基于磁珠-适配体的高效液相色谱-串联质谱法检测食品中的黄曲霉毒素B1

doi:10.11944/j.issn.1000-0518.2020.11.200125

摘要/Abstract

摘要： 本文构建了特异性识别黄曲霉毒素B₁(AFB₁)的磁珠-适配体,并与高效液相色谱-串联质谱联用(LC-MS/MS),建立食品中AFB₁的定量检测方法。利用碳二亚胺盐酸盐(EDC)活化法,将羧基磁珠进行活化。活化后的羧基磁珠与5'端氨基修饰的适配体进行孵育结合,通过酰胺反应将适配体共价连接在羧基磁珠表面,固定在磁珠表面的适配体作为捕捉探针将样品提取液中的AFB₁分离,通过LC-MS/MS对AFB₁进行定性和定量分析。检测结果表明:AFB₁在浓度0.25～25 ng/mL呈良好的线性关系,相关系数R²=0.999,定量检出限为0.25 ng/mL,回收率达到80.3%～92.5%,相对标准偏差(RSD)低于8%。该方法操作简单、快速便捷、可痕量地检测AFB₁,所制备的磁珠-适配体可重复利用,为定量检测AFB₁提供了另一种技术支持。

关键词: 磁珠, 适配体, 高效液相色谱-串联质谱, 黄曲霉毒素B₁

Abstract: In this study, a magnetic bead-aptamer that specifically recognizes aflatoxin B₁ (AFB₁) was constructed and coupled with high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) to establish a quantitative detection method for AFB₁ in food. Carboxy magnetic beads were activated by carbodiimide hydrochloride (EDC) and incubated with 5'terminal amino modified aptamers. The aptamers were covalently linked to the surface of the carboxy magnetic beads by amidation reaction and used as capture probes to separate AFB₁ in the extracted sample. The qualitative and quantitative analysis of AFB₁ was carried out by LC-MS/MS. The results show that the method has a good linear relationship at the concentration of 0.25~25 ng/mL with the correlation coefficient R²=0.999, the quantitative detection limit is 0.25 ng/mL, the recovery is 80.3%~92.5%, and the relative standard deviation (RSD) is less than 8%. The method is simple, fast and convenient, and can detect trace AFB₁. The magnetic bead aptamer can be reused, which provides another technical support for the quantitative detection of AFB₁.

Key words: magnetic beads, aptamers, LC-MS/MS, aflatoxin B₁

中图分类号:

O652.6

古鑫宇, 郎乐, 王建伟, 赵丽辉. 基于磁珠-适配体的高效液相色谱-串联质谱法检测食品中的黄曲霉毒素B₁[J]. 应用化学, 2020, 37(11): 1324-1332.

GU Xinyu, LANG Le, WANG Jianwei, ZHAO Lihui. Determination of Aflatoxin B₁ in Food by LC-MS/MS Method Based on Magnetic Bead-Aptamer[J]. Chinese Journal of Applied Chemistry, 2020, 37(11): 1324-1332.

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基于磁珠-适配体的高效液相色谱-串联质谱法检测食品中的黄曲霉毒素B₁

Determination of Aflatoxin B₁ in Food by LC-MS/MS Method Based on Magnetic Bead-Aptamer

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