Abstract

METHOD DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF ISONIAZID, ETHAMBUTOL HYDROCHLORIDE AND RIFAMPICIN IN BULK AND COMBINED TABLETS DOSAGE FORMS

Jincy Anna Valson* and Dr. Snehalatha Boddu

ABSTRACT

A simple, accurate, precise, rapid, selective and reproducible reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous estimation of Isoniazid, Ethambutol Hydrochloride and Rifampicin in tablet formulation. Good chromatographic separation was achieved isocratically using a Prontosil C18 column (250 x 4.6mm, 5μm) and mobile phase consisting of acetonitrile: 0.02M sodium dihydrogen phosphate buffer (60:40) with 1.5ml of Triethyl Amine , adjusted to pH 6.5 with Orthophosphoric acid, at flow rate 1ml/min. The first method of these three drugs which involves absorbance measurement at 208nm The retention time of Isoniazid, Ethambutol Hydrochloride and Rifampicin and was found to be 2.463min, 2.767min and 3.650min respectively. Linearity was obtained in the range of 8-48 μg/ml,18-63 μg/ml and 32-112 μg/ml respectively. The correlation coefficient for calibration curve of all three peaks was found to be 0.9999. The recoveries of Isoniazid, Ethambutol Hydrochloride and Rifampicin in pharmaceutical preparation were all greater than 98% and their relative standard deviations were not more than 2.0%. The developed methods were validated according to ICH guidelines.

Keywords: Isoniazid, Rifampicin, Ethambutol Hydrochloride, RP-HPLC, Validation