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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 rapid and effective method for the determination of biogenic amines in food. Methods Immobilized diamine oxidase was used to make enzyme column as identification element of biosensor. Microfluidic chip was combined with chemiluminescence instrument as detection element. With luminol-potassium ferricyanide as chemiluminescence system, Analytical method to detect biogenic amines in food. Results When the concentrations of putrescine, histamine and tyramine were in the range of 3 ~ 500, 2 ~ 100 and 3 ~ 200μmol / L, the linear regression equations were Y = 17.448X + 408.93, Y = 88.329X + 997.13, Y = 45.762 X + 1728.2, the correlation coefficients were 0.9966,0.9937,0.9907, the detection limits were 0.7,0.5,0.5μmol / L. In addition, the performance of the sensor was evaluated in this study. The results showed that under the optimal conditions, the solutions containing putrescine, histamine and tyramine were measured seven times respectively, the RSDs were less than 8%, the precision was better; with putrescine as substrate, continuous access to luminol And 7 times of potassium ferricyanide solution, RSD = 1.66%, showing good stability; the immobilized enzyme was stored in phosphate buffer and enzyme activity was measured every 10 days for 70 days. The immobilized diamine oxidase activity was only reduced by 15% within 2 months and the storage time was longer. The sensor was used to detect the total amount of biogenic amines in pork, crucian carp and wine. The recoveries of all three amines in the samples were In 90% ~ 94%, the recovery rate is higher, suitable for detection of total biogenic amines. Conclusion The comprehensive performance of this sensor is good and it is suitable for the rapid detection of the total amount of biogenic amines in food.