1、Designation:E124703 Designation: E1247 12Standard 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 rev
2、ision, 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 for detecting the presence of fluorescence in obj
3、ect-color specimens.NOTE 1Since the addition of fluorescing agents (colorants, whitening agents, etc.) is often intentional by the manufacturer of a material, informationon the presence or absence of fluorescent properties in a specimen may often be obtained from the maker of the material.1.2 This p
4、ractice requires the use of a spectrophotometer that both irradiates the specimen over the wavelength range from 340to 700 nm and allows the spectral distribution of illumination on the specimen to be altered as desired.1.3 Within the above limitations, this practice is general in scope rather than
5、specific as to instrument or material.1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibilityof the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatorylimi
6、tations prior to use.2. Referenced Documents2.1 ASTM Standards:2D2244 Practice for Calculation of Color Tolerances and Color Differences from Instrumentally Measured Color CoordinatesE284 Terminology of AppearanceE308 Practice for Computing the Colors of Objects by Using the CIE SystemE313 Practice
7、for Calculating Yellowness and Whiteness Indices from Instrumentally Measured Color CoordinatesE991 Practice for Color Measurement of Fluorescent Specimens Using the One-Monochromator MethodE1164 Practice for Obtaining Spectrometric Data for Object-Color EvaluationE1331 Test Method for Reflectance F
8、actor and Color by Spectrophotometry Using Hemispherical GeometryE1348 Test Method for Transmittance and Color by Spectrophotometry Using Hemispherical GeometryE1349 Test Method for Reflectance Factor and Color by Spectrophotometry Using Bidirectional (45:0 or 0:45) GeometryE2152 Practice for Comput
9、ing the Colors of Fluorescent Objects from Bispectral Photometric DataE2153 Practice for Obtaining Bispectral Photometric Data for Evaluation of Fluorescent Color3. Terminology3.1 The definitions in Terminology E284, Practices E991, E2152, and E2153 are applicable to this practice.4. Significance an
10、d Use4.1 Several standards, including Practices E991, E1164, and Test Methods E1331, E1348 and E1349, require either the presenceor absence of fluorescence exhibited by the specimen for correct application. This practice provides spectrophotometric proceduresfor identifying the presence of fluoresce
11、nce in materials.4.2 This practice is applicable to all object-color specimens, whether opaque, translucent, or transparent, meeting therequirements for specimens in the appropriate standards listed in 2.1. Translucent specimens should be measured by reflectance,with a standard non-fluorescent backi
12、ng material, usually but not necessarily black, placed behind the specimen duringmeasurement.4.3 This practice requires the use of a spectrophotometer in which the spectral distribution of the illumination on the specimen1This practice is under the jurisdiction of ASTM Committee E12 on Color and App
13、earance and is the direct responsibility of Subcommittee E12.05 on Fluorescence.Current edition approved Jan. 10, 2003. Published March 2003. Originally approved in 1988. Last previous edition approved in 2000 as E124792(2000). DOI:10.1520/E1247-03.Current edition approved July 1, 2012. Published Se
14、ptember 2012. Originally approved in 1988. Last previous edition approved in 2003 as E1247 03 which waswithdrawn January 2012 and reinstated in July 2012. DOI: 10.1520/E1247-12.2For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at serviceastm.org.
15、For Annual Book of ASTM Standardsvolume information, refer to the standards Document Summary page on the ASTM website.1This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Becauseit ma
16、y not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current versionof the standard as published by ASTM is to be considered the official document.Copyright ASTM International, 100 Barr Harb
17、or Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.can be altered by the user in one of several ways. The modification of the illumination can either be by the insertion of opticalfilters between the illuminating source and the specimen, without interfering with the detection of
18、the radiation from the specimen,or by interchange of the illuminating and detecting systems of the instrument or by scanning of both the illuminating energy anddetection output as in the two-monochromator method.4.4 The confirmation of the presence of fluorescence is made by the comparison of spectr
19、al curves, color difference, or singleparameter difference such as DY between the measurements.NOTE 2In editions of E1247 - 92 and earlier, the test of fluorescence was the two sets of spectral transmittances or radiance factor (reflectance factors)differ by 1 % of full scale at the wavelength of gr
20、eatest difference.4.5 Either bidirectional or hemispherical instrument geometry may be used in this practice. The instrument must be capable ofproviding either broadband (white light) irradiation on the specimen or monochromatic irradiation and monochromatic detection.4.6Either bidirectional or hemi
21、spherical instrument geometry may be used in this practice. The instrument must be capable ofproviding either broadband (white light) irradiation on the specimen or monochromatic irradiation and monochromatic detection.4.6 This practice describes methods to detect the presence of fluorescence only.
22、It does not address the issue of whether thefluorescence makes a significant or insignificant contribution to the colorimetric properties of the specimen for any givenapplication. The user must determine the practical significance of the effect of fluorescence on the color measurement.5. Instrumenta
23、l Requirements5.1 This practice requires instrumentation meeting the following requirements.5.1.1 The instrument source shall provide sufficient irradiation energy at the sample port to excite fluorescent emission, ifpresent.5.1.2 The instrument must provide one of the following illumination/viewing
24、 combinations:5.1.2.1 Monochromatic illumination and monochromatic viewing (that is, a two-monochromator spectrophotometer sometimescalled a bispectrometer or spectrofluorimeter).5.1.2.2 Polychromatic illumination and monochromatic viewing.5.1.2.3 Reversible illumination/viewing to allow both polych
25、romatic illumination with monochromatic viewing and mono-chromatic illumination with polychromatic viewing.5.1.3 The instrument and associated computer software shall allow the standardization/calibration of the instrument using usermodified standardization/calibration values, which is a requirement
26、 for using any of the filter methods described in this practice.NOTE 3Repeatable and accurate application of this practice requires specialized instrumentation. Some commercial one-monochromatorspectrometers are limited in their ability to allow for the insertion of optical filters and re-standardiz
27、ation with the filter in place as required in thisprocedure.6. Procedures6.1 There are three general types of procedures to detect the presence of fluorescence instrumentally. Each has its advantagesand shortcomings depending on the wavelength and intensity of the fluorescent emission and the instru
28、mentation available to theuser.6.2 Two-Monochromator Method: This method requires a colorimetric measuring instrument that is equipped with two separatemonochromators: the first, the illumination monochromator, irradiates the specimen with monochromatic light and the second, theviewing monochromator
29、, analyzes the radiation leaving the specimen. A two-dimensional array of bispectral photometric valuesis obtained by setting the illumination monochromator at a series of fixed wavelengths () in the illumination band of the specimen,and for each , using the viewing monochromator to record readings
30、for each wavelength (l) in the specimens viewing range.The resulting array, once properly corrected, is known as the Donaldson matrix, and the value of each element (,l) of this arrayis the Donaldson radiance factor (D(,l) ). The reflection values are confined to the diagonal of the matrix, and thes
31、e diagonalvalues are equal to the spectral reflectance factor of the specimen. Therefore, the presence of fluorescence is demonstrated bynon-zero off-diagonal elements. The measurement procedures for this method are given in detail in Practice E2153.6.3 Filter Methods: Filter methods follow the gene
32、ral procedure of making a measurement of spectral radiance factor using aspectrometer with broad band illumination, then adding one or more filters to remove the fluorescence-excitation energy andmeasuring the spectral radiance factor under the modified illumination. The comparison of the resulting
33、spectral curves shows thepresence or absence of fluorescence. If the exclusion of the excitation energy results in a difference in the remaining part of thecurve, fluorescence is present and must be considered in the measurement procedures. If no difference is found, then fluorescenceis not an issue
34、 in the measurement of that specimen.6.3.1 UV-Blocking MethodThis procedure is typically used for detecting the presence of optical brighteners, such as in whitepaper and textiles.6.3.1.1 Calibrate the instrument as required by the manufacturer. (See Practice E1164 and the appropriate test method fo
35、r theinstrument geometry.)NOTE 4Since the measurement will be used to detect fluorescence, it should be considered that fluorescence might be present, therefore the calibrationprocedure should include adjusting the instruments illuminator to conform as closely as possible to D65 including the UV reg
36、ion of the spectrum. Insome commercial instruments this may be accomplished by calibrating by whiteness index or the UV profile.E1247 1226.3.1.2 Measure the specimen, obtaining either a table or a graph of spectral transmittance or reflectance factor versuswavelength.6.3.1.3 Insert a long-wavelength
37、 bandpass filter between the illuminating source and the specimen. Select the cutoff wavelengthof the filter according to the color of the specimen using the recommendation in Table 1 as a guide.(a) For spectrophotometers equipped for illumination by means of an integrating sphere, the filter must b
38、e placed between theilluminating source and the illumination entrance port of the sphere for reflectance measurement. For transmittance measurement,the filter must be placed between the illuminating source and the specimen.(b) For spectrophotometers equipped for illumination by means of bidirectiona
39、l geometry, the filter must be placed between theilluminating source and the specimen.6.3.1.4 Repeat the calibration in accordance with 6.3.1 modifying the calibration values to be 0 below the cutoff of the filter.6.3.1.5 Repeat the measurement in accordance with 6.3.1.2.NOTE 5This method employing
40、only one cut-off filter is most commonly used when measuring white materials where optical brightening issuspected.6.3.2 Fluorescence-Weakening Method: In the fluorescence-weakening method two different bandpass filters are used and threemeasurements are compared (1). One filter is chosen to remove
41、all the fluorescence-exciting wavelengths (fluorescence-killingfilter), and the second filter is chosen to remove incident illumination about 20 to 40 nm shorter than the first filter(fluorescence-weakening filter). Use the procedure in 6.3.1.1 and 6.3.1.2 for the measurement without any filter in p
42、lace. Then usethe procedures in 6.3.1.3-6.3.1.5 for the measurements with each of the filters. Refer to the referenced literature for complete detailsof the application of this method.6.3.3 Filter Reduction Method: Several linear long bandpass filters are placed, one at a time, in the light path bet
43、ween the sourceand the specimen. Usually 3 to 5 filters are enough to estimate the reflected radiance factor (2). The same procedure is used tomeasure the specimen with each filter in place, following steps 6.3.1.1-6.3.1.5. The difference between the mapped reflectedradiance factor and the unfiltere
44、d measurement reveals the presence or absence of fluorescence. Refer to the referenced literaturefor complete details of the application of this method.6.3.4 Adjustment Method: In this method several narrow bandpass filters are placed in the optical path between the source andthe specimen one at a t
45、ime. This produces a series of readings which is used to determine the total radiance factor is a waysomewhat analogous to an abridged two-monochromator instrument (3), (4). Again the difference between the reflectance and thetotal radiance curves indicates the presence or absence of fluorescence. F
46、ollow the procedure in 6.3.1.1-6.3.1.5 for themeasurements with each filter. Refer to the referenced literature for complete details of the application of this method.6.3.5 Serial Filter Method: (5) This method is a more general case of the filter reduction method and may, with suitablecalibration,
47、be equivalent to the two-monochromator method. In the filter reduction method 3 to 5 filters in the region of suspectedfluorescence are used. In this method 10 to 12 filters are used to measure the entire visible spectrum. Follow the procedure in6.3.1.1-6.3.1.5 for measurements with each filter. The
48、n examine the difference between the curves. Refer to the referencedliterature for complete details of the application of this method.6.4 Two-Mode Method: The two-mode method also compares the results of two measurements. However in this case, insteadof using a filter to exclude the excitation energ
49、y, the procedure relies on the fact that the fluorescence will show up as increasedvalues at the emission wavelengths when in the mode involving polychromatic illumination, but not necessarily so when in themode involving monochromatic illumination. The two spectral curves will always have different shapes when there is fluorescence(6),(7). Therefore, instruments in which the position of the source and detector can be switched can be used to detect the presenceof fluorescence.6.4.1 Set the instrument for polychromatic illumination and calibrate it, following the instru
