Faculty of Science and Technology, Free University of Bolzano-Bozen, 39100 Bolzano, Italy.
Biosensors (Basel). 2020 Apr 10;10(4):35. doi: 10.3390/bios10040035.
Heterocyclic amine histamine is a well-known foodborne toxicant (mostly linked to "scombroid poisoning") synthesized from the microbial decarboxylation of amino acid histidine. In this work, we report the fabrication of a flexible screen-printed immunosensor based on a silver electrode coated with single-walled carbon nanotubes (SWCNTs) for the detection of histamine directly in fish samples. Biosensors were realized by first spray depositing SWCNTs on the working electrodes and by subsequently treating them with oxygen plasma to reduce the unwanted effects related to their hydrophobicity. Next, anti-histamine antibodies were directly immobilized on the treated SWCNTs. Histamine was detected using the typical reaction of histamine and histamine-labeled with horseradish peroxidase (HRP) competing to bind with anti-histamine antibodies. The developed immunosensor shows a wide linear detection range from 0.005 to 50 ng/mL for histamine samples, with a coefficient of determination as high as 98.05%. Average recoveries in fish samples were observed from 96.00% to 104.7%. The biosensor also shows good selectivity (less than 3% relative response for cadaverine, putrescine, and tyramine), reproducibility, mechanical and time stability, being a promising analytical tool for the analysis of histamine, as well as of other food hazards.
杂环胺组胺是一种众所周知的食源毒性物质（主要与“鲭鱼中毒”有关），由氨基酸组氨酸的微生物脱羧合成。在这项工作中，我们报告了一种基于涂有单壁碳纳米管 (SWCNT) 的银电极的柔性丝网印刷免疫传感器的制造，用于直接在鱼样中检测组胺。生物传感器首先通过在工作电极上喷涂 SWCNT 来实现，然后用氧等离子体处理它们以减少与它们的疏水性相关的不良影响。接下来，将抗组胺抗体直接固定在处理过的 SWCNT 上。通过组胺与辣根过氧化物酶（HRP）标记的组胺的典型反应来检测组胺，竞争与抗组胺抗体结合。开发的免疫传感器对组胺样品的线性检测范围从 0.005 到 50ng/mL 很宽，决定系数高达 98.05%。在鱼样中观察到平均回收率为 96.00%至 104.7%。该生物传感器还表现出良好的选择性（腐胺、尸胺和酪胺的相对响应小于 3%）、重现性、机械和时间稳定性，是分析组胺以及其他食品危害物的有前途的分析工具。
