ASTM D7597-09

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
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Designation: D7597 – 09
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. A number 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 MPA in 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) (referred to collectively as organophosphonates in this
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-
analytes are qualitatively and quantitatively determined by this
priate safety and health practices and determine the applica-
method. This method adheres to single reaction monitoring
bility of regulatory limitations prior to use.
(SRM) mass spectrometry.
1.2 This test method has been developed in support of the
2. Referenced Documents
National Homeland Security Research Center, US EPA by
2.1 ASTM Standards:
Region 5 Chicago Regional Laboratory.
D1129 Terminology Relating to Water
1.3 The values stated in SI units are to be regarded as
D1193 Specification for Reagent Water
standard. No other units of measurement are included in this
D2777 Practice for Determination of Precision and Bias of
standard.
Applicable Test Methods of Committee D19 on Water
1.4 The detection verification level (DVL) and reporting
D3856 Guide for Good Laboratory Practices in Laborato-
range for the organophosphonates are listed in Table 1.
ries Engaged in Sampling and Analysis of Water
1.4.1 The DVL is required to be at a concentration at least
D3694 Practices for Preparation of Sample Containers and
three times below the reporting limit (RL) and have a signal/
for Preservation of Organic Constituents
noise ratio greater than 3:1. Fig. 1 displays the signal/noise
D5847 Practice for Writing Quality Control Specifications
ratios at the DVLs for the organophosphonates in the ESI
for Standard Test Methods for Water Analysis
positive mode and Fig. 2 in the ESI negative mode.
E2554 Practice for Estimating and Monitoring the Uncer-
1.4.2 The reporting limit is the concentration of the Level 1
tainty of Test Results of a Test Method in a Single
calibration standard as shown in Table 2 for the organophos-
Laboratory Using a Control Sample Program
phonates except for MPAin the ESI negative mode which is at
2.2 Other Documents:
Level 2 due to not meeting the DVL criteria at the lower
This test method is under the jurisdiction of ASTM Committee D19 on Water
andisthedirectresponsibilityofSubcommitteeD19.06onMethodsforAnalysisfor For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Organic Substances in Water. contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Current edition approved Dec. 1, 2009. Published January 2010. DOI: 10.1520/ Standards volume information, refer to the standard’s Document Summary page on
D7597-09. the ASTM website.
Copyright ©ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA19428-2959, United States.
D7597 – 09
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. 1 Example ESI Positive Mode SRM Chromatograms Signal/Noise Ratios
EPApublication SW-846 Test Methods for Evaluating Solid 3.1.3 organophosphonates, n—inthistestmethod,diisopro-
Waste, Physical/Chemical Methods pyl methylphosphonate (DIMP), ethyl hydrogen dimethylami-
dophosphate (EHDMAP), ethyl methylphosphonic acid
3. Terminology (EMPA), isopropyl methylphosphonic acid (IMPA), meth-
ylphosphonic acid (MPA) and pinacolyl methylphosphonic
3.1 Definitions:
acid (PMPA).
3.1.1 detection verification level (DVL), n—a concentration
3.2 Abbreviations:
that has a signal/noise ratio greater than 3:1 and is at least 3
3.2.1 ND—non-detect
times below the reporting limit (RL).
3.1.2 reporting limit (RL), n—the concentration of the
4. Summary of Test Methods
lowest-level calibration standard used for quantification.
4.1 This is a performance-based method and modifications
are allowed to improve performance.
4.2 For organophosphonate analysis, samples are shipped to
Available from United States Environmental Protection Agency (EPA), Ariel
the lab between 0°C and 6°C and analyzed within 1 day of
Rios Bldg., 1200 Pennsylvania Ave., NW, Washington, DC 20460, http://
www.epa.gov. collection. In the lab, the samples are spiked with surrogate,
D7597 – 09
FIG. 2 Example 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
filtered using a syringe-driven Millex HV PVDF filter unit and ous and were used as chemical weapon agents. Organophos-
phate pesticides are usually not persistent in the environ-
analyzed directly by LC/MS/MS.
,
4 5
ment.
4.3 Theorganophosphonatesandthesurrogates;diisopropyl
5.2 This test method is for the analysis of selected
methylphosphonate-D , pinacolyl methylphosphonic acid-
organophosphorous-based chemical weapon agent degradation
C and methylphosphonic acid-D are identified by retention
6 3
products from Sarin (GB), Soman (GD), Tabun (GA) and VX.
time and one SRM transition. The target analytes and surro-
This method has been investigated for use with reagent and
gates are quantitated using the SRM transitions utilizing an
surface water.
external calibration. The final report issued for each sample
lists the concentration of each organophosphonate target com-
6. Interferences
pound and each surrogate recovery.
6.1 Method interferences may be caused by contaminants in
solvents, reagents, glassware and other apparatus producing
5. Significance and Use
discrete artifacts or elevated baselines. All of these materials
5.1 Organophosphate pesticides affect the nervous system
by disrupting the enzyme that regulates acetylcholine, a neu-
Additional information about organophosphate pesticides is available on the
rotransmitter. They were developed during the early 19th
Internet at http://www.epa.gov (2009).
century, but their effects on insects, which are similar to their 5
Additional information about chemical weapon agents is available on the
effects on humans, were discovered in 1932. Some are poison- Internet at http://www.opcw.org (2009).
D7597 – 09
TABLE 3 Multi-Laboratory Recovery Data in Reagent Water
Bias Precision
ESI Spike Conc.
Mean Min Max Overall SD Pooled Overall Pooled
Analyte # Results # Labs
Mode (ppb)
Recovery Recovery Recovery (%) within-lab RSD (%) within-lab
(%) (%) (%) 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
are demonstrated to be free from interferences by analyzing 7. Apparatus
laboratory reagent blanks under the same conditions as
7.1 LC/MS/MS System
samples.
7.1.1 Liquid Chromatography (LC) System—A complete
6.2 All glassware is washed in hot water with a detergent,
LC system is needed in order to analyze samples. A system
rinsed in hot water followed by distilled water. The glassware
that is capable of performing at the flows, pressures, controlled
is then dried and heated in an oven at 250°C for 15 to 30
temperatures, sample volumes and requirements of the stan-
minutes. All glassware is subsequently cleaned with acetone,
dard may be used.
then methanol.
7.1.2 Analytical Column-Waters—Atlantis dC18, 150 mm
6.3 All reagents and solvents should be pesticide residue
3 2.1 mm, 3 µm particle size, or equivalent.
purity or higher to minimize interference problems.
6.4 Matrix interferences may be caused by contaminants
that are co-extracted from the sample. The extent of matrix
AWatersAlliance High Performance Liquid Chromatography (HPLC) System
interferences can vary considerably from sample source de-
was used to develop this test method. The multi-laboratory study included Agilent
pending on variations of the sample matrix. and Waters LC systems.
D7597 – 09
7.1.3 Tandem Mass Spectrometer (MS/MS) System—A 8.14.1 DIMP-D14 represents deuterium labeled diisopropyl
MS/MS system capable of MRM analysis. A system that is methylphosphonate where the two isopropyl moieties contain
capable of performing at the requirements in this standard may all H.
be used. 8.15 Methylphosphonic acid-D (MPA-D3, Unlabeled CAS
# 993-13-5).
7.2 Filtration Device
8.15.1 MPA-D3 represents deuterium labeled methylphos-
7.2.1 Hypodermic syringe—Alocktipglasssyringecapable
phonic acid where the methyl moiety contains all H.
of holding a Millex HV Syringe Driven Filter Unit PVDF 0.45
µm (Millipore Corporation, Catalog # SLHV033NS) or similar 8.16 Pinacolyl methylphosphonic acid- C (PMPA-13C6,
Unlabeled CAS # 616-52-4).
may be used.
8.16.1 PMPA-13C6 represents C labeled pinacolyl meth-
7.2.1.1 A25-mLlock tip glass syringe size is recommended
ylphosphonic where all the trimethylpropyl carbon atoms are
since a 25-mL sample size is used in this test method.
uniformly labeled C.
7.2.2 Filter—Millex HV Syringe Driven Filter Unit PVDF
0.45 µm (Millipore Corporation, Catalog # SLHV033NS) or
9. Hazards
similar may be used.
9.1 Normal laboratory safety applies to this method. Ana-
lysts should wear safety glasses, gloves, and lab coats when
8. Reagents and Materials
working in the lab.Analysts should review the Material Safety
8.1 Purity of Reagents—High-performance liquid chroma-
Data Sheets (MSDS) for all reagents used in this method.
tography (HPLC) pesticide residue analysis and spectropho-
tometry grade chemicals shall be used in all tests. Unless
10. Sampling
indicated otherwise, it is intended that all reagents shall
10.1 Sampling—Grab samples must be collected in
conform to the Committee on Analytical Reagents of the
$25-mL pre-cleaned amber glass bottles with Teflon-lined
American Chemical Society. Other reagent grades may be
caps demonstrated to be free of interferenc
...