1、Designation: D 5315 04Standard Test Method forDetermination of N-Methyl-Carbamoyloximes andN-Methylcarbamates in Water by Direct Aqueous InjectionHPLC with Post-Column Derivatization1This standard is issued under the fixed designation D 5315; the number immediately following the designation indicate
2、s 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 (e) indicates an editorial change since the last revision or reapproval.1. Scope1.1 This is a high-performance liquid chromatogra
3、phic(HPLC) test method applicable to the determination of certainn-methylcarbamoyloximes and n-methylcarbamates in groundwater and finished drinking water (1)2. This test method isapplicable to any carbamate analyte that can be hydrolyzed toa primary amine. The following compounds have been vali-dat
4、ed using this test method:AnalyteChemical Abstract ServicesRegistry NumberAAldicarb 116-06-3Aldicarb sulfone 1646-88-4Aldicarb sulfoxide 1646-87-3Baygon 114-26-1Carbaryl 63-25-2Carbofuran 1563-66-23-Hydroxycarbofuran 16655-82-6Methiocarb 2032-65-7Methomyl 16752-77-5Oxamyl 23135-22-0_ANumbering syste
5、m of Chemical Abstracts, Inc.1.2 This test method has been validated in a collaborativeround-robin study (2) and estimated detection limits (EDLs)have been determined for the analytes listed in 1.1 (Table 1).Observed detection limits may vary between ground waters,depending on the nature of interfer
6、ences in the sample matrixand the specific instrumentation used.1.3 This test method is restricted to use by, or under thesupervision of, analysts experienced in both the use of liquidchromatography and the interpretation of liquid chromato-grams. Each analyst should demonstrate an ability to genera
7、teacceptable results with this test method using the proceduredescribed in 12.3.1.4 When this test method is used to analyze unfamiliarsamples for any or all of the analytes listed in 1.1, analyteidentifications should be confirmed by at least one additionalqualitative technique.1.5 The values state
8、d in SI units are to be regarded as thestandard.1.6 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 reg
9、ulatory limitations prior to use. Additional guid-ance on laboratory safety is available and suitable referencesfor the information are provided (3-5).2. Referenced Documents2.1 ASTM Standards:3D 1129 Terminology Relating to WaterD 1192 Specification for Equipment for Sampling Waterand Steam in Clos
10、ed Conduits4D 1193 Specification for Reagent WaterD 2777 Practice for Determination of Precision and Bias ofApplicable Methods of Committee D19 on WaterD 3370 Practices for Sampling Water from Closed Conduits1This test method is under the jurisdiction of ASTM Committee D19 on Waterand is the direct
11、responsibility of Subcommittee D19.06 on Methods forAnalysis forOrganic Substances in Water.Current edition approved June 1, 2004. Published June 2004. Originallyapproved in 1992. Last previous edition approved in 1998 as D 5315 92 (1998).2The boldface numbers in parentheses refer to the references
12、at the end of thistest method.3For 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.4Withdrawn.TABLE 1 Relative Ret
13、ention Times for the Primary andConfirmation Columns and EDLs for the 10 CarbamatePesticidesAnalyteRetention Time (minutes)PrimaryAConfirmationBEDLCAldicarb 27.0 21.4 1.0Aldicarb sulfone 15.2 12.2 2.0Aldicarb sulfoxide 15.0 17.5 2.0Baygon (Propoxur) 29.6 23.4 1.0Carbaryl 30.8 25.4 2.0Carbofuran 29.3
14、 24.4 1.53-Hydroxycarbofuran 23.3 19.0 2.0Methiocarb 34.9 28.6 4.0Methomyl 18.4 14.8 0.50Oxamyl 17.4 14.6 2.0APrimary column250 by 4.6 mm inside diameter Altex Ultrasphere ODS, 5m.BConfirmation column250 by 4.6 mm inside diameter Supelco LC-1, 5 m.CEstimated method detection limit in micrograms per
15、litre.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.D 3694 Practices for Preparation of Sample Containers andfor Preservation of Organic ConstituentsE 682 Practice for Liquid Chromatography Terms and Re-lationships2.2 U.S. Environm
16、ental Protection Agency Standard:EPA Method 531.1, Revision 3.0, USEPA, EMSL-Cincinnati, 19895EPA Method 531.2, Revision 1.0, USEPA, EMSL-Cincinnati, 200163. Terminology3.1 DefinitionsFor definitions of water terms used in thistest method, refer to Terminology D 1129. For definitions ofother terms u
17、sed in this test method, refer to Practice E 682.3.2 Definitions of Terms Specific to This Standard:3.2.1 calibration standard (CAL)a solution preparedfrom the primary dilution standard solution and stock standardsolutions of the internal standards and surrogate analytes. CALsolutions are used to ca
18、librate the instrument response withrespect to analyte concentration.3.2.2 field duplicates (FD1 and FD2)two separatesamples collected at the same time, placed under identicalcircumstances, and treated exactly the same throughout fieldand laboratory procedures. Analyses of FD1 and FD2 providea measu
19、re of the precision associated with sample collection,preservation, and storage, as well as with laboratory proce-dures.3.2.3 field reagent blank (FRB)reagent water placed in asample container in the laboratory and treated in all respects asa sample, including being exposed to sampling site conditio
20、ns,storage, preservation, and all analytical procedures. The pur-pose of the FRB is to determine whether method analytes orother interferences are present in the field environment.3.2.4 internal standarda pure analyte(s) added to a solu-tion in known amount(s) and used to measure the relativerespons
21、es of other analytes and surrogates that are componentsof the same solution. The internal standard must be an analytethat is not a sample component.3.2.5 laboratory duplicates (LD1 and LD2)two samplealiquots taken in the analytical laboratory and analyzed sepa-rately with identical procedures. Analy
22、ses of LD1 and LD2provide a measure of the precision associated with laboratoryprocedures, but not with sample collection, preservation, orstorage procedures.3.2.6 laboratory-fortified blank (LFB)an aliquot of re-agent water to which known quantities of the test methodanalytes are added in the labor
23、atory. The LFB is analyzedexactly as a sample is; its purpose is to determine whether themethodology is in control and whether the laboratory iscapable of making accurate and precise methods at the requiredtest method detection limit.3.2.7 laboratory-fortified sample matrix (LFM)an aliquotof an envi
24、ronmental sample to which known quantities of thetest method analytes are added in the laboratory. The LFM isanalyzed exactly as a sample is; its purpose is to determinewhether the sample matrix contributes bias to the analyticalresults. The background concentrations of the analytes in thesample mat
25、rix must be determined in a separate aliquot and themeasured values in the LFM corrected for background concen-trations.3.2.8 laboratory performance check solution (LPC)a so-lution of method analytes, surrogate compounds, and internalstandards used to evaluate the performance of the instrumentsystem
26、 with respect to a defined set of method criteria.3.2.9 laboratory reagent blank (LRB)an aliquot of re-agent water treated exactly the same as a sample, includingbeing exposed to all glassware, equipment, solvents, reagents,internal standards, and surrogates that are used with othersamples. The LRB
27、is used to determine whether methodanalytes or other interferences are present in the laboratoryenvironment, the reagents, or the apparatus.3.2.10 primary dilution standard solutiona solution ofseveral analytes prepared in the laboratory from stock standardsolutions and diluted as necessary to prepa
28、re calibrationsolutions and other necessary analyte solutions.3.2.11 quality control sample (QCS)a sample matrixcontaining test method analytes or a solution of test methodanalytes in a water miscible solvent that is used to fortify wateror environmental samples. The QCS is obtained from a sourceext
29、ernal to the laboratory and is used to check the laboratoryperformance with externally prepared test materials.3.2.12 stock standard solutiona concentrated solutioncontaining a single certified standard that is a method analyte,or a concentrated solution of a single analyte prepared in thelaboratory
30、 with an assayed reference compound. Stock stan-dard solutions are used to prepare primary dilution standards.3.2.13 surrogate analytea pure analyte(s), which is ex-tremely unlikely to be found in any sample, and which is addedto a sample aliquot in known amount(s) before extraction. It ismeasured w
31、ith the same procedures used to measure othersample components. The purpose of a surrogate analyte is tomonitor the method performance with each sample.4. Summary of Test Method4.1 The water sample is filtered, and a 200 to 400-Laliquotis injected onto a reverse phase HPLC column. Separation ofthe a
32、nalytes is achieved using gradient elution chromatogra-phy. After elution from the HPLC column, the analytes arehydrolyzed with sodium hydroxide (2.0 g/L NaOH) at 95C.The methylamine formed during hydrolysis is reacted witho-phthalaldehyde (OPA) and 2-mercaptoethanol to form ahighly fluorescent deri
33、vative that is detected by a fluorescencedetector (5).4.2 This method is applicable to any carbamte analyte thatcan be hydrolyzed to a primary amine, not necessarily methy-lamine.5. Significance and Use5.1 N-methylcarbamates and n-methylcarbomoyloximes areused in agriculture as insecticides and herb
34、icides. They aresometimes found in both surface and ground waters and can betoxic to animals and plants at moderate to high concentrations.5Published by the U.S. Environmental Protection Agency, EnvironmentalMonitoring and Support Laboratory, Cincinnati, OH 45268, 1989.6Published by the U.S. Environ
35、mental Protection Agency, EnvironmentalMonitoring and Support Laboratory, Cincinnati, OH 45268, 2001.D5315042The manufacturing precursors and degradation products maybe equally as hazardous to the environment.6. Interferences6.1 Test method interferences may be caused by contami-nants in solvents, r
36、eagents, glassware, and other sample pro-cessing apparatuses that lead to discrete artifacts or elevatedbaselines in liquid chromatograms. Specific sources of con-tamination have not been identified.All reagents and apparatusmust be routinely demonstrated to be free of interferencesunder the analysi
37、s conditions by running laboratory reagentblanks in accordance with 12.2.6.1.1 Glassware must be cleaned scrupulously. Clean allglassware as soon as possible after use by rinsing thoroughlywith the last solvent used in it.6.1.2 After drying, store glassware in a clean environmentto prevent any accum
38、ulation of dust or other contaminants.Store the glassware inverted or capped with aluminum foil.6.1.3 The use of high-purity reagents and solvents helps tominimize interference problems.6.2 Interfering contamination may occur when a samplecontaining low concentrations of analytes is analyzed imme-di
39、ately after a sample containing relatively high concentrationsof analytes. A preventive technique is between-sample rinsingof the sample syringe and filter holder with two portions ofwater. Analyze one or more laboratory method blanks afteranalysis of a sample containing high concentrations of ana-l
40、ytes.6.3 Matrix interference may be caused by contaminantspresent in the sample. The extent of matrix interference willvary considerably from source to source, depending upon thewater sampled. Positive analyte identifications must be con-firmed using the alternative conformational columns, or LC/MS.
41、6.4 The quality of the reagent water used to prepare stan-dards and samples must conform to D1193, especially in TOCcontent. High reagent water TOC causes a deterioration ofcolumn selectivity, baseline stability, and analyte sensitivity.6.5 Eliminate all sources of airborne primary amines, espe-cial
42、ly ammonia, which are absorbed into the mobile phases andeffect sensitivity.7. Apparatus7.1 Sampling Equipment:7.1.1 Sample Bottle, 60-mLscrew cap glass vials7and caps8equipped with a PTFE-faced silicone septa. Prior to use, washthe vials and septa as described in 6.1.1.7.2 Filtration Apparatus:7.2.
43、1 Macrofiltration Device, to filter derivatization solu-tions and mobile phases used in HPLC. It is recommended that47-mm, 0.45-m pore size filters be used.97.2.2 Microfiltration Device, to filter samples prior to HPLCanalysis. Use a 13-mm filter holder10and 13-mm diameter,0.2-m polyester filters.11
44、7.3 Syringes and Valves:7.3.1 Hypodermic Syringe, 10 mL, glass, with Luer-Lok12tip.7.3.2 Syringe Valve, three-way.137.3.3 Syringe Needle, 7 to 10 cm long, 17-gage, blunt tip.7.3.4 Micro Syringes, various sizes.7.4 Miscellaneous:7.4.1 Solution Storage Bottles, amber glass, 10 to 15-mLcapacity with TF
45、E-fluorocarbon-lined screw cap.7.5 High-Performance Liquid Chromatograph (HPLC):7.5.1 HPLC System,14capable of injecting 200 to 1000-Laliquots and performing ternary linear gradients at a constantflow rate. A data system is recommended for measuring peakareas. Table 2 lists the retention times obser
46、ved for test method7Sample bottle vial, Pierce No. 13075, available from Pierce Chemical Co., 3747N. Meridian Rd., Rockford, IL 61101, or equivalent.8Sample bottle cap, Pierce No. 12722, available from Pierce Chemical Co., 3747N. Meridian Rd., Rockford, IL 61101, or equivalent.9Millipore Type HA, 0.
47、45 m for water, and Millipore Type FH, 0.5 m fororganics, available from Millipore Corp., 80 Ashby Rd., Bedford, MA 01730, orequivalent.10Millipore stainless steel XX300/200, available from Millipore Corp., 80AshbyRd., Bedford, MA 01730, or equivalent.11Nucleopore 180406, available from Costar Corp.
48、, 1 Alewife Center, Cam-bridge, MA 02140, or equivalent.12Luer-Lok connectors are available from most laboratory suppliers.13Hamilton HV3-3, available from Hamilton Co., P.O. Box 10030, Reno, NV89502, or equilivalent.14Consult HPLC manufacturers operation manuals for specific instructionsrelating to
49、 the equipment.TABLE 2 Retention Times for Method Analytes Retention TimeAAnalyte PrimaryBConfirmationCConfirmationDMinutesAldicarb sulfoxide 6.80 17.5Aldicarb sulfone 7.77 12.2Oxamyl 8.20 14.6Methomyl 8.94 14.83-Hydroxycarbofuran 13.65 19Aldicarb 16.35 21.4Baygon (Propoxur) 18.86 24.4Carbofuran 19.17 23.4Carbaryl 20.29 25.4Methiocarb 24.74 28.6BDMC 25.28 .AColumns and analytical conditions are described in 7.5.2, 7.5.3.BBeckman Ultasphere ODS.CSupelco LC-1.DWaters Carbamate Analysis Column using ternary gradient conditions.D5315043analytes us
