dip Designation: PS 47 - 95 -eiycan SOCIETY FOR TESTING AN MATERIALS 100 Bar Har. Wet Canaan, PA TD ered rom te ras Back of ASTM Stns, Copy ASTM Provisional Standard Test Method for Screening, Quantification, and Characterization of Total Polychlorinated Biphenyls by Room Temperature Phosphorescence’* “Ths standard is issued under the fted designation PS 47; the umber immediatly following the designation indicates the year of orginal adoption. 1. Scope . 1.1 This provisional test method provides a simple and rapid method for screening, quantifying, or characterizing total polychlorinated biphenyls (PCBs) by room temperature phosphorescence (RTP) spectroscopy for environmental samples, with determination of PCBs in the range of 50 to 5000 ng/mL of water and 50 to 5000 ng/g of sediment. 1.2 This test method is applicable to PCBs present in samples extracted from either water or contaminated sedi ‘ment, This test method is applicable for field screening purposes. 1.3 Provisional standards? achieve limited consensus through approval of the sponsoring subcommittee. 1.4 This staadard 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 appro- priate safety and health practices and determine the applica- bility of regulatory limitations prior to use. 2. Referenced Documents 2.1 ASTM Standards 1129 Terminology Relating to Water? 1193 Specification for Reagent Water! 1D 3370 Practices for Sampling Water from Closed Con- duits E 131 Terminology Relating to Molecular Spectroscopy* E 388 Test Method for Spectral Bandwidth and Wave- length Accuracy of Fluorescence Spectrometers* 3. Terminology 3.1 Definitions —For definitions of terms used in this test method, refer to Terminologies D 1129 and E 131 4, Summary of Test Method 4.1. This provisional test method consists of RTP analysis of dilute solutions of PCBs in appropriate solvents (sce Section 8) after extraction from water of sediment. The test ‘method requires an initial qualitative characterization step involving both fixed excitation RTP emission and synchro- nous RTP scanning of the samples in order to select an appropriate calibration mixture of PCBs to be selected which "This provisonl tet method is under the D19 on “Methods for Analysis for Organi Suttances in Water ‘Curent edition approved Dec, 27, 1995. Polished A 2 Provisional standards exist fortwo years subsequent tothe approval date 3 annual Book of ASTM Standards, Vol 11.01 4+ Anmual Book of ASTM Standards, Vol 0306. is characteristic of the predominant PCB commercial mix- ture present. Intensities of peak maxima of appropriate emission spectra are then used to develop calibration curves for quantification. 4.2 Weathering of PCBs has not been determined to pose a problem to the analysis of PCBs by this test method, however, site specific standards could be taken and analyzed by gas chromatography (GC) or other methods and com- pared to this test method. 5. Significance and Use 5.1 This provisional test method is designed for character- ization and rapid screening of commercial PCB mixtures (for example, Aroclor®),> including environmental samples, ¢i ther waterbome or extracted from sediment 5.2 The PCB mixture of interest is first characterized by its RTP emission and synchronous RTP scanning spectra, Intensities of peak maxima of appropriate RTP emission spectra are then used to set up suitable calibration curves as a function of concentration. Further discussion of RTP tech- niques, as applied to the characterization and quantification of complex mixtures can be found in the journal artices.6-* 6. Interferences 6.1 The luminescence spectra may be distorted or quanti- fication may be affected if the sample is contaminated with appreciable amounts of other chemicals that exhibit phos- phorescence with similar or appreciable phosphorescence yields in the same spectral regions as PCBs and phosphores- cence lifetime range. For example, the paper background (phosphorescence signal exhibited by filter paper when excited by the same wavelength of light as the sample) has a broad emission centered at 480 nm, 6.2 The effects of potential interferants, such as poly: aromatic hydrocarbons (PAHs) can be minimized by uti- lizing background subtraction if the interferant is known to be present. By taking an RTP measurement of the PAH interferant that has no PCB and subtracting this signal from the sample signal, the interferant signal can be eliminated. Also, if PAHs are suspected as interferants, it would be 5 PCBs were manufactured inthe United Sates by the Monsanto Co, under the tradename Aroclor and by otber companies worldwide under ater tradenames. Production in the United States ceed in 1977, The Aroclor tradename was applied to a class of products consisting ofchornsed aromatic compounds. PCB ‘Aroclor products are listed in Table. ‘*VoDinh, T, Room Temperature Phasphorimary, Wily, NY, 198, 1 Pal, A, Watts, W., Caraway, J, and VoDinh, T. Anaytis, Vat 20,1999, py, 4s, * Watts, W, Pal, A. Ford, L, Mille, G.H., VoDinh, T, Eastwood, D. and Lidhere, R, Application Speciromeiy, Vel 4S, 1992, pp. 1238Gp Ps 47 possible to treat the sample with sulfuric acid since PAHs are more rapidly degraded by H,SO, than PCBs. Fluorescenée from PAHs or other fluorescing compounds is quenched by use of a heavy metal quencher (lead or thallium). If larger amounts of fluorescing compounds sufficient to distort or ‘mask the luminescence spectrum of PCBs or to interfere with ‘quantification are observed or suspected, a phosphorimeter should be used as described in Table 2. Polar Muorescing materials found in soil such as humic acids are eliminated when cyclohexane (rather than ethanol) is used as a solvent. 6.3 The following possible interferants have been checked in preliminary experiments and were not found to signifi cantly interfere at concentrations of at least 100 fold higher than concentrations of PCB: 2,4,6-trichlorophenol, 2,4,5- trichlorophenol, pentachlorophenol, _trichloroethyiene, CHC), CCly, CHCl, and DDT. It is always the responsi bility of the analyst to check the analytical method against possible interferences in unknown or complex matrices. Nore 1—-Storage of samples in improper containers, such as plastics ‘other than TFE-fuorocarbon, may result in contamination, Nore 2—All“spectroquality” solvents may not have sufficiently low RTP emission, Solvent lots vary in content of impurities giving RTP and may increase with storage time, even for unopened bottles. Nore 3—This test method is normally used without an internal standard due to possible RTP interference by the internal standard, 7. Apparatus’ 7.1 Luminescence Spectrometer—Consult manufacturer's instrument manuals for specific operating instructions. It shall be equipped with either a phosphoroscope or a pulsed excitation with gated detection. An instrument recording in ‘the spectral range of 250 to 650 nm is required for both excitation and emission spectrum measurements and ca- pable of scanning both monochromators at a constant speed ‘with a constant wavelength offset between them for synchro- nous scanning. The instrument shall meet the specifications in Table 2. 7.2 Excitation Source—A continuous (150 W) or pulsed (9.9 W) xenon lamp or other sources having sufficient intensity throughout the ultraviolet and visible regions can be used, 7.3 RTP Sample Holder—Standard sample holders wete fabricated to hold disks of filter paper substrate 0.6 cm in diameter. An example of a sample cell holder is show Fig. 1. Other commercial sample holders allowing front- surface measurements may also be used. 74 Data Recording System—Preferably, the instrament shall be interfaced to a suitable computer system compatible with the instrument and with suitable software for spectral data manipulation. Alternatively, use ofa strip chart or X-Y TABLE 1 Chlorine Content of Aroclor™ Preparations “Area Number Average Mali eee) XO at ciper Maleate Wolght wa 205-215 115 182 1232 315025 204 2 1202 2 310 261 1268 “a 390 238 1254 56 495, ser 1260 60 630 ae 1282 615-625 680 389 1268 o 870 42 * Manufactre’'sepocfcains (Monsanto Chemical Go). TABLE 2 Specifications for Phosphorescence Spectrometers for PCB RTP Method xalaion monechvomaton 32 amo baor triason menolrorata: #2 rm o better ‘8.20 or 85 responce or quart ‘exciton monectvomate apectl andonss ‘of 15 ore ‘emission monockometo: special bardpast ol 15 ols ‘maximum bancpasses for both ‘monechyomators at leet 15 rm wavelength interval between exctaton and ‘omission monoctrometers = 175 om ‘sed exctation with gated detection eter olay (0.18 ms) or cntruous wave ‘exctnton wity mechanical phosphoroscope ‘Mowing mansuremenis of Matinee down fotm cress “Use ofa heavy atom quencher elinstes most Rooressonce inlereronces, ‘uc 93 PAHS, wo enough fer Sowening But see dscisslon under urknowne, Sypetrongus scanning Prosphorescance requrerants (Prforabie for quantcaton”) recorder with a response time of one second or less for full scale deflection is acceptable, 7.5 Sample Spotting Device~Microsyringe or micro- pipette with disposable tips (I to $ pL). 76 Glass Bottles or Flasks, with 20 to 25 mL capacity. Use low-actinic glass or wrap in aluminum foil to minimize photodegradation. 7.7 Filter Paper Disk Maker—Filter paper disk-maker (for example, standard 0.6 cm diameter hole puncher) may bbe used for conveniently cutting filter paper substrates. 7.8 Dry Air Supply—A warm dry air supply is necessary for RTP measurement. ow /SEMPLE 6 porting wer ELMER ore 7S FIG. 1 Example of ATP Cell Holder and Experimental Procedure) ps 47 8. Reagents and Materials 8.1 Purity of Reagents—Spectroquality grade reagents shall be used in all instances unless stated othe 8.2 Purity of Water—Double distilled water shall be used. Reference to water means Type IV water conforming to Specification D 1193. 8.3 Cyclohexane—Highest purity commercially available. 8.4 Ethanol—Spectroquality, 200 proof, not denatured. 8.5 Heavy-Atom Salt Til) Acetate—Highest purity com- mercially available, Pb(II) acetate may be used as an alterha- tive to Ti() acetate. Prepare a thallium acetate (1M) solution in a 50/50 (v/v) ethanol/water solvent. If preferred, lead acetate shall be prepared in the same manner. 8.6 Sodium Dodecyl Sulfate (SDS)—Highest purity com- mercially available. A 1% SDS solution (w/v) in water is used as the enhancer. 8.7 PCB Standards—Aroclot 1221, 1232, 1242, 1248, 1254, and 1260 PCB standards require dilution in spec- ‘troquality ethanol or cyclohexane. The initial concentration for the PCB mixture used in standards is prepared at 5000 nesmL. 88 Filter Paper—Schleicher and Schuell 2043A, if avail~ able, or equivalent. (Note that luminescence background of the filter paper must be checked. Not all filter papers may sive the same enhancement in phosphorescence intensity for PCBs.) 9. Sampling and Sample Preparation 9.1 Collect the sample in accordance with Practices D 3370, as applicable. 9.2 Extraction of PCBs. 9.2.1 Sediment—Collect a representable sediment sample in a clean low-actinic glass container and dry in an oven (30 to 40°C) for 72 h. Heating at higher temperatures may cause lighter PCBs to be lost. However, taking more than one ‘weighing of the soil of each sample will maintain consistency between samples. It is more important to ensure that samples are treated in a similar manner. Grind the sample to a fine powder so that itis easy to extract, Place one gram of ground soil in a microcentrifuge tube and add 0.5 mL of cyclo- hexane or ethanol. Vortex the tube so that the cyclohexane ‘or ethanol infiltrates the soil. Place the tube in an ultrasonic ‘bath for 30 min, Centrifuge the sample at 2000 rpm for 20 min then collect the supernatant. (Ifit is desired to eliminate the drying stage or if humic acids are of concern as an interferent, cyclohexane is the preferred solvent.) 9.2.2 Water—Collect 30 mL of a representable aqueous sample in a clean low-actinic glass container. Add 5 mL of cyclohexane to the sample and shake for 2 min. Collect the top layer of the extract in a 10-mL. volumetric flask with a It is important to collect most of the extract to maximize recovery. Repeat to perform a second extraction, combine the two extracts and dilute to 10 mL with cyclohexane. 9.3 Prepare samples as described and store in clean bottles. Samples may be subject to photodegradation so the storage procedure shall include covering containers with aluminum foil. important when handling the materials to use extreme caution, Safety glasses and gloves must be used to avoid physical contact with the PCBs as well as with thallium acetate. To ‘minimize health and safety concerns, it may be necessary to substitute ‘thallium acetate with lead acetate, although the RTP signal will be reduced by a factor of approximately 3. 10. Preparation of Apparatus 10.1 Set up the phosphorescence spectrometer and cali- brate according to the manufacturer's instructions and practices or Test Method E 388. For example, use the signal to noise value of the water Raman band to set the sensitivity gain of the detector. Calibration procedures shall include checking wavelength accuracy using known spectral refer- fence standards such as the emission lines of the xenon light source or a low-pressure mercury lamp. Allow the instru- ment electronics and light source to stabilize. The instrument shall meet the specifications of Table 2 with fixed or variable slits capable of covering the range of spectral resolution specified in the method and capable of scanning both ‘monochromators synchronously as well as individually. The instrument and software shall be capable of using a wave- Tength offset of 175 nm. LL. Procedure 11.1 Sample Holder Preparation: 11.1.1 Clean front face sample holder ultrasonically in a mixture of ethanol and acetone. Follow by rinsing with reagent water prior to u: 11.1.2 On a 0.6 cm diameter filter paper mounted on a front face sample holder, spot 2.5 iL of heavy-atom salt solution and allow to dry. Spot 2.5 yiL of extract. As an improved method, spot 2.5 iL of SDS before application of extract and allow to dry. The addition of SDS provides three to fourfold intensity enhancement. 11.1.3 When the sample is placed in the instrument, use a warm dry air supply to flush the sample chamber to av quenching of PCB phosphorescence due to water vapor. 11.2. Emission Scans—In general, a single emission scan with an excitation at 270-280 (15 nm bandpass) shall be sufficient for analyzing PCB mixtures in a given sample Repeat a solvent blank (either ethanol or cyclohexane, depending on sample preparation) following each scan of the unknown sample. The RTP excitation spectra of Aroclor 1254 using the emission wavelength at 490 nm is shown in Fig, 2. Experiments indicate that commercial PCB mixtures exhibit excitation maxima between 260 and 280 nm. 11.3 Synchronous Scans—For synchronous RTP, both excitation and emission monochromators are locked to- gether and scanned simultaneously, The excitation and ion slits shall both be set to a bandpass of 15 nm or less and 8d shall be 175 nm. Starting at an excitation mono- chromator setting of 225 nm and an emission monochroma- tor setting of 400 nm (Ad = 400 ~ 225 = 175 nm), scan the two monochromators simultaneously from 400 to 600 nm while recording the phosphorescence intensity asa function of emission wavelength. The average wavelength difference ‘between the excitation and emission maxima has been found to be 175 nm, (See Figs. 3 and 4.) 11.4 Prepare a calibration curve for the analyte of interest (for example, Aroclor 1221) at the beginning of the study. Prepare several concentrations (ranging from 1 to S000 ng/mL) by serial dilution of stock solution and measure with the same parameters as above. Dilute the concentrationsqh) ps a7 sco ery tty ts “asa 00 Synchronous RTP of Arocior™ 1254 + SDS + TIOAC Woven om FIG. 2 RTP Excitation Spectrum of Arocior™ 1254 FIG, 3. Synchronous ATP of Arocior™ 1232 + SDS + TIOAE sufficiently to ensure a linear relationship between RTP signal intensity and concentration. The working concentra- tion shall be high enough to produce a good RTP signal. Use the peak height of Amex (0 generate the calibration curve after background subtraction. (See Fig. 5. 11.5 Determine the concentration of the diluted unknown sample solution by referring the intensity to the calibration curve. ye -fateiea = 22terecr RZ + 050s 180 200 300 fouenty ine FIG. 5 Calibration Curve of Aroctor™ 1224 12. Calculation 12.1. Soil—Calculate concentration of the original extract sample as follows: concentration in wee = C_(¥,/M,) wo where: -oncentration from calibration curve, yg/mL, weight of solid that was extracted, g, and V, = volume of diluted extract, mL. 12.2 Water—Calculate concentration of the original ex- tract sample as follows: TABLE 3. ReproductMity Study on Aroclor™ 1221 In Soll E Conmueton Wet AIP ‘Reg Net Standard Repro- Concentaton Net ATP Sgrat ‘Slal Sonam ———81/ 907.8 ra songime §——137.10/.10 " ee 138 so0orngim. —1494/11.89/1260 1282concentratios be/mL. cM) @ n from calibration curve, yg/mL, V, = volume of diluted extract, mL, and V; = volume of water that sample was extracted from, mL. 13, Quality Control 13.1. Calibrate the fluorescence spectrometer frequently to check the wavelength accuracy with an appropriate mercury or other tine source. 13.2 Run a laboratory fortified blank to check complete system 513.3 Measure solvent blanks with each sample measure- ‘ment to check the purity of the solvent and the cleanliness of the sample holders. At low concentrations it may be neces- sary to subtract out solvent blanks for accurate quantifica- tion. This is done by performing phosphorescence measure- ‘ments of the solvent using the same parameters as with the sample. The blank is then subtracted. Treat sample and standard spectra in the same manner. 13.4 For each set of samples, measure one sample in Uiplicate using separate aliquots of the same sample extract. For each set of samples, carry one sample through the entire sample extraction, preparation, and analysis procedure in triplicate. A reproducibility study was performed and results, are shown in Table 3. i) ps 47 13.5 Check recoveries by extracting the same material with a second aliquot of solvent. Where the amount of PCB materials extracted in the second aliquot exceeds a certain amount (15 to 30 %) depending on desired accuracy, com- bine the two aliquots and perform a third extraction. 14. Precision and Bias 14,1 The precision and bias of this test method need to be established. Based on preliminary single laboratory results, the precision of the overall method is estimated to be about 30%, The precision of the phosphorescence measurement itself is of the ordgr of 15 %. Reference materials shall be selected which have similar phosphorescence spectra and similar phosphorescence intensities for the same weight of PCB mixture. The precision and bias of the test method in water will be established in a laboratory round robin. 14.2 Preliminary measurements of samples extracted from sediment have been performed. Extensive evaluation in a sediment matrix will be developed at a later time. 15, Keywords 15.1 PCB quantification; phosphorescence emission: polychlorinated biphenyls (PCBs); room temperature phos- phorescence (RTP); ultraviolet-visible luminescence or phds- phorescence Tho American Socey fr Testing and Materials takes no poston respecting the vay of any patent igh asserted a connection ‘wan any tom mentioned inthis standard. Users of his standard we expressly advised that datrminain of the volo of ay uh Pte righs, andthe rk of intingement of such rg, are etal the ow respons