Determination of catechin isomers in human plasma subsequent to green tea ingestion using chiral capillary electrophoresis with a high-sensitivity cell.

A simple and reliable analytical electrophoretic method using chiral capillary electrophoresis (CCE) with a high-sensitivity cell of special design has been established for simultaneous determination of (+)-catechin (C) and (-)-epicatechin (EC) in aqueous and human plasma media. The application of a capillary with high-sensitivity cell has led to an improvement of 10-fold and 5-fold time-corrected peak area over a standard cell and a capillary with bubble cell, respectively. Analysis has involved the electrophoretic separation of C and EC in less than 4.0 min at 210 nm. The running buffer consist of 50.0 mmol L(-1) borate buffer with 1.0 mmol L(-1) beta-cyclodextrin at pH 8.5. CCE system has been proved for its intended use by applying procedure starting from calibration of CE instrument into validation of all experimental parameters. The resolution between catechin isomers under optimal conditions has been found to be more than 3.0. The detection limits of C and EC have been calculated to be 3.2 and 1.0 ng mL(-1), respectively. Good linearity has been obtained with correlation coefficient (r(2)) ranging between 0.995 and 0.996 at 99% confidence level (CL). Application of the proposed method to human plasma after ingestion of green tea has successfully been achieved and has statistically been proved. The unchanged amounts of C and EC in plasma were about 17.4 and 1.8% of the administered dose after 2 h of starting tea ingestion. The detection limits of C and EC in human plasma at 210 nm were 4.1 and 1.5 ng mL(-1), respectively.