Development and validation of RP-HPLC method for analytical characterization of the anabolic steroid Methenolone acetate in food supplements

Nutritional supplements are concentrated sources of nutrients, vitamins and minerals with a nutritional or physiological effect, the purpose of which is to supplement the normal nutritional balance. A necessity of the health risk associated with their unregulated usage requires development of suitable fast and precise methods for their evaluation. As an answer to this a novel RP-HPLC method for analytical characterization of the anabolic steroid Methenolone acetate in food supplement was developed. The method is based on separation and evaluation performed on an analytical column Kintex 5µm EVO C18 (100 × 4.6 mm) with mobile phase acetonitrile:water = 60:40, v/v; flow rate of 1.0 mL/min and UV-detection at 240 nm and column temperature, at 25 °C. The method has proved to be specific, linear, accurate and precise. The method identified a presence of both Methenolone and Methenolone acetate in the evaluated sample.