Multi-laboratory study of the analysis and kinetics of stanozolol and its metabolites in treated calves†

Abstract

The European Union banned the use of anabolic steroids for cattle fattening in 1988. Analytical techniques able to detect trace amounts of the parent drugs and their metabolites are mandatory for the control of abuse. Stanozolol (Stan) is an anabolic steroid that is often found in injection sites and cocktails. However, it has never been detected in tissues (kidney fat, meat) or excreta (urine, faeces) taken during regulatory inspection. The difference between the structure of Stan and the other steroids (a pyrazole ring fused to the androstane ring system) is probably the cause of this phenomenon. In the multi-laboratory study described here, veal calves were treated with intramuscular doses of Stan. In the excreta of these calves the presence, absence and/or concentration of Stan and of its major metabolites 16ß-hydroxystanozolol and 3A-hydroxystanozolol were determined. For the determination of these analytes the different laboratories used different extraction and clean-up procedures and also evaluated different analytical techniques such as GC-MS (negative chemical ionization) and LC-MS-MS. The aim of this investigation was to explore which analyte should be validated for veterinary inspection purposes.

Detection of stanozolol and its metabolites in equine urine by liquid chromatography?electrospray ionization ion trap mass spectrometry

“…These results contrast sharply with the ion trap CID mass spectra of stanozolol and 16␤-hydroxystanozolol published by De Brabander et al [7]. Mück and Henion's intense m/z 81 peaks in each case are below the low-mass cut-off value and are not detected.…”

“…Recently, a number of workers have reported the detection of stanozolol and/or 16␤-hydroxystanozolol at parts per billion levels in bovine urine by electrospray ionization (ESI) ion trap LC-MS [7][8][9]. As this approach is applicable to a wide range of potential stanozolol metabolites with excellent sensitivity and with no requirement for chemical derivatization of the target analytes, we decided to investigate its applicability to equine urine.…”

Urine Fingerprints of Stanozolol Treated Horses by Liquid Chromatography High Resolution Mass Spectrometry

“…For example, stanozolol is behaving differently than other anabolic steroids mainly analyzed by GC/MS. The need for derivatization when using GC-MS was a negative factor in the detection of 16β-OH stanozolol [17] but for the detection of 3'-hydroxystanozolol, it can be sufficient [18]. The detection power of the stanozolol metabolites gave better results using LC/MS.…”

Detection of stanozolol in the urine of athletes at a pg level: The possibility of passive exposure

“…However, it has never been detected in sample tissues (kidney fat, meat) or excreta (urine, faeces) obtained during regular inspection. The difference between the structure of stanozolol and the other steroids (a pyrazole ring fused to the androstane ring system) is probably the cause of this phenomenon (5). The European Union banned the use of anabolic steroids in general for cattle fattening in 1988.…”