Detection of stanozolol in hair by negative ion chemical ionization mass spectrometry.
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 degrees 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-trimethylsilylhepta-fluorobutryamide (MSHFBA) and trimethylsilylimidazole (TSIM) (1000:20, v/v) was added, and the mixture heated at 80 degrees C for 5 minutes. After cooling to room temperature, N-methyl-bisheptafluorobutyramide (MBHFBA) was added and the mixture heated at 80 degrees C for 30 min. The derivatized extracts were analyzed on a Finnigan MATTM 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 x 0.2-mm i.d.; 33-microns 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.
Utrecht Institute of Pharmaceutical Sciences (UIPS), Utrecht University, Department of Analysis and Toxicology, The Netherlands. K.M.Hold@far.ruu.nl
K M
KM
Höld
D G
DG
Wilkins
D J
DJ
Crouch
D E
DE
Rollins
R A
RA
Maes
D000577:Amides / Q000737:chemistry; D045930:Anabolic Agents / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D005260:Female; D005466:Fluorocarbons; D008401:Gas Chromatography-Mass Spectrometry; D006197:Hair / Q000737:chemistry*; D007093:Imidazoles / Q000737:chemistry; D007202:Indicators and Reagents / Q000737:chemistry; D007274:Injections, Intraperitoneal; D008297:Male; D051381:Rats; D012015:Reference Standards; D015203:Reproducibility of Results; D012972:Sodium Hydroxide / Q000737:chemistry; D018629:Spectrometry, Mass, Secondary Ion; D013197:Stanozolol / Q000008:administration & dosage / Q000032:analysis*; D013696:Temperature; D014297:Trimethylsilyl Compounds / Q000737:chemistry
D000577:Amides; D045930:Anabolic Agents; D005466:Fluorocarbons; D007093:Imidazoles; D007202:Indicators and Reagents; C103252:N-methyl-N-trimethylsilylheptafluorobutyramide; C103253:N-methyl-bis(heptafluorobutyramide); D014297:Trimethylsilyl Compounds; C054025:1-(trimethylsilyl)-1H-imidazole; D013197:Stanozolol; D012972:Sodium Hydroxide