1、Designation: E 1247 03Standard Practice forDetecting Fluorescence in Object-Color Specimens bySpectrophotometry1This standard is issued under the fixed designation E 1247; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of
2、 last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) 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-color speci-me
3、ns.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 practice requires th
4、e 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 specific as to instrum
5、ent 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 and health practices and determine the applica-bility of regulatory limitations prior to use
6、.2. Referenced Documents2.1 ASTM Standards:D 2244 Practice for Calculation of Color Tolerances andColor Differences from Intrumentally Measured ColorCoordinates2E 284 Terminology of Appearance2E 308 Practice for Computing the Colors of Objects byUsing the CIE System2E 313 Practice for Calculating Ye
7、llowness and WhitenessIndices from Instrumentally Measured Color Coordinates2E 991 Practice for Color Measurement of FluorescentSpecimens2E 1164 Practice for Obtaining Spectrophotometric Data forObject-Color Evaluation2E 1331 Test Method for Reflectance Factor and Color bySpectrophotometry Using Hem
8、ispherical Geometry2E 1348 Test Method for Transmittance and Color by Spec-trophotometry Using Hemispherical Geometry2E 1349 Test Method for Reflectance Factor and Color bySpectrophotometry Using Bidirectional Geometry2E 2152 Practice for Computing the Colors of FluorescentObjects from Bispectral Ph
9、otometric Data2E 2153 Practice for Obtaining Bispectral Photometric Datafor Evaluation of Fluorescent Color23. Terminology3.1 The definitions in Terminology E 284, Practices E 991,E 2152, and E 2153 are applicable to this practice.4. Significance and Use4.1 Several standards, including Practices E 9
10、91, E 1164,and Test Methods E 1331, E 1348 and E 1349, require eitherthe presence or absence of fluorescence exhibited by thespecimen for correct application. This practice provides spec-trophotometric procedures for identifying the presence offluorescence in materials.4.2 This practice is applicabl
11、e 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 standard non-fluorescent backing material, usually butnot necessarily black,
12、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. Themodification of the illumination can either be by the insertionof optical
13、filters between the illuminating source and thespecimen, without interfering with the detection of the radia-tion from the specimen, 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-monochrom
14、ator method.1This practice is under the jurisdiction of ASTM Committee E 12 on Color andAppearance and is the direct responsibility of Subcommittee E12.05 on Fluores-cence.Current edition approved Jan. 10, 2003. Published March 2003. Originallyapproved in 1988. Last previous edition approved in 2000
15、 as E 1247 92 (2000).2Annual Book of ASTM Standards, Vol 06.01.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.4.4 The confirmation of the presence of fluorescence ismade by the comparison of spectral curves, color difference, orsing
16、le parameter difference such as DY between the measure-ments.NOTE 2In editions of E 1247 - 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 bidir
17、ectional 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 Either bidirectional or hemispherical instrument geom-etry m
18、ay 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.5. Instrumental Requirements5.1 This practice requires instrumentation meeting the fol-lowing requirements.5.1.1
19、 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 followingillumination/viewing combinations:5.1.2.1 Monochromatic illumination and monochromaticviewing (that is, a two-monochr
20、omator spectrophotometersometimes called a bispectrometer or spectrofluorimeter).5.1.2.2 Polychromatic illumination and monochromaticviewing.5.1.2.3 Reversible illumination/viewing to allow both poly-chromatic illumination with monochromatic viewing andmonochromatic illumination with polychromatic v
21、iewing.5.1.3 The instrument and associated computer softwareshall allow the standardization/calibration of the instrumentusing user modified standardization/calibration values, whichis a requirement for using any of the filter methods describedin this practice.NOTE 3Repeatable and accurate applicati
22、on of this practice requiresspecialized instrumentation. Some commercial one-monochromator spec-trometers are limited in their ability to allow for the insertion of opticalfilters and re-standardization with the filter in place as required in thisprocedure.6. Procedures6.1 There are three general ty
23、pes of procedures to detect thepresence of fluorescence instrumentally. Each has its advan-tages and shortcomings depending on the wavelength andintensity of the fluorescent emission and the instrumentationavailable to the user.6.2 Two-Monochromator Method: This method requires acolorimetric measuri
24、ng instrument that is equipped with twoseparate monochromators: the first, the illumination monochro-mator, irradiates the specimen with monochromatic light andthe second, the viewing monochromator, analyzes the radiationleaving the specimen. A two-dimensional array of bispectralphotometric values i
25、s obtained by setting the illuminationmonochromator at a series of fixed wavelengths ()intheillumination band of the specimen, and for each , using theviewing monochromator to record readings for each wave-length (l) in the specimens viewing range. The resulting array,once properly corrected, is kno
26、wn as the Donaldson matrix,and the value of each element (,l) of this array is theDonaldson radiance factor (D(,l). The reflection values areconfined to the diagonal of the matrix, and these diagonalvalues are equal to the spectral reflectance factor of thespecimen. Therefore, the presence of fluore
27、scence is demon-strated by non-zero off-diagonal elements. The measurementprocedures for this method are given in detail in PracticeE 2153.6.3 Filter Methods: Filter methods follow the general pro-cedure of making a measurement of spectral radiance factorusing a spectrometer with broad band illumina
28、tion, then addingone or more filters to remove the fluorescence-excitationenergy and measuring the spectral radiance factor under themodified illumination. The comparison of the resulting spectralcurves shows the presence or absence of fluorescence. If theexclusion of the excitation energy results i
29、n 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 in the measurementof that specimen.6.3.1 UV-Blocking MethodThis procedure is typicallyused for detecting the pres
30、ence of optical brighteners, such asin white paper and textiles.6.3.1.1 Calibrate the instrument as required by the manu-facturer. (See Practice E 1164 and the appropriate test methodfor the instrument geometry.)NOTE 4Since the measurement will be used to detect fluorescence, itshould be considered
31、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 region ofthe spectrum. In some commercial instruments this may be accomplishedby calibrating by whiteness index or th
32、e 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 filter between theilluminating source and the specimen. Select the cutoff wave-length of the filter according to th
33、e 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 placedbetween the illuminating source and the illumination entranceport of the sphere for reflectance measurement. For trans
34、mit-tance measurement, the filter must be placed between theilluminating source and the specimen.TABLE 1 Edge-Position and Emission WavelengthsSample ColorEdge-PositionWavelength, nmMinimum EmissionWavelength, nmWhite or blue 440 400Green 510 480Yellow 540 480Orange 620 550Red 650 560E1247032(b) For
35、 spectrophotometers equipped for illumination bymeans of bidirectional 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
36、measurement in accordance with 6.3.1.2.NOTE 5This method employing only 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 and
37、three measurements are compared (1). One filter is chosen toremove all 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
38、 in 6.3.1.1 and 6.3.1.2 for the measurement without anyfilter in place. Then use the procedures in 6.3.1.3-6.3.1.5 for themeasurements with each of the filters. Refer to the referencedliterature for complete details of the application of this method.6.3.3 Filter Reduction Method: Several linear long
39、 band-pass filters are placed, one at a time, in the light path betweenthe 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 diffe
40、rence between themapped reflected radiance factor and the unfiltered 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
41、 in the optical path between thesource and the specimen one at a time. 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 r
42、adiance curves indicates thepresence or absence of fluorescence. Follow 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
43、the filter reduction method and may, withsuitable calibration, be equivalent 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 proced
44、ure in 6.3.1.1-6.3.1.5 for measurements witheach filter. Then examine the difference between the curves.Refer to the referenced literature for complete details of theapplication of this method.6.4 Two-Mode Method: The two-mode method also com-pares the results of two measurements. However in this ca
45、se,instead of using a filter to exclude the excitation energy, theprocedure 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 illu
46、mina-tion. The two spectral curves will always have different shapeswhen 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 and
47、calibrate it, following the instrument manufacturers instruc-tions. (See Practice E 1164 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 instrumen
48、t for monochromatic illuminationand calibrate it in a manner similar to that given in 6.3.1.6.4.4 Measure the specimen in accordance with 6.3.2.7. Interpretation of Results7.1 The confirmation of the presence of fluorescence ismade by examining the Donaldson matrix or by the compari-son of spectral
49、curves at the wavelength of maximum devia-tion, color difference, or single parameter difference such asDY or Whiteness Index (WI) between the measurements. Ifyou have used the two-monochromator method follow step 7.2or 7.5, or both. If you are using the comparison of spectralcurves at the wavelength of maximum deviation follow step7.3 for the filter methods (6.3) or 7.4 for the two-mode method(6.4). If you are using a color difference calculation or singleparameter difference use step 7.5.7.2 Using the two-monochromator method, examine theDonaldson matrix. Since the refle
