Standard Test Method for Determination of N-Methyl-Carbamoyloximes and N-Methylcarbamates in Water by Direct Aqueous Injection HPLC with Post-Column Derivatization

SIGNIFICANCE AND USE
N-methylcarbamates and n-methylcarbomoyloximes are used in agriculture as insecticides and herbicides. They are sometimes found in both surface and ground waters and can be toxic to animals and plants at moderate to high concentrations. The manufacturing precursors and degradation products may be equally as hazardous to the environment.
SCOPE
1.1 This is a high-performance liquid chromatographic (HPLC) test method applicable to the determination of certain n-methylcarbamoyloximes and n-methylcarbamates in ground water and finished drinking water (1) . This test method is applicable to any carbamate analyte that can be hydrolyzed to a primary amine. The following compounds have been validated using this test method:

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Publication Date
30-Apr-2011
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ASTM D5315-04(2011) - Standard Test Method for Determination of N-Methyl-Carbamoyloximes and N-Methylcarbamates in Water by Direct Aqueous Injection HPLC with Post-Column Derivatization
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation:D5315 −04 (Reapproved 2011)
Standard Test Method for
Determination of N-Methyl-Carbamoyloximes and
N-Methylcarbamates in Water by Direct Aqueous Injection
HPLC with Post-Column Derivatization
This standard is issued under the fixed designation D5315; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 1.4 When this test method is used to analyze unfamiliar
samples for any or all of the analytes listed in 1.1, analyte
1.1 This is a high-performance liquid chromatographic
identifications should be confirmed by at least one additional
(HPLC) test method applicable to the determination of certain
qualitative technique.
n-methylcarbamoyloximes and n-methylcarbamates in ground
water and finished drinking water (1) . This test method is
1.5 The values stated in SI units are to be regarded as
applicable to any carbamate analyte that can be hydrolyzed to
standard. No other units of measurement are included in this
a primary amine. The following compounds have been vali-
standard.
dated using this test method:
1.6 This standard does not purport to address all of the
Chemical Abstract Services
safety concerns, if any, associated with its use. It is the
A
Analyte Registry Number
responsibility of the user of this standard to establish appro-
Aldicarb 116-06-3
Aldicarb sulfone 1646-88-4
priate safety and health practices and determine the applica-
Aldicarb sulfoxide 1646-87-3
bility of regulatory limitations prior to use. Additional guid-
Baygon 114-26-1
ance on laboratory safety is available and suitable references
Carbaryl 63-25-2
Carbofuran 1563-66-2
for the information are provided(3-5).
3-Hydroxycarbofuran 16655-82-6
Methiocarb 2032-65-7
2. Referenced Documents
Methomyl 16752-77-5
Oxamyl 23135-22-0
2.1 ASTM Standards:
________________
D1129Terminology Relating to Water
A
Numbering system of Chemical Abstracts, Inc.
D1192Guide for Equipment for Sampling Water and Steam
in Closed Conduits (Withdrawn 2003)
1.2 This test method has been validated in a collaborative
D1193Specification for Reagent Water
round-robin study (2) and estimated detection limits (EDLs)
D2777Practice for Determination of Precision and Bias of
have been determined for the analytes listed in 1.1 (Table 1).
Applicable Test Methods of Committee D19 on Water
Observed detection limits may vary between ground waters,
D3370Practices for Sampling Water from Closed Conduits
depending on the nature of interferences in the sample matrix
D3694Practices for Preparation of Sample Containers and
and the specific instrumentation used.
for Preservation of Organic Constituents
1.3 This test method is restricted to use by, or under the
E682Practice for Liquid Chromatography Terms and Rela-
supervision of, analysts experienced in both the use of liquid
tionships
chromatography and the interpretation of liquid chromato-
2.2 U.S. Environmental Protection Agency Standard:
grams. Each analyst should demonstrate an ability to generate
EPA Method 531.1, Revision 3.0, USEPA, EMSL-
acceptable results with this test method using the procedure
Cincinnati, 1989
described in 12.3.
1 3
This test method is under the jurisdiction ofASTM Committee D19 on Water For referenced ASTM standards, visit the ASTM website, www.astm.org, or
andisthedirectresponsibilityofSubcommitteeD19.06onMethodsforAnalysisfor contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Organic Substances in Water. Standards volume information, refer to the standard’s Document Summary page on
Current edition approved May 1, 2011. Published June 2011. Originally the ASTM website.
approved in 1992. Last previous edition approved in 2004 as D5315–04. DOI: The last approved version of this historical standard is referenced on
10.1520/D5315-04R11. www.astm.org.
2 5
The boldface numbers in parentheses refer to the references at the end of this Published by the U.S. Environmental Protection Agency, Environmental
test method. Monitoring and Support Laboratory, Cincinnati, OH 45268, 1989.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D5315−04 (2011)
TABLE 1 Relative Retention Times for the Primary and
3.2.6 laboratory-fortified blank (LFB)—an aliquot of re-
Confirmation Columns and EDLs for the 10 Carbamate
agent water to which known quantities of the test method
Pesticides
analytes are added in the laboratory. The LFB is analyzed
Retention Time (minutes)
exactly as a sample is; its purpose is to determine whether the
Analyte
A B C
Primary Confirmation EDL
methodology is in control and whether the laboratory is
Aldicarb 27.0 21.4 1.0
capableofmakingaccurateandprecisemethodsattherequired
Aldicarb sulfone 15.2 12.2 2.0
test method detection limit.
Aldicarb sulfoxide 15.0 17.5 2.0
Baygon (Propoxur) 29.6 23.4 1.0
3.2.7 laboratory-fortified sample matrix (LFM)—an aliquot
Carbaryl 30.8 25.4 2.0
of an environmental sample to which known quantities of the
Carbofuran 29.3 24.4 1.5
3-Hydroxycarbofuran 23.3 19.0 2.0
test method analytes are added in the laboratory. The LFM is
Methiocarb 34.9 28.6 4.0
analyzed exactly as a sample is; its purpose is to determine
Methomyl 18.4 14.8 0.50
whether the sample matrix contributes bias to the analytical
Oxamyl 17.4 14.6 2.0
A
results. The background concentrations of the analytes in the
Primary column—250 by 4.6 mm inside diameterAltex Ultrasphere ODS, 5 µm.
B
Confirmation column—250 by 4.6 mm inside diameter Supelco LC-1, 5 µm.
samplematrixmustbedeterminedinaseparatealiquotandthe
C
Estimated method detection limit in micrograms per litre.
measuredvaluesintheLFMcorrectedforbackgroundconcen-
trations.
3.2.8 laboratory performance check solution (LPC)—a so-
EPA Method 531.2, Revision 1.0, USEPA, EMSL-
lution of method analytes, surrogate compounds, and internal
Cincinnati, 2001
standards used to evaluate the performance of the instrument
system with respect to a defined set of method criteria.
3. Terminology
3.2.9 laboratory reagent blank (LRB)—an aliquot of re-
3.1 Definitions—For definitions of water terms used in this
agent water treated exactly the same as a sample, including
test method, refer to Terminology D1129. For definitions of
being exposed to all glassware, equipment, solvents, reagents,
other terms used in this test method, refer to Practice E682.
internal standards, and surrogates that are used with other
3.2 Definitions of Terms Specific to This Standard:
samples. The LRB is used to determine whether method
3.2.1 calibrationstandard(CAL)—asolutionpreparedfrom
analytes or other interferences are present in the laboratory
the primary dilution standard solution and stock standard
environment, the reagents, or the apparatus.
solutionsoftheinternalstandardsandsurrogateanalytes.CAL
3.2.10 primary dilution standard solution—a solution of
solutions are used to calibrate the instrument response with
severalanalytespreparedinthelaboratoryfromstockstandard
respect to analyte concentration.
solutions and diluted as necessary to prepare calibration
3.2.2 fieldduplicates(FD1andFD2)—twoseparatesamples
solutions and other necessary analyte solutions.
collected at the same time, placed under identical
3.2.11 quality control sample (QCS)—a sample matrix con-
circumstances, and treated exactly the same throughout field
taining test method analytes or a solution of test method
and laboratory procedures.Analyses of FD1 and FD2 provide
analytesinawatermisciblesolventthatisusedtofortifywater
a measure of the precision associated with sample collection,
or environmental samples. The QCS is obtained from a source
preservation, and storage, as well as with laboratory proce-
external to the laboratory and is used to check the laboratory
dures.
performance with externally prepared test materials.
3.2.3 field reagent blank (FRB)—reagent water placed in a
3.2.12 stock standard solution—a concentrated solution
samplecontainerinthelaboratoryandtreatedinallrespectsas
a sample, including being exposed to sampling site conditions, containing a single certified standard that is a method analyte,
or a concentrated solution of a single analyte prepared in the
storage, preservation, and all analytical procedures. The pur-
pose of the FRB is to determine whether method analytes or laboratory with an assayed reference compound. Stock stan-
dard solutions are used to prepare primary dilution standards.
other interferences are present in the field environment.
3.2.4 internal standard—a pure analyte(s) added to a solu- 3.2.13 surrogate analyte—a pure analyte(s), which is ex-
tion in known amount(s) and used to measure the relative
tremelyunlikelytobefoundinanysample,andwhichisadded
responsesofotheranalytesandsurrogatesthatarecomponents to a sample aliquot in known amount(s) before extraction. It is
of the same solution. The internal standard must be an analyte
measured with the same procedures used to measure other
that is not a sample component. sample components. The purpose of a surrogate analyte is to
monitor the method performance with each sample.
3.2.5 laboratory duplicates (LD1 and LD2)—two sample
aliquots taken in the analytical laboratory and analyzed sepa-
4. Summary of Test Method
rately with identical procedures. Analyses of LD1 and LD2
provide a measure of the precision associated with laboratory
4.1 Thewatersampleisfiltered,anda200to400-µLaliquot
procedures, but not with sample collection, preservation, or
is injected onto a reverse phase HPLC column. Separation of
storage procedures.
the analytes is achieved using gradient elution chromatogra-
phy. After elution from the HPLC column, the analytes are
hydrolyzed with sodium hydroxide (2.0 g/L NaOH) at 95°C.
Published by the U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Cincinnati, OH 45268, 2001. The methylamine formed during hydrolysis is reacted with
D5315−04 (2011)
7 8
o-phthalaldehyde (OPA) and 2-mercaptoethanol to form a 7.1.1 Sample Bottle,60-mLscrewcapglassvials andcaps
highly fluorescent derivative that is detected by a fluorescence equipped with a PTFE-faced silicone septa. Prior to use, wash
detector (5). the vials and septa as described in 6.1.1.
4.2 This method is applicable to any carbamte analyte that 7.2 Filtration Apparatus:
can be hydrolyzed to a primary amine, not necessarily meth- 7.2.1 Macrofiltration Device, to filter derivatization solu-
ylamine. tionsandmobilephasesusedinHPLC.Itisrecommendedthat
47-mm, 0.45-µm pore size filters be used.
5. Significance and Use 7.2.2 Microfiltration Device,tofiltersamplespriortoHPLC
analysis. Use a 13-mm filter holder and 13-mm diameter,
5.1 N-methylcarbamatesandn-methylcarbomoyloximesare
0.2-µm polyester filters.
used in agriculture as insecticides and herbicides. They are
sometimesfoundinbothsurfaceandgroundwatersandcanbe 7.3 Syringes and Valves:
toxic to animals and plants at moderate to high concentrations. 7.3.1 Hypodermic Syringe, 10 mL, glass, with Luer-Lok
The manufacturing precursors and degradation products may tip.
7.3.2 Syringe Valve, three-way.
be equally as hazardous to the environment.
7.3.3 Syringe Needle, 7 to 10 cm long, 17-gage, blunt tip.
7.3.4 Micro Syringes, various sizes.
6. Interferences
6.1 Test method interferences may be caused by contami- 7.4 Miscellaneous:
7.4.1 Solution Storage Bottles, amber glass, 10 to 15-mL
nants in solvents, reagents, glassware, and other sample pro-
cessing apparatuses that lead to discrete artifacts or elevated capacity with TFE-fluorocarbon-lined screw cap.
baselines in liquid chromatograms. Specific sources of con-
7.5 High-Performance Liquid Chromatograph (HPLC):
taminationhavenotbeenidentified.Allreagentsandapparatus 14
7.5.1 HPLC System, capable of injecting 200 to 1000-µL
must be routinely demonstrated to be free of interferences
aliquots and performing ternary linear gradients at a constant
under the analysis conditions by running laboratory reagent
flow rate. A data system is recommended for measuring peak
blanks in accordance with 12.2.
areas.Table2liststheretentiontimesobservedfortestmethod
6.1.1 Glassware must be cleaned scrupulously. Clean all
analytesusingthecolumnsandanalyticalconditionsdescribed
glassware as soon as possible after use by rinsing thoroughly
below.
with the last solvent used in it.
7.5.2 Column 1 (Primary Column), 250 mm long by
6.1.2 After drying, store glassware in a clean environment
4.6-mminsidediameter,stainlesssteel,packedwith5-µmC-18
to prevent any accumulation of dust or other contaminants.
material. Mobile phase is established at 1.0 mL/min as a
Store the glassware inverted or capped with aluminum foil.
linear gradient from 15:85 methanol: water to 100% methanol
6.1.3 The use of high-purity reagents and solvents helps to
in 32 min. Data presented in this test method were obtained
minimize interference problems. 16
using this column.
7.5.3 Column 2 (Alternative Column), 250 mm long by
6.2 Interfering contamination may occur when a sample
containing low concentrations of analytes is analyzed imme- 4.6-mm inside diameter, stainless steel, packed with 5-µm
silica beads coated with trimethylsilyl. Mobile phase is
diatelyafterasamplecontainingrelativelyhighconcentrations
of analytes.Apreventive technique is between-sample rinsing established at 1.0 mL/min as a linear gradient from 15:85
methanol: water to 100% methanol in 32 min.
of the sample syringe and filter holder with two portions of
water. Analyze one or more laboratory method blanks after 7.5.4 Column 3 (Alternative Column, used for EPA 531.2
validation), 150 mm long by 3.9 mm inside diameter, stainless
analysis of a sample containing high concentrations of ana-
lytes.
Samplebottlevial,PierceNo.13075,availablefromPierceChemicalCo.,3747
6.3 Matrix interference may be caused by contaminants
N. Meridian Rd., Rockford, IL 61101, or equivalent.
present in the sample. The extent of matrix interference will 8
Samplebottlecap,PierceNo.12722,availablefromPierceChemicalCo.,3747
vary considerably from source to source, depending upon the N. Meridian Rd., Rockford, IL 61101, or equivalent.
Millipore Type HA, 0.45 µm for water, and Millipore Type FH, 0.5µ m for
water sampled. Positive analyte identifications must be con-
organics, available from Millipore Corp., 80 Ashby Rd., Bedford, MA 01730, or
firmed using the alternative conformational columns, or LC/
equivalent.
MS.
MilliporestainlesssteelXX300/200,availablefromMilliporeCorp.,80Ashby
Rd., Bedford, MA 01730, or equivalent.
6.4 The quality of the re
...

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