4.5 Article

An improved method for the determination of green and black tea polyphenols in biomatrices by high-performance liquid chromatography with coulometric array detection

Journal

ANALYTICAL BIOCHEMISTRY
Volume 279, Issue 2, Pages 164-169

Publisher

ACADEMIC PRESS INC
DOI: 10.1006/abio.2000.4487

Keywords

catechins; theaflavins; polyphenols; green tea; black tea; plasma; saliva; HPLC; coulochem detection

Funding

  1. NCI NIH HHS [CA 56673] Funding Source: Medline
  2. NIEHS NIH HHS [ES05022] Funding Source: Medline

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Tea polyphenols are strong antioxidants and are believed to have beneficial health effects, However, the blood and tissue levels of these compounds are not well characterized because of a lack of suitable analytical methods for the biological resolution of these compounds. Previously, we developed methods for the analysis of three green tea catechins. Now we report an improved method for the measurement of the levels of the different catechins and theaflavins in biological fluids and tissues. The method includes digestion of the plasma, urine, or tissue samples with beta-D-glucuronidase and sulfatase, followed by extraction with ethyl acetate and subsequent separation by reversed-phase high-performance liquid chromatography (HPLC). The polyphenols are identified on the basis of their retention times, spectral analysis, and electrochemical behavior across an array of electrodes. In a single HPLC run, it is possible to determine the major catechins and theaflavins as well as some of the catechin metabolites. The detection limits for catechins and theaflavins are from 5 to 10 ng/ml of saliva, plasma, or urine. (C) 2000 Academic Press.

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