基于间接竞争原理的流式微球技术快速检测麦芽中赭曲霉毒素A

引用本文: 肖昌彬, 刘秋桃, 豆小文, 杨美华, 孔维军, 万丽. 基于间接竞争原理的流式微球技术快速检测麦芽中赭曲霉毒素A[J]. 分析化学, 2016, 44(4): 625-632. doi: 10.11895/j.issn.0253-3820.151007 shu

Citation: XIAO Chang-Bin, LIU Qiu-Tao, DOU Xiao-Wen, YANG Mei-Hua, KONG Wei-Jun, WAN Li. Rapid Detection of Ochratoxin A in Malt by Cytometric Bead Array Based on Indirect Competition Principle[J]. Chinese Journal of Analytical Chemistry, 2016, 44(4): 625-632. doi: 10.11895/j.issn.0253-3820.151007 shu

基于间接竞争原理的流式微球技术快速检测麦芽中赭曲霉毒素A

肖昌彬 ^1,2, ,
刘秋桃 ^2,3, ,
豆小文 ^2, ,
杨美华 ^2, ,
孔维军 ^{2,
,} ,
万丽 ^{1,
,}

1.
¹ 成都中医药大学药学院, 成都 611137;
2.
² 中国医学科学院北京协和医科大学, 药用植物研究所, 北京 100193;
3.
³ 辽宁医学院药学院, 锦州 121001

通讯作者: 孔维军, 万丽; 孔维军, 万丽

基金项目:
本文系国家自然科学基金(Nos.81473346,81274072) (Nos.81473346,81274072)

协和新星人才支持计划资助

摘要: 建立了一种快速、灵敏检测中药麦芽中赭曲霉毒素A(Ochratoxin A,OTA)含量的间接竞争流式微球方法。麦芽样品经60%甲醇/PBS提取,加入20%甲醇/PBS溶液稀释5倍后,离心取上清液制备样品。在编码荧光微球上偶联牛血清白蛋白-OTA(BSA-OTA)复合物,与样品中OTA竞争结合抗OTA特异性抗体,然后加入FITC-IgG孵育,离心洗涤后,用流式细胞仪检测微球表面的平均荧光强度,实现样品中OTA的准确定性定量测定。本方法检测OTA的半数抑制浓度IC₅₀为1.20 ng/mL,相关系数R²=0.9892,检出限(IC₁₀)为0.12 ng/mL,加样回收率为93.9%-97.4%,RSD<3.6%。16份实际麦芽样品中检测到2个阳性样品,OTA的最高含量为3.83μg/kg,未超出欧盟的OTA限量标准,与液相色谱-串联质谱(LC-MS/MS)法检测结果相一致。本方法简单、快速、灵敏、可靠,可拓展用于其它复杂基质中多种真菌毒素的定性与定量检测。

关键词:

English

Rapid Detection of Ochratoxin A in Malt by Cytometric Bead Array Based on Indirect Competition Principle

1.
¹ Pharmacy College, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China;
2.
² Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China;
3.
³ Pharmacy College, Liaoning Medical University, Jinzhou 121001, China

Corresponding author: 孔维军, 万丽; 孔维军, 万丽

Received Date: 23 December 2015
Fund Project:
本文系国家自然科学基金(Nos.81473346,81274072)

协和新星人才支持计划资助

Abstract: A cytometric bead array(CBA) method based on indirect competition mode was developed for the sensitive and rapid detection of ochratoxin A(OTA) in malt. The malt samples were extracted by 60% methanol/PBS and the extracts were diluted five times by 20% methanol/PBS, then the supernatant was collected to prepare sample solution for analysis. The fluorescence microsphere surface was labeled with bovine serum albumin-OTA(BSA-OTA) to compete with OTA in the sample for anti-OTA monoclonal antibody(mAb). Then FITC-IgG was added to bind with the captured mAb on the microsphere. Finally, the mean fluorescence intensity of FITC on the surface of microsphere was detected by a BD FACSCalibur analyzer for accurate qualitative and quantitative analysis of OTA. The results showed that the half inhibitory concentration(IC₅₀) was 1.20 ng/mL with a correlation coefficient(R²) of 0.9892, and the limit of detection(LOD) for OTA was 0.12 ng/mL. The average recoveries in malt samples were 93.9%-97.4% with the relative standard deviations(RSDs) less than 3.6% at three spiked levels. Sixteen malt samples were analyzed and OTA was found in two samples with the contents less than 3.83μg/kg, which was lower than the maximum permitted residue level(5.0μg/kg) proposed by the European Union. All the positive samples were confirmed by LC-MS/MS. In this study, the CBA technique based on indirect competition was developed for the first time for successful detection of OTA in malt samples. This method was easy, rapid, sensitive and reliable, and exhibited high potential in the qualitative and quantitative determination of multiple mycotoxins in other complex matrices.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

基于间接竞争原理的流式微球技术快速检测麦芽中赭曲霉毒素A

通讯作者: 孔维军, 万丽; 孔维军, 万丽

English

Rapid Detection of Ochratoxin A in Malt by Cytometric Bead Array Based on Indirect Competition Principle

Corresponding author: 孔维军, 万丽; 孔维军, 万丽

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于间接竞争原理的流式微球技术快速检测麦芽中赭曲霉毒素A

通讯作者: 孔维军, 万丽; 孔维军, 万丽

English

Rapid Detection of Ochratoxin A in Malt by Cytometric Bead Array Based on Indirect Competition Principle

Corresponding author: 孔维军, 万丽; 孔维军, 万丽

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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