A highly sensitive method for quantifying gallocatechin gallate and its epimer using a catechin-specific peptide

Takahisa Miyamoto, Youko Mekada, Masahiro Kurahachi, Mai Umeno, Motokazu Nakayama, Naofumi Shigemune, Takashi Tsugukuni, Hajime Tokuda, Hirofumi Tachibana, Ken-ichi Honjoh

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We have developed a method for quantifying gallocatechin gallate (GCg) and epigallocatechin gallate (EGCg) using a catechin-binding peptide (part of the 67-kDa laminin receptor). Using micro titer plates, we investigated various conditions, including the quantifiable range of EGCg concentrations, the optimal concentration of the catechin-binding peptide, and the optimal reaction conditions. In this microplate assay, after each well was coated with bovine serum albumin, sample containing GCg and EGCg was added at pH 8.0, and allowed to stand at 37 °C for 2 h. After washing, biotinylated-peptide solution was added at 1 μg mL -1 and allowed to react for 1 h at 37 °C. Each well was added with streptavidin-horseradish peroxidase conjugate, followed by chromogenic reaction for 25 min at room temperature. After the reaction, absorbance was measured at 405 nm. Our method is capable of quantifying EGCg in the range of approximately 0.1-2.0 mg L -1 with a high degree of sensitivity and a high correlation (R 2 = 0.98) between EGCg concentration and absorbance. The method was specific to GCg and EGCg and seems capable of estimating GCg and EGCg contents in the presence of other catechin compounds. The method is simple and highly sensitive for quantitative GCg and EGCg measurement that requires no special equipment or operation and can measure multiple samples simultaneously.

Original languageEnglish
Pages (from-to)162-166
Number of pages5
JournalFood Control
Volume29
Issue number1
DOIs
Publication statusPublished - Jan 1 2013

Fingerprint

Catechin
epigallocatechin
catechin
peptides
Peptides
methodology
Laminin Receptors
Streptavidin
epigallocatechin gallate
gallocatechin gallate
absorbance
Horseradish Peroxidase
Bovine Serum Albumin
streptavidin
laminin
bovine serum albumin
Equipment and Supplies
Temperature
washing
ambient temperature

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science

Cite this

A highly sensitive method for quantifying gallocatechin gallate and its epimer using a catechin-specific peptide. / Miyamoto, Takahisa; Mekada, Youko; Kurahachi, Masahiro; Umeno, Mai; Nakayama, Motokazu; Shigemune, Naofumi; Tsugukuni, Takashi; Tokuda, Hajime; Tachibana, Hirofumi; Honjoh, Ken-ichi.

In: Food Control, Vol. 29, No. 1, 01.01.2013, p. 162-166.

Research output: Contribution to journalArticle

Miyamoto, Takahisa ; Mekada, Youko ; Kurahachi, Masahiro ; Umeno, Mai ; Nakayama, Motokazu ; Shigemune, Naofumi ; Tsugukuni, Takashi ; Tokuda, Hajime ; Tachibana, Hirofumi ; Honjoh, Ken-ichi. / A highly sensitive method for quantifying gallocatechin gallate and its epimer using a catechin-specific peptide. In: Food Control. 2013 ; Vol. 29, No. 1. pp. 162-166.
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