Abstract

DEVELOPMENT AND VALIDATION OF STABILITY-INDICATING RP-UPLC METHOD FOR SIMULTANEOUS ESTIMATION OF THIOCOLCHICOSIDE AND KETOPROFEN IN COMBINED DOSAGE FORM

Balan Paramasivam, Kannappan Nagappan

Abstract

A stability indicating RP-UPLC method was developed and validated for the simultaneous determination of Thiocolchicoside (TCC) 8mg and Ketoprofen (KTP) 100mg in tablet dosage form. The chromatographic separation was Carried out by Thermo Scientific UPLC Instrument, Accela 1250 Pump, auto sampler with PDA detector, using Thermo Scientific hypersil gold C18 column, (50 x 2.1mm) particle size 1.9μm using 0.005M KH2PO4 buffer, methanol and acetonitrile in the ratio of 40:52:08, pH was adjusted to 3 with ortho phosphoric acid as mobile phase at a flow rate of 250 μl/min with the detection at 258nm. The run time of TCC and KTP was about 0.59 and 1.01 minutes respectively. The detector response is linear from 4.8-7.2 μg/ml and 64-96 μg/ml concentrations for TCC and KTP respectively. The linear regression equation was found to be y = 14654x-100.29 (r2 = 0.9999) for TCC and y= 4098.3x +325.38 (r2 = 0.9999) for KTP. The detection limit and quantification limit was 0.19 μg and 0.58 μg for TCC and 0.4 μg and 1.3 μg for KTP. The percentage of assay for TCC and KTP was about 99.90% and 99.71% respectively. The stability indicating capability was established by forced degradation experiments. The method was satisfactorily validated as per the ICH guidelines.

Keywords: Thiocolchicoside; Ketoprofen; Stability indicating; Method development; RP-UPLC; Validation.