In the course of investigations on equestrian supplemental products for the presence of doping substances, two products were found to contain forbidden substances. As reported earlier a plant extract (Mexican cactus extract) named “Energy 5” contained the anabolic androgenic steroids (AAS) stanozolol, 17β-hydroxy-17α-methyl-5α-androstane-3β-ol (3β,5α-THMT) as well as mestanolone not declared on the label. In the present study, a product called “Super Kalm Paste” was tested. Analysis by gas chromatography - mass spectrometry (GC-MS) revealed that the preparation contained the class I anti-arrhythmics quinine (trade names KinidinTM, Durules) and cinchonine. The samples were prepared according to a sample preparation procedure established for anabolic steroids in nutritional supplements for humans. The sample treatment comprised the extraction and purification of the analytes as well as the chemical conversion with N-methyl-N-trimethylsilyl-trifluoracetamide (MSTFA) to yield the trimethylsilyl (TMS)-derivatives. To verify whether the administration of such products could lead to positive doping tests, a pilot excretion study on “Energy 5” was conducted with two geldings, and urine samples were collected. Gas chromatography - high resolution mass spectrometry (GC-HRMS) after solid phase extraction and mixed derivatisation has demonstrated the presence of the stanozolol metabolite 16β-hydroxy-stanozolol in urine samples after “Energy 5” application.

After the appearance of steroid contaminations in nutritional supplements for humans a plant essence for the treatment of horses containing anabolic steroids has been made out. However, this product claims to be an alternative to anabolic steroids. Two charges of the oily plant extract (Mexican Cactus Extract) were prepared according to a sample preparation procedure established for anabolic steroids in nutritional supplements for humans. The sample treatment comprised the extraction and purification of the analytes as well as the chemical conversion with N-methyl-N-trimethylsilyltrifluoracetamide (MSTFA) to yield the trimethylsilyl-(TMS)-derivatives of the steroids. The analysis by gas chromatography / mass spectrometry (GC / MS) revealed that both samples contained the androgenic anabolic steroids (AAS) stanozolol, 17beta-hydroxy-17alpha-methyl-5alpha-androstane-3beta-ol (5alpha-THMT) as well as mestanolone without being declared on the label. The concentration of stanozolol which has been identified as the main component of the detected steroids was estimated in the range of 10 mug per ml of the oily plant extract.