1、Designation: D8025 16Standard Test Method forDetermination of Select Pesticides in Water by MultipleReaction Monitoring Liquid Chromatography Tandem MassSpectrometry1This standard is issued under the fixed designation D8025; the number immediately following the designation indicates the year oforigi
2、nal 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 a method for analysis of selectedpesticide
3、s in a water matrix by filtration followed with liquidchromatography/electrospray ionization tandem mass spec-trometry analysis. The samples are prepared in 20 % methanol,filtered, and analyzed by liquid chromatography/tandem massspectrometry. This method was developed for an agriculturalrun-off stu
4、dy, not for low level analysis of pesticides indrinking water. This method may be modified for lower levelanalysis. The analytes are qualitatively and quantitativelydetermined by this method. This method adheres to multiplereaction monitoring (MRM) mass spectrometry.1.2 A full collaborative study to
5、 meet the requirements ofPractice D2777 has not been completed. This standard containssingle-operator precision and bias based on single-operatordata. Publication of standards that have not been fully validatedis done to make the current technology accessible to users ofstandards, and to solicit add
6、itional input from the user com-munity.1.3 A reporting limit check sample (RLCS) is analyzedduring every batch to ensure that if an analyte was present ina sample at or near the reporting limit it would be positivelyidentified and accurately quantitated within set quality controllimits.Amethod detec
7、tion limit (MDL) study was not done forthis method, the method detection limits would be much lowerthan the reporting limits in this method and would be irrel-evant. A RLCS was determined to be more applicable for thisstandard. If this method is adapted to report much lower or nearthe MDL then a MDL
8、 study would be warranted.1.4 UnitsThe values stated in SI units are to be regardedas standard. No other units of measurement are included in thisstandard.1.5 The Reporting Range for the target analytes are listed inTable 1.1.5.1 The reporting limit in this test method is the minimumvalue below whic
9、h data are documented as non-detects. Thereporting limit is calculated from the concentration of the Level1 calibration standard as shown in Table 6 after taking intoaccount an 8 mL water sample volume and a final dilutedsample volume of 10 mL (80 % water/20 % methanol).1.6 This standard does not pu
10、rport 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 regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D
11、1129 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 LaboratoriesEngaged in Analysis of WaterD4841 Practice for Estimation of Holding Time
12、 for WaterSamples Containing Organic and Inorganic 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 Do
13、cument:EPA publication SW-846 Test Methods for Evaluating SolidWaste, Physical/Chemical Methods33. Terminology3.1 Definitions: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
14、 in Water.Current edition approved June 1, 2016. Published August 2016. DOI: 10.1520/D8025-16.2For 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
15、 Summary page onthe ASTM website.3Available 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.htmCopyright ASTM International, 100 Barr Harbor Drive, PO Box C700, We
16、st Conshohocken, PA 19428-2959. United States13.1.1 For definitions of terms used in this standard, refer toTerminology D1129.3.2 Definitions of Terms Specific to This Standard:3.2.1 batch QC, nall the quality control samples andstandards included in an analytical procedure.3.2.2 independent referen
17、ce material, IRM, na material ofknown purity and concentration obtained either from theNational Institute of Standards andTechnology (NIST) or otherreputable supplier.3.2.2.1 DiscussionThe IRM shall be obtained from adifferent lot of material than is used for calibration.3.2.3 reporting limit, RL, n
18、the minimum concentrationbelow which data are documented as non-detects.3.2.4 reporting limit check sample, RLCS, na sample usedto ensure that if the analyte was present at the reporting limit,it would be confidently identified.3.3 Acronyms:3.3.1 CCC, nContinuing Calibration Check3.3.2 CRW, nChicago
19、 River Water3.3.3 IC, nInitial Calibration3.3.4 LC, nLiquid Chromatography3.3.5 LCS/LCSD, nLaboratory Control Sample/Laboratory Control Sample Duplicate3.3.6 MDL, nMethod Detection Limit3.3.7 MeOH, nMethanol3.3.8 mM, nmillimolar,110-3moles/L3.3.9 MRM, nMultiple Reaction Monitoring3.3.10 MS/MSD, nMat
20、rix Spike/Matrix Spike Duplicate3.3.11 NA, adjNot Available3.3.12 ND, nnon-detect3.3.13 P however, this standard is intended to beperformance based and alternative operating conditions can beused to perform this method provided data quality objectivesare attained.4.2 For pesticide analysis, samples
21、are shipped to the lab onice and analyzed within 14 days of collection.Asample (8 mL)is transferred to an amber VOA vial, an isotopically labeledpesticide surrogate mix is added to all samples followed by apesticide spike solution which is added only to the ReportingLimit Check Samples, Laboratory C
22、ontrol and Matrix Spikesamples before the addition of methanol. Then 2 mL ofmethanol is added to each sample and hand shaken or vortexedfor 1 minute. The samples are then filtered through a PTFEmembrane syringe driven filter unit and then analyzed byLC/MS/MS. All concentrations reported only to the
23、reportinglimit.TABLE 1 Reporting RangeAnalyte Reporting Ranges, (ng/L)2,4-D 25010 000Acetochlor 25010 000Alachlor 25010 000Aldicarb 25010 000Atrazine 62.52 500Desethylatrazine 62.52 500Desisopropylatrazine 1255 000Azoxystrobin 31.21 250Bentazon 25010 000Carbaryl 25010 000Chlorpyrifos 25010 000Clopyr
24、alid 25 0001 000 000Clothianidin 62.52 500Diazinon 62.52 500Dicamba 12 500500 000Fipronil 25010 000Imidacloprid 62.52 500Malathion 1255 000Methomyl 25010 000Metolachlor 62.52 500Metribuzin 1255 000Picloram 6 250250 000Propiconazole 62.52 500Simazine 62.52 500Tebuconazole 62.52 500Thiamethoxam 62.52
25、500Triclopyr 1 2505 000TABLE 2 Gradient Conditions for Neutral Liquid ChromatographyTime (min) Flow (mL/min)Percent95 % Water: 5 % MethanolPercent MethanolPercent200 mM Ammonium Formate(95 % Water: 5 % Methanol)00.39 01.5 0.3 95 0 590 09512 0.3 0 95 513 0.3 95 0 516 0.3 95 0 5D8025 1624.3 The analys
26、is of the sample requires two separate analy-sis methods, one using the LC gradient conditions in Table 2with the Methanol/Water/Ammonium Formate and the secondusing the LC gradient conditions in Table 3 with the Methanol/Water/Formic acid. Each analysis set is to be analyzed sepa-rately in two diff
27、erent complete sample sequences whichincludes the exact same samples and may even include thesame calibration level standards. The only analytes reportedfrom the formic acid run conditions are 2,4-D, Clopyralid,Dicamba, Picloram, Triclopyr, 2,4-D (Ring-D3) and Dicamba-D3, the rest of the analytes ar
28、e reported from the ammoniumformate analysis run.4.4 The pesticides are identified by comparing the singlereaction monitoring (SRM) transition and its confirmatorySRM transitions if correlated to the known standard SRMtransition (Tables 4 and 5) and quantitated utilizing an externalcalibration. The
29、final report issued for each sample lists theconcentration of pesticides, if detected, or RL, if not detected,in ng/L and surrogate recovery.5. Significance and Use5.1 Pesticides may be used in various agricultural andhousehold products. These products may enter waterways atlow levels through run-of
30、f or misuse near water resources.Hence, there is a need for quick, easy and robust method todetermine pesticide concentration in water matrices for under-standing the sources and concentration levels in affected areas.5.2 This method has been single-laboratory validated inreagent water and surface w
31、aters (Tables 12-14).6. Interferences6.1 All glassware is washed in hot water with detergent andrinsed 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 subsequently rinsed or sonicated, orboth, with acetone, n-pr
32、opanol, acetonitrile, or a combinationthereof.6.2 All reagents and solvents should be pesticide residuepurity or higher to minimize interference problems.6.3 Matrix interferences may be caused by contaminants inthe sample. The extent of matrix interferences can varyconsiderably depending on variatio
33、ns in the sample matrices.7. Apparatus7.1 LC/MS/MS System:7.1.1 Liquid Chromatography System4A complete LCsystem is required in order to analyze samples, this shouldinclude a sample injection system, an autosampler, a solventpumping system capable of mixing solvents, a sample com-partment capable of
34、 maintaining required temperature and atemperature controlled column compartment.ALC system thatis capable of performing at the flows, pressures, controlledtemperatures, sample volumes, and requirements of the stan-dard shall be used.7.1.2 Analytical Column5A reverse phase C18 particlecolumn was use
35、d 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 usedand need to be monitored.7.2 Tandem Mass Spectrometer System6AMS/MS systemcapable of multiple reaction monitoring (MRM) analysis orany
36、 system that is capable of meeting the requirements in thisstandard shall be used. Electrospray ionization is utilized forthis standard.7.3 Adjustable Volume Pipettes10, 20, 100 and 1000 Land 5 and 10 mL.7.3.1 DiscussionAny pipette may be used providing thedata generated meets the performance of the
37、 standard.7.3.2 Pipette TipsPolypropylene pipette tips free of re-lease agents or low retention coating of various sizes.7.4 Class A Volumetric Glassware.7.5 Filtration Device:7.5.1 Hypodermic SyringeA luer-lock tip glass syringecapable of holding a syringe driven filter unit.7.5.2 A10 mLLock Tip Gl
38、ass Syringe size is recommendedsince a 10 mL prepared sample size is used in this test method.If a smaller volume syringe is used, do not wash out the syringe4AWatersAcquity (a trademark of the Waters Corporation, Milford, MA) UPLCH-Class System, or equivalent, has been found suitable for use.5AWate
39、rsAcquity (a trademark of the Waters Corporation, Milford, MA) UPLCBEH C18, 2.1100 mm and 1.7 m particle size column, or equivalent, has beenfound suitable for use. It was used to develop this test method and generate theprecision and bias data presented in Section 16.6A Waters Xevo (a trademark of
40、the Waters Corporation, Milford, MA) TQ-Striple quadrupole mass spectrometer, or equivalent, has been found suitable for use.TABLE 3 Gradient Conditions for Acidic Liquid ChromatographyTime (min) Flow (mL/min)Percent95 % Water: 5 % MethanolPercent MethanolPercent4 % Formic Acid(95 % Water: 5 % Metha
41、nol)00.39 01.5 0.3 95 0 560 0959010 0.3 95 0 513 0.3 95 0 5D8025 163TABLE 4 SRM Ions and Analyte-Specific Mass Spectrometer ParametersChemicalPrimary/ConfirmatoryMRM TransitionESI ModeNeg/PosCone(V)Collision(eV)2,4-DAPrimary 218.9160.9 Neg 20 14Confirmatory 220.9162.9 14Acetochlor Primary 270.1224.1
42、 Pos 20 10Confirmatory 270.1148 18Alachlor Primary 270.1162 Pos 30 12Confirmatory 270.1238.1 18Aldicarb Primary 21388.9 Pos 20 16Confirmatory 213116 12Atrazine Primary 216.1174 Pos 20 18Confirmatory 216.195.9 24Desethylatrazine Primary 188146 Pos 30 16Confirmatory 18878.8 22Desisopropylatrazine Prim
43、ary 17496 Pos 30 16Confirmatory 17478.8 16Azoxystrobin Primary 404.2372.2 Pos 20 14Confirmatory 404.2344.2 24Bentazon Primary 239197 Pos 20 20Confirmatory 239175 20Carbaryl Primary 202.1145 Pos 10 10Confirmatory 202.1127 25Chlorpyrifos Primary 350197.9 Pos 30 20Confirmatory 350322 11ClopyralidAPrima
44、ry 189.9145.9 Neg 5 10Confirmatory 191.9147.9 10Clothianidin Primary 250169 Pos 20 15Confirmatory 250131.9 11Diazinon Primary 305.1169 Pos 30 20Confirmatory 305.1153 20DicambaAPrimary 218.9174.9 Neg 10 8Confirmatory 220.9176.9 8Fipronil Primary 435330 Pos 30 15Confirmatory 435318 23Imidacloprid Prim
45、ary 256.1209.1 Pos 30 17Confirmatory 256.1175 23Malathion Primary 331.1127 Pos 20 12Confirmatory 331.1285 6Methomyl Primary 16387.9 Pos 10 10Confirmatory 163105.9 11Metolachlor Primary 284.1252.1 Pos 30 16Confirmatory 284.1176.1 25Metribuzin Primary 215.1187.1 Pos 30 16PicloramAPrimary 240.9196.9 Ne
46、g 10 11Confirmatory 238.9194.9 11Propiconazole Primary 342.1158.9 Pos 30 22Confirmatory 342.1205 17Simazine Primary 202132 Pos 40 17Confirmatory 202124 17Tebuconazole Primary 308.270 Pos 40 20Confirmatory 308.2125 28Thiamethoxam Primary 292.1211.1 Pos 20 12Confirmatory 292.1131.9 20TriclopyrAPrimary
47、 253.9195.9 Neg 10 12Confirmatory 253.9217.9 Neg 10 6Surrogates2,4-D (Ring-D3)APrimary 221.9163.8 Neg 20 14Atrazine (ethyl-D5) Primary 221.1179 Pos 20 17Desethylatrazine (iso-propyl-D7) Primary 195146.9 Pos 20 18Desisopropylatrazine (ethyl-D5) Primary 179100.9 Pos 30 18Bentazon -D7 Primary 246.1182
48、Pos 20 20Carbofuran (Ring-13C6) Primary 228.1171 Pos 20 12Clothianidin -D3 Primary 253131.9 Pos 20 15Diazinon (Diethyl-D10) Primary 315.2170 Pos 30 20Dicamba -D3APrimary 223.9179.9 Neg 10 8Imidacloprid -D4 Primary 260.1213.1 Pos 30 15Methomyl (Acetohydroxamate-13C2 15N) Primary 16690.8 Pos 10 8Simaz
49、ine (Diethyl-D10) Primary 212.1134 Pos 40 18Tebuconazole (tert-Butyl-D9) Primary 317.269.9 Pos 40 20Thiamethoxam -D3 Primary 295214.1 Pos 30 12AIndicates analyzed under acidic LC conditions.D8025 164TABLE 5 SRM Ions, Retention times and SRM Ion RatiosChemicalPrimary/ConfirmatoryMRMTransitionRetentionTimeMinutesPrimary/ConfirmatorySRM Area Ratio2,4-DAPrimary 218.9160.9 7.6 1.5Confirmatory 220.9162.9Acetochlor Primary 270.1224.1 10.3 2.5Confirmatory 270.11
