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Steroid profile analysis and UGT2B17 genotyping of the same urine sample to determine testosterone abuse

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Abstract

When testing a urine sample for testosterone abuse, a ratio of testosterone glucuronide (T) to epitestosterone glucuronide (ET) of 4.0 or above is considered suspicious. A degree of variation, however, has been observed in T/ET ratio between individuals from both the same and different ethnic backgrounds. The majority of this variation might be due to UGT2B17 deletion genotype (UGT2B17 deletion-type). The aim of this study was to investigate the use of the same urine sample for the analysis of T/ET ratio and UGT2B17 deletion-type. Japanese men were deletion-typed via a UGT2B17 copy number assay using DNA from blood. Urinary T and ET levels were determined using gas chromatography–mass spectrometry before (n = 112) and after a testosterone injection (n = 25). Basal T level and the increase in T/ET ratio after injection were dependent on UGT2B17 deletion-type, being lower in subjects with deletion (del/del) than nondeletion (ins/del or ins/ins) genotype. UGT2B17 deletion-typing was first performed using DNA from urine cryopreserved for 1–1.5 years (n = 66). The concentration of DNA required for discrimination between the deletion and nondeletion genotype by copy number assay was more than 0.1 ng/ml urine. Discrimination was possible in 94.0 % of urine samples (5–7 ml each). These findings show that T/ET ratio and UGT 2B17 deletion-type can be analyzed exclusively via urine samples, removing the need for the collection of other samples, such as blood or buccal cells. The combination of T/ET ratio and UGT 2B17 deletion-type may help inform decisions regarding a genotype-specific T/ET cutoff ratio.

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Acknowledgments

This study was entirely supported by a grant from the Japan Sport Council.

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There are no financial or other relations that could lead to a conflict of interest.

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Authors and Affiliations

  1. Anti-doping Research Laboratory, Japan Chemical Analysis Center, 6-39-4 Minami Senju, Arakawa-ku, Tokyo, 116-0003, Japan

    Kimiko Aoki, Hirofumi Shinohara, Hiroyuki Tanaka & Makoto Ueki

  2. Nihon Pharmaceutical University, 10281 Komuro, Inamachi, Kitaadachi-gun, Saitama, 362-0806, Japan

    Kimiko Aoki

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  1. Kimiko Aoki
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  2. Hirofumi Shinohara
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  3. Hiroyuki Tanaka
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  4. Makoto Ueki
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Correspondence to Kimiko Aoki.

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Aoki, K., Shinohara, H., Tanaka, H. et al. Steroid profile analysis and UGT2B17 genotyping of the same urine sample to determine testosterone abuse. Forensic Toxicol 32, 194–200 (2014). https://doi.org/10.1007/s11419-013-0220-7

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  • DOI: https://doi.org/10.1007/s11419-013-0220-7

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