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Development and validation of a subcritical fluid extraction and high performance liquid chromatography assay for medroxyprogesterone in aquatic products

Han, Yuqian, Ma, Qinchuan, Lu, Jie, Xue, Yong, Xu, Jie, Xue, Changhu
Journal of chromatography 2012 v.897 pp. 90-93
high performance liquid chromatography, medroxyprogesterone, response surface methodology, temperature, wavelengths
A simple, rapid and sensitive method was developed for the determination of medroxyprogesterone in aquatic products by extraction with subcritical 1,1,1,2-tetrafluoroethane (R134a) and high performance liquid chromatography (HPLC). A response surface methodology (RSM) was adopted to optimise extraction pressure, temperature and co-solvent volume. The optimum extraction conditions predicted within the experimental ranges were as follows: pressure, 3MPa; temperature, 25°C; and co-solvent volume, 6ml. The analysis was carried out on Zorbax SB-C₁₈ column (4.6mm×150mm, 5μm) with the mobile phase acetonitrile–water (55:45, v/v), flow rate 1.0ml/min, temperature 30°C and wavelength 240nm. Good linearity of detection was obtained for medroxyprogesterone between concentrations of 50–250ng/ml, r²=0.999. The method was validated using samples fortified with medroxyprogesterone at levels of 10, 30 and 50ng/g, the mean recovery exceeds 90%, and the RSD values were less than 10%.