Analytical Sciences

Simultaneous Doping Analysis of Main Urinary Metabolites of Anabolic Steroids in Horse by Ion-trap Gas Chromatography-Tandem Mass Spectrometry

Masayuki YAMADA,* Sugako ARAMAKI,* Masahiko KUROSAWA,* Isao KIJIMA-SUDA,* Koichi SAITO,** and Hiroyuki NAKAZAWA**

*Laboratory of Racing Chemistry, 1731-2 Tsuruta, Utsunomiya, Tochigi 320-0851, Japan
**Department of Analytical Chemistry, Hoshi University, 2-4-41 Ebara, Tokyo 142-8501, Japan

The use of anabolic steroids in racehorses is strictly regulated. We have developed a method for the simultaneous analysis of 11 anabolic steroids: fluoxymesterone, 17α-methyltestosterone, mestanolone, methandienone, methandriol, oxymetholone, boldenone, furazabol, methenolone, nandrolone, and stanozolol, for possible application to a doping test in racehorses. We selected 15 kinds of target substances for a doping test from the main metabolites of these anabolic steroids, and established a method for simultaneous analysis. Urine was hydrolyzed and subjected to solid-phase extraction. Then, the residue from the extracts was derivatized by trimethylsilylation. The derivatized samples were subjected to ion-trap gas chromatography-tandem mass spectrometry, and their mass chromatograms and product ion spectra were obtained. The limit of detection of the target substances was 5 - 50 ng/mL, and the mean recovery and coefficient of variation were 71.3 - 104.8% and 1.1 - 9.5%, respectively.

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