ASTM D4763-06(2020)
(Practice)Standard Practice for Identification of Chemicals in Water by Fluorescence Spectroscopy
Standard Practice for Identification of Chemicals in Water by Fluorescence Spectroscopy
SIGNIFICANCE AND USE
5.1 This practice is useful for detecting and identifying (or determining the absence of) 90 chemicals with relatively high fluorescence yields (see Table 1). Most commonly, this practice will be useful for distinguishing single fluorescent chemicals in solution, simple mixtures or single fluorescing chemicals in the presence of other nonfluorescing chemicals. Chemicals with high fluorescence yields tend to have aromatic rings, some heterocyclic rings or extended conjugated double-bond systems. Typical chemicals included on this list include aromatics, substituted aromatics such as phenols, polycyclic aromatic hydrocarbons (PAH’s), some pesticides such as DDT, polychlorinated biphenyls (PCB’s), some heterocyclics, and some esters, organic acids, and ketones.
5.2 With appropriate separatory techniques (HPLC, TLC, and column chromatography) and in some cases, special detection techniques (OMA’s and diode arrays), this practice can be used to determine these 90 chemicals even in complex mixtures containing a number of other fluorescing chemicals. With the use of appropriate excitation and emission wavelengths and prior generation of calibration curves, this practice could be used for quantitation of these chemicals over a broad linear range.
5.3 Fluorescence is appropriately a trace technique and at higher concentrations (greater than 10 to 100 ppm) spectral distortions usually due to self-absorption, or inner-filter effects but sometimes ascribed to fluorescence quenching, may be observed. These effects can usually be eliminated by diluting the solution. Detection limits can be lowered following identification by using broader slit widths, but this may result in spectral broadening and distortion.
5.4 This practice assumes the use of a corrected spectrofluorometer (that is, one capable of producing corrected fluorescence spectra). On an uncorrected instrument, peak shifts and spectral distortions and changes in peak ratios may be noted. An uncorrected spect...
SCOPE
1.1 This practice allows for the identification of 90 chemicals that may be found in water or in surface layers on water. This practice is based on the use of room-temperature fluorescence spectra taken from lists developed by the U.S. Environmental Protection Agency and the U.S. Coast Guard (1). Ref (1) is the primary source for these spectra. This practice is also based on the assumption that such chemicals are either present in aqueous solution or are extracted from water into an appropriate solvent.2
1.2 Although many organic chemicals containing aromatic rings, heterocyclic rings, or extended conjugated double-bond systems have appreciable quantum yields of fluorescence, this practice is designed only for the specific compounds listed. If present in complex mixtures, preseparation by high-performance liquid chromatography (HPLC), column chromatography, or thin-layer chromatography (TLC) would probably be required.
1.3 If used with HPLC, this practice could be used for the identification of fluorescence spectra generated by optical multichannel analyzers (OMA) or diode-array detectors.
1.4 For simple mixtures, or in the presence of other nonfluorescing chemicals, separatory techniques might not be required. The excitation and emission maximum wavelengths listed in this practice could be used with standard fluorescence techniques (see Refs (2-6)) to quantitate these ninety chemicals once identification had been established. For such uses, generation of a calibration curve, to determine the linear range for use of fluorescence quantitation would be required for each chemical. Examination of solvent blanks to subtract or eliminate any fluorescence background would probably be required.
1.5 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 practi...
General Information
Relations
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:D4763 −06 (Reapproved 2020)
Standard Practice for
Identification of Chemicals in Water by Fluorescence
Spectroscopy
This standard is issued under the fixed designation D4763; 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 responsibility of the user of this standard to establish appro-
priate safety, health, and environmental practices and deter-
1.1 This practice allows for the identification of 90 chemi-
mine the applicability of regulatory limitations prior to use.
cals that may be found in water or in surface layers on water.
1.6 This international standard was developed in accor-
This practice is based on the use of room-temperature fluores-
dance with internationally recognized principles on standard-
cence spectra taken from lists developed by the U.S. Environ-
ization established in the Decision on Principles for the
mental Protection Agency and the U.S. Coast Guard (1). Ref
Development of International Standards, Guides and Recom-
(1) is the primary source for these spectra. This practice is also
mendations issued by the World Trade Organization Technical
based on the assumption that such chemicals are either present
Barriers to Trade (TBT) Committee.
in aqueous solution or are extracted from water into an
appropriate solvent.
2. Referenced Documents
1.2 Although many organic chemicals containing aromatic
2.1 ASTM Standards:
rings, heterocyclic rings, or extended conjugated double-bond
D1129 Terminology Relating to Water
systems have appreciable quantum yields of fluorescence, this
D1193 Specification for Reagent Water
practice is designed only for the specific compounds listed. If
E131 Terminology Relating to Molecular Spectroscopy
present in complex mixtures, preseparation by high-
E275 Practice for Describing and Measuring Performance of
performance liquid chromatography (HPLC), column
Ultraviolet and Visible Spectrophotometers
chromatography, or thin-layer chromatography (TLC) would
probably be required.
3. Terminology
1.3 If used with HPLC, this practice could be used for the
3.1 Definitions—For definitions of terms used in this
identification of fluorescence spectra generated by optical
practice, refer to Terminology D1129, Specification D1193,
multichannel analyzers (OMA) or diode-array detectors.
and definitions under the jurisdiction of Committee E13 such
1.4 For simple mixtures, or in the presence of other non-
as Terminology E131 and Practice E275.
fluorescing chemicals, separatory techniques might not be
4. Summary of Practice
required. The excitation and emission maximum wavelengths
listed in this practice could be used with standard fluorescence
4.1 This practice uses well tested fluorescence techniques to
techniques(seeRefs (2-6))toquantitatetheseninetychemicals
detect and identify (or determine the absence of) 90 chemicals
once identification had been established. For such uses, gen-
that have relatively high fluorescence yields. Table 1 lists for
eration of a calibration curve, to determine the linear range for
each chemical an appropriate solvent (either cyclohexane,
use of fluorescence quantitation would be required for each
water, methyl or ethyl alcohol, depending on solubility), a
chemical. Examination of solvent blanks to subtract or elimi-
suggested excitation wavelength for maximum sensitivity, a
nate any fluorescence background would probably be required.
wavelength corresponding to the emission maximum, the
1.5 This standard does not purport to address all of the number of fluorescence peaks and shoulders, the width (full
safety concerns, if any, associated with its use. It is the width at half of the maximum emission intensity) of the
strongest fluorescence peak and the detection limit for the
experimental conditions given. Detection limits could be
This practice is under the jurisdiction of ASTM Committee D19 on Water and
lowered, following identification, by using broader slit widths.
is the direct responsibility of Subcommittee D19.06 on Methods for Analysis for
Organic Substances in Water.
Current edition approved Dec. 15, 2020. Published December 2020. Originally
approved in 1988. Last previous edition approved in 2012 as D4763 – 06 (2012). For referenced ASTM standards, visit the ASTM website, www.astm.org, or
DOI: 10.1520/D4763-06R20. contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
The boldface numbers in parentheses refer to a list of references at the end of Standards volume information, refer to the standard’s Document Summary page on
this standard. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D4763−06 (2020)
A list of corrected fluorescence spectra for the chemicals 5. Significance and Use
included in this practice are also available (1).
5.1 This practice is useful for detecting and identifying (or
4.2 Identification of the sample is made by comparison of
determining the absence of) 90 chemicals with relatively high
the obtained spectra with information in Table 1 and by direct
fluorescenceyields(seeTable1).Mostcommonly,thispractice
visual comparison of appropriate spectra with positions of
willbeusefulfordistinguishingsinglefluorescentchemicalsin
principal peaks in agreement to 62 nm and ratios of peak
solution,simplemixturesorsinglefluorescingchemicalsinthe
heights in agreement to 610 % if corrected spectrofluorom-
presence of other nonfluorescing chemicals. Chemicals with
eters are used.
high fluorescence yields tend to have aromatic rings, some
4.3 Spectral distortions due to self-absorption or fluores-
heterocyclic rings or extended conjugated double-bond sys-
cence quenching or dimer formation may occur at higher
tems.Typicalchemicalsincludedonthislistincludearomatics,
concentrations (for example, 100 ppm or µg/mL). If this is
substituted aromatics such as phenols, polycyclic aromatic
suspected, the solution should be diluted and additional fluo-
hydrocarbons (PAH’s), some pesticides such as DDT, poly-
rescence spectra generated. If a suspected chemical is not
chlorinated biphenyls (PCB’s), some heterocyclics, and some
detected on excitation at the appropriate wavelength, it usually
esters, organic acids, and ketones.
can be assumed that it is not present above the detection limit,
barring interference effects due to absorption or quenching that
can usually be anticipated.
TABLE 1 Summary of Experimental Parameters and Results
Number Detection
Concentra- WHM, Shoulder
em
Chemical Code Solvent λ ,nm λ ,nm of Limit λ , nm Comments
exc DL
max
tions, ppm nm Number
Peaks (DL), ppm
Acenaphthene ACN 1.03 CH 290 323 4 . 3 0.001 290
Acetone ACT 227 CH 290 410 1 . . 212 290
Acridine ACR 96 CH 285/355 386/422 4/2 . 2/0 . .
ACR 9.6 ETOH 290/355 357/415 2/2 . 1/1 0.02/0.04 290/355
Aniline ANL 15.5 CH 280 316 1 . . 0.037 280
Anthracene ATH 1.03 CH 355 378 4 . 1 0.001 355
ATH 1.55 ETOH 355 380 4 . 1 0.001 355
Aroclor 1242 PC4 131 CH 270 317 2 35 1 0.3 270
1254 PC5 129 CH 270 317 2 36 1 2 270
Atrazine ATZ 369 CH 290 350 1 . . 300 290
Azinphosmethyl AZP 112 CH 350 410 2 60 . 10 350
AZP 122 ETOH 340 420 2 80 . 4 340
Benz(a)anthracene BAT 1.1 CH 280 386 4 . 1 0.003 280
Benzene BNZ 79 CH 250 279 3 24 1 2/4 250/265
Benzonitrile BZN 9.9 CH 260 287 2 28 1 0.1/0.1 260/270
Benzo(a)pyrene BAP 0.088 CH 370 405 6 . 2 0.002 370
Benzyl alcohol BAL 99 CH 250 284 2 27 1 0.1/0.1 250/260
Benzyl amine BZM 118 CH 250 283 1 27 2 3/2 250/260
Benzyl triethylam- BMA 210 H O 250 280 1 28 . 59 250
monium chloride
Bisphenol A BPA 10.5 ETOH 270 304 1 30 1 0.04/0.02 270/285
Brucine BRU 13.5 ETOH 280 327 1 56 . 2/2 280/295
O-tert-Butylphenol BOP 21 CH 265 295 1 30 1 0.1/0.1 265/275
p-tert-Butylphenol BTP 17.5 CH 260 295 1 31 1 0.6/0.4 260/280
Carbaryl CBY 1.0 CH 285 335 2 36 2 0.01 285
Carnauba wax WCA 63.5 CH 260 310 1 64 . 42 260
Castor oil OCA 390 ETOH 290 328 1 43 2 20 290
OCA 286 CH 280/320 . 1 . . 180/300 280/320
Catechol CTC 8.7 H O 265 310 1 46 . 0.4/0.2 265/280
4-Chloroaniline CAP 17.2 CH 290 328 1 36 1 0.2 290
1-Chloronaphthalene CNA 11.3 CH 290 328 3 34 4 0.1 290
p-Chlorophenol CPN 101 CH 260 305 1 30 . 1/0.1 260/285
Chlorpyrifos (Duraban) DUR 25.3 CH 280 326 1 52 . 1/0.5 280/295
p-Chlorotoluene CTN 23.8 CH 265 288 1 29 3 1/0.8 265/275
p-Chloro-o-toluidine COT 25 CH 290 328 1 39 1 0.09 300
Chrysene CRY 1.0 CH 270 383 5 . . 0.002 270
Coconut oil OCC 286 CH 290 330 . . . 100 290
Cod liver oil OCL 323 CH 260/280 320/320 1/1 150 . 260,140 260,280
330 500 1 65 330
Copper naphthenate CNN 98 CH 260 326 1 60 3 3/1 260/280
Cottonseed oil OCS 305 CH 280/320 320/380 . . . 165,300 280,320
Coumaphos COU 11.4 CH 320 377 1 74 . 0.3 320
o-Cresol CRO 12.0 CH 265 293 1 30 1 0.04 280
p-Cresol CRP 10.3 CH 265 299 1 30 . 0.03 280
Cumene CUM 101 CH 250 283 2 28 1 3 250
p-Cymene CMP 11.8 CH 260 285 1 28 2 0.4/0.2 260/270
DDD DDD 61.0 CH 240 294 1 30 2 4 240
DDT DDT 87 CH 245 291 2 28 2 7 245
D4763−06 (2020)
TABLE1 Continued
Number Detection
Concentra- WHM, Shoulder
em
Chemical Code Solvent λ ,nm λ ,nm of Limit λ , nm Comments
exc DL
max
tions, ppm nm Number
Peaks (DL), ppm
1,2,5,6- DBA 0.015 CH 300 396 4 . 2 0.001 300
Dibenzanthracene
Dicamba DIC 22.2 H O 310 420 1 70 . 0.9 310
Dichlorobenil DIB 108 CH 285 312 1 30 . 0.6 285
2,4-Dichlorophenoxy- DCA 159 CH 270 310 1 46 1 30 270
acetic acid
Diethylbenzene DEB 100 CH 255 283 1 28 2 0.2/0.1 255/270
Diethylene glycol DEG 202 CH 265 310 2 . . 202 265
Diethylphthalate DEP 145/289 CH 260/280 300/320 1/1 . . . 280
2,4-Dimethylphenol DMH 10.5 CH 265 300 1 31 1 0.2/0.04 265/280
3,5-Dimethylphenol DPM 10.5 CH 265 295 1 28 1 0.07/0.03 265/280
Diphenylamine DAM 11.2 CH 290 333 1 37 2 . 290 photochemical
1.2 CH 290 333 1 37 2 . 290 change
Diphenyldichlorosilane DDS 157 CH 260 285 2 30 . 3/2 260/270
Diquat dibromide DQD 35.5 H O 310 348 1 41 1 0.055 310
Dodecylbenzene DDB 116 CH 250 285 3 30 . * 250 * strong impurity
116 CH 220 285 3 30 . 13.6 220
Dowtherm A DTH 10.8 CH 260 305 2 33 2 0.035 260
Ethylbenzene ETB 103 CH 250 283 2 26 . 3.1/1.5 250/260
Fluoranthene FLA 1.0 CH 360 465 2 91 3 0.005 360
Gallic acid GLA 103 H O 290 346 1 77 . 0.70 290
Hydroquinone HDQ 1.1 H O 290 326 1 38 1 0.025 290
Indene IND 175 CH 260 309 2 32 3 0.12 260
Lard OLD 340 CH 270 330 . . . . 270
OLD 287 CH 280 330 1 . . . 280
Linseed oil OLS 355 CH 300 418 1 105 . 32 300
Methoxychlor MOC 95 CH 270 299 1 30 1 1.3/0.8 270,280
Methylaniline MAN 10.8 CH 290 325 1 35 . 0.01 290
Methyl isobutyl MIK 358 CH 290 400 1 . . . 290
ketone
Methyl styrene MSR 105 CH 255 307 1 35 2 0.12 255
Naphthalene NPT 10.5 CH 280 323 2 24 3 0.02 280
1-Naphthylamine NAD 1.85 CH 325 377 1 55 1 0.0012 325
Nonyl phenol NNP 17.1 CH 265 298 1 28 . 0.09 265
Olive oil OOL 237 CH 260 320 1 . . . 360
OOL 290 CH 310 . . . . . 310
Palm oil OPM 300 CH 260 320 1 60 . 218 260
CH 350 500 1 140 . 300 350
Peanut oil OPN 249 CH 260,290 120,320 1 . . . .
Phenol PHN 11.9 CH 265 288 1 30 2 0.011/0.007 265/275
Phenyl ether DPE 20.4 CH 265 291 1 36 1 0.10 265
Phthalic acid PHA 97 H O 280 330 1 100 . 84 280
PHA 228 H O 270 340 1 100 . 114 270
Piperazine PPZ 235 CH 280 350 1 . . . .
Polyethoxylated non- PEN 9.5 CH 265 297 1 30 . 0.08/0.03 265/280
ylphenol 17
Pyrogallol PGA 152 H O 270 335 1 86 1 30 270
Quinoline QNL 113 ETOH 275 321 5 . 2 . . photolyzes
113 ETOH 355 420 1 70 0 . . photolyzes
95 CH 275 336 3 . 2 0.37 275 photolyzes
95 CH 350 . 2 57 1 . .
Resorcinol RSC 10.1 H O 265 303 1 39 1 0.135/0.05 265/280
Salicylic acid SLA 1.5 H O 300 409 1 64 . 0.005 300
Sodium dodecylben- SDB 90 CH 290 347 1 52 2 0.90 290
zenesulfonate
Soya bean oil OSB 290 CH 270,320 . . . . 0.300 270,320
Styrene STY 1.1 CH 270 306 2 32 2 0.03 270
Tanaic acid TNA 13 H O 280 340 1 100 . 0.63 280
1,2,3,4-Tetrahydro- THN 12.3 CH 260 284 1 27 2 0.21/0.13 260/270
naphthalene
p-Toluidine TLI 14.1 CH 290 325 1 34 . 0.03 290
Toluene TOL 107 CH 250 284 2 27 1 2.1/1.6 250/215
p-Toluene sulfonic acid TAP 120 H O 260 285 1 28 1 2.1/1.5 260/265
Tricresylphosphate TCP 123 CH 260 288 1 66 1 0.55/0.35 260/270
1,3,5-Triethylbenzene TEB 122 CH 250 292 1 28 3 12.5/1.5 250/270
Turpentive TPT 301 CH 260 283 1 34 3 31/13 260/270
Undecylbenzene UDB 87.3 CH 250 284 2 33 2 6.0 250
Uranyl nitrate UAN 61.0 H O 290 520 3 56 2 6.1/10.5 290/330
m-Xylene XLM 114 CH 260 285 1 28 1 2.0/1.4 260/270
o-Xylene XLO 92 CH 260 285 1 30 . 1.5/1.3 260/270
D4763−06 (2020)
5.2 With appropriate separatory techniques (HPLC, TLC, teristics over the 250 to 700 nm spectral range. For example,
and column chromatography) and in some cases, special tubes with an S-20 response, should be used.
detection techniques (OMA’s and diode arrays), this practice
7.2 Fluorescence Cells—Standard fluorescence cells,
can be used to determine these 90 chemicals even in complex
fluorescence-free fused silica cells with a 10-mm path length.
mixtures containing a number of other fluorescing chemicals.
7.3 Recorder—Strip chart or x-y recorder.
With the use of appropriate excitation and emission wave-
lengths and prior generation of calibration curves, this practice
7.4 Weighing Pans—Aluminum, disposable.
could be used for quantitation of these chemicals over a broad
linear range.
8. Reagents
5.3 Fluorescence is appropriately a trace technique and at
8.1 Purity of Reagents—Spectroquality grade chemicals
higher concentrations (greater than 10 to 100 ppm) spectral
shall be used in all tests. Spectroquality solvents required may
distortions usually due to self-absorption, or inner-filter effects
include cyclohexane, methanol, and ethanol. Purity of solvents
but sometimes ascribed to fluorescence quenching, may be
should be checked on running solvent blanks. Anthracene and
observed. These effects can usually be eliminated by diluting
other appropriate PAH’s may be required to check spectral
the solution. Detection limits can be lowered following iden-
corrections (see Ref (1)).
tification by using broader slit widths, but this may result in
8.2 Purity of Water—Unlessotherwiseindicated,references
spectral broadening and distortion.
to water shall be understood to mean reagent water
...
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