Comparison of analytical approaches for the detection of oral testosterone undecanoate administration in men.
IRMS
antidoping
serum analysis
steroid esters
steroid profile
Journal
Drug testing and analysis
ISSN: 1942-7611
Titre abrégé: Drug Test Anal
Pays: England
ID NLM: 101483449
Informations de publication
Date de publication:
11 Jan 2024
11 Jan 2024
Historique:
revised:
14
12
2023
received:
30
10
2023
accepted:
18
12
2023
medline:
11
1
2024
pubmed:
11
1
2024
entrez:
11
1
2024
Statut:
aheadofprint
Résumé
For antidoping laboratories, the determination of an illicit testosterone (T) administration in urine samples remains a difficult process as it requires the determination of the exogenous origin by carbon isotope ratios (CIRs) of testosterone and its metabolites. As a complement to the urinary analysis, targeting testosterone esters (e.g. testosterone undecanoate [TU]) in serum samples by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) could represent a simpler approach compared with isotope ratio mass spectrometry (IRMS). These two approaches both lead to the direct detection of the administration of exogenous T but with a difference in effort and complexity of the analysis. To compare the detection window obtained with the two strategies, serum and the corresponding urine samples collected from an administration study with oral TU were analysed. Results showed that, at all timepoints where the intact TU was detected in serum, the CIRs of urinary steroids were also not in agreement with an endogenous origin. IRMS analysis required more effort but resulted in slightly longer detection windows than the ester analysis. Finally, this comparison study showed that, in the presence of a suspicious urinary steroid profile, the LC-MS/MS steroid esters analysis in the corresponding serum samples can be very helpful. If steroid esters are not detected, the IRMS analysis can then be conducted on the urine sample afterwards. Overall, the combination of matrices might facilitate the detection of prohibited T administration in sports, especially for athletes with naturally low T/E ratios.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : World Anti-Doping Agency
ID : 21A07RN
Informations de copyright
© 2024 John Wiley & Sons Ltd.
Références
Piper T, Geyer H, Haenelt N, Huelsemann F, Schaenzer W, Thevis M. Current insights into the steroidal module of the athlete biological passport. Int J Sports Med. 2021;42(10):863-878. doi:10.1055/a-1481-8683
Sottas PE, Robinson N, Rabin O, Saugy M. The athlete biological passport. Clin Chem. 2011;57(7):969-976. doi:10.1373/clinchem.2011.162271
WADA. Technical Document - TD2023APMU 2023; https://www.wada-ama.org/sites/default/files/2022-11/td2023apmu_eng_final.pdf. Accessed 28.11.2023.
Becchi M, Aguilera R, Farizon Y, Flament MM, Casabianca H, James P. Gas chromatography/combustion/isotope-ratio mass spectrometry analysis of urinary steroids to detect misuse of testosterone in sport. Rapid Commun Mass Spectrom. 1994;8(4):304-308. doi:10.1002/rcm.1290080404
Zhang Y, Tobias HJ, Sacks GL, Brenna JT. Calibration and data processing in gas chromatography combustion isotope ratio mass spectrometry. Drug Test Anal. 2012;4(12):912-922. doi:10.1002/dta.394
Cawley AT, Flenker U. The application of carbon isotope ratio mass spectrometry to doping control. J Mass Spectrom. 2008;43(7):854-864. doi:10.1002/jms.1437
Strahm E, Emery C, Saugy M, Dvorak J, Saudan C. Detection of testosterone administration based on the carbon isotope ratio profiling of endogenous steroids: international reference populations of professional soccer players. Br J Sports Med. 2009;43(13):1041-1044. doi:10.1136/bjsm.2009.058669
Brooker L, Cawley A, Drury J, Edey C, Hasick N, Goebel C. Stable carbon isotope ratio profiling of illicit testosterone preparations - domestic and international seizures. Drug Test Anal. 2014;6(10):996-1001. doi:10.1002/dta.1533
Cawley A, Collins M, Kazlauskas R, et al. Stable isotope ratio profiling of testosterone preparations. Drug Test Anal. 2010;2(11-12):557-567. doi:10.1002/dta.175
Hullstein I, Sagredo C, Hemmersbach P. Carbon isotope ratios of nandrolone, boldenone, and testosterone preparations seized in Norway compared to those of endogenously produced steroids in a Nordic reference population. Drug Test Anal. 2014;6(11-12):1163-1169. doi:10.1002/dta.1745
Piper T, Thevis M. Investigations in carbon isotope ratios of seized testosterone and boldenone preparations. Drug Test Anal. 2022;14(3):514-518. doi:10.1002/dta.3120
WADA. Technical Document - TD2022IRMS 2022; https://www.wada-ama.org/sites/default/files/2022-01/td2022irms_v1.0_final_eng_0_0.pdf. Accessed 09.08.2023.
de la Torre X, Colamonici C, Curcio D, Molaioni F, Botre F. A comprehensive procedure based on gas chromatography-isotope ratio mass spectrometry following high performance liquid chromatography purification for the analysis of underivatized testosterone and its analogues in human urine. Anal Chim Acta. 2012;756:23-29. doi:10.1016/j.aca.2012.10.013
Piper T, Emery C, Thomas A, Saugy M, Thevis M. Combination of carbon isotope ratio with hydrogen isotope ratio determinations in sports drug testing. Anal Bioanal Chem. 2013;405(16):5455-5466. doi:10.1007/s00216-013-6949-3
Lalonde K, Barber A, Ayotte C. Two-dimensional high performance liquid chromatography purification of underivatized urinary testosterone and metabolites for compound-specific stable carbon isotope analysis. Drug Test Anal. 2021;13(3):558-570. doi:10.1002/dta.2964
Honesova L, Van Eenoo P, Polet M. A uniform sample preparation procedure for gas chromatography combustion isotope ratio mass spectrometry for all human doping control relevant anabolic steroids using online 2/3-dimensional liquid chromatography fraction collection. Anal Chim Acta. 2021;1168:338610. doi:10.1016/j.aca.2021.338610
Casilli A, Piper T, de Oliveira FA, et al. Optimization of an online heart-cutting multidimensional gas chromatography clean-up step for isotopic ratio mass spectrometry and simultaneous quadrupole mass spectrometry measurements of endogenous anabolic steroid in urine. Drug Test Anal. 2016;8(11-12):1204-1211. doi:10.1002/dta.2119
Strahm E, Mullen JE, Garevik N, et al. Dose-dependent testosterone sensitivity of the steroidal passport and GC-C-IRMS analysis in relation to the UGT2B17 deletion polymorphism. Drug Test Anal. 2015;7(11-12):1063-1070. doi:10.1002/dta.1841
Ponzetto F, Mehl F, Boccard J, et al. Longitudinal monitoring of endogenous steroids in human serum by UHPLC-MS/MS as a tool to detect testosterone abuse in sports. Anal Bioanal Chem. 2016;408(3):705-719. doi:10.1007/s00216-015-9185-1
Salamin O, Nicoli R, Langer T, et al. Longitudinal evaluation of multiple biomarkers for the detection of testosterone gel administration in women with normal menstrual cycle. Drug Test Anal. 2022;14(5):833-850. doi:10.1002/dta.3040
Equey T, Salamin O, Ponzetto F, et al. Longitudinal profiling of endogenous steroids in blood using the athlete biological passport approach. J Clin Endocrinol Metab. 2023;108(8):1937-1946. doi:10.1210/clinem/dgad085
Langer T, Salamin O, Nicoli R, Grabherr S, Kuuranne T, Musenga A. A comprehensive UHPLC-MS/MS method for the analysis of endogenous and exogenous steroids in serum for anti-doping purposes. Drug Test Anal. 2022;14(11-12):1904-1919. doi:10.1002/dta.3379
Makvandi B, Pohanka A, Bergstrom M, et al. Detection of anabolic androgenic steroids in serum samples. Drug Test Anal. 2023;15(6):678-688. doi:10.1002/dta.3476
Forsdahl G, Erceg D, Geisendorfer T, et al. Detection of testosterone esters in blood. Drug Test Anal. 2015;7(11-12):983-989. doi:10.1002/dta.1914
Van Renterghem P, Viaene W, Van Gansbeke W, et al. Validation of an ultra-sensitive detection method for steroid esters in plasma for doping analysis using positive chemical ionization GC-MS/MS. J Chromatogr B Analyt Technol Biomed Life Sci. 2020;1141:122026. doi:10.1016/j.jchromb.2020.122026
Gray BP, Teale P, Pearce CM. Analysis of methyloxime derivatives of intact esters of testosterone and boldenone in equine plasma using ultra high performance liquid chromatography tandem mass spectrometry. Drug Test Anal. 2011;3(4):206-213. doi:10.1002/dta.237
Solheim SA, Levernaes MCS, Morkeberg J, et al. Stability and detectability of testosterone esters in dried blood spots after intramuscular injections. Drug Test Anal. 2021;14(11-12):1926-1937. doi:10.1002/dta.3030
de la Torre X, Iannone M, Botre F. Improving the detection of anabolic steroid esters in human serum by LC-MS. J Pharm Biomed Anal. 2021;194:113807. doi:10.1016/j.jpba.2020.113807
Forsdahl G, Vatne HK, Geisendorfer T, Gmeiner G. Screening of testosterone esters in human plasma. Drug Test Anal. 2013;5(11-12):826-833. doi:10.1002/dta.1560
Jing J, Shan Y, Liu Z, et al. Automated online dried blood spot sample preparation and detection of anabolic steroid esters for sports drug testing. Drug Test Anal. 2022;14(6):1040-1052. doi:10.1002/dta.3226
Badoud F, Boccard J, Schweizer C, Pralong F, Saugy M, Baume N. Profiling of steroid metabolites after transdermal and oral administration of testosterone by ultra-high pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. J Steroid Biochem Mol Biol. 2013;138:222-235. doi:10.1016/j.jsbmb.2013.05.018
WADA. Technical Document - TD2022EAAS 2021; https://www.wada-ama.org/sites/default/files/2022-01/td2021eaas_final_eng_v_2.0.pdf. Accessed 09.08.2023.
WADA. Technical Document - TD2022MRPL. 2022; https://www.wada-ama.org/sites/default/files/resources/files/td2022mrpl_v1.0_final_eng.pdf
Geyer H, Fussholler G, Haenelt N, et al. The ratio 5aAdiol/E as indicator for the detection of testosterone doping in athletes with naturally low T/E - a case study. In: Thevis M, Geyer H, Mareck U, eds. Proceedings of the Manfred Donike Workshop: 38th Cologne Workshop on Dope Analysis 9th to 14th Feb 2020. Sportverlag Strauß; 2020:90-93.
Thigpen AE, Silver RI, Guileyardo JM, Casey ML, McConnell JD, Russell DW. Tissue distribution and ontogeny of steroid 5 alpha-reductase isozyme expression. J Clin Invest. 1993;92(2):903-910. doi:10.1172/JCI116665
Mullen J, Borjesson A, Hopcraft O, et al. Sensitivity of doping biomarkers after administration of a single dose testosterone gel. Drug Test Anal. 2018;10(5):839-848. doi:10.1002/dta.2341
Nair VS, Husk J, Miller GD, van Eenoo P, Crouch A, Eichner D. Evaluation of longitudinal steroid profiling with the ADAMS adaptive model for detection of transdermal, intramuscular, and subcutaneous testosterone administration. Drug Test Anal. 2020;12(10):1419-1431. doi:10.1002/dta.2885
Schulze J, Suominen T, Bergstrom H, Ericsson M, Bjorkhem Bergman L, Ekstrom L. Urinary steroid profile in relation to the menstrual cycle. Drug Test Anal. 2021;13(3):550-557. doi:10.1002/dta.2960
Peng S-H, Segura J, Farré M, González JC, de la Torre X. Plasma and urinary markers of oral testosterone undecanoate misuse. Steroids. 2002;67(1):39-50. doi:10.1016/S0039-128X(01)00128-3
Piper T, Haenelt N, Fussholler G, Geyer H, Thevis M. Sensitive detection of testosterone and testosterone prohormone administrations based on urinary concentrations and carbon isotope ratios of androsterone and etiocholanolone. Drug Test Anal. 2021;13(11-12):1835-1851. doi:10.1002/dta.3168
Piper T, Emery C, Saugy M. Recent developments in the use of isotope ratio mass spectrometry in sports drug testing. Anal Bioanal Chem. 2011;401(2):433-447. doi:10.1007/s00216-011-4886-6
Piper T, Mareck U, Geyer H, et al. Determination of 13C/12C ratios of endogenous urinary steroids: method validation, reference population and application to doping control purposes. Rapid Commun Mass Spectrom. 2008;22(14):2161-2175. doi:10.1002/rcm.3601
WADA. Laboratory Guidelines - Steroids in Blood for the ABP 2023; https://www.wada-ama.org/sites/default/files/2023-07/2023_laboratory_guidelines_quantification_endogenous_steroids_blood_final_v1_0.pdf. Accessed 09.08.2023.
Nair VS, Sharpe K, Husk J, et al. Evaluation of blood parameters by linear discriminant models for the detection of testosterone administration. Drug Test Anal. 2021;13(7):1270-1281. doi:10.1002/dta.3017
Houwing NS, Maris F, Schnabel PG, Bagchus WM. Pharmacokinetic study in women of three different doses of a new formulation of oral testosterone undecanoate. Andriol Testocaps Pharmacotherapy. 2003;23(10):1257-1265. doi:10.1592/phco.23.12.1257.32707
Okano M, Shiomura S. Effectiveness of blood steroidal passport markers for detecting testosterone abuse in Asians. Drug Test Anal. 2023;1-9. doi:10.1002/dta.3588
Forsdahl G, Ostreicher C, Koller M, Gmeiner G. Carbon isotope ratio determination and investigation of seized testosterone preparations. Drug Test Anal. 2011;3(11-12):814-819. doi:10.1002/dta.373