Title

開發組織胺之微流體生物感測器

Translated Titles

Development of Microfluidic Biosensor for Monitoring Histamine

Authors

薛純婷

Key Words

微流體生物感測器 ; 組織胺 ; 二胺氧化酶山葵過氧化酵素 ; microfluidic biosensor ; histamine ; diamine oxidase ; horseradish peroxidase

PublicationName

中興大學食品暨應用生物科技學系所學位論文

Volume or Term/Year and Month of Publication

2011年

Academic Degree Category

碩士

Advisor

蔡碩文

Content Language

繁體中文

Chinese Abstract

生物死亡後,體內游離胺基酸經細菌性脫羧反應可產生生物胺,視為水產品重要安全指標,其中以容易引發過敏反應之組織胺最具代表性,美國FDA (Food and Drug Administration) 建議其含量不得超過50 ppm,然而無色無味之特性,使組織胺難以即時檢測。本研究將 diamine oxidase (DAO) 和horseradish peroxidase (HRP) 耦合,固定於單一微流體系統內,以催化組織胺轉換成可視判讀之訊號,以期未來能以檢測水產品滴液之方式,監控水產品質。使用光學檢測法以耦合酵素組合,結果顯示使用0.864 U/mL DAO和1.024 U/mL HRP,搭配1 mM 4-AAP /68.8 mM phenol,可偵測4-40 ppm組織胺,並具良好線性關係 (r2= 0.9784)。在微流體毛細管中,利用1%海藻酸鈉,將1.2 U/mL DAO、8.53 U/mL HRP與0.83 mM 4-AAP /57.3mM phenol,固定於微流體感測器之毛細管系統中,於樣品導入後一小時內,即可將10-100 ppm無色組織胺,轉換為肉眼易區別之色澤變化,於實際樣品應用上,微流體感測系統可以由目視法偵測不同濃度之組織胺 (11.65-40.89 ppm),經與官方分析方法比較得知,具高可靠性。本研究成功開發微流體檢測系統,具簡易操作、快速、高專一性、非破壞性、無需外加動力、少量樣品與分析試劑消耗及無需昂貴分析儀器等優點,可提供品管人員或消費者,另一便利分析組織胺之替代方案,具有應用於食品安全早期預警之潛力。

English Abstract

Biogenic amine produced by microbial decarboxylation of free amino acid is regarded as a significant indicator for the quality and safety of marine products. One of the representative biogenic amine is histamine that may result in allergic reaction. Food and Durg Administration (FDA) set the defect action level of histamine in 50 ppm. However, it is difficult to examine colorless and odorless histamine on the spot. The objective of this research is to couple diamine oxidase (DAO) and horseradish peroxidase (HRP) in a single microfluidic biosensor. The coupling enzymatic system can convert colorless histamine to visible dye signal. For purpose of monitoring freshness and quality in the future, histamine of sample drip was analyzed by coupling enzymatic system. Experimental results show that optimal condition of coupling enzyme system using 1 mM 4-aminoantipyrine (4AAP)/ 68.8 mM phenol as substrate for DAO and HRP was 0.864 U/mL and 1.024 U/mL, respectively. Linear response range of histamine concentration was from 4 to 40 ppm. ( r2 = 0.9784 ). The 1.2 U/mL DAO/8.53 U/mL HRP/0.83 mM 4-AAP was immobilized in the capillary by using 1% sodium alginate to fabricate the biosensor. Histamine (10-100 ppm) converted to visible dye signal was observed by naked eye after sample injected to biosensor in one hour. The biosensor could detect histamine of real sample from 11.65 to 40.89 ppm and be comparable with official method. In this study, we successfully developed a microfluidic biosensor to provide a simple, rapid, high specificific, non-destructive, sampling without pump, less sample and reagent consuming,and without need of expensive instruments method. It could offer technicians or consumers an alternative way to assure food safety.

Topic Category 農業暨自然資源學院 > 食品暨應用生物科技學系所
生物農學 > 生物科學
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Times Cited
  1. 丁懿敏(2012)。以耦合酵素系統於微流體晶片中快速偵測組織胺。中興大學食品暨應用生物科技學系所學位論文。2012。1-60。