USGS-NWQL: O-4434-12: Steroid Hormones in Unfiltered Water by GC-MS/MS

Summary
Analytes
Revision
Data and Sites

Official Method Name

Determination of Steroid Hormones and Related Compounds in Unfiltered Water by Solid-Phase Extraction, Derivatization, and Gas Chromatography with Tandem Mass Spectrometry

Current Revision

2012

Media

WATER

Instrumentation

Gas Chromatography with Tandem Mass Spectrometry Detection

Method Subcategory

Organic

Method Source

USGS-NWQL

Citation

Foreman, W.T., Gray, J.L., ReVello, R.C., Lindley, C.E., Losche, S.A., and Barber, L.B., 2012, Determination of steroid hormones and related compounds in filtered and unfiltered water by solid-phase extraction, derivatization, and gas chromatography with tandem mass spectrometry: U.S. Geological Survey Techniques and Methods, book 5, chap. B9, 118 p.

Brief Method Summary

This analytical method developed by and implemented at the U.S. Geological Survey National Water Quality Laboratory determines a suite of 20 steroid hormones and related compounds in filtered water (using laboratory schedule 2434) and in unfiltered water (using laboratory schedule 4434). USGS report TM 5-B9 documents the procedures and initial performance data for the method and provides guidance on application of the method and considerations of data quality in relation to data interpretation. The analytical method determines 6 natural and 3 synthetic estrogen compounds, 6 natural androgens, 1 natural and 1 synthetic progestin compound, and 2 sterols: cholesterol and 3--coprostanol. These two sterols have limited biological activity but typically are abundant in wastewater effluents and serve as useful tracers. Bisphenol A, an industrial chemical used primarily to produce polycarbonate plastic and epoxy resins and that has been shown to have estrogenic activity, also is determined by the method. The samples undergo chemical derivatization with activated N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) before GC/MS/MS analysis.

Scope and Application

This report describes the analytical method for the determination of selected steroid hormones and other compounds in unfiltered water (USGS method number O–4434–12). The report includes a brief outline of procedures to use for field collection, processing, and shipment of the water samples to the NWQL. The report also summarizes a set of validation studies, including spike recovery experiments in reagent water and three field matrices that are anticipated to be submitted frequently for analysis: a surface-water sample, a primary WWTP effluent sample, and a secondary WWTP effluent sample. This report also presents the results of a multi-concentration detection level determination, results of sample and extract holding-time experiments, and results from custom analysis of samples in a variety of aqueous matrices. Method data-reporting procedures are addressed, including those applied to laboratory and field-based quality-control (QC) and quality-assurance (QA) samples. Note: all isotope-dilution standards (IDSs) data presented in this report are absolute IDS recoveries obtained for the isotope.

Applicable Concentration Range

Interferences

Sample contamination is a concern because some of the hormone compounds are biogenic and can be present on human skin or might be used as personal-care products. Others analytes are synthetic hormone pharmaceuticals that are in common use. Avoid contamination of the samples by avoiding consumption or contact with such materials immediately before and during sample collection and processing procedures. Protective gloves (for example, nitrile gloves) must be worn at all times. Samples are easily contaminated with bisphenol A, see report for which to avoid laboratory tissue or other paper products during field or laboratory sample manipulations of these samples.

Quality Control Requirements

Collect field-submitted quality control samples. Batch sequence QC considerations. Equipment calibration. The report gives a lengthy description of several specifics.

Sample Handling

Filter sample; add 50 mg of ascorbic acid per 500 mL of water if halogenation treatment of the water is used, chill at collection site, ship at 4^oC. Leave air space in 500mL sample. Use high-density polyethylene bottles/caps if no residual chlorine.

Maximum Holding Time

Varies by analyte, some analytes degrade rapidly after collection; therefore, ship immediately to the lab.

Relative Cost

Greater than $400

Sample Preparation Methods

This method has 20 analytes associated with it.

Analyte	Detection Level	Bias	Precision
11-Ketotestosterone(564-35-2) 11-Ketotestosterone 11-Oxotestosterone Androst-4-ene-3,11-dione, 17-hydroxy-, (17beta)- Ketotestosterone	2.000 ng/L	100% Rec	29.10 % RSD
17alpha-Estradiol(57-91-0) 1,3,5-Estratriene-3,17alpha-diol 13beta-Methyl-1,3,5(10)-gonatriene-3,17alpha-diol 17-Epiestradiol 17alpha-Estradiol 17alpha-Estradiol (8CI) 17alpha-Oestradiol 3,17-Dihydroxyestratriene 3,17alpha-Dihydroxyestra-1,3,5(10)-triene 3,17alpha-Dihydroxyoestra-1,3,5(10)-triene Alfatradiol Epiestradiol Epiestrol Estra-1,3,5(10)-triene-3,17alpha-diol NSC 20293 Oestra-1,3,5(10)-triene-3,17alpha-diol alpha-Estradiol	0.800 ng/L	103% Rec	12.90 % RSD
17beta-Estradiol(50-28-2) (+)-3,17beta-Estradiol 13beta-Methyl-1,3,5(10)-gonatriene-3,17beta-ol 17beta-Estradiol 17beta-Oestradiol 3,17-Epidihydroxyestratriene 3,17beta-Dihydroxyestra-1,3,5(10)-triene 3,17beta-Estradiol Absorlent Matrix Aerodiol Altrad Aquadiol Bardiol Climaderm Estra-1,3,5(10)-triene-3,17-diol (17beta)-(9CI) Estradiol (8CI) NSC 9895 Oestra-1,3,5(10)-triene-3,17beta-diol Theelin, dihydro- beta-Estradiol	0.800 ng/L	102% Rec	13.00 % RSD
3beta-Coprostanol(360-68-9) 3beta-Coprostanol 3beta-Hydroxy-5beta-cholestanol 5beta-Cholestan-3beta-ol (8CI) Cholestan-3-ol, (3beta,5beta)- (9CI) Coprostan-3beta-ol Coprostanol Coprosterol Koprosterin Koprosterol NSC 18175 NSC 5060 Stercorin	200.000 ng/L	96% Rec	14.30 % RSD
Androstenedione(63-05-8) 17-Ketotestosterone 3,17-Dioxoandrost-4-ene 4-Androstene-3,17-dione ADSD Androstenedione Fecundin NSC 9563 SKF 2170 delta-4-Androstene-3,17-dione	0.800 ng/L	98% Rec	8.00 % RSD
Androsterone(53-41-8) 3-Epihydroxyetioallocholan-17-one 3alpha-Hydroxy-17-androstanone 3alpha-Hydroxy-5alpha-androstan-17-one 5alpha-Androsterone Androkinin Androsterone Androtin Atromide ICI NSC 9898 U 60366 cis-Androsterone	0.800 ng/L	97% Rec	21.00 % RSD
Bisphenol A(80-05-7) 2,2-Bis(4-hydroxyphenyl)propane 2,2-Bis(p-hydroxyphenyl)propane 2,2-Di(4-hydroxyphenyl)propane 4,4'-(1-Methylethylidene)bisphenol 4,4'-Isopropylidenediphenol 4,4'-Methylethylidendbisphenol B 0494 BPA 154 BPA 157 Bis(p-hydroxyphenyl)propane Bisphenol A Dian Diano Diphenylolpropane HT 3082 Hidorin F 285 Ipognox 88 Isopropylidenebis(4-hydroxybenzene) NSC 1767 NSC 17959 Parabis Phenol, 4,4'-(1-methylethylidene)bis- (9CI) p,p'-Bisphenol A p,p'-Dihydroxydiphenylpropane p,p'-Isopropylidenediphenol	100.000 ng/L	120% Rec	12.90 % RSD
Cholesterol(57-88-5) (-)-Cholesterol 3beta-Hydroxycholest-5-ene 5:6-Cholesten-3beta-ol Cholest-5-en-3-ol (3beta)- (9CI) Cholest-5-en-3beta-ol Cholesterin Cholesterol Cholesterol (8CI) Cholesteryl alcohol Dythol Lidinit Lidinite NSC 8798 Provitamin D delta5-Cholesten-3beta-ol	200.000 ng/L	83% Rec	10.10 % RSD
Diethylstilbestrol(56-53-1) 3,4-bis(p-Hydroxyphenyl)-3-hexene 4,4'-(1,2-Diethyl-1,2-ethenediyl)bisphenol (E) 4,4'-Dihydroxy-.alpha.,.beta.-diethylstilbene Agostilben Antigestil Bufon DES Diestryl Diethylstilbesterol, Diethylstilbestrol Diethylstilbestrol Domestrol Phenol, 4,4'-(1,2-diethyl-1,2-ethenediyl)bis, Phenol, 4,4'-(1,2-diethyl-1,2-ethenediyl)bis, (E)- Sexocretin Stilbenediol,alpha,alpha'-diethyl-4,4'- Stilbestrol Synthoestrin Tylosterone Vagestrol alpha, alpha'-Diethylstilbenediol trans-Diethylstilbestrol	0.800 ng/L	95% Rec	6.60 % RSD
Dihydrotestosterone(521-18-6) 17beta-Hydroxy-5alpha-androstan-3-one 5-alpha-Androstan-3-one, 17-beta-hydroxy- Anaboleen Anabolex Andractim Androlone Androstan-17beta-ol-3-one Androstanolone Cristerona MB DHT Dihydrotestosterone Neodrol Proteina Protona Stanaprol Stanolon Stanolone	4.000 ng/L	98% Rec	20.50 % RSD
Epitestosterone(481-30-1) 17-alpha-Hydroxyandrost-4-en-3-one 17-alpha-Testosterone Androst-4-en-3-one, 17-alpha-hydroxy- EPI Epitestosterone Isotestosterone	2.000 ng/L	98% Rec	16.60 % RSD
Equilenine(517-09-9) (+)-Equilenin 3-Hydroxyestra-1,3,5(10),6,8-pentaen-17-one Equilenin (6CI) Equilenine Estra-1,3,5,7,9-pentaen-17-one, 3-hydroxy- (8CI,9CI) NSC 9901 d-Equilenin	2.000 ng/L	94% Rec	15.20 % RSD
Equilin(474-86-2) 1,3,5,7-Estratetraen-3-ol-17-one 3-Hydroxyestra-1,3,5(10),7-tetraen-17-one 7-Dehydroestrone Equilin Equilin (6CI,7CI) NSC 10971	8.000 ng/L	91% Rec	17.40 % RSD
Estriol(50-27-1) 13beta-Methyl-1,3,5(10)-gonatriene-3,16alpha,17beta-triol 16alpha,17beta-Estriol 16alpha-Hydroxyestradiol Aacifemine Estra-1,3,5(10)-triene-3,16,17-triol, (16alpha,17beta)- Estriol Estriol (8CI) Follicular hormone hydrate Oestriol Overstin Ovestin Ovestrion Stiptanon Synapause Theelol Thulol Tridestrin Trihydroxyestrin Triodurin Triovex	2.000 ng/L	88% Rec	28.40 % RSD
Estrone(53-16-7) (+)-Estrone; delta1,3,5(10)-Estratrien-3-ol-17-one 1,3,5(10)-Estratrien-3-ol-17-one 3-Hydroxy-17-keto-estra-1,3,5-triene 3-Hydroxyestra-1,3,5(10)-trien-17-one 3-Hydroxyestra-1,3,5(10)-triene-17-one 3-Hydroxyoestra-1,3,5(10)-trien-17-one Aquacrine Cristallovar Crystogen Destrone Endofolliculina Estra-1,3,5(10)-trien-17-one, 3-hydroxy- (9CI) Estrone Estrone (8CI) Follicular hormone Glandubolin Ketohydroxyestrin NSC 9699 Oestrin Ovex Perlatan Solliculin Theelin Thelestrin Thelykinin Thynestron Tokokin Unden WAY 164397 Wynestron	0.800 ng/L	103% Rec	5.60 % RSD
Ethynylestradiol(57-63-6) 17-ethynyl- 17alpha-Ethinyl-17beta-estradiol 17alpha-Ethynylestra-1,3,5(10)-triene-3,17beta-diol 19-Nor-17alpha-pregna-1,3,5(10)-trien-20-yne-3,17beta-diol Ethynylestradiol NSC 10973 neo-Estrone	0.800 ng/L	99% Rec	7.10 % RSD
Mestranol(72-33-3) 17-alpha-19-Norpregna-1,3,5(10)-trien-20-yn-17 17-alpha-19-Norpregna-1,3,5(10)-trien-20-yn-17-ol, 3 ... 17-alpha-Ethynylestradiol 3-methyl ether 19-Norpregna-1,3,5(10)-trien-20-yn-17-ol, 3-methoxy- ... 3-Methoxy-19-nor-17-alpha-pregna-1,3,5(10)-trien-20- ... Devocin Inostral Menophase Mestranol Norquen Ovastol	0.800 ng/L	98% Rec	7.50 % RSD
Norethindrone(68-22-4) 19-Nor-17-alpha-pregn-4-en-20-yn-3-one, 17-hydroxy- Ciclovulan Noresthisterone Norethindrone Norgestin Orlest Ortho-Novum SC 4640 Synphase Utovlar	0.800 ng/L	98% Rec	7.70 % RSD
Progesterone(57-83-0) 17alpha-Hydroxy-6alpha-methylpregn-4-ene-3,20-dione Agolutin Corlutina Gesterol Gestron Glanducorpin Gynlutin Gynoluton Luteol Lutin Pregn-4-ene-3,20-dione Progesterone Progestosol Syngesterone	8.000 ng/L	91% Rec	36.10 % RSD
Testosterone(58-22-0) 17-beta-Hydroxy-delta(sup 4)-androsten-3-one AndroGel Andrusol Homosterone Mertestate Orquisteron Testobase Testosterone Virosterone	1.600 ng/L	98% Rec	14.50 % RSD

Precision Descriptor Notes:	Accuracy: Fortification level was 10 nanograms per liter (ng/L) for 17 analytes, 100 ng/L for bisphenol A, 320 or 1,000 ng/L for 3-beta-coprostanol, and 1,000 ng/L for cholesterol, assuming a 0.5-liter sample volume. Isotope-dilution standards were fortified at 100 ng/L, except cholesterol-d7, which was fortified at 10,000 ng/L. Precision: Fortification level was 25 nanograms per liter (ng/L) for 17 analytes, 250 ng/L for bisphenol A, 800 or 2,500 ng/L for 3-beta-coprostanol, and 2,500 ng/L for cholesterol assuming a 0.5-L sample volume. Isotope-dilution standards were fortified at 100 ng/L, except cholesterol-d7, which was fortified at 10,000 ng/L.
Detection Level Note:	Detection levels were derived statistically by fortifying reagent water at six different levels (0.1 to 4 ng/L) and range from about 0.4 to 4 ng/L for 16 analytes. Interim reporting levels applied to analytes in this report range from 0.8 to 8 ng/L. Bisphenol A and the sterols (cholesterol and 3-beta-coprostanol) were consistently detected in laboratory and field blanks. The minimum reporting levels were set at 100 ng/L for bisphenol A and at 200 ng/L for the two sterols to prevent any bias associated with the presence of these compounds in the blanks. A minimum reporting level of 2 ng/L was set for 11-ketotestosterone to minimize false positive risk from an interfering siloxane compound emanating as chromatographic-column bleed, from vial septum material, or from other sources at no more than 1 ng/L.

Precision Descriptor Notes:

Accuracy: Fortification level was 10 nanograms per liter (ng/L) for 17 analytes, 100 ng/L for bisphenol A, 320 or 1,000 ng/L for 3-beta-coprostanol, and 1,000 ng/L for cholesterol, assuming a 0.5-liter sample volume. Isotope-dilution standards were fortified at 100 ng/L, except cholesterol-d7, which was fortified at 10,000 ng/L. Precision: Fortification level was 25 nanograms per liter (ng/L) for 17 analytes, 250 ng/L for bisphenol A, 800 or 2,500 ng/L for 3-beta-coprostanol, and 2,500 ng/L for cholesterol assuming a 0.5-L sample volume. Isotope-dilution standards were fortified at 100 ng/L, except cholesterol-d7, which was fortified at 10,000 ng/L.

Detection Level Note:

Detection levels were derived statistically by fortifying reagent water at six different levels (0.1 to 4 ng/L) and range from about 0.4 to 4 ng/L for 16 analytes. Interim reporting levels applied to analytes in this report range from 0.8 to 8 ng/L. Bisphenol A and the sterols (cholesterol and 3-beta-coprostanol) were consistently detected in laboratory and field blanks. The minimum reporting levels were set at 100 ng/L for bisphenol A and at 200 ng/L for the two sterols to prevent any bias associated with the presence of these compounds in the blanks. A minimum reporting level of 2 ng/L was set for 11-ketotestosterone to minimize false positive risk from an interfering siloxane compound emanating as chromatographic-column bleed, from vial septum material, or from other sources at no more than 1 ng/L.

Revision	PDF/Link
2012

The Water Quality Portal contains sample results from sites that use this method.

Download WQP data

Select data:

Sites only
Sample results only

File format:

Comma-separated
Tab-separated
MS Excel 2007+

Sites where this method was used

Loading Sites ...

USGS-NWQL: O-4434-12: Steroid Hormones in Unfiltered Water by GC-MS/MS

WHO WE ARE

LEGAL