevantstandard,thisparameterisreferredtoasspectralopticaldensityontransmittance(“opticaldensity”).
optical density: seeabsorbance
Visible range: partof theoptical spectrum thatstretches from380nmto780nmofthewavelengthrangeofelectromagneticra-diation.Thisrangeisgenerallyreferredtoaslight.Thisistheonlyrangeinwhichthehumaneyecan‘see’electromagneticradiation.
spectral resolution: Thisreferstoameasuringsystem’sabilitytoseparateindividualwavelengthranges.
spectral bandwidth: Wavelengthrangethatappearswithacontinuum at the exit slit when the monochromator is ex-posed to irradiation. Spectral bandwidth is determined bythebandwidthofemittedradiationwherethelighthasrea-chedhalfthemaximumintensity.
spectral optical density on transmittance:seeabsorbance
transmittance (t): When light falls on or passes through asample,thequantityofabsorbedlightisequaltothedifferencebetweentheoriginal intensity I0and the intensity Iafter inter-action with the sample. This is because part of the irradiatedlightistransferredtothemolecules,causingthebeamtohaveasmalleroutputwhenitexitsthesample.Theextenttowhichlight is absorbed is determined by the principles of the Beer-Lambertlaw.Theamountofabsorbedlightcanbeexpressedastransmittance (see definition) or absorbance. Transmittance isnormallyexpressedasafractionof1orasapercentage,andisdefinedasfollows:T=I/I0or%T=(I/I0)*100.
ultraviolet range (uV range):alsoknownasUVradiation,thisistheshort-wavepartoftheopticalradiationspectrum.UVradiati-onhasawavelengthrangeof100nmto380nm.
Wavelength: Wavelength isthedistancebetweentwo identical,adjacentcorrespondingpointsofthesamewavephaseatacer-tainpointintime.
standards and best practice in absorption spectrometry;edited by C. Burgess and T. Frost UVSg, ISBN 0-632-05313-5BlackwellService
qualitätssicherung in der analytischen chemie;WernerFunk,VeraDammann,gerhildDonnevert;ISBN-10:3-527-31112-2;Verlag:WILeY-VCH
ISO/IeC guide 98-3:2008; evaluation of measurement data – Guide to the expression of uncertainty in measurement NISTSpecialpublication260-54StandardReferenceMaterials:certification and use of acidic potassium dichromate solutions as an ultraviolet absorbance standard – sRm 935
NIST Special publication 260-116 Standard ReferenceMaterials: Glass filters as a standard Reference material for spectrophotometry – selection, preparation, certification, and use of sRm 930 and sRm 1930
NIST Special publication 260-102: Standard ReferenceMaterials:holmium oxide solution Wavelength standard from 240 to 640 nm – sRm 2034
european pharmacopoeia (ph.eur.)
dKd3
share your experience of our
products by writing to: [email protected]
type mateRial WaVelenGth nm aRticle-no.
Glass filter for testing the wavelength accuracy
666-f1 HolmiumOxideglassFilterF1 279;361;453;536;638 666f1-339
666-f7W DidymiumglassFilterF7W 329;472;512;681;875 666f7W-323
Glass filter for testing the photometric accuracy
666-f2 NeutralDensityglassFilterF2(Nominalvalueoftheabsorption0.25)
440;465;546,1;590;635 666f2-39
666-f201 NeutralDensityglassFilterF201(Nominalvalueoftheabsorption0.3)
440;465;546,1;590;635 666f201-39
666-f3 NeutralDensityglassFilterF3(Nominalvalueoftheabsorption0.5)
440;465;546,1;590;635 666f3-38
666-f4 NeutralDensityglassFilterF4(Nominalvalueoftheabsorption1.0)
440;465;546,1;590;635 666f4-37
666-f202 NeutralDensityglassFilterF202(Nominalvalueoftheabsorption1.5)
440;465;546,1;590;635 666f202-36
666-f203 NeutralDensityglassFilterF203(Nominalvalueoftheabsorption2.0)
440;465;546,1;590;635 666f203-36
666-f7a NeutralDensityglassFilterF7a(Nominalvalueoftheabsorptionapprox.0.5–1.0)
270;280;297;320;340 666f7a-323
Glass filter for testing the wavelength accuracy and the photometric accuracy
666-f7 DidymiumglassFilterF7 a:270;280;297;320;340W:329;472;512;681;875
666f7-323
empty filter mount
666-f0 aluminumframe 666f0-71
type consistinG of WaVelenGth nm aRticle-no.
sets for testing the wavelength accuracy and the photometric accuracy
666-s000 CompleteglassFilterSet:F1,F2,F3,F4,F0
a:440;465;546,1;590;635W:279;361;453;536;638
666s000
666-s001 glassFilterSet:F3,F4,F7
a:270;280;297;320;340;440;465;546,1;590;635W:329;472;512;681;875
666s001
666-s002 glassFilterSet:F2,F3,F4 a:440;465;546,1;590;635 666s002
666-s004 glassFilterSet:F201,F202,F203 a:440;465;546,1;590;635 666s004
666-s300 glassFilterSet:F301,F303,F390(abs:0.04;2.5;3.0)
a:440;465;546,1;590;635 666s300
666-R113666-R013
5048
9. product overvIew
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Glass filters with dakks calibration certificates liquid filters with dakks calibration certificates liquid filters with dakks calibration certificates
667-UV 1
667-UV100
667-UV 60
667-UV305
667-UV 6
667-UV200
667-UV 5667-UV 12 667-UV 14667-UV 600667-UV 9
Complete Set 667-UV003
typ inhalt WellenlänGe nm aRtiKel-nR.
flüssigfilter zur Überprüfung der fotometrischen Genauigkeit
667-uV20 20mgKaliumdichromatinHClO4(0,25abs) 235;257;313;350 667020
667-uV40 40mgKaliumdichromatinHClO4(0,5abs) 235;257;313;350 667040
667-uV60 60mgKaliumdichromatinHClO4(0,75abs) 235;257;313;350 667060
667-uV80 80mgKaliumdichromatinHClO4(1,0abs) 235;257;313;350 667080
667-uV0100 100mgKaliumdichromatinHClO4(1,25abs) 235;257;313;350 6670100
667-uV600 600mgKaliumdichromatinHClO4(1,0abs) 430 667600
667-uV14 perchlorsäure(Referenzfilter) 235;257;313;350 667014
667-uV301 Filter-SetfürUV-Bereich:UV60,UV14 235;257;313;350 667301
667-uV304 Filter-SetfürVis-Bereich:UV600,UV14 430 667304
667-uV305 Filter-SetfürUV/Vis-Bereich:UV60,UV600,UV14 235;257;313;350;430 667305
flüssigfilter-set zur Überprüfung der linearität der absorption
667-uV307 Filter-Set:UV20,UV40,UV60,UV80,UV100,UV14 235;257;313;350 667307
flüssigfilter zur Überprüfung der Wellenlängengenauigkeit
667-uV5 Holmiumoxidinperchlorsäure 241;287;361;536;640 667005
667-uV400 Filter-Set:UV05,UV14 241;287;361;536;640 667400
flüssigfilter zur Überprüfung von streulicht
667-uV1 KaliumchloridinReinstwasser 200(cut-off) 667001
667-uV10 NatriumiodidinReinstwasser 259(cut-off) 667010
667-uV11 NatriumnitritinReinstwasser 385(cut-off) 667011
667-uV12 Reinstwasser(Referenzfilter) 198;200;300;400 667012
667-uV100 Filter-Set:UV1,UV12 200(cut-off) 667100
667-uV101 Filter-Set:UV10,UV12 259(cut-off) 667101
667-uV102 Filter-Set:UV11,UV12 385(cut-off) 667102
667-uV103 Filter-Set:UV1,UV10,UV11,UV12 200;259;385(cut-off) 667103
a:WavelengthsforabsorbanceW:WavelengthsforwavelengthaccuracyS:WavelengthsforstraylightR:Wavelengthsforspectralresolution *withHellmaanalyticscalibrationcertificate
Reference plates for qualifying microplate readers with dakks calibration certificates
666-F0 666-F1 666-F2 666-F3 666-F4 666-F203666-F202666-F201 666-F7
Complete Set 666-S000 SingleFilters
a:WavelengthsforabsorbanceW:Wavelengthsforwavelengthaccuracy
type content WaVelenGth nm aRticle-no.
liquid filter for testing the photometric accuracy
667-uV20 20mgpotassiumdichromateinHClO4(0.25abs) 235;257;313;350 667020
667-uV40 40mgpotassiumdichromateinHClO4(0.5abs) 235;257;313;350 667040
667-uV60 60mgpotassiumdichromateinHClO4(0.75abs) 235;257;313;350 667060
667-uV80 80mgpotassiumdichromateinHClO4(1.0abs) 235;257;313;350 667080
667-uV0100 100mgpotassiumdichromateinHClO4(1.25abs) 235;257;313;350 6670100
667-uV600 600mgpotassiumdichromateinHClO4(1.0abs) 430 667600
667-uV14 perchloricacid(HClO4),referencefilter 235;257;313;350 667014
667-uV301 FilterSetforUVrange:UV60,UV14 235;257;313;350 667301
667-uV304 FilterSetforVisrange:UV600,UV14 430 667304
667-uV305 FilterSetforUV/Visrange:UV60,UV600,UV14 235;257;313;350;430 667305
liquid filter set for testing the linearity of the absorption
667-uV307 Filter-Set:UV20,UV40,UV60,UV80,UV100,UV14 235;257;313;350 667307
liquid filter for testing the wavelength accuracy
667-uV5 Holmiumoxideinperchloricacid 241;287;361;536;640 667005
667-uV400 FilterSet:UV05,UV14 241;287;361;536;640 667400
liquid filter for testing to stray light
667-uV1 potassiumchlorideinpurewater 200(cut-off) 667001
667-uV10 Sodiumiodideinpurewater 259(cut-off) 667010
667-uV11 Sodiumnitriteinpurewater 385(cut-off) 667011
667-uV12 purewater(referencefilter) 198;200;300;400 667012
667-uV100 FilterSetUV-100:UV1,UV12 200(cut-off) 667100
667-uV101 FilterSetUV-101:UV10,UV12 259(cut-off) 667101
667-uV102 FilterSetUV-102:UV11,UV12 385(cut-off) 667102
667-uV103 FilterSetUV-103:UV1,UV10,UV11,UV12 200;259;385(cut-off) 667103
667-uV104 FilterSetUV-104:UV10,UV11,UV12 259;385(cut-off) 667104
liquid filter for testing the resolution
667-uV6* Tolueneinn-hexane Scan:265–270 667006
667-uV9* n-hexane(referencefilter) Scan:265–270 667009
667-uV200* FilterSetUV-200:UV6,UV9 Scan:265–270 667200
a:WavelengthsforabsorbanceW:WavelengthsforwavelengthaccuracyS:WavelengthsforstraylightR:Wavelengthsforspectralresolution *withHellmaanalyticscalibrationcertificate
type content WaVelenGth nm aRticle-no.
liquid filter – set according to usp 851
667-uV004 F0:aluminumframe,F2:NeutralDensityglassfilter(0.25abs),F3:NeutralDensityglassfilter(0.5abs),F4:NeutralDensitiyglassfilter(1.0abs),UV60:60mgpotassiumdichromateinHClO4UV14:perchloricacid(HClO4,referencefilter,
UV5:Holmiumoxideinperchloricacid
a:440;465;546.1;590;635a:440;465;546.1;590;635a:440;465;546.1;590;635a:235;257;313;350a:235;257;313;350W:241;250;278;287;333;345;361;385;416;452;468;485;536;640
667004
complete filter set for testing the photometer according to ph.eur.
667-uV003 CompleteFilterSet:UV1,UV12,UV6,UV9,UV60,UV600,UV14,UV5
a:235;257;313;350;430W:241;287;361;536;640S:200(cut-off)R:Scan265–270
667003
type usaGe mateRialNominalvalueofabsorption
WaVelenGth nm
aRticle-no.
666-R013 tocheckphotometricaccuracy NeutralDensityglassFilterNg11(0.25),Ng5(0.5),Ng4(1.0),Ng3(1.5),(2.5)
405,450,490,650 666R013
666-R113 tocheckphotometricaccuracyandwavelengthaccuracy
NeutralDensityglassFilterNg5(0.5),Ng4(1.0),Ng3(1.5),(2.0)HolmiumOxideglassfilter
405;450;490;650279;361;453;536;638
666R113
new
Subject to change without notice 064-501-PB05-725//GB//09//2015 Printed in Germany
High Precision in Spectro-Optics
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notes