Validated Degradation studies for the estimation of Pyrazinamide, Ethambutol, Isoniazid and Rifampacin in a fixed dose combination by UPLC

Abstract

Objectives: The intended study was to develop simple, reproducible, reliable and less time consuming analytical methods for estimation of pyrazinamide, ethambutol ,isoniazid and rifampicin in FDC by UPLC and its validation Materials and Methods: RP-UPLC method for estimation of Pyrazinamide, Isoniazid, Ethambutol and Rifampicin has been developed and validated for determination of compounds in commercial tablet dosage form. The compounds were well separated isocratically on a X Bridge 3x50mm, 3.7 µm C18 column using a mobile phase consisting of Triethylamine and Potassium Dihydrogen Ortho Phosphate pH – 7.4) [Sol A]: Methanol and Acetonitrile in a ratio of 85:15 v/v [ Sol B] in the ratio 90:10,0.5 ml/min flow rate with PDA detector. Results and Conclusions: Retention time for Pyrazinamide, Isoniazid, Ethambutol and Rifampicin was found to be 2.012, 3.120, 4.026, 5.926 min respectively. The method was validated in accordance with ICH guidelines. The study showed that the reverse phased liquid chromatography was sensitive and selective for detecting Pyrazinamide, Isoniazid, Ethambutol, Rifampicin using the single mobile phase.