Evaluation of antioxidant activity of ten compounds in different tea samples by means of an on-line HPLC-DPPH assay

Tea (Camellia sinensis), a well-known traditional beverage in China, has drawn growing attention due to its various benefits to health. In this study, an on-line assay of coupling high performance liquid chromatography separation with 1,1-diphenyl-2-picrylhydrazyl free radical reaction system (HPLC-DPPH) was applied to evaluate the antioxidant activity of different teas. Twelve main chromatographic peaks were detected in tea, and they were identified as gallic acid, 3-galloyl-quinic acid, theobromine, (-)-gallocatechin, (-)-epigallocatechin, caffeine, procyanidin dimer, (-)-epicatechin, (-)-epigallocatechin gallate, (-)-gallocatechin gallate, 1,2,6-tri-galloyl-glucose and (-)-epicatechin gallate by comparing their retention times and DAD spectra with standard compounds respectively or with reference to our previous study. DPPH assay showed that ten out of twelve investigated compounds have free radical scavenging activity, and their contents were determined or estimated. Trolox equivalent antioxidant capacities (TEACs) of the ten active components were also determined. EGCG was the most potent antioxidant with a TEAC value of 5.822. Catechin components were the major contributors to the antioxidant activity of tea; they decreased during tea fermentation and led to the reduction of antioxidant activity. Different tea samples were scientifically classified and their antioxidant activities were comprehensively evaluated by on-line HPLC-DPPH assays coupled with principal component analysis and hierarchical clustering analysis. The results indicated that the combination of the on-line HPLC-DPPH assay, principal component analysis and hierarchical clustering analysis could be suitable for the bioactivity assessment and variety characterization of tea and its derived products. Additionally, a simple and rapid method for simultaneous capture of chemical quantitative analysis and bioactivity evaluation by determining contents of the bioactive compounds was firstly established. This method could provide a comprehensive evaluation of tea. (C) 2013 Published by Elsevier Ltd.