Determination by High Performance Liquid Chromatography of Fluoxymesterone, Methyltestosterone and Testosterone in Adulterated Chinese Herbal Preparations

Yoe-Ray Ku, Ming-Jer Tsai and Kuo-Ching Wen*

National Laboratories of Foods and Drugs, Department of Health, Executive Yuan, Republic of China

ABSTRACT

   A Chinese herbal preparation was assayed simultaneously by high performance liquid chromatography on an ODS column for the presence of adulterated three androgens, fluoxymesterone, methyltestosterone and testosterone. This chromatographic method was carried out using isocratic elution with water and acetonitrile (3:2). Calibration graphs of the three adulterants were constructed in the range 10-160μg/ml and their correlation coefficients were in the range 0.9997-0.9999. When these three androgens were added to Ba-Jen-Tang,the recoveries were 87.7% for fluoxymesterone, 99.3% for methyltestosterone and 91.1 % for testosterone. The relative standard deviations for intraday and interday analyses were 0.31-2.01 % and 0.95-5.11 % (n=6), respectively. Four kinds of capsules labeled snake penis (Samples A-D) were consigned during Aug. 1995 and Jul. 1996. The methyltestosterone in the samples was assayed by the HPLC method described above. The recoveries of methyltestosterone in samples A, B, C and D were 95.4, 83.3, 92.7 and 88.8%, respectively. The content of methyltestosterone in these capsules ranged from 1.5 to 4.0% (7.5-19.7 mg in each capsule). Caffeine, diprophylline and three water soluble vitamins (ascorbic acid, nicotinamide and thiamine) were also detected in Samples A and B, and thiamine was detected in Samples C and D. The identity of these chemicals was confirmed by TLC and UV.

Key words： HPLC, Chinese herbal preparation, fluoxymesterone, methyltestosterone, testosterone, adulterant, androgen