Development and Validation of HPTLC Method for Simultaneous Determination of Reserpine and Arjunolic acid in Tensowert Tablet

摘要

Simultaneous quantitative estimation of two biologically active compounds reserpine and arjunolic acid in Tensowert tablet was performed using high-performance thin-layer chromatography. TLC aluminium plates precoated with silica gel 60F-254 (0.2 mm thickness were used. The sample were dissolve in methanol and linear ascending development was carried out in twin trough glass chamber saturated with mobile phase Toluene:Ethyl-acetate:Diethylamine: Glacial acetic acid (6.5:5.0:1.5:0.5 v/v/v/v) and densitometric determination of these compounds was carried out by TLC scanner (CAMAG) at 254 nm in reflectance/absorbance mode. The Rf value of reserpine and arjunolic acid was found to be 0.42±0.03 and 0.14±0.02, respectively. Linearity was found to be in the concentration range of 200 ng to 1600 ng for both reserpine and arjunolic acid. The linear regression data for the calibration plots showed a good linear relationship with r2=0.998 and 0.995 for reserpine and arjunolic acid, respectively. According to the ICH guideline the method was validate for accuracy, precision, recovery, robustness and ruggedness. The reserpine and arjunolic acid contents quantified from herbal formulation (Tensowert tablet) were found well within limits. Statistical analysis of the data showed that the method is reproducible and selective for the estimation of reserpine and arjunolic acid.