OPTIMIZATION AND VALIDATION OF RP-HPLC/UV DETECTION FOR SEVERAL COMPOUNDS SIMULTANEOUSLY IN SEMI-PURIFIED EXTRACT OF WHITE TEA

Abstract

Modification of separation, detection, and quantification of several compounds simultaneously in white tea leaf extract using HPLC aims to obtain easy, fast, precise, and accurate analysis procedures. An HPLC optimization strategy was devised that allowed simultaneous determination of the effectiveness of separation and quantification of gallic acid, theobromine, theophylline, catechin, caffeine and epigallocatechin gallate. The compounds were separated and isocratically on a LiChrocart-C18 column. The optimal conditions for separation were determined using central composite design (CCD) with a solvent consisting of acetonitrile: methanol: 0.005% orthophosphoric acid (10:10:80 v/v/v) at a 0.90 mL/min flow rate. Validation procedures for modified analysis methods have been carried out following the terms and conditions. All validation points meet the requirements set by the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use. The validation results were successfully applied to a white tea leaf extract. CCD is very effective for HPLC system optimization and was efficient for the simultaneous analysis of several compounds in white tea extract.