RP-HPLC METHOD DEVELOPMENT AND VALIDATION USINGEXPERIMENTAL DESIGN APPROACH FOR SIMULTANEOUS ESTIMATION OF THIOCOLCHICOSIDE AND DEXKETOPROFEN WITHFORCED DEGRADATION STUDIES

Abstract

Introduction: In the present work, sensitive RP-HPLC method been developed for the
quantitative estimation of Thiocolchicoside (THC) and Dexketoprofen (DKP) combined dosage
form.
Material & methods: Determination of THC and DKP was carried on a reverse phase C18
(250×4.6mm, 5µ) column using a mobile phase consisting of Methanol: Sodium Phosphate Buffer
(70:30 v/v) pH 4.5, Flow rate of 1.0 ml/min and the detection was carried out at 280 nm.
Result: The linearity was found to be in the range of 5-40 μg/ml and 30-240 μg/ml with
(r2=0.9991, and r2=0.9992) for THC and DKP respectively. The sharp peaks obtained were having
clear baseline separation with a retention time of 3.20 min and 8.40 min for THC and DKP
respectively.
Discussion: The forced degradation studies performed in acidic, basic, oxidative, photolytic and
thermal conditions at different time intervals. The method was validated as per the International
Conference on Harmonization (ICH) guidelines as well as for the ROBUSTNESS studies the
Quality by Design approach used based on 3-Level Factorial Design of Experiment. On the basis
of Designs the three chromatographic parameters (Flow Rate, pH and Mobile phase composition)
were changed and study were carried out with effects on Retention time and Peak Area.
Conclusion: The proposed validated method was successfully used for the quantitative analysis of
commercially available dosage form.