NOVEL METHOD DEVELOPMENT AND VALIDATION OF RP-HPTLC METHOD FOR THYMOQUINONE AND CURCUMIN

Abstract

A method for the rapid, linear, precise, and robust quantitative analysis (RP-HPTLC) has been developed and validated. This method was used to separate the components of  thymoquinone and curcumin, on a precoated, silica gel, aluminium plate with a 60F-254 stationary phase, consisting of a mobile phase composed of a 16:4 ratio of  toluene: acetic acid. The detection wavelength of the mobile phase was chosen to be 260 nm, and the detection wavelength of the detection phase was 417 nm, with the Rf value for curcumin being found to be 0.41 and for thymoquinone to be 0.55. The method was able to accurately and linearly measure the concentration range from 100-350 nd/band. The method was validated according to the ICH guideline for specificity, linearity and accuracy, as well as for reproducibility and robustness.