1、Designation: D7597 092Standard Test Method forDetermination of Diisopropyl Methylphosphonate, EthylHydrogen Dimethylamidophosphate, EthylMethylphosphonic Acid, Isopropyl Methylphosphonic Acid,Methylphosphonic Acid and Pinacolyl MethylphosphonicAcid in Water by Liquid Chromatography/Tandem MassSpectr
2、ometry1This standard is issued under the fixed designation D7597; the number immediately 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 () ind
3、icates an editorial change since the last revision or reapproval.1NOTEThis test method was changed editorially in February 2012.2NOTEAdded research report footnote to Section 16 editorially in June 2013.1. Scope1.1 This procedure covers the determination of diisopropylmethylphosphonate (DIMP), ethyl
4、 hydrogen dimethylami-dophosphate (EHDMAP), ethyl methylphosphonic acid(EMPA), isopropyl methylphosphonic acid (IMPA), methyl-phosphonic acid (MPA) and pinacolyl methylphosphonic acid(PMPA) (referred to collectively as organophosphonates in thistest method) in surface water by direct injection using
5、 liquidchromatography (LC) and detected with tandem mass spec-trometry (MS/MS) using electrospray ionization (ESI). Theseanalytes are qualitatively and quantitatively determined by thismethod. This method adheres to single reaction monitoring(SRM) mass spectrometry.1.2 This test method has been deve
6、loped by US EPARegion5 Chicago Regional Laboratory (CRL).1.3 The values stated in SI units are to be regarded asstandard. No other units of measurement are included in thisstandard.1.4 The detection verification level (DVL) and reportingrange for the organophosphonates are listed in Table 1.1.4.1 Th
7、e DVL is required to be at a concentration at leastthree times below the reporting limit (RL) and have a signal/noise ratio greater than 3:1. Fig. 1 displays the signal/noiseratios at the DVLs for the organophosphonates in the ESIpositive mode and Fig. 2 in the ESI negative mode.1.4.2 The reporting
8、limit is the concentration of the Level 1calibration standard as shown in Table 2 for the organophos-phonates except for MPAin the ESI negative mode which is atLevel 2 due to not meeting the DVL criteria at the lowerconcentration level. The DVL for MPA in the ESI negativemode is at 20 g/L, which for
9、ces a raised reporting limit.However, the multi-laboratory validation required a spike of alltarget analytes at Level 1 concentrations. The mean recoveryfor MPA in the ESI negative mode at this level was 98.7 % asshown in Table 3. If your instruments sensitivity can meet therequirements in this test
10、 method, MPA may have a 50 g/Lreporting limit.1.5 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 practices and determine the applica-bility of regul
11、atory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D1129 Terminology Relating to WaterD1193 Specification for Reagent WaterD2777 Practice for Determination of Precision and Bias ofApplicable Test Methods of Committee D19 on WaterD3856 Guide for Management Systems in Laboratori
12、esEngaged in Analysis of WaterD3694 Practices for Preparation of Sample Containers andfor Preservation of Organic ConstituentsD5847 Practice for Writing Quality Control Specificationsfor Standard Test Methods for Water Analysis1This test method is under the jurisdiction of ASTM Committee D19 on Wate
13、rand is the direct responsibility of Subcommittee D19.06 on Methods forAnalysis forOrganic Substances in Water.Current edition approved Dec. 1, 2009. Published January 2010. DOI: 10.1520/D7597-09E02.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service
14、 at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1E2554 Practice for Estimating and Monitoring the
15、 Uncer-tainty of Test Results of a Test Method Using ControlChart Techniques2.2 Other Documents:EPA publication SW-846 Test Methods for Evaluating SolidWaste, Physical/Chemical Methods33. Terminology3.1 Definitions:3.1.1 detection verification level (DVL), na concentrationthat has a signal/noise rat
16、io greater than 3:1 and is at least 3times below the reporting limit (RL).3.1.2 reporting limit (RL), nthe concentration of thelowest-level calibration standard used for quantification.3.1.3 organophosphonates, nin this test method, diisopro-pyl methylphosphonate (DIMP), ethyl hydrogen dimethylami-d
17、ophosphate (EHDMAP), ethyl methylphosphonic acid(EMPA), isopropyl methylphosphonic acid (IMPA), methyl-phosphonic acid (MPA) and pinacolyl methylphosphonic acid(PMPA).3.2 Abbreviations:3.2.1 NDnon-detect4. Summary of Test Methods4.1 This is a performance-based method and modificationsare allowed to
18、improve performance.3Available from United States Environmental Protection Agency (EPA), ArielRios Bldg., 1200 Pennsylvania Ave., NW, Washington, DC 20460, http:/www.epa.gov.TABLE 1 Detection Verification Level and Reporting RangeAnalyte ESI Mode DVL (g/L) Reporting Range (g/L)Diisopropyl methylphos
19、phonate Positive 1 5150Ethyl hydrogen dimethylamidophosphate Negative 0.25 5150Ethyl hydrogen dimethylamidophosphate Positive 0.25 5150Ethyl methylphosphonic acid Negative 5 501500Ethyl methylphosphonic acid Positive 5 501500Isopropyl methylphosphonic acid Negative 10 501500Isopropyl Methylphosphoni
20、c acid Positive 5 501500Methylphosphonic acid Negative 20 1001500Methylphosphonic acid Positive 10 501500Pinacolyl methylphosphonic acid Negative 5 501500Pinacolyl methylphosphonic acid Positive 5 501500FIG. 1 Example ESI Positive Mode SRM Chromatograms Signal/Noise RatiosD7597 09224.2 For organopho
21、sphonate analysis, samples are shipped tothe lab between 0C and 6C and analyzed within 1 day ofcollection. In the lab, the samples are spiked with surrogate,filtered using a syringe-driven Millex HV PVDF filter unit andanalyzed directly by LC/MS/MS.4.3 The organophosphonates and the surrogates; diis
22、opropylmethylphosphonate-D14, pinacolyl methylphosphonic acid-13C6and methylphosphonic acid-D3are identified by retentiontime and one SRM transition. The target analytes and surro-gates are quantitated using the SRM transitions utilizing anexternal calibration. The final report issued for each sampl
23、elists the concentration of each organophosphonate target com-pound and each surrogate recovery.5. Significance and Use5.1 Organophosphate pesticides affect the nervous systemby disrupting the enzyme that regulates acetylcholine, a neu-rotransmitter. They were developed during the early 19thcentury,
24、 but their effects on insects, which are similar to theireffects on humans, were discovered in 1932. Some are poison-ous and were used as chemical weapon agents. Organophos-phate pesticides are usually not persistent in theenvironment.4,55.2 This test method is for the analysis of selectedorganophos
25、phorous-based chemical weapon agent degradationproducts from Sarin (GB), Soman (GD), Tabun (GA) and VX.This method has been investigated for use with reagent andsurface water.6. Interferences6.1 Method interferences may be caused by contaminants insolvents, reagents, glassware and other apparatus pr
26、oducingdiscrete artifacts or elevated baselines. All of these materialsare demonstrated to be free from interferences by analyzinglaboratory reagent blanks under the same conditions assamples.4Additional information about organophosphate pesticides is available on theInternet at http:/www.epa.gov (2
27、009).5Additional information about chemical weapon agents is available on theInternet at http:/www.opcw.org (2009).FIG. 2 Example ESI Negative Mode SRM Chromatograms Signal/Noise RatiosTABLE 2 Concentrations of Calibration Standards (PPB)Analyte/Surrogate LV 1 LV 2 LV 3 LV 4 LV 5 LV 6 LV 7Diisopropy
28、l methylphosphonate 5 10 20 35 50 100 150Ethyl hydrogen dimethylamidophosphate 5 10 20 35 50 100 150Ethyl methylphosphonic acid 50 100 200 350 500 1000 1500Isopropyl methylphosphonic acid 50 100 200 350 500 1000 1500Methylphosphonic acid 50 100 200 350 500 1000 1500Pinacolyl methylphosphonic acid 50
29、 100 200 350 500 1000 1500DIMP-D14 (Surrogate) 5 10 20 35 50 100 150PMPA-13C4 (Surrogate) 25 50 100 175 250 500 750MPA-D3 (Surrogate) 25 50 100 175 250 500 750D7597 09236.2 All glassware is washed in hot water with a detergent,rinsed in hot water followed by distilled water. The glasswareis then dri
30、ed and heated in an oven at 250C for 15 to 30minutes. All glassware is subsequently cleaned with acetone,then methanol.6.3 All reagents and solvents should be pesticide residuepurity or higher to minimize interference problems.6.4 Matrix interferences may be caused by contaminantsthat are co-extract
31、ed from the sample. The extent of matrixinterferences can vary considerably from sample source de-pending on variations of the sample matrix.7. Apparatus7.1 LC/MS/MS System7.1.1 Liquid Chromatography (LC) SystemAcomplete LCsystem is needed in order to analyze samples.6Asystem that iscapable of perfo
32、rming at the flows, pressures, controlledtemperatures, sample volumes and requirements of the stan-dard may be used.6A Waters Alliance High Performance Liquid Chromatography (HPLC) Systemwas used to develop this test method. The multi-laboratory study included Agilentand Waters LC systems.TABLE 3 Mu
33、lti-Laboratory Recovery Data in Reagent WaterAnalyteESIModeSpikeConc.(ppb)#Results# LabsBias PrecisionMeanRecovery(%)MinRecovery(%)MaxRecovery(%)Overall SD(%)Pooledwithin-labSD (%)OverallRSD (%)Pooledwithin-labRSD (%)DIMP Pos 5 16 4 95.1 65.8 136.0 17.6 19.4 21.3 22.4DIMP Pos 10 19 5 98.2 80.0 121.0
34、 6.1 5.5 13.6 13.8DIMP Pos 25 26 6 102.9 74.4 128.0 5.6 5.7 14.5 14.1DIMP Pos 125 22 5 96.6 80.4 120.0 4.4 4.5 11.0 11.4DIMP-D14 Pos 25 86 6 102.6 54.8 127.6 9.8 9.5 11.2 10.9EHDMAP Neg 5 12 3 57.5 0.0 220.0 31.4 22.3 71.1 123.6EHDMAP Neg 10 16 4 47.1 0.0 178.0 13.3 10.8 66.8 142.0EHDMAP Neg 25 22 5
35、 84.1 54.0 141.2 6.9 6.6 22.9 27.3EHDMAP Neg 125 18 4 87.4 64.2 141.6 5.0 5.1 25.4 29.1EHDMAP Pos 5 16 4 77.0 0.0 134.2 7.4 8.0 51.4 66.8EHDMAP Pos 10 20 5 70.0 0.0 143.0 7.7 10.0 53.6 76.5EHDMAP Pos 25 26 6 89.6 60.0 128.8 5.2 6.2 23.5 26.2EHDMAP Pos 125 22 5 87.5 59.0 123.2 5.2 6.0 23.9 27.3EMPA N
36、eg 50 16 4 110.1 74.8 170.6 22.4 17.2 25.6 23.3EMPA Neg 100 20 5 108.3 87.7 175.0 11.1 8.7 24.8 22.9EMPA Neg 250 26 6 104.8 82.0 122.6 6.1 5.5 11.8 11.3EMPA Neg 1250 22 5 101.5 87.2 126.4 8.4 8.1 11.2 11.0EMPA Pos 50 16 4 95.4 77.6 122.8 12.9 13.3 13.1 13.7EMPA Pos 100 20 5 96.0 61.4 132.5 9.5 9.5 1
37、5.9 16.6EMPA Pos 250 26 6 99.7 70.0 133.2 5.9 5.4 18.2 18.2EMPA Pos 1250 21 5 93.9 84.0 108.4 2.7 3.0 7.7 8.2IMPA Neg 50 16 4 88.0 56.6 140.4 23.7 26.0 23.5 26.7IMPA Neg 100 20 5 88.0 68.5 118.0 12.9 14.3 13.4 15.2IMPA Neg 250 26 6 98.1 72.8 144.0 13.1 11.9 19.2 19.6IMPA Neg 1250 21 5 90.7 73.1 103.
38、0 5.5 6.1 8.7 9.6IMPA Pos 50 16 4 98.3 47.8 139.6 19.2 20.5 27.2 27.7IMPA Pos 100 19 5 95.4 72.3 120.5 9.8 10.3 12.4 13.0IMPA Pos 250 26 6 97.0 79.2 188.4 7.4 7.6 10.9 11.2IMPA Pos 1250 21 5 91.3 70.4 115.5 5.1 5.2 11.4 12.5MPA Neg 50 16 4 98.7 3.3 175.0 14.2 25.3 60.5 61.3MPA Neg 100 20 5 100.0 41.
39、9 142.0 8.9 9.2 30.8 30.8MPA Neg 250 26 6 99.5 66.0 124.5 7.6 7.6 14.5 14.5MPA Neg 1250 22 5 102.7 81.8 130.5 10.5 9.9 12.4 12.1MPA Pos 50 16 4 68.3 9.8 139.6 13.4 20.3 36.6 53.6MPA Pos 100 20 5 80.5 48.4 149.7 14.0 12.6 26.8 33.3MPA Pos 250 26 6 91.7 33.9 153.7 8.0 7.8 31.8 34.7MPA Pos 1250 22 5 95
40、.8 31.8 208.2 12.6 8.3 43.4 45.3MPA-D3 Neg 250 84 6 111.2 57.2 190.8 16.2 12.5 30.0 26.9MPA-D3 Pos 250 68 5 104.4 58.4 151.8 14.3 14.0 18.0 17.3PMPA Neg 50 15 4 87.8 77.6 124.4 8.4 8.2 13.8 15.7PMPA Neg 100 19 5 91.6 83.6 98.2 2.8 3.0 4.4 4.8PMPA Neg 250 26 6 101.0 77.2 123.8 5.0 4.8 12.1 12.0PMPA N
41、eg 1250 22 5 99.2 84.8 126.5 4.9 4.8 12.3 12.4PMPA Pos 50 16 4 90.8 60.8 148.8 15.8 16.1 25.6 28.2PMPA Pos 100 15 4 95.2 86.8 114.0 3.8 4.0 7.9 8.3PMPA Pos 250 20 5 103.8 85.2 136.1 4.5 3.8 15.3 14.7PMPA Pos 1250 12 3 99.8 88.8 117.5 5.2 5.0 7.5 7.5PMPA-13C6 Neg 250 83 6 99.5 74.8 128.3 9.4 9.2 11.1
42、 11.1D7597 09247.1.2 Analytical Column-WatersAtlantis dC18, 150 mm 2.1 mm, 3 m particle size, or equivalent.7.1.3 Tandem Mass Spectrometer (MS/MS) SystemAMS/MS system capable of MRM analysis.7A system that iscapable of performing at the requirements in this standard maybe used.7.2 Filtration Device7
43、.2.1 Hypodermic syringeAlock tip glass syringe capableof holding a Millex HV Syringe Driven Filter Unit PVDF 0.45m (Millipore Corporation, Catalog # SLHV033NS) or similarmay be used.7.2.1.1 A25-mL lock tip glass syringe size is recommendedsince a 25-mL sample size is used in this test method.7.2.2 F
44、ilterMillex HV Syringe Driven Filter Unit PVDF0.45 m (Millipore Corporation, Catalog # SLHV033NS) orsimilar may be used.8. Reagents and Materials8.1 Purity of ReagentsHigh-performance liquid chroma-tography (HPLC) pesticide residue analysis and spectropho-tometry grade chemicals shall be used in all
45、 tests. Unlessindicated otherwise, it is intended that all reagents shallconform to the Committee on Analytical Reagents of theAmerican Chemical Society.8Other reagent grades may beused provided they are first determined they are of sufficientlyhigh purity to permit their use without affecting the a
46、ccuracy ofthe measurements.8.2 Purity of WaterUnless otherwise indicated, referencesto water shall be understood to mean reagent water conformingto Type 1 of Specification D1193. It must be demonstrated thatthis water does not contain contaminants at concentrationssufficient to interfere with the an
47、alysis.8.3 GasesUltrapure nitrogen and argon.8.4 Acetonitrile (CAS # 75-05-8).8.5 Methanol (CAS # 67-56-1).8.6 Acetone (CAS # 67-64-1).8.7 Formic acid (95%, CAS # 64-18-6).8.8 Diisopropyl methylphosphonate (DIMP, CAS # 1445-75-6).8.9 Ethyl hydrogen dimethylamidophosphate (EHDMAP,CAS # 2632-86-2).8.1
48、0 Ethyl methylphosphonic acid (EMPA, CAS # 1832-53-7).8.11 Isopropyl methylphosphonic acid (IMPA, CAS # 1832-54-8).8.12 Methylphosphonic acid (MPA, CAS # 993-13-5).8.13 Pinacolyl methylphosphonic acid (PMPA, CAS # 616-52-4).8.14 Diisopropyl methylphosphonate-D14(DIMP-D14, Un-labeled CAS # 1445-75-6)
49、.8.14.1 DIMP-D14 represents deuterium labeled diisopropylmethylphosphonate where the two isopropyl moieties containall2H.8.15 Methylphosphonic acid-D3(MPA-D3, Unlabeled CAS# 993-13-5).8.15.1 MPA-D3 represents deuterium labeled methylphos-phonic acid where the methyl moiety contains all2H.8.16 Pinacolyl methylphosphonic acid-13C6(PMPA-13C6,Unlabeled CAS # 616-52-4).8.16.1 PMPA-13C6 represents13C labeled pinacolyl meth-ylphosphonic where all the trimethylpropyl carbon atoms areuniformly labeled13C.9. Hazards9.1 Normal laboratory safety
