한국식품과학회지 (Korean Journal of Food Science and Technology)

한국식품과학회 (Korean Society of Food Science and Technology)
권호 표지

포도종실 에탄올 추출물에 함유된 (+)-카테킨, (-)-에피카테킨의 모세관 전기영동법에 의한 분석

Capillary Electrophoretic Method for the Determination of (+)-Catechin, (-)-Epicatechin in Grape Seed Ethanol Extract

  • 최원균 (한경대학교 식품생물공학과 및 식품생물산업연구소) ;
  • 정양섭 (한경대학교 식품생물공학과 및 식품생물산업연구소) ;
  • 정하열 (한경대학교 식품생물공학과 및 식품생물산업연구소)
  • Choi, One-Kyun (Department of Food Science & Biotechnology and Food & Bio-industrial Research Center, Hankyong National University) ;
  • Chung, Yang-Seop (Department of Food Science & Biotechnology and Food & Bio-industrial Research Center, Hankyong National University) ;
  • Chung, Ha-Yull (Department of Food Science & Biotechnology and Food & Bio-industrial Research Center, Hankyong National University)
  • 발행 : 2005.08.31
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초록

포도종실추출물에 함유되어 강력한 항산화 작용을 나타내는 성분인 proanthocyanin의 구성 성분이면서 에피머인 (+)-catechin과 (-)-epicatechin을 CE법을 이용하여 분석하였다. CE법을 이용한 catechin류 분석의 최적 조건은 fused silica capillary$(20cm{\times}50{\mu}m)$, current $56{\mu}A$, voltage 10 kV, phosphate/borate buffer(pH 6.0), 온도 $20^{\circ}C$로서 (+)-catechin과 (-)-epicatechin을 6분 이내에 매트릭스의 방해없이 분석할 수 있었다. 비색법, HPLC법과 CE분석법의 특징을 비교해 보면 비색법은 (+)-catechin과 (-)-epicatechin합한 총 함량은 알 수 있으나 두성분의 분리는 할 수 없었다. 또한 HPLC방법은 표준품은 두 성분이 잘 분리되었으나 시료 분석 시 proanthocyanin을 가수분해하기 위해 첨가한 산으로 인해 시료가 강산성을 띠어 컬럼 손상되는 문제가 있어 직접 분석이 불가능하여 시료를 희석해야 했으며, 희석을 하여 분석할 경우에도 많은 잡 피크들로 정량적인 결과를 얻기에 용이하지 않았다. 한편 CE법의 경우 산 가수분해한 시료를 별다른 처리 없이도 다른 성분의 방해 없이 분석할 수 있었다. 분석소요시간의 경우 비색법은 실험단계가 복잡하여 많은 시간이 걸렸으며 HPLC는 15분 그리고 CE는 6분 이내에 분석이 가능하였다. 재현성과 직선성을 보면 HPLC와 CE의 방법이 모두 양호하였고 검출한계에 있어서는 CE법이 0.035mg/L로서 검출감도가 매우 우수하였다. 이와 같이 CE법은 포도종실에 존재하는 (+)-catechin과 (-)-epicatechln을 분석하는데 방법의 용이성, 분석결과의 정확성에 있어서 기존 방법인 비색법과 HPLC 방법보다 우수한 결과를 나타내었다.

Capillary electrophoresis (CE) method was developed to determine (+)-catechin and (-)-epicatechin contents in grape seed ethanol extract. CE separation was achieved using 100 mM phosphate and borate buffer at pH 6.0 as background electrolyte and fused silica capillary with 50 microns x 375 microns O.D. (effective length 20.0cm) maintained at $25^{\circ}C$. The applied voltage was 10kV, and detection was performed by DAD at 210 nm, Two catechins were well separated within 6 min with repeatability of <0.8% RSD for migration time and <2.0% RSD for peak area, and correlation coefficients higher than 0.994 were obtained from 58.0 to 174.0 mg/L with detection limit of 0.035 mg/L. Separated compounds were successfully determined. CE method was easy to handle and showed good reproducibility. CE method was compared with conventional coloring and HPLC methods, and main advantages of CE method were low amount of sample required, simple pre-sample treatment, good recovery rate, and short analysis time.

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