1、Designation: D7597 16Standard Test Method forDetermination of Diisopropyl Methylphosphonate, EthylHydrogen Dimethylamidophosphate, EthylMethylphosphonic Acid, Isopropyl Methylphosphonic Acid,Methylphosphonic Acid and Pinacolyl MethylphosphonicAcid in Water by Liquid Chromatography/Tandem MassSpectro
2、metry1This 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 () indi
3、cates an editorial change since the last revision or reapproval.1. Scope1.1 This procedure covers the determination of diisopropylmethylphosphonate (DIMP), ethyl hydrogen dimethylami-dophosphate (EHDMAP), ethyl methylphosphonic acid(EMPA), isopropyl methylphosphonic acid (IMPA), methyl-phosphonic ac
4、id (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 electrospray ionization (ESI). Theseanalytes are quali
5、tatively 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 units are to be regarded asstandard. No other units of
6、 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 limit (RL) and have a signal/noise ratio greater than 3:
7、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 for the organophos-phonates except for MPAin the ESI nega
8、tive 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 spike of alltarget analytes at Level 1 concentrations. The
9、 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 address all of thesafety concerns, if any, associated w
10、ith 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 Terminology Relating to WaterD1193 Specification for Reagen
11、t 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 andfor Preservation of Organic ConstituentsD5847 Practic
12、e 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 Using ControlChart Techniques1This test method is under the jurisdiction of ASTM Committee D19 on Waterand is the dir
13、ect responsibility of Subcommittee D19.06 on Methods forAnalysis forOrganic Substances in Water.Current edition approved Feb. 1, 2016. Published May 2016. Originallyapproved in 2009. Last previous edition approved in 2009 as D7597 091. DOI:10.1520/D7597-16.2For referenced ASTM standards, visit the A
14、STM 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.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. Unit
15、ed States12.2 Other Documents:3EPA Publication SW-846 Test Methods for Evaluating SolidWaste, Physical/Chemical Methods3. Terminology3.1 Definitions:3.1.1 For definitions of terms used in this standard, refer toTerminology D1129.3.2 Definitions of Terms Specific to This Standard:3.2.1 detection veri
16、fication level, DVL, na concentrationthat has a signal/noise ratio greater than 3:1 and is at least 3times below the reporting limit (RL).3.2.2 independent reference material, IRM, na material ofknown purity and concentration obtained either from theNational Institute of Standards andTechnology (NIS
17、T) or otherreputable supplier. The IRM shall be obtained from a differentlot of material than is used for calibration.3.2.3 organophosphonates, nin this test method, diisopro-pyl methylphosphonate (DIMP), ethyl hydrogen dimethylami-dophosphate (EHDMAP), ethyl methylphosphonic acid(EMPA), isopropyl m
18、ethylphosphonic acid (IMPA), methyl-phosphonic acid (MPA) and pinacolyl methylphosphonic acid(PMPA).3.3 Acronyms:3.3.1 CCC, nContinuing Calibration Check3.3.2 IC, nInitial Calibration3.3.3 LC, nLiquid Chromatography3Available from United States Environmental Protection Agency (EPA), ArielRios Bldg.,
19、 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 methylphosphonate Positive 1 5150Ethyl hydrogen dimethylamidophosphate Negative 0.25 5150Ethyl hydrogen dimethylami
20、dophosphate Positive 0.25 5150Ethyl methylphosphonic acid Negative 5 501500Ethyl methylphosphonic acid Positive 5 501500Isopropyl methylphosphonic acid Negative 10 501500Isopropyl Methylphosphonic acid Positive 5 501500Methylphosphonic acid Negative 20 1001500Methylphosphonic acid Positive 10 501500
21、Pinacolyl methylphosphonic acid Negative 5 501500Pinacolyl methylphosphonic acid Positive 5 501500FIG. 1 Example ESI Positive Mode SRM Chromatograms Signal/Noise RatiosD7597 1623.3.4 LCS/LCSD, nLaboratory Control Sample/Laboratory Control Sample Duplicate3.3.5 MDL, nMethod Detection Limit3.3.6 MeOH,
22、 nMethanol3.3.7 mM, nmillimolar,110-3moles/L3.3.8 MRM, nMultiple Reaction Monitoring3.3.9 MS/MSD, nMatrix Spike/Matrix Spike Duplicate3.3.10 NA, adjNot Available3.3.11 ND, nnon-detect3.3.12 P diisopropylmethylphosphonate-D14, pinacolyl methylphosphonic acid-13C6and methylphosphonic acid-D3are identi
23、fied by retentionFIG. 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 7Diisopropyl methylphosphonate 5 10 20 35 50 100 150Ethyl hydrogen dimethylamidophosphate 5 10 20 35 50 100 1
24、50Ethyl 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 50 100 200 350 500 1000 1500DIMP-D14 (Surrogate) 5 10 20 35 50 100 150PMPA-13C4 (Surrogate) 25 50 10
25、0 175 250 500 750MPA-D3 (Surrogate) 25 50 100 175 250 500 750D7597 163time 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
26、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, but their effects on insects, which are similar to
27、 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 selectedorganophosphorous-based chemical weapon agent degradation4Add
28、itional 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).TABLE 3 Multi-Laboratory Recovery Data in Reagent WaterAnalyteESIModeSpikeConc.(p
29、pb)#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 6.1 5.5 13.6 13.8DIMP Pos 25 26 6 102.9 74.4 128.0 5.6 5.7 14.5 14.1DI
30、MP 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 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
31、.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 Neg 50 16 4 110.1 74.8 170.6 22.4 17.2 25.6 23.3EMPA Neg 100 20 5 108.3
32、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 15.9 16.6EMPA Pos 250 26 6 99.7 70.0 133.2 5.9 5.4 18.2 18.2EMPA Pos 125
33、0 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 73.1 103.0 5.5 6.1 8.7 9.6IMPA Pos 50 16 4 98.3 47.8 139.6 19.2 20.5 27.2 27.7IM
34、PA 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 41.9 142.0 8.9 9.2 30.8 30.8MPA Neg 250 26 6 99.5 66.0 124.5 7.6 7.6 14.5
35、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 95.8 31.8 208.2 12.6 8.3 43.4 45.3MPA-D3 Neg 250 84 6 111.2 57.2 190.8 16
36、.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 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.
37、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 11.1D7597 164products from Sarin (GB), Soman (GD), Tabun (GA) and VX.T
38、his 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 these materialsare demonstrated to be free from
39、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 glassware is subsequent
40、ly 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 vary considerably from
41、sample source de-pending on variations of the sample matrix.7. Apparatus7.1 LC/MS/MS System:7.1.1 Liquid Chromatography (LC) SystemAcomplete LCsystem is needed in order to analyze samples.6This shouldinclude a sample injection system, a solvent pumping systemcapable of mixing solvents, a sample comp
42、artment capable ofmaintaining required temperature and a temperature controlledcolumn compartment.Asystem that is capable of performing atthe flows, pressures, controlled temperatures, sample volumesand requirements of the standard may be used.7.1.2 Analytical Column7A reverse phase C18 particlecolu
43、mn was used to develop this test method.Any column thatachieves adequate resolution may be used. The retention timesand order of elution may change depending on the column thatis used and need to be monitored.7.1.3 Tandem Mass Spectrometer (MS/MS) SystemAMS/MS system capable of MRM analysis.8A syste
44、m that iscapable of performing at the requirements in this standard maybe used.7.2 Filtration Device:7.2.1 Hypodermic SyringeA luer-lock tip glass syringecapable of holding syringe-driven filter unit.7.2.1.1 A25-mL lock tip glass syringe size is recommendedsince a 25-mL sample size is used in this t
45、est method.7.2.2 Filter Unit9A PVDF filter units were used to filterthe samples.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
46、 intended that all reagents shallconform to the Committee on Analytical Reagents of theAmerican Chemical Society.10Other 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 o
47、f 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 analysis.8.3 GasesUltrapure nitrogen and
48、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).8.10 Ethyl methylphosphonic acid (EMPA, C
49、AS # 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).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 cont
