Journal Article

Detection of Stanozolol in Hair by Negative Ion Chemical Ionization Mass Spectrometry

Karin M. Höld, Diana G. Wilkins, Dennis J. Crouch, Douglas E. Rollins and Robert A. Maes

in Journal of Analytical Toxicology

Volume 20, issue 6, pages 345-349
Published in print October 1996 | ISSN: 0146-4760
e-ISSN: 1945-2403 | DOI:
Detection of Stanozolol in Hair by Negative Ion Chemical Ionization Mass Spectrometry

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Stanozolol is an anabolic androgenic steroid occasionally abused by athletes. A sensitive, specific, and reproducible method for the quantitative determination of stanozolol in hair has been developed. After the addition of stanozolol-d3 as the internal standard, hair samples (10–25 mg) were digested with 2 mL of 1N NaOH at 65°C for at least 2 h. Digest solutions were then extracted using solid-phase extraction. The eluents were evaporated, a mixture of N-methyl-N-trimethylsilylheptafluorobutryamide (MSHFBA) and trimethylsilylimidazole (TSIM) (1000:20, v/v) was added, and the mixture heated at 80°C for 5 minutes. After cooling to room temperature, N-methyl-bisheptafluorobutyramide (MBHFBA) was added and the mixture heated at 80°C for 30 min. The derivatized extracts were analyzed on a Finnigan MAT™ 4500 mass spectrometer in the negative chemical ionization mode. Chromatographic separation was achieved with helium carrier gas on a HP-1 capillary column (15 m × 0.2-mm i.d.; 33-µm film thickness). The assay was capable of reliably quantitating 50 pg/mg of stanozolol and was linear to 2500 pg/mg. Intra-assay precision was 13.2% at 50 pg/mg and 6.6% at 2500 pg/mg. Interassay precision was 13.7% at 50 pg/mg and 6.1% at 2500 pg/mg. This method has been applied to the analysis of stanozolol incorporated into rat hair. Male Long-Evans rats were given stanozolol 20 mg/kg intraperitoneally once daily for 3 days. The mean concentrations of stanozolol in the rat hair collected on day 14 were 362.4 ± 332.4 pg/mg in pigmented hair and 90.0 ± 46.9 pg/mg in nonpigmented hair. These data demonstrate that stanozolol is incorporated preferentially into pigmented hair.

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Subjects: Medical Toxicology ; Toxicology (Non-medical)

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