METHOD DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF THIOCOLCHICOSIDE AND DICLOFENAC IN BULK AND ITS PHARMACEUTICAL FORMULATIONS

Abstract

A rapid and sensitive Reverse Phase High Performance Liquid Chromatographic [RP-HPLC] method was developed for the estimation of Thiocolchicoside and Aceclofenac 1,2,3 in pure and its tablet dosage forms. The method was validated as per International Conference on Harmonization [ICH] guidelines 35 . A C18 column (250×4.6mm, 5μm) was used with a mobile phase containing a mixture of potassium phosphate monohydrate buffer (pH-5.0): Acetonitrile: Methanol in the ratio of 40:20:40 % v/v. The analysis was performed with run time of 6 minutes at a flow rate of 1ml/min. The Thiocolchicoside and Aceclofenac was monitored at 263nm with UV detection and Thiocolchicoside and Aceclofenac was eluted at 2.8 min and 4.2 min. The method was linear (r2 =0.999) at concentration ranging from 7.5 to 25μg/ml for Thiocolchicoside and 100-300µg/ml for Aceclofenac, precise (intra-day relative standard deviation [RSD] and inter-day RSD values < 1.0%), accurate (99.3 to 100.9 for Thiocolchicoside and 100.1 to 100.6 for Aceclofenac), specific and robust. Detection limit of 0.82 for Thiocolchicoside and 8.73 μg/ml for Aceclofenac. Similarly quantification limits were 2.49 for Thiocolchicoside and 26.45 for Aceclofenac μg/ml, estimated from linearity by regression respectively. The results showed that the proposed method is suitable for the precise, accurate and rapid determination of Thiocolchicoside and Aceclofeanc in bulk, its combined dosage forms.