1、Designation: D 7485 09Standard Test Method forDetermination of Nonylphenol, p-tert-Octylphenol,Nonylphenol Monoethoxylate and Nonylphenol Diethoxylatein Environmental Waters by Liquid Chromatography/TandemMass Spectrometry1This standard is issued under the fixed designation D 7485; the number immedi
2、ately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.1. Scope1.1 This t
3、est method covers the determination of nonylphe-nol (NP), nonylphenol ethoxylate (NP1EO), nonylphenol di-ethoxylate (NP2EO), and octylphenol (OP), extracted fromwater utilizing solid phase extraction (SPE), separated usingliquid chromatography (LC) and detected with tandem massspectrometry (MS/MS).
4、These compounds are qualitativelyand quantitatively determined by this method. This methodadheres to single reaction monitoring (SRM) mass spectrom-etry.1.2 The method detection limit (MDL) and reporting limit(RL) for NP, NP1EO, NP2EO, and OP are listed in Table 1.1.3 The values stated in SI units a
5、re to be regarded asstandard. No other units of measurement are included in thisstandard.1.4 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practice
6、s and determine the applica-bility of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D 1129 Terminology Relating to WaterD 1193 Specification for Reagent WaterD 2777 Practice for Determination of Precision and Bias ofApplicable Test Methods of Committee D19 on WaterD
7、3694 Practices for Preparation of Sample Containers andfor Preservation of Organic ConstituentsD 3856 Guide for Good Laboratory Practices in Laborato-ries Engaged in Sampling and Analysis of WaterD 5847 Practice for Writing Quality Control Specificationsfor Standard Test Methods for Water AnalysisD
8、5905 Practice for the Preparation of Substitute Wastewa-ter2.2 Other DocumentsThe Code of Federal Regulations 40 CFR Part 136, Appen-dix B33. Terminology3.1 Nonylphenol (NP) is a mixture of branchedp-nonylphenol isomers. Commercial NP is produced by thereaction of phenol with commercial nonene. Comm
9、ercialnonene is not simply a linear C9H18alpha olefin; it is acomplex mixture of predominantly nine-carbon olefins, calledpropylene trimer, containing no linear isomers. This synthesisresults in a mixture of various branched nonylphenol isomersrather than a discrete chemical structure. The branched
10、nonylgroup is positioned predominantly in the para position on thephenol ring.3.2 OP represents octylphenol. Commercial octylphenol isproduced by the reaction of phenol and diisobutylene toproduce predominantly the 4-(1,1,3,3-tetramethylbutyl)phenolisomer.3.3 NP1EO represents branched nonylphenol mo
11、noethoxy-late.3.4 NP2EO represents branched nonylphenol diethoxylate.1This test method is under the jurisdiction of ASTM Committee D19 on Waterand is the direct responsibility of Subcommittee D19.06 on Methods forAnalysis forOrganic Substances in Water.Current edition approved March 1, 2009. Publish
12、ed March 2009.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3Available from U.S. Government Printing Office
13、 Superintendent of Documents,732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http:/www.access.gpo.gov.TABLE 1 MDL and Reporting LimitsAnalyteMDLA(ng/L)Reporting RangeB(ng/L)NP 33 100-2000NP1EO 9 100-2000NP2EO 9 100-2000OP 24 100-2000AMDL Determined Following The Code of Federal Regulat
14、ions, 40 CFR Part136, Appendix B.BLowest Point of the Reporting Range is Calculated from the LV 1 Concentra-tion Calibration Standard in Table 4.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.3.5 n-NP2EO represents normal straight c
15、hain nonylphenoldiethoxylate. n-NP2EO is used in this method as a surrogate. Itis not produced commercially and is not expected to be foundin environmental waters.3.6 2-Bromo-4-(1,1,3,3-tetramethylbutyl)phenol (Br-OP) isused in this method as a surrogate. It is not producedcommercially and is not ex
16、pected to be found in environmentalwaters. It was reported that compounds in highly chlorinatedbromide rich wastewaters could potentially interfere with theBr-OP surrogate. If this interference is encounteredn-nonylphenol is suggested as an alternative surrogate.3.7 Unitsparts per trillion (ng/L, pp
17、t), parts per billion(g/L, ppb), parts per million (mg/L, ppm)3.8 Environmental water shall refer to water tested usingthis method. See Section 5.4. Summary of Test Method4.1 This is a performance-based method and modificationsare allowed to improve performance.4.2 For NP, NP1EO, NP2EO, and OP analy
18、sis, solid phaseextraction is used to extract water samples.4.2.1 Solid Phase Extraction250 milliliter volume ofsample adjusted to pH 2 is extracted using a solid phaseextraction cartridge. The acetonitrile/water extract is concen-trated to a volume of 1.0 mL, and then analyzed by LC/MS/MSoperated i
19、n the multiple reaction monitoring (MRM) mode.4.3 The target compounds are identified by retention timeand SRM transition and are quantitated using the SRMtransition of the target compounds utilizing external calibra-tion. The final report issued for each sample lists the concen-tration of NP, NP1EO
20、, NP2EO, and OP.5. Significance and Use5.1 NP and OP have been shown to have toxic effects inaquatic organisms. The source of NP and OP is prominentlyfrom the use of common commercial surfactants. The mostwidely used surfactant is nonylphenol ethoxylate (NPEO)which has an average ethoxylate chain le
21、ngth of nine. Theethoxylate chain is readily biodegraded to form NP1EO,NP2EO, nonylphenol carboxylate (NPEC) and, under anaero-bic conditions, NP. NP will also biodegrade, but may bereleased into environmental waters directly at trace levels. Thismethod has been investigated and is applicable for en
22、viron-mental waters, including seawater.6. Interferences6.1 Method interferences may be caused by contaminants insolvents, reagents, glassware and other apparatus producingdiscrete artifacts or elevated baselines. All of these materialsare routinely demonstrated to be free from interferences byanaly
23、zing laboratory reagent blanks under the same conditionsas the samples.6.2 All glassware is scrupulously cleaned. All glassware iswashed in hot water with detergent such as powderedAlconox,Deto-Jet, Luminox, or Citrojet, rinsed in hot water and rinsedwith distilled water. The glassware is then dried
24、 and heated inan oven at 250C for 15 to 30 minutes. All glassware issubsequently cleaned with acetone and methanol. Detergentscontaining alkylphenolic compounds must not be used.6.3 All reagents and solvents should be of pesticide residuepurity or higher to minimize interference problems.6.4 Matrix
25、interferences may be caused by contaminantsthat are co-extracted from the sample. The extent of matrixinterferences can vary considerably from sample source tosample source, depending on variations of the sample matrix.7. Apparatus7.1 LC/MS/MS System:7.1.1 Liquid Chromatography SystemA complete Liqu
26、idChromatography system is needed in order to analyze samples.AWaters (registered trademark) ACQUITY Ultra PerformanceLiquid Chromatography (UPLC (trademark) System was usedto develop this test method, but a different system that iscapable of performing at the flows, pressures, controlledtemperature
27、s, sample volumes and requirements of the stan-dard may be used.7.1.2 Analytical ColumnWatersACQUITY UPLC(trademark) HSS T3, 1.8 mm, 2.1 3 50 mm column orequivalent.7.1.3 Tandem Mass Spectrometer (MS/MS) SystemAMS/MS system capable of MRM analysis. A Waters QuattroPremier XE mass spectrometer was us
28、ed to develop this testmethod, but another system that is capable of performing at therequirements in the standard may be used.7.2 SPE Vacuum Manifold SystemSupelco Visiprep solidphase extraction vacuum manifold or similar may be utilized.7.3 Organic solvent evaporation device.8. Reagents and Materi
29、als8.1 Purity of ReagentsHigh Performance Liquid Chroma-tography (HPLC) pesticide residue analysis and spectropho-tometry grade chemicals shall be used in all tests. Unlessindicated otherwise, it is intended that all reagents shallconform to the specifications of the Committee on AnalyticalReagents
30、of the American Chemical Society.4Other reagentgrades may be used provided it is first ascertained that they areof sufficiently high purity to permit their use without affectingthe accuracy of the measurement.58.2 Purity of WaterUnless otherwise indicated, referencesto water shall be understood to m
31、ean reagent water conformingto Type I of Specification D 1193. It must be demonstrated thatthis water does not contain contaminants at concentrationssufficient to interfere with the analysis.8.3 GasesUltrapure nitrogen and argon.8.4 Acetonitrile (CAS # 75-05-8).8.5 Methanol (CAS # 67-56-1).4Reagent
32、Chemicals, American Chemical Society Specifications, AmericanChemical Society, Washington, DC. For Suggestions on the testing of reagents notlisted by the American Chemical Society, see Annual Standards for LaboratoryChemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeiaand Nat
33、ional Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,MD.5Two sources of the alkylphenol standards are: Cambridge Isotope Laboratories,50 Frontage Road, Andover, MA 01810-5413 and Accustandard, Inc., 125 MarketStreet, New Haven, CT 06513. If you are aware of alternative suppliers, p
34、leaseprovide this information to ASTM International Headquarters. Your comments willreceive careful consideration at a meeting of the responsible technical committee,1which you may attend.D74850928.6 Isopropanol (CAS # 67-63-0).8.7 Acetone (CAS # 67-64-1).8.8 Branched nonylphenol monoethoxylate (NP1
35、EO) avail-able as a high purity custom standard.8.9 Branched nonylphenol diethoxylate (NP2EO) availableas a high purity custom standard.8.10 Nonylphenol, NP, 95 % para isomer (CAS # 84852-15-3).8.11 Octylphenol, OP, 99 + % 4-(1,1,3,3-tetramethylbutyl)phenol (CAS # 140-66-9).8.12 Concentrated HCl (CA
36、S # 7647-01-0).8.13 Ammonium Acetate (CAS # 631-61-8) (ACS ReagentGrade or Better).8.14 n-Nonylphenol diethoxylate (n-NP2EO).8.15 n-Nonylphenol (suggested alternate surrogate, ifneeded).8.16 2-Bromo-4-(1,1,3,3-tetramethylbutyl)phenol (Br-OP).8.17 Solid Phase Extraction CartridgesSep-Pak (regis-tered
37、 trademark) Vac (500 mg) tC18 Cartridges or equivalent.NOTE 1Alkylphenols have been found in SPE cartridges therefore itis advisable that the cartridges be lot certified alkylphenol free. Glasscartridges should have a much lower risk of alkylphenol contamination.9. Hazards9.1 Normal laboratory safet
38、y applies to this method. Ana-lysts should wear safety glasses, gloves and lab coats whenworking with acids.Analysts should review the Material SafetyData Sheets (MSDS) for all reagents used in this method.10. Sample Collection, Preservation, and Storage10.1 SamplingGrab samples must be collected in
39、 250 mLamber glass bottles. This must be done in order to allow for therinsing of the bottle with acidified water and acidified 10 %methanol/water in order to get complete transfer of the sampleinto the SPE cartridge and extraction process. Alkylphenolstend to adsorb to glassware and rinsing will al
40、low optimumrecoveries. Conventional sampling practices should be fol-lowed. Refer to Guide D 3856 and Practices D 3694. Auto-matic sampling equipment should be as free as possible ofTygon tubing and other potential sources of contamination.NOTE 2Pre-cleaned bottles demonstrated to be free of interfe
41、rencesmay be used.10.2 PreservationAdjust sample to pH 2 with concen-trated HCl at time of collection. Store samples between 0Cand 6C from the time of collection until extraction. Extract thesample within 14 days of collection and completely analyzewithin 14 days of extraction.10.3 Sample extracts m
42、ay be stored in sealed glass contain-ers at Product)Octylphenol neg 6.77 40 25 205.2 133NP2EO pos 7.71 20 12 326.3 183.2NP1EO pos 7.78 15 10 282.3 127.1Nonylphenol neg 7.82 40 30 219.2 133Br-OP (surrogate) neg 7.73 35 25 283.1 78.8n-NP2EO (surrogate) pos 8.62 20 17 326.3 88.9FIG. 1 Example SRM Chrom
43、atograms and Number of Scans per PeakTABLE 4 Concentrations of Calibration Standards (PPB)Analyte/Surrogate LV 1 LV 2 LV 3 LV 4 LV 5 LV 6 LV 7 LV 8NP 25 50 75 125 200 250 350 500NP1EO 25 50 75 125 200 250 350 500NP2EO 25 50 75 125 200 250 350 500Octylphenol 25 50 75 125 200 250 350 500Br-OP 25 50 75
44、 125 200 250 350 500n-NP2EO 25 50 75 125 200 250 350 500D7485094recording initial component weights carefully when workingwith the pure materials, and correctly carrying the weightsthrough the dilution calculations.12.2.1 Prepare stock standard Solution A (Level 8) byadding to a 100 mL volumetric fl
45、ask individual acetonitrile/methanol solutions of the following: 20 L of NP, OP, NP1EO,NP2EO, Br-OP and n-NP2EO each at 2500 ppm, dilute to 100mL with 75 % water/25 % acetonitrile. The 2500 ppm indi-vidual stock solutions are made in predominantly acetonitrilewith methanol added to ensure solubility
46、. Nonylphenol 2500ppm stock may require up to a 50 % concentration of methanol,the OP, NP1EO, NP2EO, Br-OP and n-NP2EO are predomi-nantly in acetonitrile. The preparation of the Level 8 standardcan be accomplished using different volumes and concentra-tions of stock solutions as is accustomed in the
47、 individuallaboratory. Depending on the stock concentrations prepared,the solubility at that concentration will have to be ensured.12.2.2 Aliquots of Solution A are then diluted with 75 %water/25 % acetonitrile to prepare the desired calibration levelsin 2 mL amber LC vials. The calibration vials mu
48、st be usedwithin 24 hours to ensure optimum results. Stock calibrationstandards are routinely replaced every six months if notpreviously discarded for QC criteria failure.12.2.3 Inject each standard and obtain a chromatogram foreach one. An external calibration is used monitoring the SRMtransition o
49、f each analyte. Calibration software is utilized toconduct the quantitation of the target analytes and surrogates.The SRM transition of each analyte is used for quantitation andconfirmation. This gives confirmation by isolating the parention, fragmenting it to the product ion fragment, and alsorelating it to the retention time in the calibration standard.12.2.4 The calibration software manual should be consultedto use the software correctly. The quantitation method is set asan external calibration using the peak areas in ppt or ppb unitsas long as the analyst is
