ASTM D7574-09
(Test Method)Standard Test Method for Determination of Bisphenol A in Environmental Waters by Liquid Chromatography/Tandem Mass Spectrometry
Standard Test Method for Determination of Bisphenol A in Environmental Waters by Liquid Chromatography/Tandem Mass Spectrometry
SIGNIFICANCE AND USE
The first reported synthesis of BPA was by the reaction of phenol with acetone by Zincke. BPA has become an important high volume industrial chemical used in the manufacture of polycarbonate plastic and epoxy resins. Polycarbonate plastic and resins are used in numerous products including electrical and electronic equipment, automobiles, sports and safety equipment, reusable food and drink containers, electrical laminates for printed circuit boards, composites, paints, adhesives, dental sealants, protective coatings and many other products.
The environmental source of BPA is predominantly from the decomposition of polycarbonate plastics and resins. BPA is not classified as bio-accumulative by the U.S. Environmental Protection Agency and will biodegrade. BPA has been reported to have adverse effects in aquatic organisms and may be released into environmental waters directly at trace levels through landfill leachate and POTW effluents. This method has been investigated for use with surface water and secondary and tertiary POTW effluent samples therefore, it is applicable to these matrices only. It has not been investigated for use with salt water or solid sample matrices.
SCOPE
1.1 This procedure covers the determination of bisphenol A (BPA) extracted from water utilizing solid phase extraction (SPE), separated using liquid chromatography (LC) and detected with tandem mass spectrometry (MS/MS). BPA is qualitatively and quantitatively determined by this method. This method adheres to multiple reaction monitoring (MRM) mass spectrometry.
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.3 The method detection limit (MDL) and reporting limit (RL) for BPA are listed in Table 1.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
TABLE 1 MDL and Reporting Range AnalyteMDLA (ng/L)Reporting RangeB (ng/L) Bisphenol A520–600
A MDL determined following the Code of Federal Regulations, 40CFR Part 136, Appendix B.
B Lowest point of the reporting range, reporting limit, is calculated from the LV 1 concentration calibration standard in Table 4. Fig. 1 displays the signal/noise ratio at the reporting limit.
General Information
Standards Content (Sample)
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: D7574 − 09
StandardTest Method for
Determination of Bisphenol A in Environmental Waters by
Liquid Chromatography/Tandem Mass Spectrometry
This standard is issued under the fixed designation D7574; 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 3.1.1 environmental water, n—effluents from secondary and
tertiary publicly owned treatment works (POTW), fresh sur-
1.1 This procedure covers the determination of bisphenolA
face and ground water.
(BPA) extracted from water utilizing solid phase extraction
(SPE), separated using liquid chromatography (LC) and de- 3.2 Abbreviations:
tected with tandem mass spectrometry (MS/MS). BPA is 3.2.1 ppt—parts per trillion, ng/L
qualitatively and quantitatively determined by this method.
4. Summary of Test Method
This method adheres to multiple reaction monitoring (MRM)
mass spectrometry. 4.1 This is a performance based method and modifications
are allowed to improve performance.
1.2 The values stated in SI units are to be regarded as
standard. No other units of measurement are included in this 4.2 Solid phase extraction is used to extract water samples.
standard. 4.2.1 Solid Phase Extraction—250 milliliter volume of
sample adjusted to pH 2 is extracted using a solid phase
1.3 The method detection limit (MDL) and reporting limit
extraction cartridge. The resulting methyl tert-butyl ether
(RL) for BPA are listed in Table 1.
(MTBE) extract is solvent exchanged into methanol, concen-
1.4 This standard does not purport to address all of the
tratedtoavolumeof0.2mL,broughttoafinalvolumeof1mL
safety concerns, if any, associated with its use. It is the
with water and then analyzed by LC/MS/MS operated in the
responsibility of the user of this standard to establish appro-
multiple reaction monitoring (MRM) mode.
priate safety and health practices and determine the applica-
4.3 The target compound, surrogate and internal standards
bility of regulatory limitations prior to use.
are identified by retention time and two SRM transitions. The
target analyte and surrogate are quantitated using the primary
2. Referenced Documents
SRM transitions utilizing internal standard calibration. The
2.1 ASTM Standards:
final report issued for each sample lists the concentration of
D1193Specification for Reagent Water
BPA and the bisphenol A (Ring-13C12) surrogate recovery.
D3694Practices for Preparation of Sample Containers and
for Preservation of Organic Constituents
5. Significance and Use
D3856Guide for Management Systems in Laboratories
5.1 The first reported synthesis of BPAwas by the reaction
Engaged in Analysis of Water
of phenol with acetone by Zincke. BPA has become an
D5847Practice for Writing Quality Control Specifications
important high volume industrial chemical used in the manu-
for Standard Test Methods for Water Analysis
facture of polycarbonate plastic and epoxy resins. Polycarbon-
D5905Practice for the Preparation of SubstituteWastewater
ate plastic and resins are used in numerous products including
electrical and electronic equipment, automobiles, sports and
3. Terminology
safetyequipment,reusablefoodanddrinkcontainers,electrical
3.1 Definitions:
laminates for printed circuit boards, composites, paints,
adhesives, dental sealants, protective coatings and many other
products.
This test method is under the jurisdiction ofASTM Committee D19 on Water
5.2 The environmental source of BPA is predominantly
andisthedirectresponsibilityofSubcommitteeD19.06onMethodsforAnalysisfor
Organic Substances in Water.
from the decomposition of polycarbonate plastics and resins.
Current edition approved Oct. 1, 2009. Published October 2009. DOI: 10.1520/
D7574-09.
2 3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or Zincke, T., 1905, “Mittheilungen aus dem chemischen Laboratorium der
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Universitat Marburg,” Justus Leibigs Annals Chemie, Vol. 343, pages 75–79.
Standards volume information, refer to the standard’s Document Summary page on Additional information about BPA is available at http://www.bisphenol-a.org
the ASTM website. (2008).
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D7574 − 09
TABLE 1 MDL and Reporting Range
7.1.1.2 Tandem Mass Spectrometer (MS/MS) System—A
A B
Analyte MDL (ng/L) Reporting Range (ng/L) MS/MS system capable of MRM analysis.
Bisphenol A 5 20–600
7.2 SPE Vacuum Manifold System—Supelco Visiprep solid
A
MDL determined following the Code of Federal Regulations, 40CFR Part 136,
phase extraction vacuum manifold or similar may be utilized.
Appendix B.
B
Lowest point of the reporting range, reporting limit, is calculated from the LV 1
7.3 Organic solvent evaporation device.
concentration calibration standard in Table 4. Fig. 1 displays the signal/noise ratio
at the reporting limit.
8. Reagents and Materials
8.1 Purity of Reagents—High Performance Liquid Chroma-
tography (HPLC) pesticide residue analysis and spectropho-
tometry grade chemicals shall be used in all tests. Unless
BPAis not classified as bio-accumulative by the U.S. Environ-
indicated otherwise, it is intended that all reagents shall
mental Protection Agency and will biodegrade. BPA has been
conform to the specifications of the Committee on Analytical
reported to have adverse effects in aquatic organisms and may
Reagents of the American Chemical Society. Other reagent
be released into environmental waters directly at trace levels
gradesmaybeused,provideditisfirstascertainedthattheyare
throughlandfillleachateandPOTWeffluents.Thismethodhas
of sufficiently high purity to permit their use without affecting
beeninvestigatedforusewithsurfacewaterandsecondaryand
theaccuracyofthemeasurement.AsourceforthelabeledBPA
tertiary POTW effluent samples therefore, it is applicable to
standards is Cambridge Isotope Laboratories, 50 Frontage
these matrices only. It has not been investigated for use with 8
Road, Andover, MA 01810-5413.
salt water or solid sample matrices.
8.2 Purity of Water—Unless otherwise indicated, references
towatershallbeunderstoodtomeanreagentwaterconforming
6. Interferences
toType I of Specification D1193. It must be demonstrated that
6.1 Methodinterferencesmaybecausedbycontaminantsin
this water does not contain contaminants at concentrations
solvents, reagents, glassware and other apparatus producing
sufficient to interfere with the analysis.
discrete artifacts or elevated baselines. The use of plastic
8.3 Gases—Ultrapure nitrogen and argon.
supplies and equipment must be avoided because they may
contain BPA.All of these materials are routinely demonstrated
8.4 Acetonitrile (CAS # 75-05-8).
to be free from interferences by analyzing laboratory reagent
8.5 Methanol (CAS # 67-56-1).
blanks under the same conditions as the samples.
8.6 2-Propanol (CAS # 67-63-0).
6.2 All glassware is washed in hot water with a detergent,
8.7 Acetone (CAS # 67-64-1).
rinsed in hot water followed by distilled water. The glassware
is then dried and heated in an oven at 250ºC for 15 to 30
8.8 Methyl tert-butyl ether (MTBE, CAS # 1634-04-4).
minutes. All glassware is subsequently cleaned with acetone,
8.9 Ammonium Hydroxide (CAS # 1336-21-6) (ACS re-
then methanol. Detergents in plastic containers that contain
agent grade or better).
BPA must not be used.
8.10 Concentrated HCl (CAS # 7647-01-0).
6.3 All reagents and solvents should be of pesticide residue
8.11 BisphenolA(BPA,2,2’-Bis(4-hydroxyphenyl)propane,
purity or higher to minimize interference problems.
CAS # 80-05-7).
6.4 Matrix interferences may be caused by contaminants
8.12 Bisphenol A (Ring-13C12) represents C labeled
that are co-extracted from the sample. The extent of matrix
bisphenol A where all ring carbon atoms are uniformly
interferences can vary considerably from sample source to
labeled C.
sample source, depending on variations of the sample matrix.
8.12.1 Bisphenol A (Ring-13C12) is used as a surrogate in
this standard.
7. Apparatus
8.13 Bisphenol A (Propane-D6) represents deuterium la-
7.1 LC/MS/MS System:
beled bisphenolAwhere the 2 methyl moieties contain all H.
7.1.1 Liquid Chromatography System—A complete liquid
chromatographysystemisneededinordertoanalyzesamples.
7.1.1.1 Columns—Column to separate instrument back-
ground (isolator): A short (2.1 × 50 mm) C8 or C18 column AWaters Quattro Premier XE mass spectrometer was used to develop this test
method. If you are aware of alternative suppliers, please provide this information to
stable at higher pH up to 12. Analytical column: ACQUITY
ASTM International Headquarters. Your comments will receive careful consider-
(trademarked) UPLC BEH C18, 1.7 µm, 2.1 × 50 mm column
ation at a meeting of the responsible technical committee, which you may attend.
or equivalent. 7
Reagent Chemicals, American Chemical Society Specifications, American
Chemical Society, Washington, DC. For Suggestions on the testing of reagents not
listed by the American Chemical Society, see Annual Standards for Laboratory
Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
A Waters ACQUITY Ultra Performance Liquid Chromatography ((trade- and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
marked) UPLC) System was used to develop this test method. If you are aware of MD.
alternative suppliers, please provide this information to ASTM International If you are aware of alternative suppliers, please provide this information to
Headquarters.Your comments will receive careful consideration at a meeting of the ASTM International Headquarters. Your comments will receive careful consider-
1 1
responsible technical committee, which you may attend. ation at a meeting of the responsible technical committee, which you may attend.
D7574 − 09
8.13.1 Bisphenol A (Propane-D6) is used as an internal theriskofhighbiaseddata.TheLCconditionsinthisstandard
standard in this method. allow for the BPAin the system to elute 0.4 minutes later than
BPA from the sample injection. The BPA concentration found
8.14 Solid Phase Extraction Cartridges—Oasis HLB 5cc
in the LC system used was below the MDLof the standard but
(200 mg) LP Glass Cartridges or equivalent.
anisolatorcolumnmustbeincorporatedtoeliminatetheriskof
NOTE 1—If plastic cartridges are used, BPAmay be found, therefore it
highbias.Anisolatorcolumnwasplacedafterthemixeroftwo
is advisable that the cartridges be lot certified BPA-free. BPAmay adhere
solvent feeds and before port 5 of the multi-port valve which
to plastic cartridges which will cause lower recoveries. Glass cartridges
contains the injection loop. No plastics are used in the system
haveamuchloweradhesivetendencytoBPAandshouldnotcontainBPA
atthispointensuringthatthefirstelutedBPApeakisonlyfrom
in the packing or support materials, therefore glass cartridges should be
used.
the BPAin the sample and not the LC system. This placement
allowsfortheimpuritiesintheLC,suchasBPA,tobetrapped
9. Hazards
bytheisolatorcolumnandeluteaftertheBPAfromthesample.
9.1 Normal laboratory safety applies to this method. Ana-
If a different LC system is used, consult with the instrument
lysts should wear safety glasses, gloves and lab coats when
manufacturer for the proper placement of the isolator column
workingwithacids.AnalystsshouldreviewtheMaterialSafety
for optimum results.
Data Sheets (MSDS) for all reagents used in this method.
11.2 LC Analytical Column—ACQUITY (trademarked)
UPLC BEH C18, 1.7 µm, 2.1 × 50 mm column or equivalent.
10. Sample Collection, Preservation, and Storage
11.3 LC Operating Conditions—Injections of all calibration
10.1 Sampling—Grab samples must be collected in 250 mL
standardsandsamplesaremadeata50µLvolumeusingafull
amber glass bottles. Rinsing of the bottle with 10% methanol
loop injection. If a 50 µLvolume loop is installed in your LC,
in water, water, and 10% methanol/2% NH OH in water is
a “full loop” mode is the preferred technique when performing
required in order to get quantitative transfer of the sample into
fast,quantitativeanalyses.Thismodeshouldbeusedwhenever
the SPE cartridge and extraction process. BPAtends to adsorb
accuracy and precision are the primary concerns. Specific
to surfaces and rinsing will allow optimum recoveries. Con-
instrument manufacturer specifications should be followed to
ventional sampling practices should be followed. Refer to
achieve maximum performance. The first sample analyzed
Guide D3856 and Practices D3694. Pre-cleaned bottles dem-
after the calibration curve is a blank to ensure there is no
onstrated to be free of interferences may be used. Automatic
carry-over. The gradient conditions for the liquid chromato-
sampling equipment should be free from plastics and tubing
graph are shown in Table 2.
thatcontainsBPAandotherpotentialsourcesofcontamination
or adhesion. 11.4 LC Sample Manager Conditions —Wash Solvents:
Weak wash is 1.2 mL of 5% methanol in water. Strong wash
10.2 Preservation—Adjust sample to pH 2 with concen-
is 1 mL of 30% acetonitrile/30% methanol/30% 2-propa-
trated HCl at time of collection. Store samples between 0ºC
nol⁄10%water.Thestrongwashsolventisneededtoeliminate
and6ºCfromthetimeofcollectionuntilextraction.Extractthe
carry-over between injections. The weak wash is used to
sample within 7 days of collection and completely analyze
remove the strong wash solvent. Specific instrument manufac-
within 14 days of extraction.
turer specifications should be followed in order to eliminate
11. Preparation of LC/MS/MS sample carry-over in the analysis of BPA. Temperatures:
Column 40°C, Sample compartment 15°C. Seal Wash: 5
11.1 LC Set Up for Liquid Chromatography BPA Isolator
minutes.
Column for Low Level BPA Analysis—BPAmay be a contami-
nantintheLCsystemduetothewidespreaduseofplasticparts 11.5 Mass Spectrometer Parameters : Your instrument may
and tubing. Incorporating an isolator column into the LC
require different settings. Variable parameters depending on
system allows for the low level analysis of BPA and reduces analyte are shown in Table 3.
TABLE 2 Gradient Conditions for Liquid Chromatography
Time Flow Percent Water/5 Percent Methanol/5
(Minutes) (µL/Minute) mmolar mmolar
NH OH NH OH
4 4
0 300 100 0
1 300 100 0
2 250 75 25
5 250 25 75
6 250 25 75
7 300 0 100
9 300 0 100
10 300 100 0
12 300 100 0
D7574 − 09
TABLE 3 Retention Times, SRM transitions, SRM Ratios and Anal
...
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