development and validation of hplc method for the determination of d-chiro inositol and myo-inositol in pharmaceutical dosage form

Polycystic ovary syndrome (pcos) is one of the most common endocrine disorders in women of reproductive age and is characterized by menstrual abnormalities, clinical or biochemical hyperandrogenism, multiple abnormal cysts and enlarge ovaries [1]. women affected by pcos often suffer of insulin resistance and of a compensatory hyperinsulinemia which put them at risk of developing several metabolic disorders. inositol is a six-carbon polyol which has been characterized as an insulin sensitizer: it exists as nine different isomers and among them d-chiro inositol and myo-inositol are the most represented and studied in physiology and physiopathology. in particular, d-chiro inositol (dci) and myo-inositol (mi) glycans administration has been reported to exert beneficial effects at metabolic, hormonal and ovarian levels.the proposed hplc method is a suitable technique for the simultaneous determination of d-chiro inositol (dci) and myo-inositol (mi) in pharmaceutical dosage form. the development and validation was performed using a liquid chromatographic system which is equipped with ri detector. the chromatographic column used for separation was hector-m nh2, 250×4.6 mm, 3µm. the mobile phase used for the separation was acetonitrile-water (75:25 % v/v). the column temperature was maintained at 55 °c and the detector at 50 °c. the injection volume was 10 µl and the flow rate was 1 ml/min. the rp-hplc method has shown good resolution between d-chiro inositol (dci) and myo-inositol (mi). the method was validated by determining system suitability, linearity, repeatability and intermediate precision, accuracy and robustness in accordance with the guideline of the international conference on harmonization (ich) [2]. the calibration curves obtained were linear (r2=0.999) over the concentration range of 0.5-3 mg/ml and 1.8-10.8 mg/ml for dci and mi respectively. the percent relative standard deviation values (%rsd) in the repeatability and intermediate precision study were less than 1%. the recovery values obtained were 99-101% and 95-96% for dci and mi respectively. the results reveal that the developed method was guileless, commercial, fast and precise analytical method for the routine quantitative determination of d-chiro inositol (dci) and myo-inositol (mi) in pharmaceutical dosage form without any interference from the excipients.