化学发光生物传感器检测食品中生物胺总量

Detection of total biogenic amines in foods by chemiluminescence biosensor

目的：建立快速地有效检测食品中生物胺总量的分析方法。方法用固定化的二胺氧化酶制成酶柱作为生物传感器的识别元件,将微流控芯片与化学发光仪结合作为检测元件,以鲁米诺-铁氰化钾作为化学发光体系,通过流动分析法来检测食品中生物胺的总量。结果当腐胺、组胺、酪胺浓度分别在3~500、2~100、3~200μmol/L时,线性回归方程分别为Y=17.448X+408.93、Y=88.329X+997.13、Y=45.762X+1728.2,相关系数分别为0.9966、0.9937、0.9907,检出限分别为0.7、0.5、0.5μmol/L。另外,本研究对传感器的性能进行了评价。结果显示,在最佳条件下对含有腐胺、组胺、酪胺的溶液分别测定7次,其RSD均小于8%,精密度较好；以腐胺为底物,连续通入鲁米诺和铁氰化钾溶液7次, RSD=1.66%,表现出良好的稳定性；将固定化酶保存在磷酸盐缓冲液中,每10 d测定一次酶活,持续70 d。2个月内固定化二胺氧化酶酶活仅降低了15%,保存时间较长；将传感器用于检测猪肉、鲫鱼和葡萄酒中的生物胺总量,样品中三种胺的添加回收率均在90%~94%之间,回收率较高,适合检测总生物胺。结论此传感器的综合性能良好,适用于食品中生物胺总量的快速检测。

Objective To establish a method for the analysis of the biogenic amines content in foods. Methods A novel micro-fluidic chip-chemiluminescence-enzyme biosensor was designed and assembled to determine the total biogenic amines content in foods. Commercial diamine oxidase was used as biological recognition element of the sensor, the micro-fluidic chip with chemiluminescence analyzer was used as a sensing element, and luminol-ferricyanide was used in chemiluminescence system. Results When the concentration of putrescine, histamine, and tyramine were 3~500, 2~100, 3~200μmol/L, the linear regression equation were Y=17.448X+408.93, Y=88.329X+997.13, Y=45.762X+1728.2, the correlation coefficient were 0.9966, 0.9937, 0.9907, and the detection limit were 0.7, 0.5, 0.5μmol/L, respectively. In addition, this study evaluated the performance of the sensor. Under optimal conditions, putrescine, histamine, tyramine solution were measured, the RSD were between 90%~94%(n=7), the precision was good; when chosing putrescine as substrate, luminol and potassium ferricyanide was entered into chemiluminescence analyzer for seven times, the RSD=1.66%, stability was good; The immobilized enzyme was stored in phosphate buffer, measured every 10 d, lasting 70 d. The activity of immobilized diamine oxidase reduced by 15% in 2 months. Using this sensor to detect the total amount of biogenic amines in pork, fish and red wine, all three recoveries of amine were more than 89%. Conclusion This sensor has good performance, and it can be used for rapid detection of biogenic amines in foods.