Journal of the East Asian Society of Dietary Life (동아시아식생활학회지)

The East Asian Society of Dietary Life (동아시아식생활학회)
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Determination of Biogenic Amines using an Amperometric Biosensor with a Carbon Nanotube Electrode and Enzyme Reactor

Carbon Nanotube 전극과 효소반응기로 구성된 Amperometric Biosensor를 이용한 Biogenic Amines 검출

  • Kim, Jong-Won (Dept. of Food Science and Nutrition, Kyungpook National University) ;
  • Jeon, Yeon-Hee (Dept. of Food Science and Nutrition, Kyungpook National University) ;
  • Kim, Mee-Ra (Dept. of Food Science and Nutrition, Center for Beautiful Aging, Kyungpook National University)
  • 김종원 (경북대학교 식품영양학과) ;
  • 전연희 (경북대학교 식품영양학과) ;
  • 김미라 (경북대학교 식품영양학과, 장수생활과학연구소)
  • Received : 2010.09.01
  • Accepted : 2010.10.05
  • Published : 2010.10.31
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Abstract

Biogenic amines are synthesized by microbial decarboxylation for the putrefaction or fermentation of foods containing protein. Although biogenic amines such as histamine, tyramine, and putrescine are required for many physiological functions in humans and animals, consumption of high amounts of biogenic amines can cause toxicological effects, including serious gastrointestinal, cutaneous, hemodynamic, and neurological symptoms. In this study, a novel amperometric biosensor wasdeveloped to detect biogenic amines. The biosensor consisted of a working electrode, a reference electrode, a counter electrode, an enzyme reactor with immobilized diamine oxidase, an injector, a peristaltic pump and a potentiostat. A working electrode was fabricated with a glassy carbon electrode (GCE) by coating functionalized multi-walled carbon nanotubes (MWCNT-$NH_2$) and by electrodepositing Prussian blue (PB) to enhance electrical conductivity. A sensor system with PB/MWCNT-$NH_2$/GCE showed linearity in the range of $0.5 {\mu}M{\sim}100 {\mu}M$ hydrogen peroxide with a detection limit of $0.5 {\mu}M$. The responses for tyramine, 2-phenylethylamine, and tryptamine were 95%, 75%, and 70% compared to that of histamine, respectively. These results imply that the biosensor system can be applied to the quantitative measurement of biogenic amines.

식품 중 잠재적 위험 물질로 작용할 수 있는 BAs의 신속 검출을 위하여 전기전도성이 우수하다고 알려진 MWCNTs의 개질, 작업 전극의 제작, 효소반응기 제작을 통해 바이오센서 시스템을 구성하고 hydrogen peroxide와 주요 BAs 물질에 대한 감응도를 측정하였다. MWCTS의 성질을 향상시키기 위해 아민기를 도입하였고, 이를 FT-IR 스펙트럼을 통해 확인하였다. 아민기를 도입한 MWCNTs를 GCE에 고착시키고 PB 막을 입힌 후 cyclic voltammetry 반응을 비교한 결과, PB/MWCNT-$NH_2$/GCE 전극이 GCE에 비해 산화 전류는 8배 이상, 전체 전류는 25배 이상의 높은 전류 흐름을 보였으며, 미세한 전류 흐름의 측정도 용이해진 것으로 나타났다. 또한, carrier buffer의 pH를 달리하여 제작된 전극의 감응 전류를 비교한 결과, pH 7.0에서 전류의 감응도가 가장 높게 측정되었다. 과산화수소를 주입하여 전극의 성능의 검증한 결과, $0.5 {\mu}M{\sim}100 {\mu}M$ 범위에서 선형관계를 보였으며, 검출 한계는 $0.5{\mu}M$로 측정되었다. Histamine 표준 용액 주입 시 $1{\mu}M{\sim}100{\mu}M$ 범위에서 선형 관계를 나타내었고, tyramine은 histamine에 비해 약 95%, 2-phenylethylamine과 tryptamine은 histamine에 비해 각각 75%, 70% 수준의 감응도를 이용하여 구성된 바이오센서 시스템이 낮은 검출 한계와 높은 수준의 반응 감도를 나타내어 BAs를 측정하는데 좋은 장치임이 확인되었다.

Keywords

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