Abstract

VALIDATED SIMULTANEOUS HPLC ASSAY FOR RIFAMPICIN, ISONIAZID AND PIPERINE FORMULATION

Sachin Shivling Bhusari*, Subendu Kumar Brahma, Avinash Narayan Chaudhary, Pravin Shridhar Wakte

ABSTRACT

A sensitive HPLC method for separation and estimation of rifampicin (RIF), piperine (PIP), an oral bioavailability enhancer of RIF and isoniazid (INH) has been developed. All the analytes were resolved at 0.45 mL/min flow rate on C18 column (2.1 x 50 mm, 3.5 μm particle size) using acetonitrile: phosphate buffer (50 mM, pH 5.0) [35:65 (v/v)]. Adequate resolution of 4.323 and 8.994 was obtained between INH, RIF and PIP. Good linearity was observed (r2 > 0.999) over the studied range of all the analytes. The intra- and inter-day accuracy and precision values for all the analytes were within the acceptable range. The LOQ for INH and RIF was 0.10μg/mL whereas that of PIP was 0.05μg/mL. The developed method was found to be robust. The method was successfully used for the quantitative analysis of marketed formulation containing INH, RIF and PIP.

Keywords: HPLC, rifampicin, piperine, isoniazid.