1、Designation: D7742 11Standard Practice forDetermination of Nonylphenol Polyethoxylates (NPnEO, 3 #n # 18) and Octylphenol Polyethoxylates (OPnEO, 2 # n #12) in Water by Single Reaction Monitoring (SRM) LiquidChromatography/ Tandem Mass Spectrometry (LC/MS/MS)1This standard is issued under the fixed
2、designation D7742; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revis
3、ion or reapproval.1. Scope1.1 This procedure covers the determination of nonylphenolpolyethoxylates (NPnEO, 3 # n # 18) and octylphenolpolyethoxylates (OPnEO, 2 # n # 12) in water by SingleReaction Monitoring (SRM) Liquid Chromatography/ TandemMass Spectrometry (LC/MS/MS) using direct injection liqu
4、idchromatography (LC) and detected with tandem mass spec-trometry (MS/MS) detection. This is a screening Practice withqualified quantitative data to check for the presence of longerchain ethoxylates in a water sample.1.1.1 All data are qualified because neat standards of eachalkylphenol ethoxylate (
5、APEO) are not available and thesynthesis and characterization of these neat standards would bevery expensive. The Igepalt Brand standards, which contain amixture of various chain lengths of the alkylphenol ethoxylates(APEOs), were used. The mixture was characterized in-houseassuming the instrument r
6、esponse at an optimum electrosprayionization cone and collision voltage for each APEO was thesame. This assumption, which may not be accurate, is used todetermine qualified amounts of each ethoxylate in the stan-dards. The n-Nonylphenol diethoxylate (n- NP2EO) surrogatewas available as a neat charac
7、terized standard, therefore, thisconcentration and recovery data was not estimated. APEOs arenot regulated by the EPA, but nonylphenol, a breakdownproduct of NPnEOs, is regulated for fresh and saltwaterdischargers. A request by a sewage treatment plant (STP) wasmade to make this Practice available t
8、hroughASTM in order toscreen for the influent or effluent from sources of APEOscoming into the STP. The interest lies in stopping the source ofthe longer chainAPEOs from entering the STP in order to meeteffluent guidelines. Based upon the above, this is a Practicerather than a Standard Method.Acompa
9、rison between samplesis possible using this Practice to determine which has a higherconcentration of APEOs.1.2 UnitsThe values stated in SI units are to be regardedas standard. No other units of measurement are included in thisstandard.1.3 The estimated screening range shown in Table 1 wascalculated
10、 from the concentration of the Level 1 and 7calibration standards shown in Table 4. These numbers arequalified, as explained in Section 1, and must be reported assuch. Figs. 1-5 show the SRM chromatograms of each analyteat the Level 1 concentration with the signal to noise (S/N) ratio.This is a scre
11、ening Practice and method detection limits are notgiven. The S/N ratio for each analyte at the Level 1 concen-tration must be at least 5:1 for adequate sensitivity. If theinstrument can not meet the criteria, the screening limit mustbe raised to an acceptable level.1.4 This standard does not purport
12、 to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D1193
13、Specification for Reagent WaterD2777 Practice for Determination of Precision and Bias ofApplicable Test Methods of Committee D19 on WaterD3856 Guide for Good Laboratory Practices in Laborato-ries Engaged in Sampling and Analysis of WaterD3694 Practices for Preparation of Sample Containers andfor Pre
14、servation of Organic ConstituentsD5847 Practice for Writing Quality Control Specificationsfor Standard Test Methods for Water AnalysisE2554 Practice for Estimating and Monitoring the Uncer-tainty of Test Results of a Test Method in a SingleLaboratory Using a Control Sample Program1This Practice is u
15、nder the jurisdiction of ASTM Committee D19 on Water andis the direct responsibility of Subcommittee D19.06 on Methods for Analysis forOrganic Substances in Water.Current edition approved June 15, 2011. Published July 2011.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orconta
16、ct ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.2.2 Other Standard:3EPA
17、 SW-846 Test Methods for Evaluating Solid Waste,Physical/Chemical Methods3. Terminology3.1 Definitions:3.1.1 Screening Limit, SL, nthe estimated concentrationof the lowest-level calibration standard used for quantificationaccounting for the sample dilution.3.1.2 Alkylphenol Ethoxylates, nin this Pra
18、ctice, nonyl-phenol polyethoxylates (NPnEO, 3#n#18) and octylphenolpolyethoxylates (OPnEO, 2 # n # 12) collectively.3.2 Abbreviations:3.2.1 ppt parts per trillion, ng/L3.2.2 mM millimolar,1x10-3moles/L3.2.3 ND non-detect4. Summary of Practice4.1 This is a performance based Practice and modifications
19、are allowed to improve performance.4.2 For APEOs analysis, samples are shipped to the labbetween 0C and 6C containing 1% formaldehyde and ana-lyzed within 7 days of collection. In the lab, an aliquot of thesample is filtered, spiked with surrogate, and analyzed directlyby LC/MS/MS.4.2.1 Field sample
20、s from sewage systems propose a chal-lenging analysis. Since this is a screening technique to deter-mine if APEOs are present, a 10-25 mL aliquot of the sampleis filtered through a PVDF syringe driven filter unit beforespiking with surrogate. It was demonstrated that similar recov-eries of the APEOs
21、 are achieved filtered and unfiltered usingPVDF filters. Filtering using PTFE filters produced muchlower recoveries. This Practice does not account for theAPEOsadhered to particulates or the sample bottle.4.3 Nonylphenol polyethoxylates (NPnEO, 3 # n # 18),octylphenol polyethoxylates (OPnEO, 2 # n #
22、 12), andn-nonylphenol diethoxylate (n-NP2EO, surrogate) are identi-fied by retention time and one SRM transition. The targetanalytes and surrogates are quantitated using the SRM transi-tion by external calibration. The final report issued for eachsample lists their qualified concentration and the s
23、urrogaterecovery.5. Significance and Use5.1 This Practice has been developed in support of the USEPA Office of Water, Office of Science and Technology by theChicago Regional Laboratory (CRL).5.2 Nonylphenol (NP) and Octylphenol (OP) have beenshown to have toxic effects in aquatic organisms. The prom
24、i-nent source of NP and OP is from common commercialsurfactants which are longer chain APEOs. The most widelyused surfactant is nonylphenol polyethoxylate (NPnEO) whichhas an average ethoxylate chain length of nine. TheAPEOs arereadily biodegraded to form NP1EO, NP2EO, nonylphenolcarboxylate (NPEC)
25、and NP. NP will also biodegrade, but maybe released into environmental waters directly at trace levels.This Practice screens for the longer chain APEOs which mayenter the STP at elevated levels and may cause a STP to violateits permitted discharge concentration of nonylphenol.6. Interferences6.1 Pra
26、ctice interferences may be caused by contaminants insolvents, reagents, glassware and other apparatus producingdiscrete artifacts or elevated baselines. All of these materials3Available from National Technical Information Service (NTIS), U.S. Depart-ment of Commerce, 5285 Port Royal Road, Springfiel
27、d, VA, 22161 or at http:/www.epa.gov/epawaste/hazard/testmethods/index.htmTABLE 1 Estimated Screening RangeAnalyte Estimated ScreeningRange (g/L)Nonylphenoltriethoxylate (NP3EO)0.73-11.6Nonylphenoltetraethoxylate (NP4EO)1.1-18.3Nonylphenolpentaethoxylate (NP5EO)1.4-22.1Nonylphenolhexaethoxylate (NP6
28、EO)1.8-28.2Nonylphenolheptaethoxylate (NP7EO)1.9-30.1Nonylphenoloctaethoxylate (NP8EO)1.8-29.2Nonylphenolnonaethoxylate (NP9EO)1.6-26.3Nonylphenoldecaethoxylate (NP10EO)1.5-24.1Nonylphenolundecaethoxylate (NP11EO)1.3-21.3Nonylphenoldodecaethoxylate (NP12EO)1.0-15.7Nonylphenoltridecaethoxylate (NP13E
29、O)0.64-10.3Nonylphenoltetradecaethoxylate (NP14EO)0.41-6.5Nonylphenolpendecaethoxylate (NP15EO)0.21-3.4Nonylphenolhexadecaethoxylate (NP16EO)0.11-1.7Nonylphenolheptadecaethoxylate (NP17EO)0.05-0.80Nonylphenoloctodecaethoxylate (NP18EO)0.023-0.4Total NPnEO 16-250Octylphenoldiethoxylate (OP2EO)0.14-2.
30、3Octylphenoltriethoxylate (OP3EO)1.4-22.2Octylphenoltetraethoxylate (OP4EO)2.2-35.2Octylphenolpentaethoxylate (OP5EO)2.9-45.8Octylphenolhexaethoxylate (OP6EO)2.6-41.9Octylphenolheptaethoxylate (OP7EO)2.5-40.4Octylphenoloctaethoxylate (OP8EO)1.8-28.8Octylphenolnonaethoxylate (OP9EO)1.1-17.6Octylpheno
31、ldecaethoxylate (OP10EO)0.62-9.9Octylphenolundecaethoxylate (OP11EO)0.26-4.2Octylphenoldodecaethoxylate (OP12EO)0.11-1.8Total OPnEO 16-250n-Nonylphenoldiethoxylate (n-NP2EO)15.6-250 (Not Estimated)D7742 112are routinely demonstrated to be free from interferences byanalyzing laboratory reagent blanks
32、 under the same conditionsas the samples.6.2 All glassware is washed in hot water with detergent suchas powderedAlconox, Deto-Jet, Luminox, or Citrojet, rinsed inhot water, and rinsed with distilled water. The glassware is thendried and heated in an oven at 250C for 15 to 30 minutes. Allglassware is
33、 subsequently cleaned with acetone and methanol.Detergents containing alkylphenolic compounds must not beused.6.3 All reagents and solvents should be of pesticide residuepurity or higher to minimize interference problems.6.4 Matrix interferences may be caused by contaminantsthat are co-extracted fro
34、m the sample. The extent of matrixinterferences can vary considerably from sample source tosample source, depending on variations of the sample matrix.7. Apparatus7.1 LC/MS/MS System7.1.1 Liquid Chromatography SystemA complete LC sys-tem is needed in order to analyze samples.4Any system that iscapab
35、le of performing at the flows, pressures, controlledtemperatures, sample volumes, and requirements of the stan-dard may be used.7.1.2 Analytical Column WatersAtlantisy dC18, 2.1 x150 mm, 3 m particle size was used to develop this Practice.Any column that achieves adequate resolution may be used.The
36、retention times and order of elution may change dependingon the column used and need to be monitored.7.1.3 Tandem Mass Spectrometer SystemA MS/MS sys-tem capable of MRM analysis.5Any system that is capable ofperforming at the requirements in this Practice may be used.7.2 Filtration Device7.2.1 Hypod
37、ermic syringeA Lock Tip Glass Syringe ca-pable of holding a Millext HV Syringe Driven Filter UnitPVDF 0.45 m or similar may be used.7.2.1.1 A 25 mL Lock Tip Glass Syringe size is recom-mended for this Practice.7.2.2 FilterMillext HV Syringe Driven Filter Unit PVDF0.45 m (Millipore Corporation, Catal
38、og # SLHV033NS) wasused to develop this Practice, any similar filter may be used.4Waters ACQUITY H-Class Ultra Performance Liquid Chromatography(UPLCt) System was used to develop this test method. All parameters in this testmethod are based on this system and may vary depending on your instrument.5A
39、 Waters Quattro Microy tandem quadrupole mass spectrometer was used todevelop this test method.All parameters in this test method are based on this systemand may vary depending on your instrument.FIG. 1 SRM Chromatograms NP3EO-NP8EOD7742 1138. Reagents and Materials8.1 Purity of ReagentsHigh Perform
40、ance Liquid Chroma-tography (HPLC) pesticide residue analysis and spectropho-tometry grade chemicals shall be used in all tests. Unlessindicated otherwise, it is intended that all reagents shallconform to the specifications of the Committee on AnalyticalReagents of the American Chemical Society.6Oth
41、er reagentgrades may be used provided it is first ascertained that they areof sufficiently high purity to permit their use without affectingthe accuracy of the measurement.8.2 Purity of WaterUnless indicated, references to watershall be understood to mean reagent water conforming to TypeI of Specifi
42、cation D1193. It must be demonstrated that thiswater does not contain contaminants at concentrations suffi-cient to interfere with the analysis.8.3 GasesUltrapure nitrogen and argon8.4 Acetonitrile (CAS # 75-05-8)8.5 Methanol (CAS # 67-56-1)8.6 2-Propanol (CAS # 67-63-0)8.7 Acetone (CAS # 67-64-1)8.
43、8 Nonylphenol pentaethoxylate mixture (several NPnEOisomer groups with an average of NP5EO, Igepalt CO-520)8.9 Nonylphenol nonaethoxylate mixture (several NPnEOisomer groups with an average of NP9EO, Igepalt CO-630)8.10 Octylphenol diethoxylate mixture (several OPnEO iso-mer groups with an average o
44、f OP2EO, Igepalt CA-210)8.11 Octylphenol pentaethoxylate mixture (several OPnEOisomer groups with an average of OP5EO, Igepalt CA-520)8.12 Formaldehyde (CAS # 50-00-0, 37 wt. % solution inwater)8.13 Ammonium Acetate (CAS # 631-61-8)8.14 n-Nonylphenol diethoxylate (n-NP2EO)9. Hazards9.1 Normal labora
45、tory safety applies to this Practice. Ana-lysts should wear safety glasses, gloves, and lab coats whenworking in the lab. Analysts should review the Material SafetyData Sheets (MSDS) for all reagents used in this Practice.10. Sampling10.1 Grab samples may be collected in 40 mL pre-cleanedamber glass
46、 vials with Teflont lined caps demonstrated to befree of interferences, larger sample sizes may be used since asubsample aliquot is only required. All samples are preservedwith 1% concentration of formaldehyde, shipped between 0Cand 6C, and stored in the laboratory between 0C and 6C.Conventional sam
47、pling Practices should be followed. Refer to6Reagent Chemicals, American Chemical Society Specifications, AmericanChemical Society, Washington, D.C. For Suggestions on the testing of reagents notlisted by the American Chemical Society, see Annual Standards for LaboratoryChemicals, BDH Ltd., Poole, D
48、orset, U.K., and the United States Pharmacopeia andNational Formulators, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,MD.FIG. 2 SRM Chromatograms NP9EO-NP14EOD7742 114Guide D3856 and Practices D3694.Automatic sampling equip-ment should be as free as possible of Tygon tubing and otherpotenti
49、al sources of contamination or cause adhesion ofAPEOs. Analyze the sample within 7 days of collection.11. Preparation of LC/MS/MS11.1 LC Chromatograph Operating Conditions411.1.1 Injection volumes of all calibration standards andsamples are made at 100 L volume. The first sample analyzedafter the calibration curve is a blank to ensure there is nocarry-over. The gradient conditions for the liquid chromato-graph are shown in Table 2.11.2 LC Sample Manager Conditions:11.2.1 Wash SolventPre-inject and post-inject wash areboth 8 seconds of 60% CH3CN/40% 2-p
