Simultaneous Estimation of Rifampicin And Isoniazid by RP-HPLC

Abstract

A New Simple, Accurate, Precise, Robust And Specific Rp-Hplc Method Has Been Developed And Validated For The Simultaneous Estimation Of Rifampicin And Isoniazid In Pure Form And Marketed Pharmaceutical Formulation. Chromatographic Analysis Of Rifampicin And Isoniazid In The Present Assay Was Carried Out Isocratically With A Symmetry C18 (4.6mm×150mm, 5.0 µm) Particle Size Column And Mobile Phase Consisting Of Methanol: Acetonitrile V/V In The Ratio Of 45:55v/V. The Flow Rate Was 1.0ml/Min. The Effluent Was Monitored At A Wavelength Of 265nm At 36ºc Temperature With A Retention Time Of Rifampicin And Isoniazid Was Found To Be 2.343min And 3.288min Respectively. The Proposed Rp-Hplc Method Was Validated According To ICH Guidelines With Respect To Specificity, Precision, Linearity And Accuracy. The Percentage Rsd For Precision And Accuracy Of The Proposed Method For Rifampicin And Isoniazid Was Found To Be Less Than 2%. The Inter-Day And Intra-Day Precisions Were Found To Be Within Limits. The Method Was Linear Over The Concentration Range Of 30- 70μg/Ml (R2 -0.999), With Limits Of Detection And Quantification Of 2.65μg/Ml & 7.95μg/Ml For Rifampicin (R2 -0.999) And The Method Was Linear Over The Concentration Range Of (10-50μg/Ml) (R2 -0.999) Limits Of Detection And Quantification Of 3.4μg/Ml & 10.2μg/Ml For Isoniazid (R 2 -0.999). The Proposed Rp-Hplc Method Was Successfully Applied For Routine Analysis Of Rifampicin And Isoniazid In Pure Form And Marketed Pharmaceutical Formulation