A SIMPLE AND SENSITIVE RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF HYDROXYCHLOROQUINE SULPHATE AND NITAZOXANIDE IN BINARY COMBINATION

The purpose of the present research was to develop a suitable, simple, precise, accurate, robust and reproducible RP-HPLC method for reliable simultaneous quantification of Hydroxychloroquine sulphate (HCQ) and Nitazoxanide (NTZ) in a synthetic mixture. The samples were assayed by the cyber lab HPLC instrument using a C18 (250mm, × 4.6mm, D., 5µm) –cyber-sil under isocratic conditions. The mobile phase consisted of Phosphate buffer (pH-3 adjusted with 1% OPA): Methanol: Acetonitrile (30:35:35 v/v/v) with a flow rate of 1.5 ml/min. The wavelength for detection was 266nm (Isoabsorptive point). The Retention time of HCQ and NTZ was 2.8 min and 9. 3min, respectively. The method was validated by determining system suitability, selectivity, sensitivity, linearity, inter-day and intra-day precision, accuracy, robustness and stability in accordance with the ICH guidelines. The calibration curve was linear over the concentration range, 2-10μg/ml for HCQ and 5-25μg/ml for NTZ respectively (n=6). The developed chromatographic method proved to be simple, precise, accurate, robust, and reproducible. Moreover, the samples showed stability at room temperature over a period of 48 h. Thus, this method would be employed for routine simultaneous quantification of Hydroxychloroquine sulphate and Nitazoxanide.