ASTM D7742-17

Standard Practice for Determination of Nonylphenol Polyethoxylates (NPnEO, 3 ≤ n ≤ 18) and Octylphenol Polyethoxylates (OPnEO, 2 ≤ n ≤ 12) in Water by Single Reaction Monitoring (SRM) Liquid Chromatography/ Tandem Mass Spectrometry (LC/MS/MS)

SIGNIFICANCE AND USE
5.1 This practice has been developed in support of the U.S. EPA Office of Water, Office of Science and Technology by the Chicago Regional Laboratory (CRL).  
5.2 Nonylphenol (NP) and Octylphenol (OP) have been shown to have toxic effects in aquatic organisms. The prominent source of NP and OP is from common commercial surfactants which are longer chain APEOs. The most widely used surfactant is nonylphenol polyethoxylate (NPnEO) which has an average ethoxylate chain length of nine. The APEOs are readily biodegraded to form NP1EO, NP2EO, nonylphenol carboxylate (NPEC) and NP. NP will also biodegrade, but may be released into environmental waters directly at trace levels. This practice screens for the longer chain APEOs which may enter the STP at elevated levels and may cause a STP to violate its permitted discharge concentration of nonylphenol.
SCOPE
1.1 This practice covers the determination of nonylphenol polyethoxylates (NPnEO, 3 ≤ n ≤ 18) and octylphenol polyethoxylates (OPnEO, 2 ≤ n ≤ 12) in water by Single Reaction Monitoring (SRM) Liquid Chromatography/ Tandem Mass Spectrometry (LC/MS/MS) using direct injection liquid chromatography (LC) and detected with tandem mass spectrometry (MS/MS) detection. This is a screening practice with qualified quantitative data to check for the presence of longer chain ethoxylates in a water sample.  
1.1.1 All data are qualified because neat standards of each alkylphenol ethoxylate (APEO) are not available and the synthesis and characterization of these neat standards would be very expensive. The Igepal2 brand standards, which contain a mixture of various chain lengths of the alkylphenol ethoxylates (APEOs), were used. The mixture was characterized in-house assuming the instrument response at an optimum electrospray ionization cone and collision voltage for each APEO was the same. This assumption, which may not be accurate, is used to determine qualified amounts of each ethoxylate in the standards. The n-Nonylphenol diethoxylate (n-NP2EO) surrogate was available as a neat characterized standard, therefore, this concentration and recovery data was not estimated. APEOs are not regulated by the EPA, but nonylphenol, a breakdown product of NPnEOs, is regulated for fresh and saltwater dischargers. A request by a sewage treatment plant (STP) was made to make this practice available through ASTM in order to screen for the influent or effluent from sources of APEOs coming into the STP. The interest lies in stopping the source of the longer chain APEOs from entering the STP in order to meet effluent guidelines. Based upon the above, this is a practice rather than a test method. A comparison between samples is possible using this practice to determine which has a higher concentration of APEOs.  
1.2 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this practice.  
1.3 The estimated screening range shown in Table 1 was calculated from the concentration of the Level 1 and 7 calibration standards shown in Table 4. These numbers are qualified, as explained in Section 1, and must be reported as such. Figs. 1-5 show the SRM chromatograms of each analyte at the Level 1 concentration with the signal to noise (S/N) ratio. This is a screening practice and method detection limits are not given. The S/N ratio for each analyte at the Level 1 concentration must be at least 5:1 for adequate sensitivity. If the instrument can not meet the criteria, the screening limit must be raised to an acceptable level.  
FIG. 1 SRM Chromatograms NP3EO-NP8EO  
FIG. 2 SRM Chromatograms NP9EO-NP14EO  
FIG. 3 SRM Chromatograms NP15EO-NP18EO and n-NP2EO  
FIG. 4 SRM Chromatograms OP2EO-OP7EO  
FIG. 5 SRM Chromatograms OP8EO-OP12EO  
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,...

General Information

• Status: Published
• Publication Date: 14-Dec-2017
• ICS: 71.100.40 - Surface active agents
• Technical Committee: D19 - Water
• Drafting Committee: D19.06 - Methods for Analysis for Organic Substances in Water

Relations

• Replaces: ASTM D7742-11 - Standard Practice for Determination of Nonylphenol Polyethoxylates (NPnEO, 3 ≤ n ≤ 18) and Octylphenol Polyethoxylates (OPnEO, 2 ≤ n ≤ n ≤ 12 in Water by Single Reaction Monitoring (SRM) Liquid Chromatography/ Tandem Mass Spe
• Effective Date: 15-Dec-2017
• Refers: ASTM D3694-96(2024) - Standard Practices for Preparation of Sample Containers and for Preservation of Organic Constituents
• Effective Date: 01-Apr-2024
• Refers: ASTM D1129-13(2020)e2 - Standard Terminology Relating to Water
• Effective Date: 01-May-2020
• Refers: ASTM E2554-18e1 - Standard Practice for Estimating and Monitoring the Uncertainty of Test Results of a Test Method Using Control Chart Techniques
• Effective Date: 01-Apr-2018
• Refers: ASTM E2554-18 - Standard Practice for Estimating and Monitoring the Uncertainty of Test Results of a Test Method Using Control Chart Techniques
• Effective Date: 01-Apr-2018
• Refers: ASTM E2554-13 - Standard Practice for Estimating and Monitoring the Uncertainty of Test Results of a Test Method Using Control Chart Techniques
• Effective Date: 01-Apr-2013
• Refers: ASTM D2777-12 - Standard Practice for Determination of Precision and Bias of Applicable Test Methods of Committee D19 on Water
• Effective Date: 15-Jun-2012
• Refers: ASTM D3856-11 - Standard Guide for Management Systems in Laboratories Engaged in Analysis of Water
• Effective Date: 15-Nov-2011
• Refers: ASTM D3694-96(2011) - Standard Practices for Preparation of Sample Containers and for Preservation of Organic Constituents
• Effective Date: 01-May-2011
• Refers: ASTM D1129-10 - Standard Terminology Relating to Water
• Effective Date: 01-Mar-2010
• Refers: ASTM D2777-08 - Standard Practice for Determination of Precision and Bias of Applicable Test Methods of Committee D19 on Water
• Effective Date: 15-Jan-2008
• Refers: ASTM E2554-07 - Standard Practice for Estimating and Monitoring the Uncertainty of Test Results of a Test Method in a Single Laboratory Using a Control Sample Program
• Effective Date: 01-May-2007
• Refers: ASTM D1129-06a - Standard Terminology Relating to Water
• Effective Date: 01-Sep-2006
• Refers: ASTM D1129-06ae1 - Standard Terminology Relating to Water
• Effective Date: 01-Sep-2006
• Refers: ASTM D2777-06 - Standard Practice for Determination of Precision and Bias of Applicable Test Methods of Committee D19 on Water
• Effective Date: 15-Aug-2006

Overview

ASTM D7742-17 is the Standard Practice for the Determination of Nonylphenol Polyethoxylates (NPnEO, 3 ≤ n ≤ 18) and Octylphenol Polyethoxylates (OPnEO, 2 ≤ n ≤ 12) in Water by Single Reaction Monitoring (SRM) Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS). Developed under the jurisdiction of ASTM Committee D19 on Water, this screening practice plays a crucial role in supporting environmental monitoring programs, especially in assessing the presence of alkylphenol ethoxylates (APEOs) in water.

The primary application of this standard is to detect and compare longer chain APEOs in both influent and effluent waters from sources such as sewage treatment plants (STPs). This helps facilities maintain compliance with discharge guidelines, particularly regarding the regulation of nonylphenol, a toxic breakdown product of NPnEOs, by the U.S. EPA.

Key Topics

• Target Compounds: The practice focusses on nonylphenol polyethoxylates (NPnEOs, chain length 3–18) and octylphenol polyethoxylates (OPnEOs, chain length 2–12)-both common breakdown products of commercial surfactants that can have toxic effects on aquatic organisms.
• Screening Practice: Instead of an exact quantification, ASTM D7742-17 offers a screening approach, qualifying relative levels of APEOs in samples, as synthesis of pure standards for all chain lengths is impractical.
• Analytical Technique: Utilizes LC/MS/MS with Single Reaction Monitoring (SRM) for sensitive detection of target analytes.
• Sample Handling: Water samples should be preserved with formaldehyde and analyzed within seven days. PVDF filters are preferred for sample filtration to maximize APEO recovery.
• Calibration and Quality Control: The standard outlines recommended calibration curve protocols, blank analysis, surrogate spiking, laboratory control samples (LCS), and matrix spike procedures to ensure data quality.
• Reporting: Results are reported as qualified concentrations due to inherent limitations in available standards and matrix effects.

Applications

• Environmental Compliance: Sewage treatment plants and industrial facilities use this standard to monitor APEO levels in water to avoid exceeding allowable nonylphenol concentrations in effluents, as regulated for both freshwater and saltwater dischargers in the United States.
• Screening Water Sources: Useful for screening both influent and effluent water for the presence of longer chain alkylphenol ethoxylates, allowing for prioritization of source identification and mitigation actions.
• Comparative Analysis: Enables identification of water samples with relatively higher concentrations of APEOs, guiding targeted interventions and source control initiatives.
• Supporting Regulatory Initiatives: Developed with input from the EPA Office of Water and the Chicago Regional Laboratory, this standard supports federal initiatives on water quality and contaminant reduction.

Related Standards

The effective implementation of ASTM D7742-17 may require reference to related ASTM and EPA standards, such as:

• ASTM D1129 – Terminology Relating to Water
• ASTM D1193 – Specification for Reagent Water
• ASTM D2777 – Practice for Precision and Bias Determination
• ASTM D3694 – Sample Container Preparation and Organic Constituent Preservation
• ASTM D5847 – Quality Control Specifications for Water Testing
• ASTM E2554 – Estimating and Monitoring Uncertainty of Test Results
• EPA SW-846 – Test Methods for Evaluating Solid Waste, Physical/Chemical Methods

Practical Value

By offering a structured, performance-based approach to screening for nonylphenol and octylphenol ethoxylates, ASTM D7742-17 helps organizations safeguard water resources against harmful surfactant residues, comply with regulatory standards, and proactively address emerging contaminants in the environment. Its adoption supports reliable comparison between samples and enhances overall water quality monitoring programs.

Keywords: ASTM D7742-17, nonylphenol polyethoxylates, octylphenol polyethoxylates, NPnEO, OPnEO, LC/MS/MS, water analysis, environmental compliance, APEO screening, alkylphenol ethoxylates, U.S. EPA, standard practice.

Standards Content (Sample)

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: D7742 − 17
Standard Practice for
Determination of Nonylphenol Polyethoxylates (NPnEO, 3#
n# 18) and Octylphenol Polyethoxylates (OPnEO, 2# n#
12) in Water by Single Reaction Monitoring (SRM) Liquid
Chromatography/ Tandem Mass Spectrometry (LC/MS/MS)
This standard is issued under the fixed designation D7742; 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 possible using this practice to determine which has a higher
concentration of APEOs.
1.1 This practice covers the determination of nonylphenol
polyethoxylates (NPnEO, 3 ≤ n ≤ 18) and octylphenol poly- 1.2 Units—The values stated in SI units are to be regarded
ethoxylates (OPnEO, 2 ≤ n ≤ 12) in water by Single Reaction asstandard.Nootherunitsofmeasurementareincludedinthis
Monitoring (SRM) Liquid Chromatography/ Tandem Mass practice.
Spectrometry (LC/MS/MS) using direct injection liquid chro-
1.3 The estimated screening range shown in Table 1 was
matography(LC)anddetectedwithtandemmassspectrometry
calculated from the concentration of the Level 1 and 7
(MS/MS) detection. This is a screening practice with qualified
calibration standards shown in Table 4. These numbers are
quantitative data to check for the presence of longer chain
qualified, as explained in Section 1, and must be reported as
ethoxylates in a water sample.
such. Figs. 1-5 show the SRM chromatograms of each analyte
1.1.1 All data are qualified because neat standards of each
attheLevel1concentrationwiththesignaltonoise(S/N)ratio.
alkylphenol ethoxylate (APEO) are not available and the
Thisisascreeningpracticeandmethoddetectionlimitsarenot
synthesisandcharacterizationoftheseneatstandardswouldbe
given. The S/N ratio for each analyte at the Level 1 concen-
very expensive. The Igepal brand standards, which contain a
tration must be at least 5:1 for adequate sensitivity. If the
mixtureofvariouschainlengthsofthealkylphenolethoxylates
instrument can not meet the criteria, the screening limit must
(APEOs), were used. The mixture was characterized in-house
be raised to an acceptable level.
assuming the instrument response at an optimum electrospray
1.4 This standard does not purport to address all of the
ionization cone and collision voltage for each APEO was the
safety concerns, if any, associated with its use. It is the
same. This assumption, which may not be accurate, is used to
responsibility of the user of this standard to establish appro-
determine qualified amounts of each ethoxylate in the stan-
priate safety, health, and environmental practices and deter-
dards. The n-Nonylphenol diethoxylate (n-NP2EO) surrogate
mine the applicability of regulatory limitations prior to use.
was available as a neat characterized standard, therefore, this
1.5 This international standard was developed in accor-
concentration and recoverydatawasnotestimated.APEOsare
dance with internationally recognized principles on standard-
not regulated by the EPA, but nonylphenol, a breakdown
ization established in the Decision on Principles for the
product of NPnEOs, is regulated for fresh and saltwater
Development of International Standards, Guides and Recom-
dischargers.Arequest by a sewage treatment plant (STP) was
mendations issued by the World Trade Organization Technical
madetomakethispracticeavailablethroughASTMinorderto
Barriers to Trade (TBT) Committee.
screen for the influent or effluent from sources of APEOs
comingintotheSTP.Theinterestliesinstoppingthesourceof
2. Referenced Documents
thelongerchainAPEOsfromenteringtheSTPinordertomeet
2.1 ASTM Standards:
effluent guidelines. Based upon the above, this is a practice
D1129Terminology Relating to Water
rather than a test method. A comparison between samples is
D1193Specification for Reagent Water
D2777Practice for Determination of Precision and Bias of
Applicable Test Methods of Committee D19 on Water
This practice is under the jurisdiction ofASTM Committee D19 on Water and
is the direct responsibility of Subcommittee D19.06 on Methods for Analysis for
Organic Substances in Water.
Current edition approved Dec. 15, 2017. Published January 2018. Originally For referenced ASTM standards, visit the ASTM website, www.astm.org, or
approved in 2011. Last previous edition approved in 2011 as D7742 – 11. DOI: contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
10.1520/D7742-17. Standards volume information, refer to the standard’s Document Summary page on
Igepal is a trademark of Rhodia Operations, Aubervilliers, CA. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D7742 − 17
TABLE 1 Estimated Screening Range
2.2 Other Standard:
Analyte Estimated Screening EPAPublication SW-846Test Methods for Evaluating Solid
Range (µg/L)
Waste, Physical/Chemical Methods
Nonylphenol 0.73–11.6
triethoxylate (NP3EO)
3. Terminology
Nonylphenol 1.1–18.3
tetraethoxylate (NP4EO)
3.1 Definitions:
Nonylphenol 1.4–22.1
3.1.1 For definitions of terms used in this standard, refer to
pentaethoxylate (NP5EO)
Nonylphenol 1.8–28.2
Terminology D1129.
hexaethoxylate (NP6EO)
Nonylphenol 1.9–30.1 3.2 Definitions of Terms Specific to This Standard:
heptaethoxylate (NP7EO)
3.2.1 alkylphenol ethoxylates, n—in this practice,nonylphe-
Nonylphenol 1.8–29.2
nol polyethoxylates (NPnEO, 3 ≤ n ≤ 18) and octylphenol
octaethoxylate (NP8EO)
Nonylphenol 1.6–26.3 polyethoxylates (OPnEO, 2 ≤ n ≤ 12) collectively.
nonaethoxylate (NP9EO)
3.2.2 screening limit, SL, n—the estimated concentration of
Nonylphenol 1.5–24.1
decaethoxylate (NP10EO) the lowest-level calibration standard used for quantification
Nonylphenol 1.3–21.3
accounting for the sample dilution.
undecaethoxylate (NP11EO)
Nonylphenol 1.0–15.7
3.3 Abbreviations:
dodecaethoxylate (NP12EO) –3
3.3.1 mM—millimolar,1×10 moles/L
Nonylphenol 0.64–10.3
tridecaethoxylate (NP13EO)
3.3.2 ND—non-detect
Nonylphenol 0.41–6.5
3.3.3 ppt—parts per trillion, ng/L
tetradecaethoxylate (NP14EO)
Nonylphenol 0.21–3.4
pendecaethoxylate (NP15EO)
4. Summary of Practice
Nonylphenol 0.11–1.7
hexadecaethoxylate (NP16EO) 4.1 This is a performance-based practice and modifications
Nonylphenol 0.05–0.80
are allowed to improve performance.
heptadecaethoxylate (NP17EO)
Nonylphenol 0.023–0.4
4.2 For APEOs analysis, samples are shipped to the lab
octodecaethoxylate (NP18EO)
between 0°C and 6°C containing 1 % formaldehyde and
Total NPnEO 16–250
analyzed within 7 days of collection. In the lab, an aliquot of
Octylphenol 0.14–2.3
diethoxylate (OP2EO)
the sample is filtered, spiked with surrogate, and analyzed
Octylphenol 1.4–22.2
directly by LC/MS/MS.
triethoxylate (OP3EO)
Octylphenol 2.2–35.2 4.2.1 Field samples from sewage systems propose a chal-
tetraethoxylate (OP4EO)
lenging analysis. Since this is a screening technique to deter-
Octylphenol 2.9–45.8
mine ifAPEOs are present, a 10–25 mL aliquot of the sample
pentaethoxylate (OP5EO)
Octylphenol 2.6–41.9 is filtered through a PVDF syringe driven filter unit before
hexaethoxylate (OP6EO)
spiking with surrogate. It was demonstrated that similar recov-
Octylphenol 2.5–40.4
eries of the APEOs are achieved filtered and unfiltered using
heptaethoxylate (OP7EO)
Octylphenol 1.8–28.8
PVDF filters. Filtering using PTFE filters produced much
octaethoxylate (OP8EO)
lowerrecoveries.ThispracticedoesnotaccountfortheAPEOs
Octylphenol 1.1–17.6
adhered to particulates or the sample bottle.
nonaethoxylate (OP9EO)
Octylphenol 0.62–9.9
4.3 Nonylphenol polyethoxylates (NPnEO, 3 ≤ n ≤ 18),
decaethoxylate (OP10EO)
Octylphenol 0.26–4.2 octylphenol polyethoxylates (OPnEO, 2 ≤ n ≤ 12), and
undecaethoxylate (OP11EO)
n-nonylphenol diethoxylate (n-NP2EO, surrogate) are identi-
Octylphenol 0.11–1.8
fied by retention time and one SRM transition. The target
dodecaethoxylate (OP12EO)
analytes and surrogates are quantitated using the SRM transi-
Total OPnEO 16–250
n-Nonylphenol 15.6–250 (Not Estimated)
tion by external calibration. The final report issued for each
diethoxylate (n-NP2EO)
sample lists their qualified concentration and the surrogate
recovery.
5. Significance and Use
5.1 This practice has been developed in support of the U.S.
D3856Guide for Management Systems in Laboratories
EPAOffice of Water, Office of Science and Technology by the
Engaged in Analysis of Water
Chicago Regional Laboratory (CRL).
D3694Practices for Preparation of Sample Containers and
5.2 Nonylphenol (NP) and Octylphenol (OP) have been
for Preservation of Organic Constituents
shown to have toxic effects in aquatic organisms. The promi-
D5847Practice for Writing Quality Control Specifications
nent source of NP and OP is from common commercial
for Standard Test Methods for Water Analysis
E2554Practice for Estimating and Monitoring the Uncer-
tainty of Test Results of a Test Method Using Control
Available from NationalTechnical Information Service (NTIS), 5301 Shawnee
Chart Techniques Rd., Alexandria, VA 22312, http://www.ntis.gov.
D7742 − 17
FIG. 1 SRM Chromatograms NP3EO-NP8EO
surfactants which are longer chain APEOs. The most widely Allglasswareissubsequentlycleanedwithacetoneandmetha-
used surfactant is nonylphenol polyethoxylate (NPnEO) which nol. Detergents containing alkylphenolic compounds must not
hasanaverageethoxylatechainlengthofnine.TheAPEOsare be used.
readily biodegraded to form NP1EO, NP2EO, nonylphenol
6.3 All reagents and solvents should be of pesticide residue
carboxylate (NPEC) and NP. NPwill also biodegrade, but may
purity or higher to minimize interference problems.
be released into environmental waters directly at trace levels.
6.4 Matrix interferences may be caused by contaminants
This practice screens for the longer chain APEOs which may
that are co-extracted from the sample. The extent of matrix
entertheSTPatelevatedlevelsandmaycauseaSTPtoviolate
interferences can vary considerably from sample source to
its permitted discharge concentration of nonylphenol.
sample source, depending on variations of the sample matrix.
6. Interferences
7. Apparatus
6.1 Practiceinterferencesmaybecausedbycontaminantsin
7.1 LC/MS/MS System:
solvents, reagents, glassware and other apparatus producing
7.1.1 Liquid Chromatography System—Acomplete LC sys-
discrete artifacts or elevated baselines. All of these materials
tem is needed in order to analyze samples. Any system that is
are routinely demonstrated to be free from interferences by
capable of performing at the flows, pressures, controlled
analyzing laboratory reagent blanks under the same conditions
temperatures, sample volumes, and requirements of the stan-
as the samples.
dard may be used.
6.2 Allglasswareiswashedinhotwaterwithdetergentsuch
7.1.2 Analytical Column—WatersAtlantis dC18, 2.1 × 150
as powderedAlconox, Det-o-Jet, Luminox, or Citrajet, rinsed
mm, 3 µm particle size was used to develop this practice.Any
in hot water, and rinsed with distilled water. The glassware is
thendriedandheatedinanovenat250ºCfor15to30minutes.
Waters ACQUITY H-Class Ultra Performance Liquid Chromatography
(UPLC) System, a trademark of the Waters Corporation, Milford, MA, was used to
develop this practice. All parameters in this practice are based on this system and
Alconox, Det-o-Jet, Luminox, and Citrajet are trademarks of Alconox, Inc., may vary depending on your instrument.
White Plains, NY. WatersAtlantis dC18 is a trademark of the Waters Corporation, Milford, MA.
D7742 − 17
FIG. 2 SRM Chromatograms NP9EO-NP14EO
column that achieves adequate resolution may be used. The 8. Reagents and Materials
retention times and order of elution may change depending on
8.1 Purity of Reagents—High Performance Liquid Chroma-
the column used and need to be monitored.
tography (HPLC) pesticide residue analysis and spectropho-
7.1.3 Tandem Mass Spectrometer System—A MS/MS sys-
tometry grade chemicals shall be used in all tests. Unless
tem capable of MRM analysis. Any system that is capable of
indicated otherwise, it is intended that all reagents shall
performing at the requirements in this practice may be used.
conform to the specifications of the Committee on Analytical
7.2 Filtration Device: Reagents of the American Chemical Society. Other reagent
7.2.1 Hypodermic syringe—Alock-tipglasssyringecapable gradesmaybeusedprovideditisfirstascertainedthattheyare
of holding a Millex HVSyringe Driven Filter Unit PVDF 0.45 of sufficiently high purity to permit their use without affecting
9,10
µm, or similar, may be used. the accuracy of the measurement.
7.2.1.1 A25–mLlock-tipglasssyringesizeisrecommended
8.2 Purity of Water—Unless indicated, references to water
for this practice.
shall be understood to mean reagent water conforming toType
7.2.2 Filter—Millex HV Syringe Driven Filter Unit PVDF
I of Specification D1193. It must be demonstrated that this
0.45 µm was used to develop this practice, any similar filter
water does not contain contaminants at concentrations suffi-
may be used.
cient to interfere with the analysis.
8.3 Gases—Ultrapure nitrogen and argon.
8 8.4 Acetonitrile (CAS # 75-05-8).
AWaters Quattro Micro tandem quadrupole mass spectrometer, a trademark of
the Waters Corporation, Milford, MA., was used to develop this practice. All
8.5 Methanol (CAS # 67-56-1).
parametersinthispracticearebasedonthissystemandmayvarydependingonyour
instrument.
The sole source of supply of the Millex HV Syringe Driven Filter Unit PVDF
0.45 µm known to the committee at this time is Millipore Corporation, Catalog # Reagent Chemicals, American Chemical Society Specifications, American
SLHV033NS. If you are aware of alternative suppliers, please provide this Chemical Society, Washington, DC. For suggestions on the testing of reagents not
information to ASTM International Headquarters. Your comments will receive listed by the American Chemical Society, see Analar Standards for Laboratory
careful consideration at a meeting of the responsible technical committee, which Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
you may attend. and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
Millex is a trademark of Merck KGAA, Darmstadt, Germany. MD.
D7742 − 17
FIG. 3 SRM Chromatograms NP15EO-NP18EO and n-NP2EO
8.6 2-Propanol (CAS # 67-63-0). strated to be free of interferences, larger sample sizes may be
used since a subsample aliquot is only required. All samples
8.7 Acetone (CAS # 67-64-1).
arepreservedwith1%concentrationofformaldehyde,shipped
8.8 Nonylphenol pentaethoxylate mixture (several NPnEO
between 0°C and 6°C, and stored in the laboratory between
isomer groups with an average of NP5EO, Igepal CO-520).
0°C and 6°C. Conventional sampling practices should be
followed. Refer to Guide D3856 and Practices D3694. Auto-
8.9 Nonylphenol nonaethoxylate mixture (several NPnEO
matic sampling equipment should be as free as possible of
isomer groups with an average of NP9EO, Igepal CO-630).
Tygon tubing and other potential sources of contamination or
8.10 Octylphenoldiethoxylatemixture(severalOPnEOiso-
causeadhesionofAPEOs.Analyzethesamplewithin7daysof
mer groups with an average of OP2EO, Igepal CA-210).
collection.
8.11 Octylphenol pentaethoxylate mixture (several OPnEO
isomer groups with an average of OP5EO, Igepal CA-520).
11. Preparation of LC/MS/MS
8.12 Formaldehyde (CAS # 50-00-0, 37 wt. % solution in 6
11.1 LC Chromatograph Operating Conditions:
water).
11.1.1 Injection volumes of all calibration standards and
8.13 Ammonium Acetate (CAS # 631-61-8). samplesaremadeat100-µLvolume.Thefirstsampleanalyzed
after the calibration curve is a blank to ensure there is no
8.14 n-Nonylphenol diethoxylate (n-NP2EO).
carry-over. The gradient conditions for the liquid chromato-
graph are shown in Table 2.
9. Hazards
9.1 Normal laboratory safety applies to this practice. Ana- 11.2 LC Sample Manager Conditions:
lysts should wear safety glasses, gloves, and lab coats when 11.2.1 Wash Solvent—Pre-inject and post-inject wash are
working in the lab.Analysts should review the Material Safety both 8 seconds of 60 % CH CN/40 % 2-propanol.
Data Sheets (MSDS) for all reagents used in this practice. 11.2.2 Purge Solvent—50 % Water/50 % CH CN.
11.2.3 Temperatures—Column, 35°C; Sample
10. Sampling
compartment, 15°C.
10.1 Grab samples may be collected in 40-mL pre-cleaned 11.2.4 Seal Wash—Solvent: 50 % CH CN /50 % Water;
amber glass vials with TFE-fluorocarbon–lined caps demon- Time: 5 minutes.
D7742 − 17
FIG. 4 SRM Chromatograms OP2EO-OP7EO
11.3 Mass Spectrometer Parameters:
The instrument is set in the electrospray positive source setting.
Capillary Voltage: 3.5 kV
11.3.1 To acquire the maximum number of data points per
Cone: Variable depending on analyte (Table 3)
SRM channel while maintaining adequate sensitivity, the tune
Extractor: 2 Volts
parameters must be optimized according to the instrument.
RF Lens: 0.1 Volts
Source Temperature: 120°C
Each peak requires at least 10 scans per peak for adequate
Desolvation Temperature: 325°C
quantitation. Variable parameters regarding retention times,
Desolvation Gas Flow: 800 L/hr
SRM transitions, and cone and collision energies are shown in Cone Gas Flow: 25 L/hr
12 13
Low Mass Resolution 1: 14.0
Table 3. Previous studies by Houde and Torrents demon-
High Mass Resolution 1: 14.0
+
strated that the [M+NH ] adducts for theAPEOs are suitable
Ion Energy 1: 0.8
for tandem mass spectrometry analysis and the fragmentation
Entrance Energy: –1
Collision Energy: Variable depending on analyte (Table 3)
wasveryreproducibleforthesetargetanalytes.Itisbesttouse
Exit Energy: 2
+ –
[M+H] or [M–H] as the precursor ion, but in this case, the
Low Mass Resolution 2: 14.0
ammonium adduct allowed for the best sensitivity and ease of High Mass resolution 2: 14.0
Ion Energy 2: 1.5
analysis in one electrospray ionization mode. Mass spectrom-
Multiplier: 650
eter parameters used in the development of this practice are –3
Gas Cell Pirani Gauge: 7.0 × 10 Torr
listed below, others may vary and require optimization: Inter-Channel Delay: 0.02 seconds
Inter-Scan Delay: 0.1 seconds
Dwell: 0.05 seconds
Houde, F., DeBlois, C., and Berryman, D., “Liquid Chromatographic-Tandem
MassSpectrometricDeterminationofNonlylphenolPolyethoxylatesandNonylphe- 12. Calibration and Standardization
nol Carboxylic Acids in Surface Water,” Journal of Chromatography A, Vol 961,
2002, pp. 245–256. 12.1 The mass spectrometer must be calibrated in accor-
Loyo-Rosales, J.E., Schmitz-Alfonso, I., Rice, C.P.,Torrents,A., “Analysis of
dance with manufacturer specifications before analysis. In
Octyl- and Nonylphenol and Their Ethoxylates in Water and Sediments by Liquid
order to obtain accurate analytical values through using this
Chromatography/Tandem Mass Spectrometry,” Analytical Chemistry, Vol 75, No.
18, September 15, 2003, pp. 4811–4817. practice, the following procedures must be followed when
D7742 − 17
FIG. 5 SRM Chromatograms OP8EO-OP12EO
TABLE 2 Gradient Conditions for Liquid Chromatography TABLE 3 Retention Times, SRM Ions, and Analyte-Specific Mass
Spectrometer Parameters
Percent 95 % Water/ Percent 95 % CH CN/
Time Flow
5%CH CN, 5 % Water,
3 SRM Mass Transition
(min) (µL/min)
5mMNH OAc 5mMNH OAc
Analyte Retention Precursor Product Cone Collision
4 4
0 300 100 0
Time (m/z) (m/z) Voltage Energy
2 300 100 0
(Minutes) (V) (eV)
5 300 70 30
NP3EO 17.4 370.2 227.2 21 14
10 200 60 40
NP4EO 17.4 414.2 271.1 19 17
15 200 25 75
NP5EO 17.4 458.2 133.2 24 22
20 200 20 80 NP6EO 17.4 502.2 133.2 29 33
25 300 15 90
NP7EO 17.4 546.2 133.2 31 25
30 300 0 100
NP8EO 17.4 590.2 133.2 32 26
35 300 0 100
NP9EO 17.4 634.3 133.2 34 28
40 300 100 0
NP10EO 17.4 678.4 133.2 36 28
NP11EO 17.4 722.5 133.2 38 30
NP12EO 17.4 766.6 133.2 39 30
NP13EO 17.4 810.6 133.2 42 31
performing the practice. Prepare all solutions in the lab using
NP14EO 17.4 854.7 133.2 43 32
Class A volumetric glassware. NP15EO 17.4 898.7 133.2 45 32
NP16EO 17.4 942.7 133.2 47 35
12.2 Calibration and Standardization—To calibrate the
NP17EO 17.4 986.7 133.2 47 40
NP18EO 17.4 1030.7 133.2 49 37
instrument, analyze seven calibration standards containing the
OP2EO 16.4 312.2 183.3 19 12
seven concentration levels of the APEOs prior to analysis as
OP3EO 16.4 356.1 227.2 20 13
shown in Table 4. A calibration stock standard solution is
OP4EO 16.4 400.1 271.1 19 16
OP5EO 16.4 444.2 315.1 18 18
prepared from standard materials or purchased as certified
OP6EO 16.4 488.2 359.1 28 18
solutions.StockstandardSolutionAcontainingtheAPEOsand
OP7EO 16.4 532.2 133.2 29 23
n-NP2EO surrogate is prepared, and aliquots of that solution
OP8EO 16.4 576.2 133.2 32 26
OP9EO 16.4 620.3 133.2 33 27
are diluted to prepare Levels 1 through 7. The following steps
OP10EO 16.4 664.4 133.2 35 28
produce standards with the concentration values shown in
OP11EO 16.4 708.4 133.2 38 30
Table 4. The analyst is responsible for recording initial OP12EO 16.4 752.5 133.2 40 30
n-NP2EO 18.8 326.2 309.2 20 9
component weights carefully when working with pure materi-
als and correctly carrying the weights through the dilution
calculations.
D7742 − 17
TABLE 4 Estimated Concentrations of Calibration Standards
setasanexternalcalibrationusingthepeakareasinpptorppb
(PPB)
units, but consistency is warranted. Concentrations may be
Analyte Level 1 Level 2 Level 3 Level 4 Level 5 Level 6 Level 7
calculated using the data system software to generate linear
NP3EO 0.73 1.46 2.91 4.66 5.82 8.78 11.64
regression or quadratic calibration curves. Forcing the calibra-
NP4EO 1.14 2.29 4.58 7.33 9.16 13.74 18.32
NP5EO 1.38 2.76 5.53 8.84 11.06 16.59 22.11 tion curve through the origin is not recommended.
NP6EO 1.76 3.52 7.04 11.26 14.08 21.12 28.16
12.2.4 Linear calibration may be used if the coefficient of
NP7EO 1.88 3.77 7.53 12.06 15.07 22.61 30.14
determination, r , is >0.98 for the analyte. The point of origin
NP8EO 1.82 3.65 7.29 11.67 14.59 21.89 29.17
NP9EO 1.64 3.29 6.57 10.51 13.14 19.71 26.28
isexcluded,andafitweightingof1/Xisusedtogiveemphasis
NP10EO 1.50 3.01 6.02 9.63 12.04 18.06 24.07
to the lower concentrations. If one of the calibration standards
NP11EO 1.33 2.67 5.33 8.53 10.67 16.01 21.33
NP12EO 0.98 1.96 3.91 6.26 7.83 11.75 15.65 otherthanthehighorlowpointcausesther ofthecurvetobe
NP13EO 0.64 1.29 2.58 4.12 5.16 7.74 10.31
<0.98,thispointmustbere-injectedoranewcalibrationcurve
NP14EO 0.41 0.82 1.63 2.61 3.26 4.89 6.52
must be regenerated. If the low or high point, or both, is
NP15EO 0.21
...

This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: D7742 − 11 D7742 − 17
Standard Practice for
Determination of Nonylphenol Polyethoxylates (NPnEO, 3 #
n # 18) and Octylphenol Polyethoxylates (OPnEO, 2 # n #
12) in Water by Single Reaction Monitoring (SRM) Liquid
Chromatography/ Tandem Mass Spectrometry (LC/MS/MS)
This standard is issued under the fixed designation D7742; 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
1.1 This procedurepractice covers the determination of nonylphenol polyethoxylates (NPnEO, 3 ≤ n ≤ 18) and octylphenol
polyethoxylates (OPnEO, 2 ≤ n ≤ 12) in water by Single Reaction Monitoring (SRM) Liquid Chromatography/ Tandem Mass
Spectrometry (LC/MS/MS) using direct injection liquid chromatography (LC) and detected with tandem mass spectrometry
(MS/MS) detection. This is a screening practice with qualified quantitative data to check for the presence of longer chain
ethoxylates in a water sample.
1.1.1 All data are qualified because neat standards of each alkylphenol ethoxylate (APEO) are not available and the synthesis
and characterization of these neat standards would be very expensive. The Igepal®Igepal Brandbrand standards, which contain
a mixture of various chain lengths of the alkylphenol ethoxylates (APEOs), were used. The mixture was characterized in-house
assuming the instrument response at an optimum electrospray ionization cone and collision voltage for each APEO was the same.
This assumption, which may not be accurate, is used to determine qualified amounts of each ethoxylate in the standards. The
n-Nonylphenol diethoxylate (n- NP2EO) (n-NP2EO) surrogate was available as a neat characterized standard, therefore, this
concentration and recovery data was not estimated. APEOs are not regulated by the EPA, but nonylphenol, a breakdown product
of NPnEOs, is regulated for fresh and saltwater dischargers. A request by a sewage treatment plant (STP) was made to make this
practice available through ASTM in order to screen for the influent or effluent from sources of APEOs coming into the STP. The
interest lies in stopping the source of the longer chain APEOs from entering the STP in order to meet effluent guidelines. Based
upon the above, this is a practice rather than a Standard Method.test method. A comparison between samples is possible using this
practice to determine which has a higher concentration of APEOs.
1.2 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this
practice.
1.3 The estimated screening range shown in Table 1 was calculated from the concentration of the Level 1 and 7 calibration
standards shown in Table 4. These numbers are qualified, as explained in Section 11,, and must be reported as such. Figs. 1-5 show
the SRM chromatograms of each analyte at the Level 1 concentration with the signal to noise (S/N) ratio. This is a screening
practice and method detection limits are not given. The S/N ratio for each analyte at the Level 1 concentration must be at least
5:1 for adequate sensitivity. If the instrument can not meet the criteria, the screening limit must be raised to an acceptable level.
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 safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.5 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.
This practice is under the jurisdiction of ASTM Committee D19 on Water and is the direct responsibility of Subcommittee D19.06 on Methods for Analysis for Organic
Substances in Water.
Current edition approved June 15, 2011Dec. 15, 2017. Published July 2011January 2018. Originally approved in 2011. Last previous edition approved in 2011 as D7742
– 11. DOI: 10.1520/D7742-11.10.1520/D7742-17.
Igepal is a trademark of Rhodia Operations, Aubervilliers, CA.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D7742 − 17
TABLE 1 Estimated Screening Range
Analyte Estimated Screening
Range (μg/L)
Nonylphenol 0.73–11.6
triethoxylate (NP3EO)
Nonylphenol 0.73–11.6
triethoxylate (NP3EO)
Nonylphenol 1.1–18.3
tetraethoxylate (NP4EO)
Nonylphenol 1.4–22.1
pentaethoxylate (NP5EO)
Nonylphenol 1.8–28.2
hexaethoxylate (NP6EO)
Nonylphenol 1.8–28.2
hexaethoxylate (NP6EO)
Nonylphenol 1.9–30.1
heptaethoxylate (NP7EO)
Nonylphenol 1.9–30.1
heptaethoxylate (NP7EO)
Nonylphenol 1.8–29.2
octaethoxylate (NP8EO)
Nonylphenol 1.8–29.2
octaethoxylate (NP8EO)
Nonylphenol 1.6–26.3
nonaethoxylate (NP9EO)
Nonylphenol 1.6–26.3
nonaethoxylate (NP9EO)
Nonylphenol 1.5–24.1
decaethoxylate (NP10EO)
Nonylphenol 1.5–24.1
decaethoxylate (NP10EO)
Nonylphenol 1.3–21.3
undecaethoxylate (NP11EO)
Nonylphenol 1.3–21.3
undecaethoxylate (NP11EO)
Nonylphenol 1.0–15.7
dodecaethoxylate (NP12EO)
Nonylphenol 1.0–15.7
dodecaethoxylate (NP12EO)
Nonylphenol 0.64–10.3
tridecaethoxylate (NP13EO)
Nonylphenol 0.64–10.3
tridecaethoxylate (NP13EO)
Nonylphenol 0.41–6.5
tetradecaethoxylate (NP14EO)
Nonylphenol 0.41–6.5
tetradecaethoxylate (NP14EO)
Nonylphenol 0.21–3.4
pendecaethoxylate (NP15EO)
Nonylphenol 0.21–3.4
pendecaethoxylate (NP15EO)
Nonylphenol 0.11–1.7
hexadecaethoxylate (NP16EO)
Nonylphenol 0.11–1.7
hexadecaethoxylate (NP16EO)
Nonylphenol 0.05–0.80
heptadecaethoxylate (NP17EO)
Nonylphenol 0.05–0.80
heptadecaethoxylate (NP17EO)
Nonylphenol 0.023–0.4
octodecaethoxylate (NP18EO)
Nonylphenol 0.023–0.4
octodecaethoxylate (NP18EO)
Total NPnEO 16–250
Octylphenol 0.14–2.3
diethoxylate (OP2EO)
Octylphenol 0.14–2.3
diethoxylate (OP2EO)
Octylphenol 1.4–22.2
triethoxylate (OP3EO)
Octylphenol 2.2–35.2
tetraethoxylate (OP4EO)
Octylphenol 2.9–45.8
pentaethoxylate (OP5EO)
Octylphenol 2.6–41.9
hexaethoxylate (OP6EO)
Octylphenol 2.6–41.9
hexaethoxylate (OP6EO)
D7742 − 17
Analyte Estimated Screening
Range (μg/L)
Octylphenol 2.5–40.4
heptaethoxylate (OP7EO)
Octylphenol 2.5–40.4
heptaethoxylate (OP7EO)
Octylphenol 1.8–28.8
octaethoxylate (OP8EO)
Octylphenol 1.8–28.8
octaethoxylate (OP8EO)
Octylphenol 1.1–17.6
nonaethoxylate (OP9EO)
Octylphenol 0.62–9.9
decaethoxylate (OP10EO)
Octylphenol 0.62–9.9
decaethoxylate (OP10EO)
Octylphenol 0.26–4.2
undecaethoxylate (OP11EO)
Octylphenol 0.26–4.2
undecaethoxylate (OP11EO)
Octylphenol 0.11–1.8
dodecaethoxylate (OP12EO)
Total OPnEO 16–250
n-Nonylphenol 15.6–250 (Not Estimated)
diethoxylate (n-NP2EO)
n-Nonylphenol 15.6–250 (Not Estimated)
diethoxylate (n-NP2EO)
2. Referenced Documents
2.1 ASTM Standards:
D1129 Terminology Relating to Water
D1193 Specification for Reagent Water
D2777 Practice for Determination of Precision and Bias of Applicable Test Methods of Committee D19 on Water
D3856 Guide for Management Systems in Laboratories Engaged in Analysis of Water
D3694 Practices for Preparation of Sample Containers and for Preservation of Organic Constituents
D5847 Practice for Writing Quality Control Specifications for Standard Test Methods for Water Analysis
E2554 Practice for Estimating and Monitoring the Uncertainty of Test Results of a Test Method Using Control Chart Techniques
2.2 Other Standard:
EPA Publication SW-846 Test Methods for Evaluating Solid Waste, Physical/Chemical Methods
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
Available from National Technical Information Service (NTIS), U.S. Department of Commerce, 5285 Port Royal Road, Springfield, VA, 22161 or at http://www.epa.gov/
epawaste/hazard/testmethods/index.htm5301 Shawnee Rd., Alexandria, VA 22312, http://www.ntis.gov.
D7742 − 17
FIG. 1 SRM Chromatograms NP3EO-NP8EO
3. Terminology
3.1 Definitions:
3.1.1 For definitions of terms used in this standard, refer to Terminology D1129.
3.2 Definitions:Definitions of Terms Specific to This Standard:
3.1.1 Screening Limit, SL, n—the estimated concentration of the lowest-level calibration standard used for quantification
accounting for the sample dilution.
3.2.1 Alkylphenol Ethoxylates,alkylphenol ethoxylates, n—in this practice, nonylphenol polyethoxylates (NPnEO, 3 ≤ n ≤ 18)
and octylphenol polyethoxylates (OPnEO, 2 ≤ n ≤ 12) collectively.
3.2.2 screening limit, SL, n—the estimated concentration of the lowest-level calibration standard used for quantification
accounting for the sample dilution.
3.3 Abbreviations:
3.2.1 ppt—parts per trillion, ng/L
–3
3.3.1 mM—millimolar, 1 × 10 moles/L
3.3.2 ND—non-detect
3.3.3 ppt—parts per trillion, ng/L
4. Summary of Practice
4.1 This is a performance-based practice and modifications are allowed to improve performance.
4.2 For APEOs analysis, samples are shipped to the lab between 0°C and 6°C containing 1 % formaldehyde and analyzed within
7 days of collection. In the lab, an aliquot of the sample is filtered, spiked with surrogate, and analyzed directly by LC/MS/MS.
4.2.1 Field samples from sewage systems propose a challenging analysis. Since this is a screening technique to determine if
APEOs are present, a 10–25 mL aliquot of the sample is filtered through a PVDF syringe driven filter unit before spiking with
surrogate. It was demonstrated that similar recoveries of the APEOs are achieved filtered and unfiltered using PVDF filters.
Filtering using PTFE filters produced much lower recoveries. This practice does not account for the APEOs adhered to particulates
or the sample bottle.
D7742 − 17
FIG. 2 SRM Chromatograms NP9EO-NP14EO
4.3 Nonylphenol polyethoxylates (NPnEO, 3 ≤ n ≤ 18), octylphenol polyethoxylates (OPnEO, 2 ≤ n ≤ 12), and n-nonylphenol
diethoxylate (n-NP2EO, surrogate) are identified by retention time and one SRM transition. The target analytes and surrogates are
quantitated using the SRM transition by external calibration. The final report issued for each sample lists their qualified
concentration and the surrogate recovery.
5. Significance and Use
5.1 This practice has been developed in support of the USU.S. EPA Office of Water, Office of Science and Technology by the
Chicago Regional Laboratory (CRL).
5.2 Nonylphenol (NP) and Octylphenol (OP) have been shown to have toxic effects in aquatic organisms. The prominent source
of NP and OP is from common commercial surfactants which are longer chain APEOs. The most widely used surfactant is
nonylphenol polyethoxylate (NPnEO) which has an average ethoxylate chain length of nine. The APEOs are readily biodegraded
to form NP1EO, NP2EO, nonylphenol carboxylate (NPEC) and NP. NP will also biodegrade, but may be released into
environmental waters directly at trace levels. This practice screens for the longer chain APEOs which may enter the STP at elevated
levels and may cause a STP to violate its permitted discharge concentration of nonylphenol.
6. Interferences
6.1 Practice interferences may be caused by contaminants in solvents, reagents, glassware and other apparatus producing
discrete artifacts or elevated baselines. All of these materials are routinely demonstrated to be free from interferences by analyzing
laboratory reagent blanks under the same conditions as the samples.
6.2 All glassware is washed in hot water with detergent such as powdered Alconox, Deto-Jet,Det-o-Jet, Luminox, or
Citrojet,Citrajet, rinsed in hot water, and rinsed with distilled water. The glassware is then dried and heated in an oven at 250ºC
for 15 to 30 minutes. All glassware is subsequently cleaned with acetone and methanol. Detergents containing alkylphenolic
compounds must not be used.
6.3 All reagents and solvents should be of pesticide residue purity or higher to minimize interference problems.
Alconox, Det-o-Jet, Luminox, and Citrajet are trademarks of Alconox, Inc., White Plains, NY.
D7742 − 17
FIG. 3 SRM Chromatograms NP15EO-NP18EO and n-NP2EO
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 sample source, depending on variations of the sample matrix.
D7742 − 17
FIG. 4 SRM Chromatograms OP2EO-OP7EO
7. Apparatus
7.1 LC/MS/MS System:
7.1.1 Liquid Chromatography System—A complete LC system is needed in order to analyze samples. Any system that is
capable of performing at the flows, pressures, controlled temperatures, sample volumes, and requirements of the standard may be
used.
7.1.2 Analytical Column– Waters—Column—Atlantis™ Waters Atlantis dC18, 2.1 × 150 mm, 3 μm particle size was used to
develop this practice. Any column that achieves adequate resolution may be used. The retention times and order of elution may
change depending on the column used and need to be monitored.
7.1.3 Tandem Mass Spectrometer System—A MS/MS system capable of MRM analysis. Any system that is capable of
performing at the requirements in this practice may be used.
7.2 Filtration Device:
7.2.1 Hypodermic syringe—A Lock Tip Glass Syringe lock-tip glass syringe capable of holding a Millex®Millex HV Syringe
9,10
Driven Filter Unit PVDF 0.45 μm, or similar, may be used.
7.2.1.1 A 25 mL Lock Tip Glass Syringe 25–mL lock-tip glass syringe size is recommended for this practice.
7.2.2 Filter—Millex®Millex HV Syringe Driven Filter Unit PVDF 0.45 μm (Millipore Corporation, Catalog # SLHV033NS)
was used to develop this practice, any similar filter may be used.
Waters ACQUITY H-Class Ultra Performance Liquid Chromatography (UPLC®) System (UPLC) System, a trademark of the Waters Corporation, Milford, MA, was
used to develop this test method. practice. All parameters in this test method practice are based on this system and may vary depending on your instrument.
Waters Atlantis dC18 is a trademark of the Waters Corporation, Milford, MA.
A Waters Quattro Micro™Micro tandem quadrupole mass spectrometer spectrometer, a trademark of the Waters Corporation, Milford, MA., was used to develop this
test method. practice. All parameters in this test method practice are based on this system and may vary depending on your instrument.
The sole source of supply of the Millex HV Syringe Driven Filter Unit PVDF 0.45 μm known to the committee at this time is Millipore Corporation, Catalog #
SLHV033NS. If you are aware of alternative suppliers, please provide this information to ASTM International Headquarters. Your comments will receive careful consideration
at a meeting of the responsible technical committee, which you may attend.
Millex is a trademark of Merck KGAA, Darmstadt, Germany.
D7742 − 17
FIG. 5 SRM Chromatograms OP8EO-OP12EO
8. Reagents and Materials
8.1 Purity of Reagents—High Performance Liquid Chromatography (HPLC) pesticide residue analysis and spectrophotometry
grade chemicals shall be used in all tests. Unless indicated otherwise, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical Society. Other reagent grades may be used
provided it is first ascertained that they are of sufficiently high purity to permit their use without affecting the accuracy of the
measurement.
8.2 Purity of Water—Unless indicated, references to water shall be understood to mean reagent water conforming to Type I of
Specification D1193. It must be demonstrated that this water does not contain contaminants at concentrations sufficient to interfere
with the analysis.
8.3 Gases—Ultrapure nitrogen and argonargon.
8.4 Acetonitrile (CAS # 75-05-8)75-05-8).
8.5 Methanol (CAS # 67-56-1)67-56-1).
8.6 2-Propanol (CAS # 67-63-0)67-63-0).
8.7 Acetone (CAS # 67-64-1)67-64-1).
8.8 Nonylphenol pentaethoxylate mixture (several NPnEO isomer groups with an average of NP5EO, Igepal® CO-520)Igepal
CO-520).
8.9 Nonylphenol nonaethoxylate mixture (several NPnEO isomer groups with an average of NP9EO, Igepal® CO-630)Igepal
CO-630).
8.10 Octylphenol diethoxylate mixture (several OPnEO isomer groups with an average of OP2EO, Igepal® CA-210)Igepal
CA-210).
Reagent Chemicals, American Chemical Society Specifications, American Chemical Society, Washington, D.C.DC. For Suggestionssuggestions on the testing of reagents
not listed by the American Chemical Society, see AnnualAnalar Standards for Laboratory Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
and National Formulators,Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville, MD.
D7742 − 17
8.11 Octylphenol pentaethoxylate mixture (several OPnEO isomer groups with an average of OP5EO, Igepal® CA-520)Igepal
CA-520).
8.12 Formaldehyde (CAS # 50-00-0, 37 wt. % solution in water)water).
8.13 Ammonium Acetate (CAS # 631-61-8)631-61-8).
8.14 n-Nonylphenol diethoxylate (n-NP2EO)(n-NP2EO).
9. Hazards
9.1 Normal laboratory safety applies to this practice. Analysts should wear safety glasses, gloves, and lab coats when working
in the lab. Analysts should review the Material Safety Data Sheets (MSDS) for all reagents used in this practice.
10. Sampling
10.1 Grab samples may be collected in 40 mL 40-mL pre-cleaned amber glass vials with Teflon® lined TFE-fluorocarbon–lined
caps demonstrated to be free of interferences, larger sample sizes may be used since a subsample aliquot is only required. All
samples are preserved with 1 % concentration of formaldehyde, shipped between 0°C and 6°C, and stored in the laboratory
between 0°C and 6°C. Conventional sampling practices should be followed. Refer to Guide D3856 and Practices D3694.
Automatic sampling equipment should be as free as possible of Tygon tubing and other potential sources of contamination or cause
adhesion of APEOs. Analyze the sample within 7 days of collection.
11. Preparation of LC/MS/MS
11.1 LC Chromatograph Operating Conditions:
11.1.1 Injection volumes of all calibration standards and samples are made at 100 μL 100-μL volume. The first sample analyzed
after the calibration curve is a blank to ensure there is no carry-over. The gradient conditions for the liquid chromatograph are
shown in Table 2.
11.2 LC Sample Manager Conditions:
11.2.1 Wash Solvent—Pre-inject and post-inject wash are both 8 seconds of 60 % CH CN/40 % 2-propanol.
11.2.2 Purge Solvent—50 % Water/50 % CH CNCN.
11.2.3 Temperatures—Column, 35°C; Sample compartment, 15°C.
11.2.4 Seal Wash—Solvent: 50 % CH CN /50 % Water; Time: 5 minutesminutes.
TABLE 2 Gradient Conditions for Liquid Chromatography
Percent 95 % Water/ Percent 95 % CH CN/
Time Flow
5 % CH CN, 5 % Water,
(min) (μL/min)
5 mM NH OAc 5 mM NH OAc
4 4
0 300 100 0
2 300 100 0
5 300 70 30
10 200 60 40
15 200 25 75
20 200 20 80
25 300 15 90
30 300 0 100
35 300 0 100
40 300 100 0
D7742 − 17
11.3 Mass Spectrometer Parameters:
11.3.1 To acquire the maximum number of data points per SRM channel while maintaining adequate sensitivity, the tune
parameters must be optimized according to the instrument. Each peak requires at least 10 scans per peak for adequate quantitation.
Variable parameters regarding retention times, SRM transitions, and cone and collision energies are shown in Table 3. Previous
12 13 +
studies by Houde and Torrents demonstrated that the [M+NH ] adducts for the APEOs are suitable for tandem mass
+ –
spectrometry analysis and the fragmentation was very reproducible for these target analytes. It is best to use [M+H] or [M–H]
as the precursor ion, but in this case, the ammonium adduct allowed for the best sensitivity and ease of analysis in one electrospray
ionization mode. Mass spectrometer parameters used in the development of this practice are listed below, others may vary and
require optimization:
The instrument is set in the electrospray positive source setting.
Capillary Voltage: 3.5 kV
Cone: Variable depending on analyte (Table 3)
Extractor: 2 Volts
RF Lens: 0.1 Volts
Source Temperature: 120°C
Desolvation Temperature: 325°C
Desolvation Gas Flow: 800 L/hr
Cone Gas Flow: 25 L/hr
Low Mass Resolution 1: 14.0
High Mass Resolution 1: 14.0
Ion Energy 1: 0.8
Entrance Energy: –1
Collision Energy: Variable depending on analyte (Table 3)
Exit Energy: 2
Low Mass Resolution 2: 14.0
High Mass resolution 2: 14.0
TABLE 3 Retention Times, SRM Ions, and Analyte-Specific Mass
Spectrometer Parameters
SRM Mass Transition
Analyte Retention Precursor Product Cone Collision
Time (m/z) (m/z) Voltage Energy
(Minutes) (V) (eV)
NP3EO 17.4 370.2 227.2 21 14
NP4EO 17.4 414.2 271.1 19 17
NP5EO 17.4 458.2 133.2 24 22
NP6EO 17.4 502.2 133.2 29 33
NP7EO 17.4 546.2 133.2 31 25
NP8EO 17.4 590.2 133.2 32 26
NP9EO 17.4 634.3 133.2 34 28
NP10EO 17.4 678.4 133.2 36 28
NP11EO 17.4 722.5 133.2 38 30
NP12EO 17.4 766.6 133.2 39 30
NP13EO 17.4 810.6 133.2 42 31
NP14EO 17.4 854.7 133.2 43 32
NP15EO 17.4 898.7 133.2 45 32
NP16EO 17.4 942.7 133.2 47 35
NP17EO 17.4 986.7 133.2 47 40
NP18EO 17.4 1030.7 133.2 49 37
OP2EO 16.4 312.2 183.3 19 12
OP3EO 16.4 356.1 227.2 20 13
OP4EO 16.4 400.1 271.1 19 16
OP5EO 16.4 444.2 315.1 18 18
OP6EO 16.4 488.2 359.1 28 18
OP7EO 16.4 532.2 133.2 29 23
OP8EO 16.4 576.2 133.2 32 26
OP9EO 16.4 620.3 133.2 33 27
OP10EO 16.4 664.4 133.2 35 28
OP11EO 16.4 708.4 133.2 38 30
OP12EO 16.4 752.5 133.2 40 30
n-NP2EO 18.8 326.2 309.2 20 9
Houde, F., DeBlois, C., and Berryman, D., “Liquid Chromatographic-Tandem Mass Spectrometric Determination of Nonlylphenol Polyethoxylates and Nonylphenol
Carboxylic Acids in Surface Water,” Journal of Chromatography A, Vol 961, 2002, pp. 245–256.
Loyo-Rosales, J.E., Schmitz-Alfonso, I., Rice, C.P., Torrents, A., “Analysis of Octyl- and Nonylphenol and Their Ethoxylates in Water and Sediments by Liquid
Chromatography/Tandem Mass Spectrometry,” Analytical Chemistry, Vol 75, No. 18, September 15, 2003, pp. 4811–4817.
D7742 − 17
Ion Energy 2: 1.5
Multiplier: 650
–3
Gas Cell Pirani Gauge: 7.0 × 10 Torr
Inter-Channel Delay: 0.02 seconds
Inter-Scan Delay: 0.1 seconds
Dwell: 0.05 seconds
The instrument is set in the electrospray positive source setting.
Capillary Voltage: 3.5 kV
Cone: Variable depending on analyte (Table 3)
Extractor: 2 Volts
RF Lens: 0.1 Volts
Source Temperature: 120°C
Desolvation Temperature: 325°C
Desolvation Gas Flow: 800 L/hr
Cone Gas Flow: 25 L/hr
Low Mass Resolution 1: 14.0
High Mass Resolution 1: 14.0
Ion Energy 1: 0.8
Entrance Energy: –1
Collision Energy: Variable depending on analyte (Table 3)
Exit Energy: 2
Low Mass Resolution 2: 14.0
High Mass resolution 2: 14.0
Ion Energy 2: 1.5
Multiplier: 650
–3
Gas Cell Pirani Gauge: 7.0 × 10 Torr
Inter-Channel Delay: 0.02 seconds
Inter-Scan Delay: 0.1 seconds
Dwell: 0.05 seconds
12. Calibration and Standardization
12.1 The mass spectrometer must be calibrated per in accordance with manufacturer specifications before analysis. In order to
obtain accurate analytical values through using this practice, the following procedures must be followed when performing the
practice. Prepare all solutions in the lab using Class A volumetric glassware.
12.2 Calibration and Standardization—To calibrate the instrument, analyze seven calibration standards containing the seven
concentration levels of the APEOs prior to analysis as shown in Table 4. A calibration stock standard solution is prepared from
standard materials or purchased as certified solutions. Stock standard solutionSolution A containing the APEOs and n-NP2EO
surrogate is prepared, and aliquots of that solution are diluted to prepare Levels 1 through 7. The following steps produce standards
TABLE 4 Estimated Concentrations of Calibration Standards
(PPB)
Analyte Level 1 Level 2 Level 3 Level 4 Level 5 Level 6 Level 7
NP3EO 0.73 1.46 2.91 4.66 5.82 8.78 11.64
NP4EO 1.14 2.29 4.58 7.33 9.16 13.74 18.32
NP5EO 1.38 2.76 5.53 8.84 11.06 16.59 22.11
NP6EO 1.76 3.52 7.04 11.26 14.08 21.12 28.16
NP7EO 1.88 3.77 7.53 12.06 15.07 22.61 30.14
NP8EO 1.82 3.65 7.29 11.67 14.59 21.89 29.17
NP9EO 1.64 3.29 6.57 10.51 13.14 19.71 26.28
NP10EO 1.50 3.01 6.02 9.63 12.04 18.06 24.07
NP11EO 1.33 2.67 5.33 8.53 10.67 16.01 21.33
NP12EO 0.98 1.96 3.91 6.26 7.83 11.75 15.65
NP13EO 0.64 1.29 2.58 4.12 5.16 7.74 10.31
NP14EO 0.41 0.82 1.63 2.61 3.26 4.89 6.52
NP15EO 0.21 0.43 0.85 1.36 1.70 2.55 3.41
NP16EO 0.11 0.21 0.43 0.68 0.86 1.29 1.71
NP17EO 0.050 0.10 0.20 0.32 0.40 0.60 0.80
NP18EO 0.023 0.045 0.090 0.14 0.18 0.27 0.36
OP2EO 0.14 0.28 0.56 0.90 1.13 1.70 2.26
OP3EO 1.39 2.77 5.55 8.88 11.10 16.65 22.20
OP4EO 2.20 4.40 8.80 14.08 17.61 26.42 35.21
OP5EO 2.86 5.72 11.45 18.32 22.90 34.35 45.80
OP6EO 2.62 5.24 10.48 16.77 20.96 31.44 41.92
OP7EO 2.52 5.05 10.09 16.15 20.19 30.29 40.38
OP8EO 1.80 3.59 7.19 11.50 14.38 21.57 28.76
OP9EO 1.10 2.20 4.40 7.04 8.79 13.19 17.59
OP10EO 0.62 1.23 2.47 3.94 4.93 7.
...