ASTM D7597-16(2017)
(Test Method)Standard Test Method for Determination of Diisopropyl Methylphosphonate, Ethyl Hydrogen Dimethylamidophosphate, Ethyl Methylphosphonic Acid, Isopropyl Methylphosphonic Acid, Methylphosphonic Acid and Pinacolyl Methylphosphonic Acid in Water by Liquid Chromatography/Tandem Mass Spectrometry
Standard Test Method for Determination of Diisopropyl Methylphosphonate, Ethyl Hydrogen Dimethylamidophosphate, Ethyl Methylphosphonic Acid, Isopropyl Methylphosphonic Acid, Methylphosphonic Acid and Pinacolyl Methylphosphonic Acid in Water by Liquid Chromatography/Tandem Mass Spectrometry
SIGNIFICANCE AND USE
5.1 Organophosphate pesticides affect the nervous system by disrupting the enzyme that regulates acetylcholine, a neurotransmitter. They were developed during the early 19th century, but their effects on insects, which are similar to their effects on humans, were discovered in 1932. Some are poisonous and were used as chemical weapon agents. Organophosphate pesticides are usually not persistent in the environment.4,5
5.2 This test method is for the analysis of selected organophosphorous-based chemical weapon agent degradation products from Sarin (GB), Soman (GD), Tabun (GA) and VX. This test method has been investigated for use with reagent and surface water.
SCOPE
1.1 This procedure covers the determination of diisopropyl methylphosphonate (DIMP), ethyl hydrogen dimethylamidophosphate (EHDMAP), ethyl methylphosphonic acid (EMPA), isopropyl methylphosphonic acid (IMPA), methylphosphonic acid (MPA) and pinacolyl methylphosphonic acid (PMPA) (referred to collectively as organophosphonates in this test method) in surface water by direct injection using liquid chromatography (LC) and detected with tandem mass spectrometry (MS/MS) using electrospray ionization (ESI). These analytes are qualitatively and quantitatively determined by this test method. This test method adheres to single reaction monitoring (SRM) mass spectrometry.
1.2 This test method has been developed by U.S. EPA Region 5 Chicago Regional Laboratory (CRL).
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.4 The detection verification level (DVL) and reporting range for the organophosphonates are listed in Table 1.
TABLE 1 Detection Verification Level and Reporting Range
Analyte
ESI Mode
DVL (μg/L)
Reporting Range (μg/L)
Diisopropyl methylphosphonate
Positive
1
5–150
Ethyl hydrogen dimethylamidophosphate
Negative
0.25
5–150
Ethyl hydrogen dimethylamidophosphate
Positive
0.25
5–150
Ethyl methylphosphonic acid
Negative
5
50–1500
Ethyl methylphosphonic acid
Positive
5
50–1500
Isopropyl methylphosphonic acid
Negative
10
50–1500
Isopropyl Methylphosphonic acid
Positive
5
50–1500
Methylphosphonic acid
Negative
20
100–1500
Methylphosphonic acid
Positive
10
50–1500
Pinacolyl methylphosphonic acid
Negative
5
50–1500
Pinacolyl methylphosphonic acid
Positive
5
50–1500
1.4.1 The DVL is required to be at a concentration at least three times below the reporting limit (RL) and have a signal/noise ratio greater than 3:1. Fig. 1 displays the signal/noise ratios at the DVLs for the organophosphonates in the ESI positive mode and Fig. 2 in the ESI negative mode.
FIG. 1 Example ESI Positive Mode SRM Chromatograms Signal/Noise Ratios
FIG. 2 Example ESI Negative Mode SRM Chromatograms Signal/Noise Ratios
1.4.2 The reporting limit is the concentration of the Level 1 calibration standard as shown in Table 2 for the organophosphonates except for MPA in the ESI negative mode which is at Level 2 due to not meeting the DVL criteria at the lower concentration level. The DVL for MPA in the ESI negative mode is at 20 μg/L, which forces a raised reporting limit. However, the multi-laboratory validation required a spike of all target analytes at Level 1 concentrations. The mean recovery for MPA in the ESI negative mode at this level was 98.7 % as shown in Table 3. If your instrument’s sensitivity can meet the requirements in this test method, MPA may have a 50 μg/L reporting limit.
TABLE 2 Concentrations of Calibration Standards (PPB)
Analyte/Surrogate
LV 1
LV 2
LV 3
LV 4
LV 5
LV 6
LV 7
Diisopropyl methylphosphonate
5
10
20
35
50
100
150
Ethyl hydrogen dimethylamidophosphate
5
10
20
35
50
100
150
Ethyl methylphosphonic acid ...
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This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation:D7597 −16 (Reapproved 2017)
Standard Test Method for
Determination of Diisopropyl Methylphosphonate, Ethyl
Hydrogen Dimethylamidophosphate, Ethyl
Methylphosphonic Acid, Isopropyl Methylphosphonic Acid,
Methylphosphonic Acid and Pinacolyl Methylphosphonic
Acid in Water by Liquid Chromatography/Tandem Mass
Spectrometry
This standard is issued under the fixed designation D7597; 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 concentration level. The DVL for MPA in the ESI negative
mode is at 20 µg/L, which forces a raised reporting limit.
1.1 This procedure covers the determination of diisopropyl
However,themulti-laboratoryvalidationrequiredaspikeofall
methylphosphonate (DIMP), ethyl hydrogen dimethylami-
target analytes at Level 1 concentrations. The mean recovery
dophosphate (EHDMAP), ethyl methylphosphonic acid
for MPAin the ESI negative mode at this level was 98.7% as
(EMPA), isopropyl methylphosphonic acid (IMPA), methyl-
shown in Table 3. If your instrument’s sensitivity can meet the
phosphonic acid (MPA) and pinacolyl methylphosphonic acid
requirements in this test method, MPA may have a 50 µg/L
(PMPA)(referredtocollectivelyasorganophosphonatesinthis
reporting limit.
test method) in surface water by direct injection using liquid
1.5 This standard does not purport to address all of the
chromatography (LC) and detected with tandem mass spec-
safety concerns, if any, associated with its use. It is the
trometry (MS/MS) using electrospray ionization (ESI). These
responsibility of the user of this standard to establish appro-
analytesarequalitativelyandquantitativelydeterminedbythis
priate safety, health, and environmental practices and deter-
test method. This test method adheres to single reaction
mine the applicability of regulatory limitations prior to use.
monitoring (SRM) mass spectrometry.
1.6 This international standard was developed in accor-
1.2 This test method has been developed by U.S. EPA
dance with internationally recognized principles on standard-
Region 5 Chicago Regional Laboratory (CRL).
ization established in the Decision on Principles for the
1.3 The values stated in SI units are to be regarded as
Development of International Standards, Guides and Recom-
standard. No other units of measurement are included in this
mendations issued by the World Trade Organization Technical
standard.
Barriers to Trade (TBT) Committee.
1.4 The detection verification level (DVL) and reporting
2. Referenced Documents
range for the organophosphonates are listed in Table 1.
1.4.1 The DVL is required to be at a concentration at least 2.1 ASTM Standards:
three times below the reporting limit (RL) and have a signal/ D1129Terminology Relating to Water
noise ratio greater than 3:1. Fig. 1 displays the signal/noise D1193Specification for Reagent Water
ratios at the DVLs for the organophosphonates in the ESI D2777Practice for Determination of Precision and Bias of
positive mode and Fig. 2 in the ESI negative mode. Applicable Test Methods of Committee D19 on Water
1.4.2 The reporting limit is the concentration of the Level 1 D3856Guide for Management Systems in Laboratories
calibration standard as shown in Table 2 for the organophos- Engaged in Analysis of Water
phonates except for MPAin the ESI negative mode which is at D3694Practices for Preparation of Sample Containers and
Level 2 due to not meeting the DVL criteria at the lower for Preservation of Organic Constituents
D5847Practice for Writing Quality Control Specifications
for Standard Test Methods for Water Analysis
This test method is under the jurisdiction ofASTM Committee D19 on Water
andisthedirectresponsibilityofSubcommitteeD19.06onMethodsforAnalysisfor
Organic Substances in Water. For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Current edition approved June 15, 2017. Published July 2017. Originally contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
approved in 2009. Last previous edition approved in 2016 as D7597 – 16. DOI: Standards volume information, refer to the standard’s Document Summary page on
10.1520/D7597-16R17. the ASTM website.
Copyright ©ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA19428-2959. United States
D7597−16 (2017)
TABLE 1 Detection Verification Level and Reporting Range
Analyte ESI Mode DVL (µg/L) Reporting Range (µg/L)
Diisopropyl methylphosphonate Positive 1 5–150
Ethyl hydrogen dimethylamidophosphate Negative 0.25 5–150
Ethyl hydrogen dimethylamidophosphate Positive 0.25 5–150
Ethyl methylphosphonic acid Negative 5 50–1500
Ethyl methylphosphonic acid Positive 5 50–1500
Isopropyl methylphosphonic acid Negative 10 50–1500
Isopropyl Methylphosphonic acid Positive 5 50–1500
Methylphosphonic acid Negative 20 100–1500
Methylphosphonic acid Positive 10 50–1500
Pinacolyl methylphosphonic acid Negative 5 50–1500
Pinacolyl methylphosphonic acid Positive 5 50–1500
FIG. 1Example ESI Positive Mode SRM Chromatograms Signal/Noise Ratios
E2554Practice for Estimating and Monitoring the Uncer- 3.2 Definitions of Terms Specific to This Standard:
tainty of Test Results of a Test Method Using Control
3.2.1 detection verification level, DVL, n—a concentration
Chart Techniques
that has a signal/noise ratio greater than 3:1 and is at least 3
2.2 Other Documents:
times below the reporting limit (RL).
EPAPublication SW-846Test Methods for Evaluating Solid
3.2.2 independent reference material, IRM, n—amaterialof
Waste, Physical/Chemical Methods
known purity and concentration obtained either from the
3. Terminology
NationalInstituteofStandardsandTechnology(NIST)orother
reputable supplier. The IRM shall be obtained from a different
3.1 Definitions:
lot of material than is used for calibration.
3.1.1 For definitions of terms used in this standard, refer to
Terminology D1129.
3.2.3 organophosphonates, n—inthistestmethod,diisopro-
pyl methylphosphonate (DIMP), ethyl hydrogen dimethylami-
dophosphate (EHDMAP), ethyl methylphosphonic acid
(EMPA), isopropyl methylphosphonic acid (IMPA), methyl-
Available from United States Environmental Protection Agency (EPA), Ariel
phosphonic acid (MPA) and pinacolyl methylphosphonic acid
Rios Bldg., 1200 Pennsylvania Ave., NW, Washington, DC 20460, http://
www.epa.gov. (PMPA).
D7597−16 (2017)
FIG. 2Example ESI Negative Mode SRM Chromatograms Signal/Noise Ratios
TABLE 2 Concentrations of Calibration Standards (PPB)
Analyte/Surrogate LV 1 LV 2 LV 3 LV 4 LV 5 LV 6 LV 7
Diisopropyl methylphosphonate 5 10 20 35 50 100 150
Ethyl hydrogen dimethylamidophosphate 5 10 20 35 50 100 150
Ethyl methylphosphonic acid 50 100 200 350 500 1000 1500
Isopropyl methylphosphonic acid 50 100 200 350 500 1000 1500
Methylphosphonic acid 50 100 200 350 500 1000 1500
Pinacolyl methylphosphonic acid 50 100 200 350 500 1000 1500
DIMP-D14 (Surrogate) 5 10 20 35 50 100 150
PMPA-13C4 (Surrogate) 25 50 100 175 250 500 750
MPA-D3 (Surrogate) 25 50 100 175 250 500 750
3.3 Acronyms: 3.3.16 QC, adj—Quality Control
3.3.1 CCC, n—Continuing Calibration Check
3.3.17 RL, n—Reporting Limit
3.3.2 IC, n—Initial Calibration
3.3.18 RSD, n—Relative Standard Deviation
3.3.3 LC, n—Liquid Chromatography
3.3.19 RT, n—Retention Time
3.3.4 LCS/LCSD, n—Laboratory Control Sample/
3.3.20 SDS, n—Safety Data Sheets
Laboratory Control Sample Duplicate
3.3.5 MDL, n—Method Detection Limit 3.3.21 SRM, n—Single Reaction Monitoring
3.3.6 MeOH, n—Methanol
3.3.22 SS, n—Surrogate Standard
–3
3.3.7 mM, n—millimolar,1×10 moles/L
3.3.23 TC, n—Target Compound
3.3.8 MRM, n—Multiple Reaction Monitoring –6
3.3.24 µM, n—micromolar,1×10 moles/L
3.3.9 MS/MSD, n—Matrix Spike/Matrix Spike Duplicate
3.3.25 VOA, n—Volatile Organic Analysis
3.3.10 NA, adj—Not Available
3.3.11 ND, n—non-detect
4. Summary of Test Method
3.3.12 P&A, n—Precision and Accuracy
4.1 This is a performance-based test method and modifica-
3.3.13 PPB, n—parts per billion tions are allowed to improve performance.
3.3.14 PPT, n—parts per trillion
4.2 Fororganophosphonateanalysis,samplesareshippedto
3.3.15 QA, adj—Quality Assurance the lab between 0°C and 6°C and analyzed within 1 day of
D7597−16 (2017)
TABLE 3 Multi-Laboratory Recovery Data in Reagent Water
Bias Precision
Spike
ESI #
Mean Min Max Pooled Pooled
Analyte Conc. # Labs
Overall SD Overall
Mode Results
Recovery Recovery Recovery within-lab within-lab
(ppb)
(%) RSD (%)
(%) (%) (%) SD (%) RSD (%)
DIMP Pos 5 16 4 95.1 65.8 136.0 17.6 19.4 21.3 22.4
DIMP Pos 10 19 5 98.2 80.0 121.0 6.1 5.5 13.6 13.8
DIMP Pos 25 26 6 102.9 74.4 128.0 5.6 5.7 14.5 14.1
DIMP Pos 125 22 5 96.6 80.4 120.0 4.4 4.5 11.0 11.4
DIMP-D14 Pos 25 86 6 102.6 54.8 127.6 9.8 9.5 11.2 10.9
EHDMAP Neg 5 12 3 57.5 0.0 220.0 31.4 22.3 71.1 123.6
EHDMAP Neg 10 16 4 47.1 0.0 178.0 13.3 10.8 66.8 142.0
EHDMAP Neg 25 22 5 84.1 54.0 141.2 6.9 6.6 22.9 27.3
EHDMAP Neg 125 18 4 87.4 64.2 141.6 5.0 5.1 25.4 29.1
EHDMAP Pos 5 16 4 77.0 0.0 134.2 7.4 8.0 51.4 66.8
EHDMAP Pos 10 20 5 70.0 0.0 143.0 7.7 10.0 53.6 76.5
EHDMAP Pos 25 26 6 89.6 60.0 128.8 5.2 6.2 23.5 26.2
EHDMAP Pos 125 22 5 87.5 59.0 123.2 5.2 6.0 23.9 27.3
EMPA Neg 50 16 4 110.1 74.8 170.6 22.4 17.2 25.6 23.3
EMPA Neg 100 20 5 108.3 87.7 175.0 11.1 8.7 24.8 22.9
EMPA Neg 250 26 6 104.8 82.0 122.6 6.1 5.5 11.8 11.3
EMPA Neg 1250 22 5 101.5 87.2 126.4 8.4 8.1 11.2 11.0
EMPA Pos 50 16 4 95.4 77.6 122.8 12.9 13.3 13.1 13.7
EMPA Pos 100 20 5 96.0 61.4 132.5 9.5 9.5 15.9 16.6
EMPA Pos 250 26 6 99.7 70.0 133.2 5.9 5.4 18.2 18.2
EMPA Pos 1250 21 5 93.9 84.0 108.4 2.7 3.0 7.7 8.2
IMPA Neg 50 16 4 88.0 56.6 140.4 23.7 26.0 23.5 26.7
IMPA Neg 100 20 5 88.0 68.5 118.0 12.9 14.3 13.4 15.2
IMPA Neg 250 26 6 98.1 72.8 144.0 13.1 11.9 19.2 19.6
IMPA Neg 1250 21 5 90.7 73.1 103.0 5.5 6.1 8.7 9.6
IMPA Pos 50 16 4 98.3 47.8 139.6 19.2 20.5 27.2 27.7
IMPA Pos 100 19 5 95.4 72.3 120.5 9.8 10.3 12.4 13.0
IMPA Pos 250 26 6 97.0 79.2 188.4 7.4 7.6 10.9 11.2
IMPA Pos 1250 21 5 91.3 70.4 115.5 5.1 5.2 11.4 12.5
MPA Neg 50 16 4 98.7 3.3 175.0 14.2 25.3 60.5 61.3
MPA Neg 100 20 5 100.0 41.9 142.0 8.9 9.2 30.8 30.8
MPA Neg 250 26 6 99.5 66.0 124.5 7.6 7.6 14.5 14.5
MPA Neg 1250 22 5 102.7 81.8 130.5 10.5 9.9 12.4 12.1
MPA Pos 50 16 4 68.3 9.8 139.6 13.4 20.3 36.6 53.6
MPA Pos 100 20 5 80.5 48.4 149.7 14.0 12.6 26.8 33.3
MPA Pos 250 26 6 91.7 33.9 153.7 8.0 7.8 31.8 34.7
MPA Pos 1250 22 5 95.8 31.8 208.2 12.6 8.3 43.4 45.3
MPA-D3 Neg 250 84 6 111.2 57.2 190.8 16.2 12.5 30.0 26.9
MPA-D3 Pos 250 68 5 104.4 58.4 151.8 14.3 14.0 18.0 17.3
PMPA Neg 50 15 4 87.8 77.6 124.4 8.4 8.2 13.8 15.7
PMPA Neg 100 19 5 91.6 83.6 98.2 2.8 3.0 4.4 4.8
PMPA Neg 250 26 6 101.0 77.2 123.8 5.0 4.8 12.1 12.0
PMPA Neg 1250 22 5 99.2 84.8 126.5 4.9 4.8 12.3 12.4
PMPA Pos 50 16 4 90.8 60.8 148.8 15.8 16.1 25.6 28.2
PMPA Pos 100 15 4 95.2 86.8 114.0 3.8 4.0 7.9 8.3
PMPA Pos 250 20 5 103.8 85.2 136.1 4.5 3.8 15.3 14.7
PMPA Pos 1250 12 3 99.8 88.8 117.5 5.2 5.0 7.5 7.5
PMPA-13C6 Neg 250 83 6 99.5 74.8 128.3 9.4 9.2 11.1 11.1
collection. In the lab, the samples are spiked with surrogate, lists the concentration of each organophosphonate target com-
filtered using a syringe-driven filter unit and analyzed directly pound and each surrogate recovery.
by LC/MS/MS.
5. Significance and Use
4.3 Theorganophosphonatesandthesurrogates;diisopropyl
methylphosphonate-D , pinacolyl methylphosphonic acid-
5.1 Organophosphate pesticides affect the nervous system
C and methylphosphonic acid-D are identified by retention
6 3
by disrupting the enzyme that regulates acetylcholine, a neu-
time and one SRM transition. The target analytes and surro-
rotransmitter. They were developed during the early 19th
gates are quantitated using the SRM transitions utilizing an
century, but their effects on insects, which are similar to their
external calibration. The final report issued for each sample
D7597−16 (2017)
effects on humans, were discovered in 1932. Some are poison- 7.1.3 Tandem Mass Spectrometer (MS/MS) System—A
ous and were used as chemical weapon agents. Organophos- MS/MS system capable of MRM analysis. A system that is
capableofperformingattherequirementsinthisstandardmay
phate pesticides are usually not persistent in the
4,5
be used.
environment.
7.2 Filtration Device:
5.2 This test method is for the analysis of selected
7.2.1 Hypodermic Syringe—A luer-lock tip glass syringe
organophosphorous-based chemical weapon agent degradation
capable of holding syringe-driven filter unit.
products from Sarin (GB), Soman (GD), Tabun (GA) and VX.
7.2.1.1 A25-mLlocktipglasssyringesizeisrecommended
Thistestmethodhasbeeninvestigatedforusewithreagentand
since a 25-mL sample size is used in this test method.
surface water.
7.2.2 Filter Unit —A PVDF filter units were used to filter
the samples.
6. Interferences
8. Reagents and Materials
6.1 Test method interferences may be caused by contami-
8.1 Purity of Reagents—High-performance liquid chroma-
nants in solvents, reagents, glassware and other apparatus
tography (HPLC) pesticide residue analysis and spectropho-
producing discrete artifacts or elevated baselines. All of these
tometry grade chemicals shall be used in all tests. Unless
materials are demonstrated to be free from interferences by
indicated otherwise, it is intended that all reagents shall
analyzing laboratory reagent blanks under the same conditions
conform to the Committee on Analytical Reagents of the
as samples.
American Chemical Society. Other reagent grades may be
6.2 All glassware is washed in hot water with a detergent,
used provided they are first determined they are of sufficiently
rinsed in hot water followed by distilled water. The glassware
highpuritytopermittheirusewithoutaffectingtheaccuracyof
is then dried and heated in an oven at 250°C for 15 to 30
the measurements.
minutes. All glassware is subsequently cleaned with acetone,
8.2 Purity of Water—Unless otherwise indicated, references
then methanol.
towatershallbeunderstoodtomeanreagentwaterconforming
6.3 All reagents and solvents should be pesticide residue toType1ofSpecificationD1193.Itmustbedemonstratedthat
this water does not contain contaminants at concentrations
purity or higher to minimize interference problems.
sufficient to interfere with the analysis.
6.4 Matrix interferences may b
...
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