QUANTITATIVE ANALYSIS OF TRIGONELLINE AND CUCURBITACIN-E WITH VALIDATED ISOCRATIC RP-HPLC METHODS FOR STANDARDIZING THE HERBAL FORMULATION: REPCHOL PREMIX

Herbal formulations are complex mixtures of medicinal plants and great efforts are necessary to guarantee a constant and optimum quality. Since pharmacological response to the herbal formulations depends on the secondary metabolites present in the herbs, it is important to evaluate the phytoconstituents present in the formulation. Standardization has become a important part for the regulation of the herbal industry with optimum quality measures.  The development of validated reverse phase - high performance liquid chromatographic (RP-HPLC) methods for the quantification of secondary metabolites trigonelline and cucurbitacin-E in herbal formulation Repchol^{®} Premix are reported. The developed methods were validated following ICH guidelines for precision, linearity, accuracy, specificity, limit of detection, limit of quantification, and robustness. The outcome of the present investigation underlines the importance of the standardization of Ayurvedic formulations. This will ensure the batch to batch consistency in quality and efficacy. The amounts of trigonelline and cucurbitacin-E in Repchol^{®} Premix a commercial herbal formulation, were successfully quantified by the developed RP-HPLC methods. The average content of these markers in different batches of the formulation was 0.079 % and 0.104 % (w/w) respectively. Our results offer new, sensitive and reliable tools for the standardization of herbal formulations containing Trigonella foenum-graecum and Citrullus colocynthis.