Development and Validation of Spectrophotometric and High-Performance Column Liquid Chromatographic Methods for the Simultaneous Determination of Dienogest and Estradiol Valerate in Pharmaceutical Preparations

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ÇAĞLAYAN M. G., PALABIYIK İ. M., Onur F.

JOURNAL OF AOAC INTERNATIONAL, cilt.93, sa.3, ss.862-868, 2010 (SCI-Expanded, Scopus)

Özet

Simultaneous determination of dienogest (DIE) and estradiol valerate (EST) in sugar-coated tablets was performed by using HPLC and spectrophotometry. In HPLC, the separation was achieved on an ACE C8 column using the mobile phase acetonitrile-NH4NO3 (0.03 M, pH 5.4; 70 + 30, v/v) at a flow rate of 2 mL/min. The detection wavelength was 280 nm, and cyproterone acetate was selected as an internal standard. The linearity range was 3.0-45.0 mu g/mL for DIE and 18.0-100.0 mu g/mL for EST. As spectrophotometric methods, two chemometric methods, principal component regression and partial least-squares, were developed. In the chemometric techniques, the concentration data matrix was prepared by using mixtures containing these drugs in methanol-water (3 + 1, v/v). The absorbance data matrix corresponding to the concentration data matrix in these methods was obtained by the measurement of absorbances in their zero-order spectra; then, the calibration was obtained by using the data matrix for the prediction of unknown concentrations of DIE and EST in their binary mixture. Working ranges were found as 2.0-24.0 mu g/mL for DIE and 20.0-270.0 mu g/mL EST in the methods. These three developed methods were validated and successfully applied to a pharmaceutical preparation, a sugar-coated tablet, and the results were compared with each other.