1、Designation: D 4763 06Standard Practice forIdentification of Chemicals in Water by FluorescenceSpectroscopy1This standard is issued under the fixed designation D 4763; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of las
2、t 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 allows for the identification of 90 chemi-cals that may be found in water or in surface layers on water.Th
3、is practice is based on the use of room-temperature fluores-cence spectra taken from lists developed by the U.S. Environ-mental Protection Agency and the U.S. Coast Guard (1).2Ref(1) is the primary source for these spectra. This practice is alsobased on the assumption that such chemicals are either
4、presentin aqueous solution or are extracted from water into anappropriate solvent.1.2 Although many organic chemicals containing aromaticrings, heterocyclic rings, or extended conjugated double-bondsystems have appreciable quantum yields of fluorescence, thispractice is designed only for the specifi
5、c compounds listed. Ifpresent in complex mixtures, preseparation by high-performance liquid chromatography (HPLC), column chroma-tography, or thin-layer chromatography (TLC) would probablybe required.1.3 If used with HPLC, this practice could be used for theidentification of fluorescence spectra gen
6、erated by opticalmultichannel analyzers (OMA) or diode-array detectors.1.4 For simple mixtures, or in the presence of other non-fluorescing chemicals, separatory techniques might not berequired. The excitation and emission maximum wavelengthslisted in this practice could be used with standard fluore
7、scencetechniques (Refs 2-6) to quantitate these ninety chemicals onceidentification had been established. For such uses, generationof a calibration curve, to determine the linear range for use offluorescence quantitation would be required for each chemical.Examination of solvent blanks to subtract o
8、r eliminate anyfluorescence background would probably be required.1.5 This standard does not purport to address the safetyconcerns, if any, associated with its use. It is the responsibilityof the user of this standard to establish appropriate safety andhealth practices and determine the applicabilit
9、y of regulatorylimitations prior to use.2. Referenced Documents2.1 ASTM Standards:3D 1129 Terminology Relating to WaterD 1193 Specification for Reagent WaterE 131 Terminology Relating to Molecular SpectroscopyE 275 Practice for Describing and Measuring Performanceof Ultraviolet, Visible, and Near-In
10、frared Spectrophotom-eters3. Terminology3.1 DefinitionsFor definitions of terms used in this prac-tice, refer to Terminology D 1129, Specification D 1193, anddefinitions under the jurisdiction of Committee E-13 such asDefinitions E 131 and Practice E 275.4. Summary of Practice4.1 This practice uses
11、well tested fluorescence techniques todetect and identify (or determine the absence of) 90 chemicalsthat have relatively high fluorescence yields. Table 1 lists foreach chemical an appropriate solvent (either cyclohexane,water, methyl or ethyl alcohol, depending on solubility), asuggested excitation
12、 wavelength for maximum sensitivity, awavelength corresponding to the emission maximum, thenumber of fluorescence peaks and shoulders, the width (fullwidth at half of the maximum emission intensity) of thestrongest fluorescence peak and the detection limit for theexperimental conditions given. Detec
13、tion limits could be low-ered, following identification, by using broader slit widths. Alist of corrected fluorescence spectra for the chemicals includedin this practice are also available (1).4.2 Identification of the sample is made by comparison ofthe obtained spectra with information in Table 1 a
14、nd by directvisual comparison of appropriate spectra with positions ofprincipal peaks in agreement to 62 nm and ratios of peakheights in agreement to 610 % if corrected spectrofluorom-eters are used.4.3 Spectral distortions due to self-absorption or fluores-cence quenching or dimer formation may occ
15、ur at higher1This practice is under the jurisdiction of ASTM Committee D19 on Water andis the direct responsibility of Subcommittee D19.06 on Methods for Analysis forOrganic Substances in Water.Current edition approved July 1, 2006. Published July 2006. Originally approvedin 1988. Last previous edit
16、ion approved in 2001 as D 4763 88 (2001).2The boldface numbers in parentheses refer to the list of references at the end ofthis practice.3For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume i
17、nformation, refer to the standards Document Summary page onthe ASTM website.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.concentrations (for example, 100 ppm or g/mL). If this issuspected, the solution should be diluted and additi
18、onal fluo-rescence spectra generated. If a suspected chemical is notdetected on excitation at the appropriate wavelength, it usuallycan be assumed that it is not present above the detection limit,barring interference effects due to absorption or quenching thatcan usually be anticipated.5. Significan
19、ce and Use5.1 This practice is useful for detecting and identifying (ordetermining the absence of) 90 chemicals with relatively highfluorescence yields (see Table 1). Most commonly, this practicewill be useful for distinguishing single fluorescent chemicals insolution, simple mixtures or single fluo
20、rescing chemicals in thepresence of other nonfluorescing chemicals. Chemicals withhigh fluorescence yields tend to have aromatic rings, someheterocyclic rings or extended conjugated double-bond sys-tems. Typical chemicals included on this list include aromatics,substituted aromatics such as phenols,
21、 polycyclic aromatichydrocarbons (PAHs), some pesticides such as DDT, poly-chlorinated biphenyls (PCBs), some heterocyclics, and someesters, organic acids, and ketones.TABLE 1 Summary of Experimental Parameters and ResultsChemical CodeConcentra-tions, ppmSolvent lexc,nm lmaxem,nmNumberof PeaksWHM,nm
22、ShoulderNumberDetectionLimit(DL), ppmlDL, nm CommentsAcenaphthene ACN 1.03 CH 290 323 4 . 3 0.001 290Acetone ACT 227 CH 290 410 1 . . 212 290Acridine ACR 96 CH 285/355 386/422 4/2 . 2/0 . .ACR 9.6 ETOH 290/355 357/415 2/2 . 1/1 0.02/0.04 290/355Aniline ANL 15.5 CH 280 316 1 . . 0.037 280Anthracene A
23、TH 1.03 CH 355 378 4 . 1 0.001 355ATH 1.55 ETOH 355 380 4 . 1 0.001 355Aroclor 12421254PC4PC5131129CHCH270270317317223536110.32270270Atrazine ATZ 369 CH 290 350 1 . . 300 290Azinphosmethyl AZP 112 CH 350 410 2 60 . 10 350AZP 122 ETOH 340 420 2 80 . 4 340Benz(a)anthracene BAT 1.1 CH 280 386 4 . 1 0.0
24、03 280Benzene BNZ 79 CH 250 279 3 24 1 2/4 250/265Benzonitrile BZN 9.9 CH 260 287 2 28 1 0.1/0.1 260/270Benzo(a)pyrene BAP 0.088 CH 370 405 6 . 2 0.002 370Benzyl alcohol BAL 99 CH 250 284 2 27 1 0.1/0.1 250/260Benzyl amine BZM 118 CH 250 283 1 27 2 3/2 250/260Benzyl triethylam-monium chlorideBMA 210
25、 H2O 250 280 1 28 . 59 250Bisphenol A BPA 10.5 ETOH 270 304 1 30 1 0.04/0.02 270/285Brucine BRU 13.5 ETOH 280 327 1 56 . 2/2 280/295O-tert-Butylphenol BOP 21 CH 265 295 1 30 1 0.1/0.1 265/275p-tert-Butylphenol BTP 17.5 CH 260 295 1 31 1 0.6/0.4 260/280Carbaryl CBY 1.0 CH 285 335 2 36 2 0.01 285Carna
26、uba wax WCA 63.5 CH 260 310 1 64 . 42 260Castor oil OCA 390 ETOH 290 328 1 43 2 20 290OCA 286 CH 280/320 . 1 . . 180/300 280/320Catechol CTC 8.7 H2O 265 310 1 46 . 0.4/0.2 265/2804-Chloroaniline CAP 17.2 CH 290 328 1 36 1 0.2 2901-Chloronaphthalene CNA 11.3 CH 290 328 3 34 4 0.1 290p-Chlorophenol CP
27、N 101 CH 260 305 1 30 . 1/0.1 260/285Chlorpyrifos (Duraban) DUR 25.3 CH 280 326 1 52 . 1/0.5 280/295p-Chlorotoluene CTN 23.8 CH 265 288 1 29 3 1/0.8 265/275p-Chloro-o-toluidine COT 25 CH 290 328 1 39 1 0.09 300Chrysene CRY 1.0 CH 270 383 5 . . 0.002 270Coconut oil OCC 286 CH 290 330 . . . 100 290Cod
28、 liver oil OCL 323 CH 260/280330320/3205001/11150 . 260,14065260,280330Copper naphthenate CNN 98 CH 260 326 1 60 3 3/1 260/280Cottonseed oil OCS 305 CH 280/320 320/380 . . . 165,300 280,320Coumaphos COU 11.4 CH 320 377 1 74 . 0.3 320o-Cresol CRO 12.0 CH 265 293 1 30 1 0.04 280p-Cresol CRP 10.3 CH 26
29、5 299 1 30 . 0.03 280Cumene CUM 101 CH 250 283 2 28 1 3 250p-Cymene CMP 11.8 CH 260 285 1 28 2 0.4/0.2 260/270DDD DDD 61.0 CH 240 294 1 30 2 4 240DDT DDT 87 CH 245 291 2 28 2 7 2451,2,5,6-Dibenzanthracene DBA 0.015 CH 300 396 4 . 2 0.001 300Dicamba DIC 22.2 H2O 310 420 1 70 . 0.9 310Dichlorobenil DI
30、B 108 CH 285 312 1 30 . 0.6 2852,4-Dichlorophenoxy-acetic acidDCA 159 CH 270 310 1 46 1 30 270Diethylbenzene DEB 100 CH 255 283 1 28 2 0.2/0.1 255/270D4763062TABLE 1 ContinuedChemical CodeConcentra-tions, ppmSolvent lexc,nm lmaxem,nmNumberof PeaksWHM,nmShoulderNumberDetectionLimit(DL), ppmlDL, nm Co
31、mmentsDiethylene glycol DEG 202 CH 265 310 2 . . 202 265Diethylphthalate DEP 145/289 CH 260/280 300/320 1/1 . . . 2802,4-Dimethylphenol DMH 10.5 CH 265 300 1 31 1 0.2/0.04 265/2803,5-Dimethylphenol DPM 10.5 CH 265 295 1 28 1 0.07/0.03 265/280Diphenylamine DAM 11.21.2CHCH29029033333311373722.290290ph
32、otochemicalchangeDiphenyldichlorosilane DDS 157 CH 260 285 2 30 . 3/2 260/270Diquat dibromide DQD 35.5 H2O 310 348 1 41 1 0.055 310Dodecylbenzene DDB 116 CH 250 285 3 30 . * 250 * strong impurity116 CH 220 285 3 30 . 13.6 220Dowtherm A DTH 10.8 CH 260 305 2 33 2 0.035 260Ethylbenzene ETB 103 CH 250
33、283 2 26 . 3.1/1.5 250/260Fluoranthene FLA 1.0 CH 360 465 2 91 3 0.005 360Gallic acid GLA 103 H2O 290 346 1 77 . 0.70 290Hydroquinone HDQ 1.1 H2O 290 326 1 38 1 0.025 290Indene IND 175 CH 260 309 2 32 3 0.12 260Lard OLD 340 CH 270 330 . . . . 270OLD 287 CH 280 330 1 . . . 280Linseed oil OLS 355 CH 3
34、00 418 1 105 . 32 300Methoxychlor MOC 95 CH 270 299 1 30 1 1.3/0.8 270,280Methylaniline MAN 10.8 CH 290 325 1 35 . 0.01 290Methyl isobutylketoneMIK 358 CH 290 400 1 . . . 290Methyl styrene MSR 105 CH 255 307 1 35 2 0.12 255Naphthalene NPT 10.5 CH 280 323 2 24 3 0.02 2801-Naphthylamine NAD 1.85 CH 32
35、5 377 1 55 1 0.0012 325Nonyl phenol NNP 17.1 CH 265 298 1 28 . 0.09 265Olive oil OOL 237 CH 260 320 1 . . . 360OOL 290 CH 310 . . . . . 310Palm oil OPM 300 CH 260 320 1 60 . 218 260CH 350 500 1 140 . 300 350Peanut oil OPN 249 CH 260,290 120,320 1 . . . .Phenol PHN 11.9 CH 265 288 1 30 2 0.011/0.007
36、265/275Phenyl ether DPE 20.4 CH 265 291 1 36 1 0.10 265Phthalic acid PHA 97 H2O 280 330 1 100 . 84 280PHA 228 H2O 270 340 1 100 . 114 270Piperazine PPZ 235 CH 280 350 1 . . . .Polyethoxylated non-ylphenolPEN 9.5 CH 265 297 1 30 . 0.08/0.0317265/280Pyrogallol PGA 152 H2O 270 335 1 86 1 30 270Quinolin
37、e QNL 113 ETOH 275 321 5 . 2 . . photolyzes113 ETOH 355 420 1 70 0 . . photolyzes95 CH 275 336 3 . 2 0.37 275 photolyzes95 CH 350 . 2 57 1 . .Resorcinol RSC 10.1 H2O 265 303 1 39 1 0.135/0.05 265/280Salicylic acid SLA 1.5 H2O 300 409 1 64 . 0.005 300Sodium dodecylben-zenesulfonateSDB 90 CH 290 347 1
38、 52 2 0.90 290Soya bean oil OSB 290 CH 270,320 . . . . 0.300 270,320Styrene STY 1.1 CH 270 306 2 32 2 0.03 270Tanaic acid TNA 13 H2O 280 340 1 100 . 0.63 2801,2,3,4-Tetrahydro-naphthaleneTHN 12.3 CH 260 284 1 27 2 0.21/0.13 260/270p-Toluidine TLI 14.1 CH 290 325 1 34 . 0.03 290Toluene TOL 107 CH 250
39、 284 2 27 1 2.1/1.6 250/215p-Toluene sulfonic acid TAP 120 H2O 260 285 1 28 1 2.1/1.5 260/265Tricresylphosphate TCP 123 CH 260 288 1 66 1 0.55/0.35 260/2701,3,5-Triethylbenzene TEB 122 CH 250 292 1 28 3 12.5/1.5 250/270Turpentive TPT 301 CH 260 283 1 34 3 31/13 260/270Undecylbenzene UDB 87.3 CH 250
40、284 2 33 2 6.0 250Uranyl nitrate UAN 61.0 H2O 290 520 3 56 2 6.1/10.5 290/330m-Xylene XLM 114 CH 260 285 1 28 1 2.0/1.4 260/270o-Xylene XLO 92 CH 260 285 1 30 . 1.5/1.3 260/2705.2 With appropriate separatory techniques (HPLC, TLC,and column chromatography) and in some cases, specialdetection techniq
41、ues (OMAs and diode arrays), this practicecan be used to determine these 90 chemicals even in complexmixtures containing a number of other fluorescing chemicals.With the use of appropriate excitation and emission wave-lengths and prior generation of calibration curves, this practicecould be used for
42、 quantitation of these chemicals over a broadlinear range.5.3 Fluorescence is appropriately a trace technique and athigher concentrations (greater than 10 to 100 ppm) spectraldistortions usually due to self-absorption, or inner-filter effectsD4763063but sometimes ascribed to fluorescence quenching,
43、may beobserved. These effects can usually be eliminated by dilutingthe solution. Detection limits can be lowered following iden-tification by using broader slit widths, but this may result inspectral broadening and distortion.5.4 This practice assumes the use of a corrected spectrof-luorometer (that
44、 is, one capable of producing corrected fluo-rescence spectra). On an uncorrected instrument, peak shiftsand spectral distortions and changes in peak ratios may benoted. An uncorrected spectrofluorometer can also be used ifappropriate data is generated on the instrument to be used.6. Interferences6.
45、1 For the identification of compounds with low fluores-cence yields and relatively high detection limits, the presenceof other chemicals with high fluorescence yields emitting in thesame spectral region, for example, anthracene, fluorescein, etc.,may interfere unless separatory techniques are employ
46、ed.6.2 Some naturally occurring fluorescing materials, such ashumic acids from leaf mold, may also interfere with theidentification of chemicals with relatively low fluorescenceyields especially at dilute concentrations of the hazardouschemicals, especially for emission in the near ultraviolet.6.3 S
47、ince light must be absorbed before being reemitted,colored solutions, or solutions with absorbances greater than0.02 at the excitation or emission wavelengths of interest willalso interfere. Such solutions usually require further dilution.6.4 Halogenated solvents and other solvents containingpossibl
48、e quenchers are not recommended for this applicationsince they may raise detection limits.7. Apparatus7.1 Scanning Fluorescence Spectrophotometer or Spectrof-luorometer, corrected to give constant emission intensityto 65 to 10 % for fluorescence spectra over the spectral rangescanned, normally from
49、220 to 600 nm. The spectral correctionshould be checked using an appropriate chemical such asanthracene for which the peak ratios of the corrected fluores-cence peaks are known. The instrument should have anappropriate excitation source such as a high-pressure xenonlamp or other continuum source with at least 150 or 250 W.Band widths should be adjustable to at least 5 nm for excitationslit widths and at least 2 nm for emission slit widths. Anappropriate photomultiplier tube with good detection charac-teristics over the 250 to 700 nm spectral range. For example,tubes with an S-2
