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基于阻抗法的食品中 3-氨基-2-恶唑烷酮残留检测免疫传感器的研制。

Development of an impedimetric immunosensor for the determination of 3-amino-2-oxazolidone residue in food samples.

机构信息

School of Pharmacy, China Pharmaceutical University, Nanjing, PR China.

出版信息

Anal Chim Acta. 2011 Nov 7;706(1):120-7. doi: 10.1016/j.aca.2011.08.018. Epub 2011 Sep 1.

DOI:10.1016/j.aca.2011.08.018
PMID:21995918
Abstract

The use of furazolidone in food animals has been banned in European Union (EU) because of its carcinogenicity and mutagenicity on human health, but its continued misuse is widespread. Therefore, there is an urgent need for a simple, reliable, and rapid method for the detection of its marker residue, 3-amino-2-oxazolidinone (AOZ), in food products. In this regard, a sensitive and reliable electrochemical method was presented to detect AOZ based on a novel label-free electrochemical impedimetric immunosensor to address this need. The immobilization of monoclonal antibody against AOZ (denoted as AOZ-McAb) on the gold electrode was carried out through a stable acyl amino ester intermediate generated by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydrosuccinimide (NHS), which could condense antibodies on the self-assembled monolayer (SAM). The detection of AOZ was performed by measuring the relative change in charge transfer resistance before and after AOZ and AOZ-McAb immunoreaction by electrochemical impedance spectroscopy (EIS). Under the optimized conditions, the relative change in charge transfer resistance was proportional to the logarithmic value of AOZ concentrations in the range of 20.0 to 1.0×10(4) ng mL(-1) (r=0.9987). Moreover, the proposed immunosensor has a high selectivity to AOZ alone with no significant response to the metabolites of other nitrofuran antibiotics, such as 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ), semicarbazide (SEM), and 1-aminohydantoin hydrochloride (AHD). This protocol has been applied to detect AOZ in food samples with satisfactory results.

摘要

在欧盟,由于 furazolidone 对人类健康具有致癌性和致突变性,已被禁止将其用于食用动物,但 furazolidone 的继续滥用仍很普遍。因此,迫切需要一种简单、可靠和快速的方法来检测其标记残留 3-氨基-2-恶唑烷酮(AOZ)在食品中的残留。在这方面,提出了一种基于新型无标记电化学阻抗免疫传感器检测 AOZ 的灵敏可靠电化学方法来满足这一需求。通过 1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDC)和 N-羟基琥珀酰亚胺(NHS)生成的稳定酰氨基酯中间体制备 AOZ 单克隆抗体(AOZ-McAb)在金电极上的固定化,该中间体能将抗体浓缩在自组装单层(SAM)上。通过电化学阻抗谱(EIS)测量 AOZ 和 AOZ-McAb 免疫反应前后的电荷转移电阻的相对变化来检测 AOZ。在优化条件下,电荷转移电阻的相对变化与 AOZ 浓度的对数在 20.0 至 1.0×10(4)ng mL(-1) 范围内呈正比(r=0.9987)。此外,该免疫传感器对 AOZ 具有高选择性,对其他硝基呋喃类抗生素的代谢物如 3-氨基-5-吗啉甲基-2-恶唑烷酮(AMOZ)、氨基脲(SEM)和 1-氨基乙内酰脲盐酸盐(AHD)无明显响应。该方案已应用于检测食品样品中的 AOZ,结果令人满意。

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