Targeted Metabolomics Approach to Detect the Misuse of Steroidal
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Steroid Profiling in Urine of Intact Glucuronidated and Sulfated
CORE Metadata, citation and similar papers at core.ac.uk Provided by Ghent University Academic Bibliography Journal of Chromatography A 1624 (2020) 461231 Contents lists available at ScienceDirect Journal of Chromatography A journal homepage: www.elsevier.com/locate/chroma Steroid profiling in urine of intact glucuronidated and sulfated steroids using liquid chromatography-mass spectrometry ∗ Laurie De Wilde , Kris Roels, Pieter Van Renterghem, Peter Van Eenoo, Koen Deventer 1 Doping Control Laboratory (DoCoLab), Ghent University (UGent), Department Diagnostic Sciences, Technologiepark 30B, B-9052 Zwijnaarde, Belgium a r t i c l e i n f o a b s t r a c t Article history: Detection of endogenous anabolic androgenic steroids (EAAS) misuse is a major challenge in doping con- Received 26 November 2019 trol analysis. Currently, a number of endogenous steroids, which constitute the steroid profile, are quanti- Revised 6 May 2020 fied using gas chromatography (GC). With this methodology, only the sum of the free and glucuronidated Accepted 10 May 2020 steroids is measured together. A dilute-and-shoot LC-MS method, which is compliant with the quality Available online 23 May 2020 requirements for measuring EAAS established by the World Anti-Doping Agency (WADA), was devel- Keywords: oped and validated containing glucuronidated and sulfated steroids in order to gain some extra infor- Doping mation and to expand the existing steroid profile. The developed method is, to the best of our knowl- Urine edge, the first method to combine both steroid glucuronides and sulfates, which is compliant with the Steroid profile quality standards of the technical document on EAAS, established by WADA. -
Development and Application of Methods for Extraction and LC/MS/MS Analysis of Sex Steroids and Conjugates from Fish Feces
Development and Application of Methods for Extraction and LC/MS/MS Analysis of Sex Steroids and Conjugates from Fish Feces By Lisa E. Peters A Thesis submitted to the Faculty of Graduate Studies of The University of Manitoba in partial fulfilment of the requirements of the degree of Doctor of Philosophy Department of Environment and Geography University of Manitoba Winnipeg Copyright © 2014 Lisa E. Peters 1 Acknowledgments I would like to start by thanking my supervisor, Dr. Gregg Tomy, for his support, patience and resources during my Ph.D. program. He took a chance when I tried to convince him that my biology background would be a great addition to his chemistry lab. My husband, Vince Palace, and I also appreciated his support and genuine happiness for us when I announced we were expecting a baby in the middle of my studies. A sincere thanks also goes to my co-supervisor, Dr. Mark Hanson, for his support on so many levels (I wouldn’t even know where to start), and for doing his best to keep me on track, which was no small task. I would also like to thank my other thesis committee members, Drs. Gary Anderson and Feiyue Wang, for their encouragement and insightful comments. Gary gave a lot of extra time to review various thesis chapters and data, and to lend his expertise during the fish surgeries. I also had the pleasure of taking his Endocrinology course, which was probably the most informative and enjoyable class of my entire university student career. I would like to thank the technical staff and students from DFO, Suzanne Mittermuller, Kerry Wautier, Alea Goodmanson, Danielle Godard, Lisa Friedrich and Kirstin Dangerfield, and my lab mates Bruno Rosenberg, Kerri Pleskach, Colin Darling, Bonnie Gemmill and Lianna Bestvater for their technical input and help during my experiments. -
Androstenedione
NTP TECHNICAL REPORT ON THE TOXICOLOGY ANd CARCINOGENESIS STUdIES OF ANdROSTENEdIONE (CAS NO. 63-05-8) IN F344/N RATS ANd B6C3F1 MICE (GAVAGE STUdIES) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 September 2010 NTP TR 560 NIH Publication No. 10-5901 National Institutes of Health Public Health Service U.S. dEPARTMENT OF HEALTH ANd HUMAN SERVICES FOREWORd The National Toxicology Program (NTP) is an interagency program within the Public Health Service (PHS) of the Department of Health and Human Services (HHS) and is headquartered at the National Institute of Environmental Health Sciences of the National Institutes of Health (NIEHS/NIH). Three agencies contribute resources to the program: NIEHS/NIH, the National Institute for Occupational Safety and Health of the Centers for Disease Control and Prevention (NIOSH/CDC), and the National Center for Toxicological Research of the Food and Drug Administration (NCTR/FDA). Established in 1978, the NTP is charged with coordinating toxicological testing activities, strengthening the science base in toxicology, developing and validating improved testing methods, and providing information about potentially toxic substances to health regulatory and research agencies, scientific and medical communities, and the public. The Technical Report series began in 1976 with carcinogenesis studies conducted by the National Cancer Institute. In 1981, this bioassay program was transferred to the NTP. The studies described in the Technical Report series are designed and conducted to characterize and evaluate the toxicologic potential, including carcinogenic activity, of selected substances in laboratory animals (usually two species, rats and mice). Substances selected for NTP toxicity and carcinogenicity studies are chosen primarily on the basis of human exposure, level of production, and chemical structure. -
Analysis of Anabolic Steroid Glucuronide and Sulfate Conjugates
Applicability of an innovative steroid-profiling method to determine synthetic growth promoter abuse in cattle Blokland, M.H.; Tricht, E.F. van; Ginkel L.A. van; Sterk, S.S. This is a "Post-Print" accepted manuscript, which has been published in “Catena” This version is distributed under a non-commencial no derivatives Creative Commons (CC-BY-NC-ND) user license, which permits use, distribution, and reproduction in any medium, provided the original work is properly cited and not used for commercial purposes. Further, the restriction applies that if you remix, transform, or build upon the material, you may not distribute the modified material. Please cite this publication as follows: Blokland, M.H.; Tricht, E.F. van; Ginkel L.A. van; Sterk, S.S. (2017) Applicability of an innovative steroid-profiling method to determine synthetic growth promoter abuse in cattle. The Journal of Steroid Biochemistry and Molecular Biology 174, p. 265-275 You can download the published version at: https://doi.org/10.1016/j.jsbmb.2017.10.007 Applicability of an innovative steroid-profiling method to determine synthetic growth promoter abuse in cattle 5 M.H. Blokland*, E.F. van Tricht, L.A van Ginkel, S.S. Sterk RIKILT Wageningen University & Research, P.O. Box 230, Wageningen, The Netherlands 10 *Corresponding author: M.H. Blokland, Tel.: +31 317 480417, E-mail: [email protected] 15 Keywords: synthetic steroids, growth promoters, cattle, UHPLC-MS/MS, steroid profiling, steroidogenesis Abstract 20 A robust LC-MS/MS method was developed to quantify a large number of phase I and phase II steroids in urine. -
The Metabolism of Androstenone and Other Steroid Hormone Conjugates in Relation to Boar Taint
The Metabolism of Androstenone and Other Steroid Hormone Conjugates in Relation to Boar Taint by Heidi M. Laderoute A Thesis presented to The University of Guelph In partial fulfillment of requirements for the degree of Master of Science in Animal and Poultry Science with Toxicology Guelph, Ontario, Canada © Heidi M. Laderoute, April, 2015 ABSTRACT THE METABOLISM OF ANDROSTENONE AND OTHER STEROID HORMONE CONJUGATES IN RELATION TO BOAR TAINT Heidi M. Laderoute Advisor: University of Guelph, 2015 Dr. E.J. Squires Increased public interest in the welfare of pigs reared for pork production has led to an increased effort in finding new approaches for controlling the unpleasant odour and flavour from heated pork products known as boar taint. Therefore, this study investigated the metabolism of androstenone and the enzymes involved in its sulfoconjugation in order to further understand the pathways and genes involved in the development of this meat quality defect. Leydig cells that were incubated with androstenone produced 3-keto- sulfoxy-androstenone, providing direct evidence, for the first time, that sulfoconjugation of this steroid does occur in the boar. In addition, human embryonic kidney cells that were overexpressed with porcine sulfotransferase (SULT) enzymes showed that SULT2A1, but not SULT2B1, was responsible for sulfoconjugating androstenone. These findings emphasize the importance of conjugation in steroid metabolism and its relevance to boar taint is discussed. ACKNOWLEDGEMENTS I would like to gratefully and sincerely thank my advisor, Dr. E. James Squires, for providing me with the opportunity to be a graduate student and for introducing me to the world of boar taint. This project would not have been possible without your guidance, encouragement, and patience over the last few years. -
Sports Drug Testing and Toxicology TOP ARTICLES SUPPLEMENT
Powered by Sports drug testing and toxicology TOP ARTICLES SUPPLEMENT CONTENTS REVIEW: Applications and challenges in using LC–MS/MS assays for quantitative doping analysis Bioanalysis Vol. 8 Issue 12 REVIEW: Current status and recent advantages in derivatization procedures in human doping control Bioanalysis Vol. 7 Issue 19 REVIEW: Advances in the detection of designer steroids in anti-doping Bioanalysis Vol. 6 Issue 6 Review For reprint orders, please contact [email protected] 8 Review 2016/05/28 Applications and challenges in using LC–MS/MS assays for quantitative doping analysis Bioanalysis LC–MS/MS is useful for qualitative and quantitative analysis of ‘doped’ biological Zhanliang Wang‡,1, samples from athletes. LC–MS/MS-based assays at low-mass resolution allow fast Jianghai Lu*,‡,2, and sensitive screening and quantification of targeted analytes that are based on Yinong Zhang1, Ye Tian2, 2 ,2 preselected diagnostic precursor–product ion pairs. Whereas LC coupled with high- Hong Yuan & Youxuan Xu** 1Food & Drug Anti-doping Laboratory, resolution/high-accuracy MS can be used for identification and quantification, both China Anti-Doping Agency, 1st Anding have advantages and challenges for routine analysis. Here, we review the literature Road, ChaoYang District, Beijing 100029, regarding various quantification methods for measuring prohibited substances in PR China athletes as they pertain to World Anti-Doping Agency regulations. 2National Anti-doping Laboratory, China Anti-Doping Agency, 1st Anding Road, First draft submitted: -
United States Patent (19) 11 Patent Number: 6,068,830 Diamandis Et Al
US00606883OA United States Patent (19) 11 Patent Number: 6,068,830 Diamandis et al. (45) Date of Patent: May 30, 2000 54) LOCALIZATION AND THERAPY OF FOREIGN PATENT DOCUMENTS NON-PROSTATIC ENDOCRINE CANCER 0217577 4/1987 European Pat. Off.. WITH AGENTS DIRECTED AGAINST 0453082 10/1991 European Pat. Off.. PROSTATE SPECIFIC ANTIGEN WO 92/O1936 2/1992 European Pat. Off.. WO 93/O1831 2/1993 European Pat. Off.. 75 Inventors: Eleftherios P. Diamandis, Toronto; Russell Redshaw, Nepean, both of OTHER PUBLICATIONS Canada Clinical BioChemistry vol. 27, No. 2, (Yu, He et al), pp. 73 Assignee: Nordion International Inc., Canada 75-79, dated Apr. 27, 1994. Database Biosis BioSciences Information Service, AN 21 Appl. No.: 08/569,206 94:393008 & Journal of Clinical Laboratory Analysis, vol. 8, No. 4, (Yu, He et al), pp. 251-253, dated 1994. 22 PCT Filed: Jul. 14, 1994 Bas. Appl. Histochem, Vol. 33, No. 1, (Papotti, M. et al), 86 PCT No.: PCT/CA94/00392 Pavia pp. 25–29 dated 1989. S371 Date: Apr. 11, 1996 Primary Examiner Yvonne Eyler S 102(e) Date: Apr. 11, 1996 Attorney, Agent, or Firm-Banner & Witcoff, Ltd. 87 PCT Pub. No.: WO95/02424 57 ABSTRACT It was discovered that prostate-specific antigen is produced PCT Pub. Date:Jan. 26, 1995 by non-proStatic endocrine cancers. It was further discov 30 Foreign Application Priority Data ered that non-prostatic endocrine cancers with Steroid recep tors can be stimulated with Steroids to cause them to produce Jul. 14, 1993 GB United Kingdom ................... 93.14623 PSA either initially or at increased levels. -
An Investigation Into the Influence of Rooibos (Aspalathus Linearis) on Androgen Metabolism in Normal and Prostate Cancer Cells
AN INVESTIGATION INTO THE INFLUENCE OF ROOIBOS (ASPALATHUS LINEARIS) ON ANDROGEN METABOLISM IN NORMAL AND PROSTATE CANCER CELLS by Therina du Toit Thesis presented in fulfilment of the requirements for the degree of Master of Science in the Faculty of Science at Stellenbosch University Supervisor: Prof AC Swart Co-supervisor: Dr K-H Storbeck March 2015 Stellenbosch University https://scholar.sun.ac.za Declaration By submitting this thesis electronically, I declare that the entirety of the work contained therein is my own, original work, that I am the sole author thereof (save to the extent explicitly otherwise stated), that reproduction and publication thereof by Stellenbosch University will not infringe any third party rights and that I have not previously in its entirety or in part submitted it for obtaining any qualification. T du Toit March 2015 Copyright © 2015 Stellenbosch University All rights reserved ii Stellenbosch University https://scholar.sun.ac.za Abstract In this study, the influence of rooibos on the catalytic activity of enzymes 17β-hydroxysteroid dehydrogenase type 3 (17βHSD3), 17β-hydroxysteroid dehydrogenase type 5 (AKR1C3), 17β-hydroxysteroid dehydrogenase type 2 (17βHSD2), 5α-reductase type 1 (SRD5A1) and 5α-reductase type 2 (SRD5A2), which catalyse prostate androgen metabolism, was investigated. The activities of both 17βHSD3 and AKR1C3 heterologously expressed in CHO-K1 and HEK293 cells were inhibited significantly by rooibos, with rooibos reducing the conversion of androstenedione (A4) and 11keto-androstenedione (11KA4) to testosterone (T) and 11keto- testosterone (11KT), respectively. The catalytic activity of 17βHSD2 towards T, 11hydroxy- testosterone (11OHT) and 11KT was also significantly inhibited by rooibos in transiently transfected HEK293 cells. -
Increased UDP-Glucuronosyltransferase Activity and Decreased Prostate Specific Antigen Production by Biochanin a in Prostate Cancer Cells Xiao-Ya Sun, Cathie A
[CANCER RESEARCH 58. 2379-2384, June 1, 1998| Increased UDP-glucuronosyltransferase Activity and Decreased Prostate Specific Antigen Production by Biochanin A in Prostate Cancer Cells Xiao-Ya Sun, Cathie A. Plouzek, James P. Henry,1 Thomas T. Y. Wang, and James M. Phang2 Laboratory of Nutritional and Molecular Regulation, National Cancer Institute-Frederick Cancer Research and Development Center. NIH. Frederick. Maryland 21702-1201 ABSTRACT Because androgen levels in the prostate may be clinically impor tant, the modulation of UDPGT and androgen metabolism by dietary Our laboratory has characterized androgen metabolism in an andro- factors is of considerable interest. Various studies have shown that gen-responsive prostate cancer cell line (LNCaP) and showed that these consumption of soybean is associated with low incidence of prostate cells accumulated intracellular testosterone primarily as glucuronidated metabolites. Using a cell-free assay with testosterone as substrate, we cancer in Asian populations (6). Flavonoids, a family of compounds showed that LNCaP had UDP-glucuronosyltransferase (UDPGT) activity. plentiful in fruits and vegetables and especially in soy products, may Because dietary factors, such as flavonoids in soy products, may reduce reduce the risk for hormone-dependent cancers (7). We undertook the the risk for hormone-dependent cancers, we studied the effects of fla current studies to examine the possible effects of flavonoids on vonoids on testosterone-UDPGT activity. LNCaP cells were exposed to testosterone-UDPGT activity and on the metabolism of testosterone in selected flavonoids for up to 6 days. The increase in UDPGT-specific prostate cancer cells. We found that UDPGT was induced by fla activity was linear over this period. -
OATP-Mediated Hepatic Uptake of Glucuronide Metabolites of Androgens
Molecular Pharmacology Fast Forward. Published on June 25, 2020 as DOI: 10.1124/mol.120.119891 This article has not been copyedited and formatted. The final version may differ from this version. MOL#119891 Title Page OATP-mediated hepatic uptake of glucuronide metabolites of androgens Cindy Yanfei Li, Anshul Gupta, Zsuzsanna Gáborik, Emese Kis, and Bhagwat Prasad* Department of Pharmaceutics, University of Washington, Seattle, WA, USA (CYL) Downloaded from Amgen Research, Department of Pharmacokinetics and Drug Metabolism, Cambridge, MA, USA 02142 (AG) molpharm.aspetjournals.org SOLVO Biotechnology, Budapest, Hungary (ZG and EK) Department of Pharmaceutical Sciences, Washington State University, Spokane, WA, USA (BP) at ASPET Journals on September 29, 2021 1 Molecular Pharmacology Fast Forward. Published on June 25, 2020 as DOI: 10.1124/mol.120.119891 This article has not been copyedited and formatted. The final version may differ from this version. MOL#119891 Running Title Page Running title: Hepatic uptake of glucuronide metabolites of androgens *To whom correspondence should be addressed: Bhagwat Prasad, Ph.D., Department of Pharmaceutical Sciences, Washington State University, 412 E Spokane Falls Blvd, Spokane, WA 99202, Email: [email protected]. Phone: 509-358-7739; Fax: 509-368-6561. Downloaded from Number of text pages: 34 Number of tables: 2 molpharm.aspetjournals.org Number of figures: 4 Number of references: 51 at ASPET Journals on September 29, 2021 Number of words in the Abstract: 246 Number of words in the Introduction: -
Analysis of 19-Norsteroids, Testosterone and Precursors Metabolites in Human Urine”
PROJECT REVIEW “Analysis of 19-Norsteroids, Testosterone and Precursors Metabolites in Human Urine” C. Ayotte (Institut Armand-Frapier (INRS), Québec, Canada), W. Schänzer (Deutsche Sporthochschule, Köln Insitut für Biochemie, Köln, Germany), M. Ueki (Tokyo Antidoping Laboratory, Japan) Detecting the use of androgenic anabolic steroids, potentially endogenous in humans, and which are prohibited substances in sport doping control programmes, still represents a major challenge to the analysts. These steroids include testosterone, its precursors androstenedione and dehydroepiandrosterone, and the 19-norsteroids equivalents, some of which are commercially available for oral self- administration. This project is aimed at applying the IRMS (isotope ratio mass spectrometry) to the detection of the “natural” testosterone and 19-nortestosterone anabolic agents. Complement of the existing GC/MS methods, the GCIC/IRMS permits the differentiation of the exogenous or endogenous origin of urinary androgens metabolites, by measuring the isotopic content of their carbon atoms. That novel applcation of an otherwise well-known technique, now requires its validation in different experienced laboratories, the determination of international reference ranges of the metabolites of endogenous origin and the documentation of the changes commonly observed following the administration of these steroids. Ultimately, one of its most direct outcomes will be to verify that although different methodological approaches and different equipment are used, the same determination is made from the analysis of common specimens. Testosterone and 19-nortestosterone (nandrolone) are potent androgenic anabolic steroids of known abuse in sport. Anabolic steroids are banned in Olympic sports for more than 20 years and since 1986, the highest number of positive cases reported are due to these two steroids. -
Sex Hormones and Metabolites
Accession # 00268796 Male Sample Report 123 A Street Sometown, CA 90266 Sex Hormones and Metabolites Ordering Physician: DOB: 1967-08-09 Collection Times: Precision Analytical Age: 50 2017-08-09 06:01AM 2017-08-09 08:01AM Gender: Male 2017-08-09 05:01PM 2017-08-09 10:01PM Category Test Result Units Normal Range Progesterone Metabolites (Urine) b-Pregnanediol Low end of range 110.0 ng/mg 75 - 400 a-Pregnanediol Low end of range 40.0 ng/mg 20 - 130 Estrogens and Metabolites (Urine) Estrone(E1) High end of range 15.3 ng/mg 4 - 16 Estradiol(E2) Above range 2.4 ng/mg 0.5 - 2.2 Estriol(E3) Within range 4.2 ng/mg 2 - 8 2-OH-E1 Within range 3.73 ng/mg 0 - 5.9 4-OH-E1 Above range 1.2 ng/mg 0 - 0.8 16-OH-E1 Within range 0.7 ng/mg 0 - 1.2 2-Methoxy-E1 Within range 2.1 ng/mg 0 - 2.8 2-OH-E2 Above range 0.61 ng/mg 0 - 0.6 4-OH-E2 Within range 0.2 ng/mg 0 - 0.3 2-Methoxy-E2 Within range 0.6 ng/mg 0 - 0.8 Androgens and Metabolites (Urine) DHEA-S Low end of range 95.0 ng/mg 30 - 1500 Androsterone Low end of range 835.0 ng/mg 500 - 3000 Etiocholanolone Below range 387.0 ng/mg 400 - 1500 Testosterone Below range 21.6 ng/mg 25 - 115 5a-DHT Low end of range 8.2 ng/mg 5 - 25 5a-Androstanediol Low end of range 52.0 ng/mg 30 - 250 5b-Androstanediol Low end of range 46.0 ng/mg 40 - 250 Epi-Testosterone Low end of range 38.1 ng/mg 25 - 115 Precision Analytical (Raymond Grimsbo, Lab Director) Male Sample Report Page 1 of 6 3138 Rivergate Street #301C FINAL REPORT CLIA Lic.