Validated HPTLC Method for Simultaneous Quantitation of Aceclofenac, Paracetamol and Thiocolchicoside in Bulk drug and Pharmaceutical Formulation

A simple high performance thin layer chromatographic method for simultaneous quantification of aceclofenac, paracetamol and thiocolchicoside in bulk and tablet dosage form was investigated. Chromatographic separation of the drugs were performed on aluminium plates precoated with silica gel 60 F254 as the stationary phase and the solvent system consisted of chloroform: methanol: ethyl acetate: glacial acetic acid (5: 2.5: 2.5: 0.1, v/v/v/v). Densitometric evaluation of the separated zones was performed at 272 nm and the method was validated. The Rf values and drug content of aceclofenac, paracetamol and thiocolchicoside were 0.52±0.03, 0.72±0.03 and 0.30 ± 0.03 and 100.37%, 100.30% and 100.43% respectively. The calibration curves of peak area versus concentration, which were linear from 50 300 ng/band of aceclofenac, 250 1500 ng/ band of paracetamol and 50 300 ng/band of thiocolchicoside, coefficient of determination (r2) was greater than 0.999. The method was validated for linearity, accuracy, precision, robustness, specificity and application for assay as per ICH guidelines. Keyword: HPTLC, Method development, Validation, Aceclofenac, Paracetamol, Thiocolchicoside, ICH guidelines