A combination of cyproterone acetate (2 mg) and ethinylestradiol (35 mg) is commonly used for the treatment of androgen-dependent acne, seborrhoea, alopecia and hirsutism in females and as oral contraceptive in patients with androgenic symptoms.2) The combination of these drugs is also used in polycystic ovary syndrome.3,4) Literature survey in well-known Pharmacoepias showed no official procedure for simultaneous determination of cyproterone acetate and ethinylestradiol in pharmaceutical preparations. Direct UV-visible spectrophotometric method is not suitable for simultaneous determination of cyproterone acetate and ethinylestradiol due to their spectral overlapping. Application of derivative technique of spectrophotometry offers a powerful tool for quantitative analysis of multi-component mixtures. [5][6][7][8][9] In the last decades, this technique has rapidly gained application in the field of pharmaceutical analysis to overcome the problem of interference, due to substances other than analytes, commonly present in pharmaceutical formulations or for combination of two or more drug substances. [10][11][12][13][14][15][16][17] Lack of any published method for simultaneous spectrophotometric determination of cyproterone acetate and ethinylestradiol, therefore, provoked us to investigate the application of derivative spectrophotometry for simultaneous determination of these compounds in pharmaceutical dosage forms using zero-crossing method.
ExperimentalMaterials and Methods Cyproterone acetate and ethinylestradiol were from Cipex (Handelsges, Wien, Austria) and obtained from Abouraihan Pharmaceutical Company (Tehran, Iran). Methanol was of analytical grade and obtained from Merck (Darmstadt, Germany).Apparatus Absorption and derivative spectra were recorded in 1 cm quartz cells using a Shimadzu UV-160 double beam UV-visible spectrophotometer (Shimadzu, Kyoto, Japan) with a fixed bandwidth (2 nm) and data processing capacity. The zero-order absorption spectra were recorded over the wavelength range 200-400 nm, against a solvent blank. The derivative spectra were obtained over the same wavelength range at different slit width (Dl). The ordinate, maximum and minimum, was adjusted to the magnitude of derivative values.Standard and Calibration Solutions Standard stock solution of cyproterone acetate was prepared by dissolving 100 mg of cyproterone acetate in 100 ml methanol. Two stock solutions of ethinylestradiol were prepared by dissolving 100 and 35 mg of ethinylesradiol in 100 ml methanol respectively. Two other solutions of ethinylestradiol (1 mg/100 ml, 3.5 mg/100 ml) were prepared from the stock solutions.Accurate volumes of the standard stock solutions were transferred into two sets of 100 ml calibrated flasks. The first series contained a constant quantity of ethinylestradiol (35 mg/100 ml) and varying concentrations of cyproterone acetate (0.5-6 mg/100 ml). The second series contained a constant amount of cyproterone acetate (2 mg/100 ml) and varying concentrations of ethinylestradiol (10-80 mg/100 ml).Ph...