化学发光生物传感器检测食品中生物胺总量
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、3~200 μmol/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、0.5 μmol/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, 3~200 μmol/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, 0.5 μmol/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.
标题：化学发光生物传感器检测食品中生物胺总量
英文标题：Detection of total biogenic amines in foods by chemiluminescence biosensor
作者：
唐晗 华中农业大学食品科技学院
王晓朋 华中农业大学食品科技学院
吴中波 华中农业大学食品科技学院
万德慧 华中农业大学食品科技学院
刘晓宇 华中农业大学食品科技学院;环境食品学教育部重点实验室
中文关键词:生物胺,二胺氧化酶,微流控芯片,酶传感器,化学发光,
英文关键词:biogenic amines,diamine oxidase,micro-fluidic chip,enzyme biosensor,chemiluminescence,
发表日期：2014-04-09
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