Validated High Performance liquid Chromatographic Method for Estimation of Thiocolchicoside, and its Application to Ex-vivo Permeation Studies for Transdermal Therapeutic Systems
P. Gayatri*1, M. Ajitha2, P. Pavan Kumar3, Y Madhusudan Rao4
A simple, accurate and sensitive high performance liquid chromatography (HPLC) method for estimation of thiocolchicoside (THC) in skin permeated samples was developed and validated. The chromatographic separation was accomplished on a C 18 column (Inertsil ODS 3V C-18, 250 × 4.6 mm, 5 μ )using a mixture of Buffer: Methanol (45:55) as mobile phase. The flow rate was fixed at 1.5 mL/min. The linearity range for the proposed method was 0.25- 50μg/mL. Photodiode array detector was set at a wavelength of 264nm to monitor the elution. The LOD and LOQ for thiocolchicoside were 0.05μg/mL and 0.125μg/mL respectively. The correlation coefficient (‘r’2) values were found to be >0.996 and % RSD values for intra-day and inter-day were found to be less than 10%. The method was validated for linearity, specificity, accuracy, precision, robustness, LOD and LOQ as per ICH guidelines and was suitable for estimation of thiocolchicoside in ex-vivo skin diffusate samples and transdermal patches.
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