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Abstract

VALIDATED HPTLC METHOD FOR SIMULTANEOUS ESTIMATION OF RIFAMPICIN, ISONIAZID AND PYRIDOXINE HYDROCHLORIDE IN COMBINED TABLET DOSAGE FORM

*Shajahan Puthusseri, Mary Mathew

ABSTRACT

This present study reports for the first time simultaneous quantification of Rifampicin, Isoniazid and Pyridoxine Hydrochloride in tablet dosage form by employing High Performance Thin Layer Chromatographic method. Chromatographic separation of the drugs were performed on aluminium plates precoated with silica gel 60 G F254 as the stationary phase and the mobile phase used was a mixture of Ethyl acetate: Methanol: Acetone: Acetic acid (5.5: 2.0: 2.0: 0.5, v/v). Densitometric evaluation of the separated zones was performed using a UV detector at 254 nm in absorbance mode. The Rf values for Rifampicin, Isoniazid and Pyridoxine Hydrochloride were found to be 0.27 0.01, 0.47 0.01 and 0.75 0.01 respectively. The calibration Curve was found to be linear between 200 to 1000 ng/spot for each drug. The method was specific because no chromatographic interferences from the tablet excipients were found. The accuracy and reliability of the method was assessed by linearity, precision (intraday % RSD and interday % RSD of Rifampicin, Isoniazid and Pyridoxine Hydrochloride) and specificity in accordance with ICH guidelines. The limits of detection and quantification were found to be 75.28 ng/spot and 88.83 ng/spot for RIF, and 80.20 ng/spot and 102.49 ng/spot for INH, and 118.46 ng/spot and 132.30 ng/spot for PDX, respectively. The developed method can be successfully employed for the simultaneous determination of RIF, INH and PDX in marketed tablet formulation.

Keywords: Simultaneous estimation, Rifampicin (RIF), Isoniazid (INH), Pyridoxine Hydrochloride (PDX), High Performance Thin Layer Chromatography (HPTLC), Densitometry, Validation.


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