1、Designation: E1247 12 (Reapproved 2017)Standard Practice forDetecting Fluorescence in Object-Color Specimens bySpectrophotometry1This standard is issued under the fixed designation E1247; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revis
2、ion, 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 practice provides spectrophotometric methods fordetecting the presence of fluorescence in object
3、-color speci-mens.NOTE 1Since the addition of fluorescing agents (colorants, whiteningagents, etc.) is often intentional by the manufacturer of a material,information on the presence or absence of fluorescent properties in aspecimen may often be obtained from the maker of the material.1.2 This pract
4、ice requires the use of a spectrophotometerthat both irradiates the specimen over the wavelength rangefrom 340 to 700 nm and allows the spectral distribution ofillumination on the specimen to be altered as desired.1.3 Within the above limitations, this practice is general inscope rather than specifi
5、c as to instrument or material.1.4 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, health, and environmental practices and deter-mine the applicability of regu
6、latory limitations prior to use.1.5 This international standard was developed in accor-dance with internationally recognized principles on standard-ization established in the Decision on Principles for theDevelopment of International Standards, Guides and Recom-mendations issued by the World Trade O
7、rganization TechnicalBarriers to Trade (TBT) Committee.2. Referenced Documents2.1 ASTM Standards:2D2244 Practice for Calculation of Color Tolerances andColor Differences from Instrumentally Measured ColorCoordinatesE284 Terminology of AppearanceE308 Practice for Computing the Colors of Objects by Us
8、ingthe CIE SystemE313 Practice for Calculating Yellowness and WhitenessIndices from Instrumentally Measured Color CoordinatesE991 Practice for Color Measurement of Fluorescent Speci-mens Using the One-Monochromator MethodE1164 Practice for Obtaining Spectrometric Data for Object-Color EvaluationE133
9、1 Test Method for Reflectance Factor and Color bySpectrophotometry Using Hemispherical GeometryE1348 Test Method for Transmittance and Color by Spec-trophotometry Using Hemispherical GeometryE1349 Test Method for Reflectance Factor and Color bySpectrophotometry Using Bidirectional (45:0 or 0:45)Geom
10、etryE2152 Practice for Computing the Colors of FluorescentObjects from Bispectral Photometric DataE2153 Practice for Obtaining Bispectral Photometric Datafor Evaluation of Fluorescent Color3. Terminology3.1 The definitions in Terminology E284, Practices E991,E2152, and E2153 are applicable to this p
11、ractice.4. Significance and Use4.1 Several standards, including Practices E991, E1164, andTest Methods E1331, E1348 and E1349, require either thepresence or absence of fluorescence exhibited by the specimenfor correct application. This practice provides spectrophoto-metric procedures for identifying
12、 the presence of fluorescencein materials.4.2 This practice is applicable to all object-color specimens,whether opaque, translucent, or transparent, meeting the re-quirements for specimens in the appropriate standards listed in2.1. Translucent specimens should be measured by reflectance,with a stand
13、ard non-fluorescent backing material, usually butnot necessarily black, placed behind the specimen duringmeasurement.4.3 This practice requires the use of a spectrophotometer inwhich the spectral distribution of the illumination on thespecimen can be altered by the user in one of several ways. Themo
14、dification of the illumination can either be by the insertion1This practice is under the jurisdiction of ASTM Committee E12 on Color andAppearance and is the direct responsibility of Subcommittee E12.05 on Fluores-cence.Current edition approved Nov. 1, 2017. Published November 2017. Originallyapprov
15、ed in 1988. Last previous edition approved in 2012 as E1247 12. DOI:10.1520/E1247-12R17.2For 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 Summa
16、ry page onthe ASTM website.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United StatesThis international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles
17、for theDevelopment of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.1of optical filters between the illuminating source and thespecimen, without interfering with the detection of the radia-tion from the specimen
18、, or by interchange of the illuminatingand detecting systems of the instrument or by scanning of boththe illuminating energy and detection output as in the two-monochromator method.4.4 The confirmation of the presence of fluorescence ismade by the comparison of spectral curves, color difference, ors
19、ingle parameter difference such as Y between the measure-ments.NOTE 2In editions of E1247 - 92 and earlier, the test of fluorescencewas the two sets of spectral transmittances or radiance factor (reflectancefactors) differ by 1 % of full scale at the wavelength of greatest difference.4.5 Either bidi
20、rectional or hemispherical instrument geom-etry may be used in this practice. The instrument must becapable of providing either broadband (white light) irradiationon the specimen or monochromatic irradiation and monochro-matic detection.4.6 This practice describes methods to detect the presenceof fl
21、uorescence only. It does not address the issue of whetherthe fluorescence makes a significant or insignificant contribu-tion to the colorimetric properties of the specimen for anygiven application. The user must determine the practicalsignificance of the effect of fluorescence on the color measure-m
22、ent.5. Instrumental Requirements5.1 This practice requires instrumentation meeting the fol-lowing requirements.5.1.1 The instrument source shall provide sufficient irradia-tion energy at the sample port to excite fluorescent emission, ifpresent.5.1.2 The instrument must provide one of the followingi
23、llumination/viewing combinations:5.1.2.1 Monochromatic illumination and monochromaticviewing (that is, a two-monochromator spectrophotometersometimes called a bispectrometer or spectrofluorimeter).5.1.2.2 Polychromatic illumination and monochromaticviewing.5.1.2.3 Reversible illumination/viewing to
24、allow both poly-chromatic illumination with monochromatic viewing andmonochromatic illumination with polychromatic viewing.5.1.3 The instrument and associated computer softwareshall allow the standardization/calibration of the instrumentusing user modified standardization/calibration values, whichis
25、 a requirement for using any of the filter methods describedin this practice.NOTE 3Repeatable and accurate application of this practice requiresspecialized instrumentation. Some commercial one-monochromator spec-trometers are limited in their ability to allow for the insertion of opticalfilters and
26、re-standardization with the filter in place as required in thisprocedure.6. Procedures6.1 There are three general types of procedures to detect thepresence of fluorescence instrumentally. Each has its advan-tages and shortcomings depending on the wavelength andintensity of the fluorescent emission a
27、nd the instrumentationavailable to the user.6.2 Two-Monochromator Method: This method requires acolorimetric measuring instrument that is equipped with twoseparate monochromators: the first, the illuminationmonochromator, irradiates the specimen with monochromaticlight and the second, the viewing mo
28、nochromator, analyzes theradiation leaving the specimen. A two-dimensional array ofbispectral photometric values is obtained by setting the illumi-nation monochromator at a series of fixed wavelengths ()inthe illumination band of the specimen, and for each , using theviewing monochromator to record
29、readings for each wave-length () in the specimens viewing range. The resulting array,once properly corrected, is known as the Donaldson matrix,and the value of each element (,) of this array is theDonaldson radiance factor (D(,). The reflection values areconfined to the diagonal of the matrix, and t
30、hese diagonalvalues are equal to the spectral reflectance factor of thespecimen. Therefore, the presence of fluorescence is demon-strated by non-zero off-diagonal elements. The measurementprocedures for this method are given in detail in PracticeE2153.6.3 Filter Methods: Filter methods follow the ge
31、neral pro-cedure of making a measurement of spectral radiance factorusing a spectrometer with broad band illumination, then addingone or more filters to remove the fluorescence-excitationenergy and measuring the spectral radiance factor under themodified illumination. The comparison of the resulting
32、 spectralcurves shows the presence or absence of fluorescence. If theexclusion of the excitation energy results in a difference in theremaining part of the curve, fluorescence is present and mustbe considered in the measurement procedures. If no differenceis found, then fluorescence is not an issue
33、in the measurementof that specimen.6.3.1 UV-Blocking MethodThis procedure is typicallyused for detecting the presence of optical brighteners, such asin white paper and textiles.6.3.1.1 Calibrate the instrument as required by the manu-facturer. (See Practice E1164 and the appropriate test methodfor t
34、he instrument geometry.)NOTE 4Since the measurement will be used to detect fluorescence, itshould be considered that fluorescence might be present, therefore thecalibration procedure should include adjusting the instruments illumina-tor to conform as closely as possible to D65 including the UV regio
35、n ofthe spectrum. In some commercial instruments this may be accomplishedby calibrating by whiteness index or the UV profile.6.3.1.2 Measure the specimen, obtaining either a table or agraph of spectral transmittance or reflectance factor versuswavelength.6.3.1.3 Insert a long-wavelength bandpass fil
36、ter between theilluminating source and the specimen. Select the cutoff wave-length of the filter according to the color of the specimen usingthe recommendation in Table 1 as a guide.(a) For spectrophotometers equipped for illumination bymeans of an integrating sphere, the filter must be placedbetwee
37、n the illuminating source and the illumination entranceE1247 12 (2017)2port of the sphere for reflectance measurement. For transmit-tance measurement, the filter must be placed between theilluminating source and the specimen.(b) For spectrophotometers equipped for illumination bymeans of bidirection
38、al geometry, the filter must be placedbetween the illuminating source and the specimen.6.3.1.4 Repeat the calibration in accordance with 6.3.1modifying the calibration values to be 0 below the cutoff of thefilter.6.3.1.5 Repeat the measurement in accordance with 6.3.1.2.NOTE 5This method employing o
39、nly one cut-off filter is mostcommonly used when measuring white materials where optical brighten-ing is suspected.6.3.2 Fluorescence-Weakening Method: In the fluorescence-weakening method two different bandpass filters are used andthree measurements are compared (1). One filter is chosen toremove a
40、ll the fluorescence-exciting wavelengths(fluorescence-killing filter), and the second filter is chosen toremove incident illumination about 20 to 40 nm shorter thanthe first filter (fluorescence-weakening filter). Use the proce-dure in 6.3.1.1 and 6.3.1.2 for the measurement without anyfilter in pla
41、ce. Then use the procedures in 6.3.1.3 6.3.1.5 forthe measurements with each of the filters. Refer to thereferenced literature for complete details of the application ofthis method.6.3.3 Filter Reduction Method: Several linear long band-pass filters are placed, one at a time, in the light path betwe
42、enthe source and the specimen. Usually 3 to 5 filters are enoughto estimate the reflected radiance factor (2). The same proce-dure is used to measure the specimen with each filter in place,following steps 6.3.1.1 6.3.1.5. The difference between themapped reflected radiance factor and the unfiltered
43、measure-ment reveals the presence or absence of fluorescence. Refer tothe referenced literature for complete details of the applicationof this method.6.3.4 Adjustment Method: In this method several narrowbandpass filters are placed in the optical path between thesource and the specimen one at a time
44、. This produces a seriesof readings which is used to determine the total radiance factoris a way somewhat analogous to an abridged two-monochromator instrument (3), (4). Again the difference be-tween the reflectance and the total radiance curves indicates thepresence or absence of fluorescence. Foll
45、ow the procedure in6.3.1.1 6.3.1.5 for the measurements with each filter. Refer tothe referenced literature for complete details of the applicationof this method.6.3.5 Serial Filter Method: (5) This method is a moregeneral case of the filter reduction method and may, withsuitable calibration, be equ
46、ivalent to the two-monochromatormethod. In the filter reduction method 3 to 5 filters in theregion of suspected fluorescence are used. In this method 10 to12 filters are used to measure the entire visible spectrum.Follow the procedure in 6.3.1.1 6.3.1.5 for measurementswith each filter. Then examine
47、 the difference between thecurves. Refer to the referenced literature for complete details ofthe application of this method.6.4 Two-Mode Method: The two-mode method also com-pares the results of two measurements. However in this case,instead of using a filter to exclude the excitation energy, thepro
48、cedure relies on the fact that the fluorescence will show upas increased values at the emission wavelengths when in themode involving polychromatic illumination, but not necessar-ily so when in the mode involving monochromatic illumina-tion. The two spectral curves will always have different shapesw
49、hen there is fluorescence(6),(7). Therefore, instruments inwhich the position of the source and detector can be switchedcan be used to detect the presence of fluorescence.6.4.1 Set the instrument for polychromatic illumination andcalibrate it, following the instrument manufacturers instruc-tions. (See Practice E1164 and the appropriate test method forthe instrument geometry.)6.4.2 Measure the specimen, obtaining either a table or agraph of spectral transmittance or reflectance factor versuswavelength.6.4.3 Set the instrument for monochromatic illuminati
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