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

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

中文摘要:

      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。

英文摘要:

      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.