Elsevier

Steroids

Volume 74, Issue 9, 15 September 2009, Pages 773-778
Steroids

Analysis of anabolic steroids in hair: Time courses in guinea pigs

https://doi.org/10.1016/j.steroids.2009.04.008Get rights and content

Abstract

Sensitive, specific, and reproducible methods for the quantitative determination of eight anabolic steroids in guinea pig hair have been developed using LC/MS/MS and GC/MS/MS. Methyltestosterone, stanozolol, methandienone, nandrolone, trenbolone, boldenone, methenolone and DHEA were administered intraperitoneally in guinea pigs. After the first injection, black hair segments were collected on shaved areas of skin. The analysis of these segments revealed the distribution of anabolic steroids in the guinea pig hair. The major components in hair are the parent anabolic steroids. The time courses of the concentrations of the steroids in hair (except methenolone, which does not deposit in hair) demonstrated that the peak concentrations were reached on days 2–4, except stanozolol, which peaked on day 10 after administration. The concentrations in hair appeared to be related to the physicochemical properties of the drug compound and to the dosage. These studies on the distribution of drugs in the hair shaft and on the time course of their concentration changes provide information relevant to the optimal time and method of collecting hair samples. Such studies also provide basic data that will be useful in the application of hair analysis in the control of doping and in the interpretation of results.

Introduction

Hair analysis is gaining increasing attention for its unique potential in the detection and control of doping [1], [2], [3], [4], [5], [6]. It permits a distinction between a single dose and long-term use, and discrimination between endogenous production and exogenous exposure. Hair analysis is an excellent complement to urine testing, and can substitute for urine in spot-checking for doping. These practical advantages make it likely that hair analysis will eventually experience wide application in the control of doping.

However, there are still some problems with doping analysis using hair analysis [1], [2], [3], [4], [5], [6], [7]. For example, high sensitivity is required for the analytical method because the concentrations of doping agents such as anabolic steroids are found in the picograms per milligram range in hair, whereas opiates or amphetamines are generally found in the nanograms per milligram range. Also, expansion of the drug screening repertoire to hair samples will require the detection and quantitation of different kinds of anabolic steroids. The interpretation of results from hair analysis must be based on further research, for example determining the time period for the appearance of analytical targets after a single-dose administration, and the relationship between detected concentration and dosage. In recent years, investigations have been ongoing, but relatively few publications are available in the field of doping analysis based on hair samples [8], [9], [10], [11], [12], [13].

Building on previous work with a focus on doping detection and control, this study was designed to determine the time courses of deposit of eight anabolic steroids in guinea pig hair, and to investigate the metabolites of anabolic steroids in hair.

Section snippets

Standards and reagents

Methandienone, trenbolone, methenolone, and stanozolol were purchased from Cerilliant (Round Rock, TX, USA). Boldenone, nandrolone, methyltestosterone were kindly provided by Dr. Ehrenstorfer (Augsburg, Germany). Dehydroepiandrosterone (DHEA), 6β-hydroxymethandienone, 3′-hydroxystanozolol, and the internal standards, D3-testosterone, D3-epitestosterone were kindly donated by the China Anti-doping Center (Beijing, China). Melanin and methanol were purchased from Sigma (Deisenhofen, Germany).

Results and discussion

A quantitative, specific assay was needed to support analysis of the steroid compounds from hair samples produced in the controlled animal experiments. The validation data has been reported in our earlier publications [15], [16]. The analytical parameters for LC/MS/MS and GC/MS/MS are shown in Table 4.

Gas chromatography/tandem mass spectrometry (GC/MS/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) generally provide superior LLOQ, sensitivity, and improved selectivity. Because

Conclusion

The sensitive LC/MS/MS and GC/MS/MS methods developed and described here were suitable for the detection and quantification of anabolic steroids in hair samples and may be useful in doping control. Studies on the time courses of the concentrations of eight anabolic steroids in guinea pig hair have demonstrated that methyltestosterone, stanozolol, methandienone, nandrolone, trenbolone, boldenone, and DHEA could incorporate into hair and distribute widely in the hair shaft. The concentrations of

Acknowledgments

The authors would like to express their gratitude to Dr. Moutian Wu, China Anti-doping agency, for his valuable discussion regarding anabolic steroids. This work has been supported by National Natural Science Foundation, PR China (No. 20475037).

References (32)

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