ASTM D7485-09

Standard Test Method for Determination of Nonylphenol, p-<span class="bdit">tert</span>-Octylphenol, Nonylphenol Monoethoxylate and Nonylphenol Diethoxylate in Environmental Waters by Liquid Chromatography/Tandem Mass Spectrometry

SIGNIFICANCE AND USE
NP and OP have been shown to have toxic effects in aquatic organisms. The source of NP and OP is prominently from the use of common commercial surfactants. The most widely used surfactant is nonylphenol ethoxylate (NPEO) which has an average ethoxylate chain length of nine. The ethoxylate chain is readily biodegraded to form NP1EO, NP2EO, nonylphenol carboxylate (NPEC) and, under anaerobic conditions, NP. NP will also biodegrade, but may be released into environmental waters directly at trace levels. This method has been investigated and is applicable for environmental waters, including seawater.
SCOPE
1.1 This test method covers the determination of nonylphenol (NP), nonylphenol ethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), and octylphenol (OP), extracted from water utilizing solid phase extraction (SPE), separated using liquid chromatography (LC) and detected with tandem mass spectrometry (MS/MS). These compounds are qualitatively and quantitatively determined by this method. This method adheres to single reaction monitoring (SRM) mass spectrometry.
1.2 The method detection limit (MDL) and reporting limit (RL) for NP, NP1EO, NP2EO, and OP are listed in Table 1.
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 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 Limits  AnalyteMDLA
(ng/L)Reporting RangeB
(ng/L) NP33100-2000 NP1EO9100-2000 NP2EO9100-2000 OP24100-2000
A MDL Determined Following The Code of Federal Regulations, 40 CFR Part 136, Appendix B.
B Lowest Point of the Reporting Range is Calculated from the LV 1 Concentration Calibration Standard in Table 4.

General Information

• Status: Historical
• Publication Date: 28-Feb-2009
• ICS: 13.060.50 - Examination of water for chemical substances
• Technical Committee: D19 - Water
• Drafting Committee: D19.06 - Methods for Analysis for Organic Substances in Water

Relations

• Referred By: ASTM F2931-19a - Standard Guide for Analytical Testing of Substances of Very High Concern in Materials and Products
• Effective Date: 01-Mar-2009

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: D7485 − 09
StandardTest Method for
Determination of Nonylphenol, p-tert-Octylphenol,
Nonylphenol Monoethoxylate and Nonylphenol Diethoxylate
in Environmental Waters by Liquid Chromatography/Tandem
Mass Spectrometry
This standard is issued under the fixed designation D7485; 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 D3856Guide for Management Systems in Laboratories
Engaged in Analysis of Water
1.1 This test method covers the determination of nonylphe-
D5847Practice for Writing Quality Control Specifications
nol (NP), nonylphenol ethoxylate (NP1EO), nonylphenol di-
for Standard Test Methods for Water Analysis
ethoxylate (NP2EO), and octylphenol (OP), extracted from
D5905Practice for the Preparation of SubstituteWastewater
water utilizing solid phase extraction (SPE), separated using
2.2 Other Documents
liquid chromatography (LC) and detected with tandem mass
The Code of Federal Regulations40 CFR Part 136,Appen-
spectrometry (MS/MS). These compounds are qualitatively
dix B
and quantitatively determined by this method. This method
adheres to single reaction monitoring (SRM) mass spectrom-
3. Terminology
etry.
3.1 Nonylphenol (NP) is a mixture of branched
1.2 The method detection limit (MDL) and reporting limit
p-nonylphenol isomers. Commercial NP is produced by the
(RL) for NP, NP1EO, NP2EO, and OP are listed in Table 1.
reaction of phenol with commercial nonene. Commercial
1.3 The values stated in SI units are to be regarded as
nonene is not simply a linear C H alpha olefin; it is a
9 18
standard. No other units of measurement are included in this
complex mixture of predominantly nine-carbon olefins, called
standard.
propylene trimer, containing no linear isomers. This synthesis
results in a mixture of various branched nonylphenol isomers
1.4 This standard does not purport to address all of the
rather than a discrete chemical structure. The branched nonyl
safety concerns, if any, associated with its use. It is the
group is positioned predominantly in the para position on the
responsibility of the user of this standard to establish appro-
phenol ring.
priate safety and health practices and determine the applica-
bility of regulatory limitations prior to use.
3.2 OP represents octylphenol. Commercial octylphenol is
produced by the reaction of phenol and diisobutylene to
2. Referenced Documents
produce predominantly the 4-(1,1,3,3-tetramethylbutyl)phenol
isomer.
2.1 ASTM Standards:
D1129Terminology Relating to Water
3.3 NP1EO represents branched nonylphenol monoethoxy-
D1193Specification for Reagent Water
late.
D2777Practice for Determination of Precision and Bias of
3.4 NP2EO represents branched nonylphenol diethoxylate.
Applicable Test Methods of Committee D19 on Water
3.5 n-NP2EO represents normal straight chain nonylphenol
D3694Practices for Preparation of Sample Containers and
for Preservation of Organic Constituents diethoxylate.n-NP2EOisusedinthismethodasasurrogate.It
is not produced commercially and is not expected to be found
in environmental waters.
This test method is under the jurisdiction ofASTM Committee D19 on Water
3.6 2-Bromo-4-(1,1,3,3-tetramethylbutyl)phenol (Br-OP) is
andisthedirectresponsibilityofSubcommitteeD19.06onMethodsforAnalysisfor
used in this method as a surrogate. It is not produced
Organic Substances in Water.
commerciallyandisnotexpectedtobefoundinenvironmental
Current edition approved March 1, 2009. Published March 2009. DOI: 10.1520/
D7485-09.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM AvailablefromU.S.GovernmentPrintingOfficeSuperintendentofDocuments,
Standards volume information, refer to the standard’s Document Summary page on 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
the ASTM website. www.access.gpo.gov.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D7485 − 09
TABLE 1 MDL and Reporting Limits
6.2 All glassware is scrupulously cleaned. All glassware is
A B
MDL Reporting Range washedinhotwaterwithdetergentsuchaspowderedAlconox,
Analyte
(ng/L) (ng/L)
Deto-Jet, Luminox, or Citrojet, rinsed in hot water and rinsed
NP 33 100-2000
with distilled water. The glassware is then dried and heated in
NP1EO 9 100-2000
an oven at 250°C for 15 to 30 minutes. All glassware is
NP2EO 9 100-2000
OP 24 100-2000
subsequently cleaned with acetone and methanol. Detergents
A
MDLDeterminedFollowingTheCodeofFederalRegulations,40CFRPart136, containing alkylphenolic compounds must not be used.
Appendix B.
B
6.3 All reagents and solvents should be of pesticide residue
Lowest Point of the Reporting Range is Calculated from the LV 1 Concentration
Calibration Standard in Table 4.
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
interferences can vary considerably from sample source to
waters. It was reported that compounds in highly chlorinated
sample source, depending on variations of the sample matrix.
bromide rich wastewaters could potentially interfere with the
Br-OP surrogate. If this interference is encountered
7. Apparatus
n-nonylphenol is suggested as an alternative surrogate.
7.1 LC/MS/MS System:
3.7 Units—parts per trillion (ng/L, ppt), parts per billion
7.1.1 Liquid Chromatography System—A complete Liquid
(µg/L, ppb), parts per million (mg/L, ppm)
Chromatographysystemisneededinordertoanalyzesamples.
3.8 Environmental water shall refer to water tested using
AWaters(registeredtrademark)ACQUITYUltraPerformance
this method. See Section 5.
LiquidChromatography(UPLC(trademark))Systemwasused
to develop this test method, but a different system that is
4. Summary of Test Method
capable of performing at the flows, pressures, controlled
4.1 This is a performance-based method and modifications temperatures, sample volumes and requirements of the stan-
are allowed to improve performance.
dard may be used.
7.1.2 Analytical Column—Waters—ACQUITY UPLC
4.2 For NP, NP1EO, NP2EO, and OP analysis, solid phase
(trademark) HSS T3, 1.8 µm, 2.1 × 50 mm column or
extraction is used to extract water samples.
equivalent.
4.2.1 Solid Phase Extraction—250 milliliter volume of
7.1.3 Tandem Mass Spectrometer (MS/MS) System—A
sample adjusted to pH 2 is extracted using a solid phase
MS/MS system capable of MRM analysis. A Waters Quattro
extraction cartridge. The acetonitrile/water extract is concen-
Premier XE mass spectrometer was used to develop this test
tratedtoavolumeof1.0mL,andthenanalyzedbyLC/MS/MS
method,butanothersystemthatiscapableofperformingatthe
operated in the multiple reaction monitoring (MRM) mode.
requirements in the standard may be used.
4.3 The target compounds are identified by retention time
7.2 SPE Vacuum Manifold System—Supelco Visiprep solid
and SRM transition and are quantitated using the SRM
phase extraction vacuum manifold or similar may be utilized.
transition of the target compounds utilizing external calibra-
tion. The final report issued for each sample lists the concen-
7.3 Organic solvent evaporation device.
tration of NP, NP1EO, NP2EO, and OP.
8. Reagents and Materials
5. Significance and Use
8.1 Purity of Reagents—High Performance Liquid Chroma-
5.1 NP and OP have been shown to have toxic effects in tography (HPLC) pesticide residue analysis and spectropho-
aquatic organisms. The source of NP and OP is prominently tometry grade chemicals shall be used in all tests. Unless
from the use of common commercial surfactants. The most indicated otherwise, it is intended that all reagents shall
widely used surfactant is nonylphenol ethoxylate (NPEO) conform to the specifications of the Committee on Analytical
which has an average ethoxylate chain length of nine. The Reagents of the American Chemical Society. Other reagent
gradesmaybeusedprovideditisfirstascertainedthattheyare
ethoxylate chain is readily biodegraded to form NP1EO,
NP2EO, nonylphenol carboxylate (NPEC) and, under anaero- of sufficiently high purity to permit their use without affecting
the accuracy of the measurement.
bic conditions, NP. NP will also biodegrade, but may be
releasedintoenvironmentalwatersdirectlyattracelevels.This
method has been investigated and is applicable for environ-
Reagent Chemicals, American Chemical Society Specifications, American
mental waters, including seawater.
Chemical Society, Washington, DC. For Suggestions on the testing of reagents not
listed by the American Chemical Society, see Annual Standards for Laboratory
6. Interferences
Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
6.1 Methodinterferencesmaybecausedbycontaminantsin
MD.
solvents, reagents, glassware and other apparatus producing
Twosourcesofthealkylphenolstandardsare:CambridgeIsotopeLaboratories,
50 Frontage Road, Andover, MA 01810-5413 and Accustandard, Inc., 125 Market
discrete artifacts or elevated baselines. All of these materials
Street, New Haven, CT 06513. If you are aware of alternative suppliers, please
are routinely demonstrated to be free from interferences by
provide this information toASTM International Headquarters.Your comments will
analyzing laboratory reagent blanks under the same conditions
receive careful consideration at a meeting of the responsible technical committee,
as the samples. which you may attend.
D7485 − 09
NOTE 2—Pre-cleaned bottles demonstrated to be free of interferences
8.2 Purity of Water—Unless otherwise indicated, references
may be used.
towatershallbeunderstoodtomeanreagentwaterconforming
toType I of Specification D1193. It must be demonstrated that 10.2 Preservation—Adjust sample to pH 2 with concen-
this water does not contain contaminants at concentrations trated HCl at time of collection. Store samples between 0ºC
sufficient to interfere with the analysis. and6ºCfromthetimeofcollectionuntilextraction.Extractthe
sample within 14 days of collection and completely analyze
8.3 Gases—Ultrapure nitrogen and argon.
within 14 days of extraction.
8.4 Acetonitrile (CAS # 75-05-8).
10.3 Sample extracts may be stored in sealed glass contain-
8.5 Methanol (CAS # 67-56-1).
ers at < 0ºC indefinitely.
8.6 Isopropanol (CAS # 67-63-0).
11. Preparation of LC/MS/MS
8.7 Acetone (CAS # 67-64-1).
11.1 LC Chromatograph Operating Conditions:
8.8 Branched nonylphenol monoethoxylate (NP1EO) avail-
11.1.1 Injectionsofallcalibrationstandardsandsamplesare
able as a high purity custom standard.
made at a 50 µLvolume using a full loop injection. If a 50 µL
8.9 Branched nonylphenol diethoxylate (NP2EO) available volume loop is installed in your LC, a “full loop” mode is the
as a high purity custom standard.
preferredtechniquewhenperformingfast,qualitativeanalyses.
Thismodeshouldbeusedwheneveraccuracyandprecisionare
8.10 Nonylphenol, NP, >95% para isomer (CAS # 84852-
the primary concerns. The first sample analyzed after the
15-3).
calibrationcurveisablanktoensurethereisnocarry-over.The
8.11 Octylphenol, OP, 99 + % 4-(1,1,3,3-
gradient conditions for the liquid chromatograph are shown in
tetramethylbutyl)phenol (CAS # 140-66-9).
Table 2.
8.12 Concentrated HCl (CAS # 7647-01-0).
11.2 LC Sample Manager Conditions :
8.13 AmmoniumAcetate (CAS # 631-61-8) (ACS Reagent 11.2.1 Wash Solvents—Weak wash is 1.2 mL of 95%
Grade or Better). water/5 % acetonitrile, Strong wash is 1 mL of 30 %
acetonitrile, 30% methanol, 30% isopropyl alcohol, 10%
8.14 n-Nonylphenol diethoxylate (n-NP2EO).
water. The strong wash solvent is needed to eliminate carry-
8.15 n-Nonylphenol (suggested alternate surrogate, if
over between injections of alkylphenol samples. The weak
needed).
wash is used to remove the strong wash solvent. Specific
instrument manufacturer specifications should be followed in
8.16 2-Bromo-4-(1,1,3,3-tetramethylbutyl)phenol (Br-OP).
order to eliminate sample carry-over in the analysis of alkyl-
8.17 Solid Phase Extraction Cartridges—Sep-Pak (regis-
phenols.
tered trademark) Vac (500 mg) tC18 Cartridges or equivalent.
11.2.2 Temperatures—Column, 30°C; Sample
NOTE 1—Alkylphenols have been found in SPE cartridges therefore it
compartment, 15°C.
is advisable that the cartridges be lot certified alkylphenol free. Glass
cartridges should have a much lower risk of alkylphenol contamination. 11.2.3 Seal Wash—5 minutes.
11.3 Mass Spectrometer Parameters :
9. Hazards
11.3.1 Your instrument may require different settings.
9.1 Normal laboratory safety applies to this method. Ana-
11.3.2 Variable parameters depending on analyte are shown
lysts should wear safety glasses, gloves and lab coats when
in Table 3.
workingwithacids.AnalystsshouldreviewtheMaterialSafety
Data Sheets (MSDS) for all reagents used in this method.
TABLE 2 Gradient Conditions for Liquid Chromatography
10. Sample Collection, Preservation, and Storage
Percent Percent
Time Flow
95 % CH CN/5 % Water 95 % Water/5 % CH CN
3 3
10.1 Sampling—Grab samples must be collected in 250 mL
(min) (µL/min)
2 mmolar NH OAc 2 mmolar NH OAc
4 4
amberglassbottles.Thismustbedoneinordertoallowforthe
0 300 0 100
rinsing of the bottle with acidified water and acidified 10%
1 300 0 100
methanol/water in order to get complete transfer of the sample
3 300 50 50
4 300 60 40
into the SPE cartridge and extraction process. Alkylphenols
6 300 70 30
tend to adsorb to glassware and rinsing will allow optimum
7 300 70 30
recoveries. Conventional sampling practices should be fol- 9 300 100 0
13 300 100 0
lowed. Refer to Guide D3856 and Practices D3694.Automatic
14 300 0 100
sampling equipment should be as free as possible of Tygon
16 300 0 100
tubing and other potential sources of contamination.
D7485 − 09
TABLE 3 Retention Times, MRM Ions, and Analyte-Specific Mass Spectrometer Parameters
SRM
Retention Time Cone Voltage Collision Energy
Analyte ESI Mode Mass Transition
(min) (Volts) (eV)
(Parent > Product)
Octylphenol neg 6.77 40 25 205.2 > 133
NP2EO pos 7.71 20 12 326.3 > 183.2
NP1EO pos 7.78 15 10 282.3 > 127.1
Nonylphenol neg 7.82 40 30 219.2 > 133
Br-OP (surrogate) neg 7.73 35 25 283.1 > 78.8
n-NP2EO (surrogate) pos 8.62 20 17 326.3 > 88.9
instrument. Fig. 1 displays a SRM chromatogram of each
The instrument is set in the Electrospray source setting.
Capillary Voltage: 3.5 kV
analyte and the numb
...