QUANTITATION OF 17-ESTRADIOL IN RABBIT PLASMA BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH FLUORESCENCE DETECTION

A new method was developed for the determination of 17 -estradiol in rabbit plasma by reversed-phase high performance liquid chromatography (RP-HPLC) with fluorescence (FL) detection. The method employed one-step extraction of 17 -estradiol from plasma matrix with a mixture of water and ethylacetate (1.9:5, v/v) using estriol as an internal standard. The mobile phase consisted of a mixture of metanol and water (70:30, v/v) flowing at a flow rate of 1.0mLmin1. Calibration curve was linear between concentration range of 1256000ngmL1. Average recovery of 17 -estradiol and the internal standard from the biological matrix was more than 94.9 and 92.5%, respectively. The intra-day and inter-day precision and accuracy were between 3.748.12 and 3.728.80%, respectively. Also, the method was successfully applied to determine of 17 -estradiol in New Zealand white rabbits.