ASTM E1247-12(2017)
(Practice)Standard Practice for Detecting Fluorescence in Object-Color Specimens by Spectrophotometry
Standard Practice for Detecting Fluorescence in Object-Color Specimens by Spectrophotometry
SIGNIFICANCE AND USE
4.1 Several standards, including Practices E991, E1164, and Test Methods E1331, E1348 and E1349, require either the presence or absence of fluorescence exhibited by the specimen for correct application. This practice provides spectrophotometric procedures for identifying the presence of fluorescence in materials.
4.2 This practice is applicable to all object-color specimens, whether opaque, translucent, or transparent, meeting the requirements for specimens in the appropriate standards listed in 2.1. Translucent specimens should be measured by reflectance, with a standard non-fluorescent backing material, usually but not necessarily black, placed behind the specimen during measurement.
4.3 This practice requires the use of a spectrophotometer in which the spectral distribution of the illumination on the specimen can be altered by the user in one of several ways. The modification of the illumination can either be by the insertion of optical filters between the illuminating source and the specimen, without interfering with the detection of 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 and detection output as in the two-monochromator method.
4.4 The confirmation of the presence of fluorescence is made by the comparison of spectral curves, color difference, or single parameter difference such as ΔY between the measurements.
Note 2: In 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 greatest difference.
4.5 Either bidirectional or hemispherical instrument geometry may be used in this practice. The instrument must be capable of providing either broadband (white light) irradiation on the specimen or monochromatic irradiation and monochromatic detection.
4.6 This practice describes methods to detect the...
SCOPE
1.1 This practice provides spectrophotometric methods for detecting the presence of fluorescence in object-color specimens.
Note 1: Since the addition of fluorescing agents (colorants, whitening agents, etc.) is often intentional by the manufacturer of a material, information on the presence or absence of fluorescent properties in a specimen may often be obtained from the maker of the material.
1.2 This practice requires the use of a spectrophotometer that both irradiates the specimen over the wavelength range from 340 to 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 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 responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
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Standards Content (Sample)
This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E1247 − 12 (Reapproved 2017)
Standard Practice for
Detecting Fluorescence in Object-Color Specimens by
Spectrophotometry
This standard is issued under the fixed designation E1247; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope E308 PracticeforComputingtheColorsofObjectsbyUsing
the CIE System
1.1 This practice provides spectrophotometric methods for
E313 Practice for Calculating Yellowness and Whiteness
detecting the presence of fluorescence in object-color speci-
Indices from Instrumentally Measured Color Coordinates
mens.
E991 Practice for Color Measurement of Fluorescent Speci-
NOTE 1—Since the addition of fluorescing agents (colorants, whitening
mens Using the One-Monochromator Method
agents, etc.) is often intentional by the manufacturer of a material,
E1164 PracticeforObtainingSpectrometricDataforObject-
information on the presence or absence of fluorescent properties in a
Color Evaluation
specimen may often be obtained from the maker of the material.
E1331 Test Method for Reflectance Factor and Color by
1.2 This practice requires the use of a spectrophotometer
Spectrophotometry Using Hemispherical Geometry
that both irradiates the specimen over the wavelength range
E1348 Test Method for Transmittance and Color by Spec-
from 340 to 700 nm and allows the spectral distribution of
trophotometry Using Hemispherical Geometry
illumination on the specimen to be altered as desired.
E1349 Test Method for Reflectance Factor and Color by
1.3 Within the above limitations, this practice is general in
Spectrophotometry Using Bidirectional (45°:0° or 0°:45°)
scope rather than specific as to instrument or material.
Geometry
E2152 Practice for Computing the Colors of Fluorescent
1.4 This standard does not purport to address all of the
Objects from Bispectral Photometric Data
safety concerns, if any, associated with its use. It is the
E2153 Practice for Obtaining Bispectral Photometric Data
responsibility of the user of this standard to establish appro-
for Evaluation of Fluorescent Color
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
3. Terminology
1.5 This international standard was developed in accor-
dance with internationally recognized principles on standard- 3.1 The definitions in Terminology E284, Practices E991,
ization established in the Decision on Principles for the E2152, and E2153 are applicable to this practice.
Development of International Standards, Guides and Recom-
4. Significance and Use
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
4.1 Several standards, including Practices E991, E1164, and
Test Methods E1331, E1348 and E1349, require either the
2. Referenced Documents
presence or absence of fluorescence exhibited by the specimen
2.1 ASTM Standards: for correct application. This practice provides spectrophoto-
metric procedures for identifying the presence of fluorescence
D2244 Practice for Calculation of Color Tolerances and
Color Differences from Instrumentally Measured Color in materials.
Coordinates
4.2 This practice is applicable to all object-color specimens,
E284 Terminology of Appearance
whether opaque, translucent, or transparent, meeting the re-
quirements for specimens in the appropriate standards listed in
2.1. Translucent specimens should be measured by reflectance,
This practice is under the jurisdiction of ASTM Committee E12 on Color and
with a standard non-fluorescent backing material, usually but
Appearance and is the direct responsibility of Subcommittee E12.05 on Fluores-
cence.
not necessarily black, placed behind the specimen during
Current edition approved Nov. 1, 2017. Published November 2017. Originally
measurement.
approved in 1988. Last previous edition approved in 2012 as E1247 – 12. DOI:
10.1520/E1247-12R17.
4.3 This practice requires the use of a spectrophotometer in
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
which the spectral distribution of the illumination on the
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
specimencanbealteredbytheuserinoneofseveralways.The
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. modification of the illumination can either be by the insertion
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E1247 − 12 (2017)
of optical filters between the illuminating source and the tages and shortcomings depending on the wavelength and
specimen, without interfering with the detection of the radia- intensity of the fluorescent emission and the instrumentation
tion from the specimen, or by interchange of the illuminating available to the user.
and detecting systems of the instrument or by scanning of both
6.2 Two-Monochromator Method: This method requires a
the illuminating energy and detection output as in the two-
colorimetric measuring instrument that is equipped with two
monochromator method.
separate monochromators: the first, the illumination
4.4 The confirmation of the presence of fluorescence is
monochromator, irradiates the specimen with monochromatic
made by the comparison of spectral curves, color difference, or
light and the second, the viewing monochromator, analyzes the
single parameter difference such as ∆Y between the measure-
radiation leaving the specimen. A two-dimensional array of
ments.
bispectral photometric values is obtained by setting the illumi-
nation monochromator at a series of fixed wavelengths (µ)in
NOTE 2—In editions of E1247 - 92 and earlier, the test of fluorescence
theilluminationbandofthespecimen,andforeachµ,usingthe
was the two sets of spectral transmittances or radiance factor (reflectance
factors) differ by 1 % of full scale at the wavelength of greatest difference. viewing monochromator to record readings for each wave-
length(λ)inthespecimen’sviewingrange.Theresultingarray,
4.5 Either bidirectional or hemispherical instrument geom-
once properly corrected, is known as the Donaldson matrix,
etry may be used in this practice. The instrument must be
and the value of each element (µ,λ) of this array is the
capable of providing either broadband (white light) irradiation
Donaldson radiance factor (D(µ,λ)). The reflection values are
on the specimen or monochromatic irradiation and monochro-
confined to the diagonal of the matrix, and these diagonal
matic detection.
values are equal to the spectral reflectance factor of the
4.6 This practice describes methods to detect the presence
specimen. Therefore, the presence of fluorescence is demon-
of fluorescence only. It does not address the issue of whether
strated by non-zero off-diagonal elements. The measurement
the fluorescence makes a significant or insignificant contribu-
procedures for this method are given in detail in Practice
tion to the colorimetric properties of the specimen for any
E2153.
given application. The user must determine the practical
6.3 Filter Methods: Filter methods follow the general pro-
significance of the effect of fluorescence on the color measure-
cedure of making a measurement of spectral radiance factor
ment.
usingaspectrometerwithbroadbandillumination,thenadding
one or more filters to remove the fluorescence-excitation
5. Instrumental Requirements
energy and measuring the spectral radiance factor under the
5.1 This practice requires instrumentation meeting the fol-
modifiedillumination.Thecomparisonoftheresultingspectral
lowing requirements.
curves shows the presence or absence of fluorescence. If the
5.1.1 The instrument source shall provide sufficient irradia-
exclusion of the excitation energy results in a difference in the
tion energy at the sample port to excite fluorescent emission, if
remaining part of the curve, fluorescence is present and must
present.
be considered in the measurement procedures. If no difference
5.1.2 The instrument must provide one of the following is found, then fluorescence is not an issue in the measurement
illumination/viewing combinations: of that specimen.
5.1.2.1 Monochromatic illumination and monochromatic
6.3.1 UV-Blocking Method—This procedure is typically
viewing (that is, a two-monochromator spectrophotometer
used for detecting the presence of optical brighteners, such as
sometimes called a bispectrometer or spectrofluorimeter).
in white paper and textiles.
5.1.2.2 Polychromatic illumination and monochromatic
6.3.1.1 Calibrate the instrument as required by the manu-
viewing.
facturer. (See Practice E1164 and the appropriate test method
5.1.2.3 Reversible illumination/viewing to allow both poly-
for the instrument geometry.)
chromatic illumination with monochromatic viewing and
NOTE 4—Since the measurement will be used to detect fluorescence, it
monochromatic illumination with polychromatic viewing.
should be considered that fluorescence might be present, therefore the
5.1.3 The instrument and associated computer software
calibration procedure should include adjusting the instrument’s illumina-
shall allow the standardization/calibration of the instrument tor to conform as closely as possible to D65 including the UV region of
the spectrum. In some commercial instruments this may be accomplished
using user modified standardization/calibration values, which
by calibrating by whiteness index or the UV profile.
is a requirement for using any of the filter methods described
in this practice.
6.3.1.2 Measure the specimen, obtaining either a table or a
graph of spectral transmittance or reflectance factor versus
NOTE 3—Repeatable and accurate application of this practice requires
wavelength.
specialized instrumentation. Some commercial one-monochromator spec-
trometers are limited in their ability to allow for the insertion of optical
6.3.1.3 Insert a long-wavelength bandpass filter between the
filters and re-standardization with the filter in place as required in this
illuminating source and the specimen. Select the cutoff wave-
procedure.
length of the filter according to the color of the specimen using
the recommendation in Table 1 as a guide.
6. Procedures
(a) For spectrophotometers equipped for illumination by
6.1 There are three general types of procedures to detect the means of an integrating sphere, the filter must be placed
presence of fluorescence instrumentally. Each has its advan- between the illuminating source and the illumination entrance
E1247 − 12 (2017)
TABLE 1 Edge-Position and Emission Wavelengths
method. In the filter redu
...
NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E1247 − 12 (Reapproved 2017)
Standard Practice for
Detecting Fluorescence in Object-Color Specimens by
Spectrophotometry
This standard is issued under the fixed designation E1247; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope E308 Practice for Computing the Colors of Objects by Using
the CIE System
1.1 This practice provides spectrophotometric methods for
E313 Practice for Calculating Yellowness and Whiteness
detecting the presence of fluorescence in object-color speci-
Indices from Instrumentally Measured Color Coordinates
mens.
E991 Practice for Color Measurement of Fluorescent Speci-
NOTE 1—Since the addition of fluorescing agents (colorants, whitening
mens Using the One-Monochromator Method
agents, etc.) is often intentional by the manufacturer of a material,
E1164 Practice for Obtaining Spectrometric Data for Object-
information on the presence or absence of fluorescent properties in a
Color Evaluation
specimen may often be obtained from the maker of the material.
E1331 Test Method for Reflectance Factor and Color by
1.2 This practice requires the use of a spectrophotometer
Spectrophotometry Using Hemispherical Geometry
that both irradiates the specimen over the wavelength range
E1348 Test Method for Transmittance and Color by Spec-
from 340 to 700 nm and allows the spectral distribution of
trophotometry Using Hemispherical Geometry
illumination on the specimen to be altered as desired.
E1349 Test Method for Reflectance Factor and Color by
1.3 Within the above limitations, this practice is general in
Spectrophotometry Using Bidirectional (45°:0° or 0°:45°)
scope rather than specific as to instrument or material.
Geometry
E2152 Practice for Computing the Colors of Fluorescent
1.4 This standard does not purport to address all of the
Objects from Bispectral Photometric Data
safety concerns, if any, associated with its use. It is the
E2153 Practice for Obtaining Bispectral Photometric Data
responsibility of the user of this standard to establish appro-
for Evaluation of Fluorescent Color
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
3. Terminology
1.5 This international standard was developed in accor-
dance with internationally recognized principles on standard- 3.1 The definitions in Terminology E284, Practices E991,
ization established in the Decision on Principles for the E2152, and E2153 are applicable to this practice.
Development of International Standards, Guides and Recom-
4. Significance and Use
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
4.1 Several standards, including Practices E991, E1164, and
Test Methods E1331, E1348 and E1349, require either the
2. Referenced Documents
presence or absence of fluorescence exhibited by the specimen
for correct application. This practice provides spectrophoto-
2.1 ASTM Standards:
D2244 Practice for Calculation of Color Tolerances and metric procedures for identifying the presence of fluorescence
in materials.
Color Differences from Instrumentally Measured Color
Coordinates
4.2 This practice is applicable to all object-color specimens,
E284 Terminology of Appearance
whether opaque, translucent, or transparent, meeting the re-
quirements for specimens in the appropriate standards listed in
2.1. Translucent specimens should be measured by reflectance,
This practice is under the jurisdiction of ASTM Committee E12 on Color and
with a standard non-fluorescent backing material, usually but
Appearance and is the direct responsibility of Subcommittee E12.05 on Fluores-
cence.
not necessarily black, placed behind the specimen during
Current edition approved Nov. 1, 2017. Published November 2017. Originally
measurement.
approved in 1988. Last previous edition approved in 2012 as E1247 – 12. DOI:
10.1520/E1247-12R17.
4.3 This practice requires the use of a spectrophotometer in
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
which the spectral distribution of the illumination on the
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
specimen can be altered by the user in one of several ways. The
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. modification of the illumination can either be by the insertion
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E1247 − 12 (2017)
of optical filters between the illuminating source and the tages and shortcomings depending on the wavelength and
specimen, without interfering with the detection of the radia- intensity of the fluorescent emission and the instrumentation
tion from the specimen, or by interchange of the illuminating available to the user.
and detecting systems of the instrument or by scanning of both
6.2 Two-Monochromator Method: This method requires a
the illuminating energy and detection output as in the two-
colorimetric measuring instrument that is equipped with two
monochromator method.
separate monochromators: the first, the illumination
4.4 The confirmation of the presence of fluorescence is
monochromator, irradiates the specimen with monochromatic
made by the comparison of spectral curves, color difference, or
light and the second, the viewing monochromator, analyzes the
single parameter difference such as ΔY between the measure-
radiation leaving the specimen. A two-dimensional array of
ments.
bispectral photometric values is obtained by setting the illumi-
nation monochromator at a series of fixed wavelengths (µ) in
NOTE 2—In editions of E1247 - 92 and earlier, the test of fluorescence
the illumination band of the specimen, and for each µ, using the
was the two sets of spectral transmittances or radiance factor (reflectance
factors) differ by 1 % of full scale at the wavelength of greatest difference. viewing monochromator to record readings for each wave-
length (λ) in the specimen’s viewing range. The resulting array,
4.5 Either bidirectional or hemispherical instrument geom-
once properly corrected, is known as the Donaldson matrix,
etry may be used in this practice. The instrument must be
and the value of each element (µ,λ) of this array is the
capable of providing either broadband (white light) irradiation
Donaldson radiance factor (D(µ,λ)). The reflection values are
on the specimen or monochromatic irradiation and monochro-
confined to the diagonal of the matrix, and these diagonal
matic detection.
values are equal to the spectral reflectance factor of the
4.6 This practice describes methods to detect the presence
specimen. Therefore, the presence of fluorescence is demon-
of fluorescence only. It does not address the issue of whether
strated by non-zero off-diagonal elements. The measurement
the fluorescence makes a significant or insignificant contribu-
procedures for this method are given in detail in Practice
tion to the colorimetric properties of the specimen for any
E2153.
given application. The user must determine the practical
6.3 Filter Methods: Filter methods follow the general pro-
significance of the effect of fluorescence on the color measure-
cedure of making a measurement of spectral radiance factor
ment.
using a spectrometer with broad band illumination, then adding
one or more filters to remove the fluorescence-excitation
5. Instrumental Requirements
energy and measuring the spectral radiance factor under the
5.1 This practice requires instrumentation meeting the fol-
modified illumination. The comparison of the resulting spectral
lowing requirements.
curves shows the presence or absence of fluorescence. If the
5.1.1 The instrument source shall provide sufficient irradia-
exclusion of the excitation energy results in a difference in the
tion energy at the sample port to excite fluorescent emission, if
remaining part of the curve, fluorescence is present and must
present.
be considered in the measurement procedures. If no difference
5.1.2 The instrument must provide one of the following is found, then fluorescence is not an issue in the measurement
illumination/viewing combinations: of that specimen.
5.1.2.1 Monochromatic illumination and monochromatic
6.3.1 UV-Blocking Method—This procedure is typically
viewing (that is, a two-monochromator spectrophotometer
used for detecting the presence of optical brighteners, such as
sometimes called a bispectrometer or spectrofluorimeter).
in white paper and textiles.
5.1.2.2 Polychromatic illumination and monochromatic
6.3.1.1 Calibrate the instrument as required by the manu-
viewing.
facturer. (See Practice E1164 and the appropriate test method
5.1.2.3 Reversible illumination/viewing to allow both poly- for the instrument geometry.)
chromatic illumination with monochromatic viewing and
NOTE 4—Since the measurement will be used to detect fluorescence, it
monochromatic illumination with polychromatic viewing.
should be considered that fluorescence might be present, therefore the
5.1.3 The instrument and associated computer software
calibration procedure should include adjusting the instrument’s illumina-
shall allow the standardization/calibration of the instrument tor to conform as closely as possible to D65 including the UV region of
the spectrum. In some commercial instruments this may be accomplished
using user modified standardization/calibration values, which
by calibrating by whiteness index or the UV profile.
is a requirement for using any of the filter methods described
in this practice.
6.3.1.2 Measure the specimen, obtaining either a table or a
graph of spectral transmittance or reflectance factor versus
NOTE 3—Repeatable and accurate application of this practice requires
wavelength.
specialized instrumentation. Some commercial one-monochromator spec-
trometers are limited in their ability to allow for the insertion of optical
6.3.1.3 Insert a long-wavelength bandpass filter between the
filters and re-standardization with the filter in place as required in this
illuminating source and the specimen. Select the cutoff wave-
procedure.
length of the filter according to the color of the specimen using
the recommendation in Table 1 as a guide.
6. Procedures
(a) For spectrophotometers equipped for illumination by
6.1 There are three general types of procedures to detect the means of an integrating sphere, the filter must be placed
presence of fluorescence instrumentally. Each has its advan- between the illuminating source and the illumination entrance
E1247 − 12 (2017)
TABLE 1 Edge-Position and Emission Wavelengths
method. In the filter reduction method 3 to 5 filters in the
Edge-Position Minimum Emission region of suspected fluorescence are used. In this method 10 to
Sample Color
Wavelength, nm Wavelength, nm
12 filters are used to measure the entire visible spectrum.
White or blue 440 400
Follow the procedure in 6.3.1.1 – 6.3.1.5 for measurements
Green 510 480
with each filter. Then examine the difference between the
Yellow 540 480
Orange 620 550 curves. Refer to the referenced literature for complete details of
Red 650 560
the 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,
port of the sphere for reflectance measurement. For transmit- instead of using a filter to exclude the excitation energy, the
tance measurement, the filter must be placed between the
procedure relies on the fact that the fluorescence will show up
illuminating source and the specimen. as increased values at the emission wavelengths when in the
(b) For spectrophotometers equipped for illumination by
mode involving polychromatic illumination, but not necessar-
means of bidirectional geometry, the filter must be placed ily so when in the mode involving monochromatic illumina-
between the illuminating source and the specimen.
tion. The two spectral curves will always have different shapes
6.3.1.4 Repeat the calibration in accordance with 6.3.1 when there is fluorescence(6),(7). Therefore, instruments in
modifying the calibration values to be 0 below the cutoff of the
which the position of the source and detector can be switched
filter. can be used to detect the presence of fluorescence.
6.3.1.5 Repeat the measurement in accordance with 6.3.1.2.
6.4.1 Set the instrument for polychromatic illumination and
calibrate it, following the instrument manufacturer’s instruc-
NOTE 5—This method employing only one cut-off filter is most
commonly used when measuring white materials where optical brighten- tions. (See Practice E1164 and the appropriate test method for
ing is suspected.
the instrument geometry.)
6.3.2 Fluorescence-Weakening Method: In the fluorescence-
6.4.2 Measure the specimen, obtaining either a table or a
weakening method two different bandpass filters are used and
graph of spectral transmittance or reflectance factor versus
three measurements are compared (1). One filter is chosen to wavelength.
remove all the fluorescence-exciting wavelengths
6.4.3 Set the instrument for monochromatic illumination
(fluorescence-killing filter), and the second filter is chosen to
and calibrate it in a manner similar to that given in 6.3.1.
remove incident illumination about 20 to 40 nm shorter than
6.4.4 Measure the specimen in accordance with 6.3.2.
the first filter (fluorescence-weakening filter). Use the proce-
dure in 6.3.1.1 and 6.3.1.2 for the measurement without any
7. Interpretation of Results
filter in place. Then use the procedures in 6.3.1.3 – 6.3.1.5 for
the measurements with each of the filters. Refer to the 7.1 The confirmation of the presence of fluorescence is
made by examining the Donaldson matrix or by the compari-
referenced literature for complete details of the application of
this method. son of spectral curves at the wavelength of maximum
deviation, color difference, or single parameter difference such
6.3.3 Filter Reduction Method: Several linear long band-
pass filters are placed, one at a time, in the light path between as ΔY or Whiteness Index (WI) between the measurements. If
you have used the two-monochromator method follow step 7.2
the source and the specimen. Usually 3 to 5 filters are enough
to estimate the reflected radiance factor (2). The same proce- or 7.5, or both. If you are using the comparison of spectral
curves at the wavelength of maximum deviation follow step
dure is used to measure the specimen with each filter i
...
This 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. Because
it may 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 version
of the standard as published by ASTM is to be considered the official document.
Designation: E1247 − 12 E1247 − 12 (Reapproved 2017)
Standard Practice for
Detecting Fluorescence in Object-Color Specimens by
Spectrophotometry
This standard is issued under the fixed designation E1247; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This practice provides spectrophotometric methods for detecting the presence of fluorescence in object-color specimens.
NOTE 1—Since the addition of fluorescing agents (colorants, whitening agents, etc.) is often intentional by the manufacturer of a material, information
on the presence or absence of fluorescent properties in a specimen may often be obtained from the maker of the material.
1.2 This practice requires the use of a spectrophotometer that both irradiates the specimen over the wavelength range from 340
to 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 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 responsibility
of the user of this standard to establish appropriate safety safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization
established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued
by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
2. Referenced Documents
2.1 ASTM Standards:
D2244 Practice for Calculation of Color Tolerances and Color Differences from Instrumentally Measured Color Coordinates
E284 Terminology of Appearance
E308 Practice for Computing the Colors of Objects by Using the CIE System
E313 Practice for Calculating Yellowness and Whiteness Indices from Instrumentally Measured Color Coordinates
E991 Practice for Color Measurement of Fluorescent Specimens Using the One-Monochromator Method
E1164 Practice for Obtaining Spectrometric Data for Object-Color Evaluation
E1331 Test Method for Reflectance Factor and Color by Spectrophotometry Using Hemispherical Geometry
E1348 Test Method for Transmittance and Color by Spectrophotometry Using Hemispherical Geometry
E1349 Test Method for Reflectance Factor and Color by Spectrophotometry Using Bidirectional (45°:0° or 0°:45°) Geometry
E2152 Practice for Computing the Colors of Fluorescent Objects from Bispectral Photometric Data
E2153 Practice for Obtaining Bispectral Photometric Data for Evaluation of Fluorescent Color
3. Terminology
3.1 The definitions in Terminology E284, Practices E991, E2152, and E2153 are applicable to this practice.
4. Significance and Use
4.1 Several standards, including Practices E991, E1164, and Test Methods E1331, E1348 and E1349, require either the presence
or absence of fluorescence exhibited by the specimen for correct application. This practice provides spectrophotometric procedures
for identifying the presence of fluorescence in materials.
This practice is under the jurisdiction of ASTM Committee E12 on Color and Appearance and is the direct responsibility of Subcommittee E12.05 on Fluorescence.
Current edition approved July 1, 2012Nov. 1, 2017. Published September 2012November 2017. Originally approved in 1988. Last previous edition approved in 20032012
as E1247 – 03 which was withdrawn January 2012 and reinstated in July 2012. DOI: 10.1520/E1247-12.12. DOI: 10.1520/E1247-12R17.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’sstandard’s Document Summary page on the ASTM website.
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E1247 − 12 (2017)
4.2 This practice is applicable to all object-color specimens, whether opaque, translucent, or transparent, meeting the
requirements for specimens in the appropriate standards listed in 2.1. Translucent specimens should be measured by reflectance,
with a standard non-fluorescent backing material, usually but not necessarily black, placed behind the specimen during
measurement.
4.3 This practice requires the use of a spectrophotometer in which the spectral distribution of the illumination on the specimen
can be altered by the user in one of several ways. The modification of the illumination can either be by the insertion of optical
filters between the illuminating source and the specimen, without interfering with the detection of 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 and
detection output as in the two-monochromator method.
4.4 The confirmation of the presence of fluorescence is made by the comparison of spectral curves, color difference, or single
parameter difference such as ΔY between the measurements.
NOTE 2—In 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 greatest difference.
4.5 Either bidirectional or hemispherical instrument geometry may be used in this practice. The instrument must be capable of
providing 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. It does not address the issue of whether the
fluorescence makes a significant or insignificant contribution to the colorimetric properties of the specimen for any given
application. The user must determine the practical significance of the effect of fluorescence on the color measurement.
5. Instrumental Requirements
5.1 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, if
present.
5.1.2 The instrument must provide one of the following illumination/viewing combinations:
5.1.2.1 Monochromatic illumination and monochromatic viewing (that is, a two-monochromator spectrophotometer sometimes
called a bispectrometer or spectrofluorimeter).
5.1.2.2 Polychromatic illumination and monochromatic viewing.
5.1.2.3 Reversible illumination/viewing to allow both polychromatic 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 user
modified standardization/calibration values, which is a requirement for using any of the filter methods described in this practice.
NOTE 3—Repeatable and accurate application of this practice requires specialized instrumentation. Some commercial one-monochromator
spectrometers are limited in their ability to allow for the insertion of optical filters and re-standardization with the filter in place as required in this
procedure.
6. Procedures
6.1 There are three general types of procedures to detect the presence of fluorescence instrumentally. Each has its advantages
and shortcomings depending on the wavelength and intensity of the fluorescent emission and the instrumentation available to the
user.
6.2 Two-Monochromator Method: This method requires a colorimetric measuring instrument that is equipped with two separate
monochromators: the first, the illumination monochromator, irradiates the specimen with monochromatic light and the second, the
viewing monochromator, analyzes the radiation leaving the specimen. A two-dimensional array of bispectral photometric values
is obtained by setting the illumination monochromator at a series of fixed wavelengths (μ) in the illumination band of the specimen,
and for each μ, using the viewing monochromator to record readings for each wavelength (λ) in the specimen’s viewing range. The
resulting array, once properly corrected, is known as the Donaldson matrix, and the value of each element (μ,λ) of this array is the
Donaldson radiance factor (D(μ,λ)). The reflection values are confined to the diagonal of the matrix, and these diagonal values are
equal to the spectral reflectance factor of the specimen. Therefore, the presence of fluorescence is demonstrated by non-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 general procedure of making a measurement of spectral radiance factor using a
spectrometer with broad band illumination, then adding one or more filters to remove the fluorescence-excitation energy and
measuring the spectral radiance factor under the modified illumination. The comparison of the resulting spectral curves shows the
presence or absence of fluorescence. If the exclusion of the excitation energy results in a difference in the remaining part of the
curve, fluorescence is present and must be considered in the measurement procedures. If no difference is found, then fluorescence
is not an issue in the measurement of that specimen.
6.3.1 UV-Blocking Method—This procedure is typically used for detecting the presence of optical brighteners, such as in white
paper and textiles.
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6.3.1.1 Calibrate the instrument as required by the manufacturer. (See Practice E1164 and the appropriate test method for the
instrument geometry.)
NOTE 4—Since the measurement will be used to detect fluorescence, it should be considered that fluorescence might be present, therefore the calibration
procedure should include adjusting the instrument’s illuminator to conform as closely as possible to D65 including the UV region of the spectrum. In
some commercial instruments this may be accomplished by calibrating by whiteness index or the UV profile.
6.3.1.2 Measure the specimen, obtaining either a table or a graph of spectral transmittance or reflectance factor versus
wavelength.
6.3.1.3 Insert a long-wavelength bandpass filter between the illuminating source and the specimen. Select the cutoff wavelength
of 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 be placed between the
illuminating 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.
(a) For spectrophotometers equipped for illumination by means of an integrating sphere, the filter must be placed between the
illuminating 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 bidirectional geometry, the filter must be placed between
the illuminating source and the specimen.
(b) For spectrophotometers equipped for illumination by means of bidirectional geometry, the filter must be placed between the
illuminating 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 5—This method employing only one cut-off filter is most commonly used when measuring white materials where optical brightening is suspected.
6.3.2 Fluorescence-Weakening Method: In the fluorescence-weakening method two different bandpass filters are used and three
measurements are compared (1). One filter is chosen to remove all the fluorescence-exciting wavelengths (fluorescence-killing
filter), 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 place. Then use
the 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
details of 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 between the source
and the specimen. Usually 3 to 5 filters are enough to estimate the reflected radiance factor (2). The same procedure is used to
measure the specimen with each filter in place, following steps 6.3.1.1 – 6.3.1.5. The difference between the mapped reflected
radiance factor and the unfiltered measurement reveals the presence or absence of fluorescence. Refer to the referenced literature
for 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 and
the specimen one at a time. This produces a series of readings which is used to determine the total radiance factor is a way
somewhat analogous to an abridged two-monochromator instrument (3), (4). Again the difference between the reflectance and the
total radiance curves indicates the presence or absence of fluorescence. Follow the procedure in 6.3.1.1 – 6.3.1.5 for the
measurements 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 suitable
calibration, be equivalent to the two-monochromator method. In the filter reduction method 3 to 5 filters in the region of suspected
fluorescence are used. In this method 10 to 12 filters are used to measure the entire visible spectrum. Follow the procedure in
6.3.1.1 – 6.3.1.5 for measurements with each filter. Then examine the difference between the curves. Refer to the referenced
literature for complete details of the application of this method.
6.4 Two-Mode Method: The two-mo
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