1、Designation: D7597 091Standard 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 2012.1. Scope1.1 This procedure covers the determination of diisopropylmethylphosphonate (DIMP), ethyl hydrogen dimethylami-dophosphate (EHDMAP), ethyl methylphosphonic acid(EMPA), isopr
4、opyl 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 liquidchromatography (LC) and detected with tandem mass spec-trometry (MS/MS) using
5、 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 developed by US EPARegion5 Chicago Regional Laboratory (CRL).1.3 The values stated in SI
6、 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 The DVL is required to be at a concentration at leastthree times below the reporting l
7、imit (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 limit is the concentration of the Level 1calibration standard as shown in Table 2 fo
8、r the organophos-phonates except for MPA in 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 forces a raised reporting limit.However, the multi-laboratory validation required a sp
9、ike 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 method, MPA may have a 50 g/Lreporting limit.1.5 This standard does not purport to
10、 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 regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D1129 Ter
11、minology 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 LaboratoriesEngaged in Analysis of WaterD3694 Practices for Preparation of Sample Containers
12、andfor Preservation of Organic ConstituentsD5847 Practice for Writing Quality Control Specificationsfor Standard Test Methods for Water AnalysisE2554 Practice for Estimating and Monitoring the Uncer-tainty of Test Results of a Test Method in a SingleLaboratory Using a Control Sample Program2.2 Other
13、 Documents: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 Dec. 1, 2009. Published February 2012. DOI: 10.1520/D7597-09E01.2For referenced
14、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.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Consh
15、ohocken, PA 19428-2959, United States.EPApublication 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 ratio greater than 3:1 and is at least 3times below the report
16、ing 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-dophosphate (EHDMAP), ethyl methylphosphonic acid(EMPA), iso
17、propyl 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 improve performance.4.2 For organophosphonate analysis, sam
18、ples are shipped tothe lab between 0C and 6C and analyzed within 1 day ofcollection. In the lab, the samples are spiked with surrogate,3Available from United States Environmental Protection Agency (EPA), ArielRios Bldg., 1200 Pennsylvania Ave., NW, Washington, DC 20460, http:/www.epa.gov.TABLE 1 Det
19、ection Verification Level and Reporting RangeAnalyte ESI Mode DVL (g/L) Reporting Range (g/L)Diisopropyl methylphosphonate Positive 1 5150Ethyl hydrogen dimethylamidophosphate Negative 0.25 5150Ethyl hydrogen dimethylamidophosphate Positive 0.25 5150Ethyl methylphosphonic acid Negative 5 501500Ethyl
20、 methylphosphonic acid Positive 5 501500Isopropyl methylphosphonic acid Negative 10 501500Isopropyl Methylphosphonic acid Positive 5 501500Methylphosphonic acid Negative 20 1001500Methylphosphonic acid Positive 10 501500Pinacolyl methylphosphonic acid Negative 5 501500Pinacolyl methylphosphonic acid
21、 Positive 5 501500FIG. 1 Example ESI Positive Mode SRM Chromatograms Signal/Noise RatiosD7597 0912filtered using a syringe-driven Millex HV PVDF filter unit andanalyzed directly by LC/MS/MS.4.3 The organophosphonates and the surrogates; diisopropylmethylphosphonate-D14, pinacolyl methylphosphonic ac
22、id-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 samplelists the concentration of each organophosphonate target c
23、om-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, but their effects on insects, which are similar to theiref
24、fects 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 the environ-ment.4,55.2 This test method is for the analysis of selectedorganophosphorous-based chemical weapon agent degradationproducts f
25、rom 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 producingdiscrete artifacts or elevated baselines. All of t
26、hese materials4Additional information about organophosphate pesticides is available on theInternet at http:/www.epa.gov (2009).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
27、 RatiosTABLE 2 Concentrations of Calibration Standards (PPB)Analyte/Surrogate LV 1 LV 2 LV 3 LV 4 LV 5 LV 6 LV 7Diisopropyl 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 methylphosph
28、onic acid 50 100 200 350 500 1000 1500Methylphosphonic acid 50 100 200 350 500 1000 1500Pinacolyl methylphosphonic acid 50 100 200 350 500 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 0913are demon
29、strated to be free from interferences by analyzinglaboratory reagent blanks under the same conditions assamples.6.2 All glassware is washed in hot water with a detergent,rinsed in hot water followed by distilled water. The glasswareis then dried and heated in an oven at 250C for 15 to 30minutes. All
30、 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-extracted from the sample. The extent of matrixinterferences can
31、 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) SystemA completeLC system is needed in order to analyze samples.6A systemthat is capable of performing at the flows, pressures, controlledtemperatures,
32、sample volumes and requirements of the stan-dard may be used.7.1.2 Analytical Column-WatersAtlantis dC18, 150 mm3 2.1 mm, 3 m particle size, or equivalent.6A Waters Alliance High Performance Liquid Chromatography (HPLC) Systemwas used to develop this test method. The multi-laboratory study included
33、Agilentand Waters LC systems.TABLE 3 Multi-Laboratory Recovery Data in Reagent WaterAnalyteESIModeSpike Conc.(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
34、 21.3 22.4DIMP Pos 10 19 5 98.2 80.0 121.0 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 17
35、8.0 13.3 10.8 66.8 142.0EHDMAP Neg 25 22 5 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
36、 5 87.5 59.0 123.2 5.2 6.0 23.9 27.3EMPA Neg 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 13.7E
37、MPA Pos 100 20 5 96.0 61.4 132.5 9.5 9.5 15.9 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
38、19.2 19.6IMPA Neg 1250 21 5 90.7 73.1 103.0 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.
39、2 25.3 60.5 61.3MPA Neg 100 20 5 100.0 41.9 142.0 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 1
40、53.7 8.0 7.8 31.8 34.7MPA Pos 1250 22 5 95.8 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
41、6 101.0 77.2 123.8 5.0 4.8 12.1 12.0PMPA Neg 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
42、 Neg 250 83 6 99.5 74.8 128.3 9.4 9.2 11.1 11.1D7597 09147.1.3 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 Device7.2.1 Hypodermic syringeAlock tip glass syringe capa
43、bleof holding a Millex HV Syringe Driven Filter Unit PVDF 0.45m (Millipore Corporation, Catalog # SLHV033NS) or similarmay be used.7.2.1.1 A 25-mL lock tip glass syringe size is recommendedsince a 25-mL sample size is used in this test method.7.2.2 FilterMillex HV Syringe Driven Filter Unit PVDF0.45
44、 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 tests. Unlessindicated otherwise, it is intended
45、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 accuracy ofthe measurements.8.2 Purity of WaterUnle
46、ss 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 analysis.8.3 GasesUltrapure nitrogen and argon.8.4 A
47、cetonitrile (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).8.10 Ethyl methylphosphonic acid (EMPA, CAS # 1832-5
48、3-7).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).8.14.1 DIMP-D14 represents deuterium labeled dii
49、sopropylmethylphosphonate 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 applies to this method. Ana-lysts should wear safety
