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

A new method was developed for the determination of 17 β-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 β-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 1.0 mL min−1. Calibration curve was linear between concentration range of 125–6000 ng mL−1. Average recovery of 17 β-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 3.74–8.12 and 3.72–8.80%, respectively. Also, the method was successfully applied to determine of 17 β-estradiol in New Zealand white rabbits.