Method Development and Validation for the Gc-fid Assay of 17 -estradiol in Pharmaceutical Preparation

A simple and rapid method for determination of 17 -estradiol in pharmaceutical preparation was developed and validated using gas chromatography with flame ionization detection (GC-FID). The solutions of standard and the sample were prepared in methanol. GC separation was performed in about 7.7 min using a 30 m x 0.32 mm I.D. (film thickness 0.25 μm) HP-5 capilary column. Nitrogen was used as carrier gas at a flow-rate of 2 ml min. After injection of the sample at inlet temperature 250 C, the temperature of the GC oven was as follows: initial temperature was 150 C, held for 1.5 min, increased to 260 C at a rate of 50 C min held for 5 min, and finally to 270 C at a rate of 10 C min and held for 3.3 min. Detector temperature is 300 C. 2 μl was injected in splitless mode. Calibration curves were linear between the concentration range 0.25-50 g ml. The method was validated for specificity, linearity, precision, accuracy and limit of quantitation. Also, the method was applied to directly and easily to the analysis of the pharmaceutical preparation (Estrofem tablet).