ASTM_E_2719_-_09_2014.pdf

Designation:E271909(Reapproved 2014)Standard Guide forFluorescenceInstrument Calibration and Qualification1This standard is issued under the fixed designation E2719;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.1.Scope1.1 This guide(1)2lists the available materials and methodsfor each type of calibration or correction for fluorescenceinstruments(spectralemissioncorrection,wavelengthaccuracy,and so forth)with a general description,the level ofquality,precision and accuracy attainable,limitations,anduseful references given for each entry.1.2 The listed materials and methods are intended for thequalification of fluorometers as part of complying with regu-latory and other quality assurance/quality control(QA/QC)requirements.1.3 Precision and accuracy or uncertainty are given at a 1 confidence level and are approximated in cases where thesevalues have not been well established.31.4 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.5 This standard does not purport to address all of thesafety concerns,if any,associated with its use.It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2.Referenced Documents2.1 ASTM Standards:4E131 Terminology Relating to Molecular SpectroscopyE388 Test Method for Wavelength Accuracy and SpectralBandwidth of Fluorescence SpectrometersE578 Test Method for Linearity of Fluorescence MeasuringSystemsE579 Test Method for Limit of Detection of Fluorescence ofQuinine Sulfate in Solution3.Terminology3.1 Definitions(2):3.1.1 absorption coeffcient(),na measure of absorptionof radiant energy from an incident beam as it traverses anabsorbing medium according to Bouguers law,I/Io=e-b,where I and Ioare the transmitted and incident intensities,respectively,and b is the path length of the beam through thesample.E1313.1.1.1 DiscussionNote that transmittance T=I/Ioandabsorbance A=log T.3.1.2 absorptivity(a),nthe absorbance divided by theproduct of the concentration of the substance and the samplepathlength,a=A/bc.E1313.1.3 Beer-Lambert law,nrelates the dependence of theabsorbance(A)of a sample on its path length(see absorptioncoeffcient,)and concentration(c),such that A=a bc.3.1.3.1 DiscussionAlso called Beers law or Beer-Lambert-Bouquer law.E1313.1.4 calibrated detector(CD),noptical radiation detectorwhose responsivity as a function of wavelength has beendetermined along with corresponding uncertainties(3).3.1.5 calibrated diffuse reflector(CR),nLambertian re-flector whose reflectance as a function of wavelength has beendetermined along with corresponding uncertainties(4).3.1.6 calibrated optical radiation source(CS),nopticalradiation source whose radiance as a function of wavelengthhas been determined along with corresponding uncertainties(5,6).3.1.7 calibration,nset of procedures that establishes therelationship between quantities measured on an instrument andthe corresponding values realized by standards.3.1.8 certified reference material(CRM),nmaterial withproperties of interest whose values and corresponding uncer-tainties have been certified by a standardizing group ororganization.E1313.1.9 certified value,nvalue for which the certifying bodyhas the highest confidence in its accuracy in that all known or1This guide is under the jurisdiction of ASTM Committee E13 on MolecularSpectroscopy and Separation Science and is the direct responsibility of Subcom-mittee E13.01 on Ultra-Violet,Visible,and Luminescence Spectroscopy.Current edition approved May 1,2014.Published June 2014.Originallyapproved in 2009.Last previous edition approved in 2009 as E271909.DOI:10.1520/E2719-09R14.2The boldface numbers in parentheses refer to the list of references at the end ofthis standard.3Certain commercial equipment,instruments,or materials are identified in thisguide to foster understanding.Such identification does not imply recommendationor endorsement by ASTM International nor does it imply that the materials orequipment identified are necessarily the best available for the purpose.4For referenced ASTM standards,visit the ASTM website,www.astm.org,orcontact ASTM Customer Service at serviceastm.org.For Annual Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 suspected sources of bias have been investigated or accountedfor by the certifying body(7).3.1.10 diffuse scatterer,nmaterial that scatters opticalradiation in multiple directions;this includes diffuse reflectors,which are often Lambertian,and scattering soluti