Simultaneous determination of seven sexual hormones in cosmetics by reversed-phase high performance liquid chromatography

An efficient method for analyzing seven sexual hormones in cosmetics, namely, estriol, estradiol-17β, estrone, testosterone, methyl-testosterone, progesterone and diethylstil-bestrol by reversed-phase high performance liquid chromatography was developed. Samples were saponified with 20 g/L sodium hydroxide. The sexual hormones were then extracted with dichloromethane-acetic acetate (40: 1, v/v) under acidic conditions (pH 3, adjusted with 1 mol/LHCl). An XTerra^™RP₁₈ column was employed and a mixture of water-methanol-acetonitrile (50: 32: 18, v/v) was used as mobile phase. The seven sexual hormones were detected at 230 nm. The average spiked recoveries for the seven sexual hormones ranged from 75. 6% to 97. 8% with relative standard deviations of 1.9% to 7.2%. The linear ranges of determination were from 5 to 50 mg/L with correlation coefficients of 0.999 9, and the limits of detection were from 3. 7 to 12 ng. The method is suitable to simultaneously assay the seven sexual hormones in cosmetics with simplicity and accuracy in a single injection.