1、Designation: D7600 16 (Reapproved 2017)Standard Test Method forDetermination of Aldicarb, Carbofuran, Oxamyl andMethomyl by Liquid Chromatography/Tandem MassSpectrometry1This standard is issued under the fixed designation D7600; the number immediately following the designation indicates the year ofo
2、riginal 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 procedure covers the determination of aldicarb,carbofur
3、an, oxamyl and methomyl (referred to collectively ascarbamates in this test method) in surface water by directinjection using liquid chromatography (LC) and detected withtandem mass spectrometry (MS/MS). These analytes are quali-tatively and quantitatively determined by this test method. Thistest me
4、thod adheres to multiple reaction monitoring (MRM)mass spectrometry.1.2 This test method has been developed by U.S. EPARegion 5 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 Detect
5、ion Verification Level (DVL) and ReportingRange for the carbamates are listed in Table 1.1.4.1 The DVL is required to be at a concentration at least3 times below the Reporting Limit (RL) and have a signal/noise ratio greater than 3:1. Fig. 1 displays the signal/noiseratios of the primary single reac
6、tion monitoring (SRM) transi-tions and Fig. 2 displays the confirmatory SRM transitions atthe DVLs for the carbamates.1.4.2 The reporting limit is the concentration of the Level 1calibration standard as shown in Table 2 for the carbamates.1.5 This standard does not purport to address all of thesafet
7、y 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.1.6 This international standard was developed in accor-dance with internatio
8、nally recognized principles on standard-ization established in the Decision on Principles for theDevelopment of International Standards, Guides and Recom-mendations issued by the World Trade Organization TechnicalBarriers to Trade (TBT) Committee.2. Referenced Documents2.1 ASTM Standards:2D1129 Term
9、inology 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 a
10、ndfor 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 Using ControlChart Techniques2.2 Other Documents:3EPA publicat
11、ion 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 carbamates, nin this test method, aldicarb,carbofuran, ox
12、amyl and methomyl collectively.3.2.2 detection verification level, DVL, na concentrationthat has a signal/noise ratio greater than 3:1 and is at least 3times below the reporting limit (RL).3.2.3 independent reference material, IRM, na material ofknown purity and concentration obtained either from th
13、e1This 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 June 15, 2017. Published July 2017. Originallyapproved in 2009. Last previous edition appr
14、oved in 2016 as D7600 16. DOI:10.1520/D7600-16R17.2For 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
15、from National Technical Information Service (NTIS), U.S. Depart-ment of Commerce, 5285 Port Royal Road, Springfield, VA, 22161 or at http:/www.epa.gov/epawaste/hazard/testmethods/index.htm.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United Stat
16、esThis international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for theDevelopment of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Tra
17、de (TBT) Committee.1National Institute of Standards andTechnology (NIST) or otherreputable supplier. The IRM shall be obtained from a differentlot of material than is used for calibration3.3 Acronyms:3.3.1 CCC, nContinuing Calibration Check3.3.2 IC, nInitial Calibration3.3.3 LC, nLiquid Chromatograp
18、hy3.3.4 LCS/LCSD, nLaboratory Control Sample/Laboratory Control Sample Duplicate3.3.5 MDL, nMethod Detection Limit3.3.6 MeOH, nMethanol3.3.7 mM, nmillimolar,1103moles/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
19、-detect3.3.12 P&A, nPrecision and Accuracy3.3.13 PPB, nparts per billion3.3.14 PPT, nparts per trillion3.3.15 QA, adjQuality Assurance3.3.16 QC, adjQuality Control3.3.17 RL, nReporting Limit3.3.18 RSD, nRelative Standard Deviation3.3.19 RT, nRetention Time3.3.20 SDS, nSafety Data Sheets3.3.21 SRM, n
20、Single Reaction Monitoring3.3.22 SS, nSurrogate Standard3.3.23 TC, nTarget Compound3.3.24 M, nmicromolar,1106moles/L3.3.25 VOA, nVolatile Organic Analysis4. Summary of Test Methods4.1 This is a performance-based method and modificationsare allowed to improve performance.4.2 For carbamate analysis, s
21、amples are shipped to the labbetween 0C and 6C and analyzed within 7 days of collection.In the lab, the samples are spiked with surrogate, filtered usinga syringe-driven filter unit and analyzed directly by LC/MS/MS.4.3 Aldicarb, carbofuran, oxamyl, methomyl, and 4-bromo-3,5-dimethylphenyl-N-methylc
22、arbamate (BDMC, surrogate)are identified by retention time and two SRM transitions. Thetarget analytes and surrogate are quantitated using the primarySRM transitions utilizing an external calibration. The finalreport issued for each sample lists the concentration ofaldicarb, carbofuran, oxamyl, meth
23、omyl and the BDMC sur-rogate recovery.5. Significance and Use5.1 The N-methyl carbamate (NMC) pesticides: aldicarb,carbaryl, carbofuran, formetanate hydrochloride, methiocarb,methomyl, oxamyl, pirimicarb, propoxur, and thiodicarb havebeen identified by EPA as working through a common mecha-nism. The
24、y affect the nervous system by reducing the ability ofthe enzyme cholinesterase. Cholinesterase inhibition was theprimary toxicological effect of regulatory concern to EPA inassessing the NMCs food, drinking water and residential risks.In most of the country, NMC residues in drinking watersources ar
25、e at levels that are not likely to contribute substan-tially to the multi-pathway cumulative exposure. Shallowprivate wells extending through highly permeable soils intoshallow, acidic ground water represent what the EPA believesto be the most vulnerable drinking water.45.2 This test method has been
26、 investigated for use withreagent and surface water for the selected carbamates: aldicarb,carbofuran, oxamyl, and methomyl.6. Interferences6.1 Method interferences may be caused by contaminants insolvents, reagents, glassware and other apparatus producingdiscrete artifacts or elevated baselines. All
27、 of these materialsare demonstrated 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 2
28、50C 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-extracted from the sample. The exten
29、t of matrixinterferences can vary considerably from 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.5This shouldinclude a sample injection system, a solvent pum
30、ping systemcapable of mixing solvents, a sample compartment capable ofmaintaining required temperature and a temperature controlledcolumn compartment.Asystem that is capable of performing at4Additional information about carbamate pesticides can be found on the Internetat http:/www.epa.gov.5A Waters
31、Alliance High Performance Liquid Chromatography (HPLC) System(a trademark of the Waters Corporation, Milford, MA), or equivalent, was foundsuitable for use. The multi-laboratory study included Agilent and Waters LCsystems.TABLE 1 Detection Verification Level and Reporting RangeAnalyte DVL (ng/L) Rep
32、orting Range (g/L)Aldicarb 100 1100Carbofuran 100 1100Oxamyl 100 1100Methomyl 100 1100D7600 16 (2017)2FIG. 1 Example Primary SRM Chromatograms Signal/Noise RatiosFIG. 2 Example Confirmatory SRM Chromatograms Signal/Noise RatiosD7600 16 (2017)3the flows, pressures, controlled temperatures, sample vol
33、umesand requirements of the standard may be used.7.1.2 Analytical Column6A C18 column was used todevelop this test method. Any column that achieves adequateresolution may be used. The retention times and order ofelution may change depending on the column that is used andneed to be monitored.7.1.3 Ta
34、ndem 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 Device:7.2.1 Hypodermic syringeA luer-lock tip glass syringecapable of holding a syringe-driven filter unit.7.2.1.1 A25-mL
35、lock tip glass syringe size is recommendedsince a 25-mL sample size is used in this test method.7.2.2 Filter unit8PVDF filter units were used to filter thesamples.8. Reagents and Materials8.1 Purity of ReagentsHigh-performance liquid chroma-tography (HPLC) pesticide residue analysis and spectropho-t
36、ometry grade chemicals shall be used in all tests. Unlessindicated otherwise, it is intended that all reagents shallconform to the Committee on Analytical Reagents of theAmerican Chemical Society.9Other reagent grades may beused provided they are first determined to be of sufficientlyhigh purity to
37、permit their use without affecting the accuracy 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 concentra
38、tionssufficient 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).8.6 Acetone (CAS # 67-64-1).8.7 Ammonium acetate (CAS # 631-61-8).8.8 Ammonium hydroxide (Concentrated, CAS # 1336-21-6).8.9 Aldicarb (CAS # 116-06-3).8.
39、10 Carbofuran (CAS # 1563-66-2).8.11 Oxamyl (CAS # 23135-22-0).8.12 Methomyl (CAS # 16752-77-5).8.13 4-Bromo-3,5-dimethylphenyl-N-methylcarbamate(BDMC, CAS # 672-99-1).8.13.1 BDMC is used as a surrogate in this standard.9. Hazards9.1 Normal laboratory safety applies to this method. Ana-lysts should
40、wear safety glasses, gloves, and lab coats whenworking in the lab. Analysts should review the Safety DataSheets (SDS) for all reagents used in this test method.10. Sampling10.1 SamplingGrab samples must be collected in 25-mLpre-cleaned amber glass bottles with Teflon-lined caps demon-strated to be f
41、ree of interferences. This test method requires a25-mL sample size per analysis. Conventional sampling prac-tices should be followed. Refer to Guide D3856 and PracticesD3694.10.2 PreservationStore samples between 0C and 6Cfrom the time of collection until analysis. Analyze the samplewithin 7 days of
42、 collection.11. Preparation of LC/MS/MS11.1 LC Chromatograph Operating Conditions:511.1.1 Injection volumes of all calibration standards andsamples are 100 L. The first sample analyzed after thecalibration curve is a blank to ensure there is no carry-over.Thegradient conditions for the liquid chroma
43、tograph are shown inTable 3.11.1.2 TemperaturesColumn, 30C; Samplecompartment, 15C.6AWaters (a trademark of the Waters Corporation, Milford, MA) XBridge C18,150 mm 2.1 mm, 3.5 m particle size, or equivalent, has been found suitable foruse.7A Waters Quattro micro API mass spectrometer (a trademark of
44、 the WatersCorporation, Milford, MA), or equivalent, was found suitable for use. Themulti-laboratory study included Applied Biosystems and Waters mass spectrom-eters.8A Millex HV Syringe Driven Filter Unit PVDF 0.45 m (MilliporeCorporation, Catalog # SLHV033NS; a trademark of the Waters Corporation,
45、Milford, MA) has been found suitable for use for this test method, any filter unitmay be used that meets the performance of this test method may be used.9Reagent Chemicals, American Chemical Society Specifications, AmericanChemical Society, Washington, DC. For Suggestions on the testing of reagents
46、notlisted by the American Chemical Society, see Annual Standards for LaboratoryChemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeiaand National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,MD.TABLE 2 Concentrations of Calibration Standards (PPB)Analyte/Sur
47、rogate LV 1 LV 2 LV 3 LV 4 LV 5 LV 6Aldicarb 1 5 10 25 50 100Carbofuran 1 5 10 25 50 100Oxamyl 1 5 10 25 50 100Methomyl 1 5 10 25 50 100BDMC (Surrogate) 2 10 20 50 100 200TABLE 3 Gradient Conditions for Liquid ChromatographyTime(min)Flow(L/min)PercentCH3CNPercent95 % Water 5%CH3CNPercent 50 mmolarNH
48、4OAc/NH4OH in95 % Water/5 % CH3CN0 300 0 95 52 300 0 95 54 300 30 65 56 300 35 60 58 300 35 60 510 300 75 20 511.5 300 75 20 512 300 95 0 518 300 95 0 520 300 0 95 523 300 0 95 5D7600 16 (2017)411.1.3 Seal WashSolvent: 50 % Acetonitrile/50 % Water;Time: 5 minutes.11.1.4 Needle WashSolvent: 50 % Acet
49、onitrile/50 % Wa-ter; Normal wash, approximately 13 second wash time.11.1.5 Autosampler PurgeThree loop volumes.11.1.6 Specific instrument manufacturer wash/purge speci-fications should be followed in order to eliminate samplecarry-over in the analysis of carbamates.11.2 Mass Spectrometer Parameters:711.2.1 In order to acquire the maximum number of datapoints per SRM channel while maintaining adequatesensitivity, the tune parameters may be optimized according toyour instrument. Each peak re
