A HPLC-UV Method for Analysis of Total Plant Extract and Catechin Fraction of Bancha Green Tea

Background: The pharmacological characterization of Bancha green tea requires a preliminary establishment of its phytochemical composition. Objective: The aim of the present study was to develop a simple, sensitive, and environmentally friendly HPLC-UV procedure for the simultaneous quantification of caffeine, (+)-catechin, epigallocatechin gallate and gallic acid in different Bancha green tea isolates. Methods: The chromatographic separation of the four phytochemicals was performed on a Hypersil GOLD aQ column with a mobile phase consisting of acetonitrile/acetic acid/bi-distilled water (8:1:91 v/v/v) in an isocratic mode of elution. The suitability of the proposed procedure was verified by achieving satisfying linearity and specificity. Results: The method was also assessed as accurate and precise, with the obtained limits of quantification of caffeine, (+)-catechin, epigallocatechin gallate, and gallic acid of 0.01, 0.30, 0.50 and 0.03 mu g/mL, respectively. As it was expected, the analytical procedure approbation showed the highest epigallocatechin gallate content (16.78% +/- 0.54) in the catechin fraction. On the other hand, the amount of caffeine and epigallocatechin gallate in the total plant extract was almost the same (4.19% +/- 0.14 and 4.12% +/- 0.44, respectively). Conclusion: A reliable HPLC-UV method for phytochemical characterization of Bancha green tea extracts has been developed. The conducted quantitate assay could be used as a basis for subsequent pharmacological in vivo and in vitro studies of Bancha green tea.