核酸适配体亲和柱净化-高效液相色谱法测定莲子中黄曲霉毒素B<sub>1</sub>

引用本文: 赵颖, 王楠, 高华龙, 郭子轩, 陆安祥, 郭晓军, 卢静华, 栾云霞. 核酸适配体亲和柱净化-高效液相色谱法测定莲子中黄曲霉毒素B₁[J]. 分析化学, 2020, 48(5): 662-669. doi: 10.19756/j.issn.0253-3820.201030 shu

Citation: ZHAO Ying, WANG Nan, GAO Hua-Long, GUO Zi-Xuan, LU An-Xiang, GUO Xiao-Jun, LU Jing-Hua, LUAN Yun-Xia. Determination of Aflatoxin B₁ in Lotus Seed by High Performance Liquid Chromatography with Aptamer Affinity Column for Purification and Enrichment[J]. Chinese Journal of Analytical Chemistry, 2020, 48(5): 662-669. doi: 10.19756/j.issn.0253-3820.201030 shu

核酸适配体亲和柱净化-高效液相色谱法测定莲子中黄曲霉毒素B₁

1.
锦州医科大学, 药学院, 锦州 121001;
2.
北京农业质量标准与检测技术研究中心, 农业农村部农产品质量安全风险评估实验室(北京), 农产品产地环境监测北京市重点实验室, 北京 100097;
3.
首都师范大学, 北京 100097
基金项目:
本文系北京市农林科学院科技创新专项建设基金项目（No.KJCX20170401）和青年基金项目（No.QNJJ201903）资助

摘要: 以特异性的核酸适配体作为识别元件，制备了黄曲霉毒素B₁ （AFB₁）的适配体亲和柱（AAC），用于AFB₁的特异性识别和富集。对AAC的载样液中有机溶剂含量、载样液的体积和淋洗体积等条件进行了优化。此AAC吸附AFB₁的柱容量达到（334.6±18.2） ng，且AAC可重复使用20次。莲子样品经AAC净化富集后，用高效液相色谱-光化学衍生荧光法检测，外标法定量，AFB₁检出限达到0.05 ng/mL，回收率为91.8%~108.6%。本方法为食品、农产品和中药等复杂基质中痕量毒素的提取富集和分析提供了新的参考，具有良好的应用前景。

关键词:

English

Determination of Aflatoxin B₁ in Lotus Seed by High Performance Liquid Chromatography with Aptamer Affinity Column for Purification and Enrichment

1.
College of Pharmacy, Jinzhou Medical University, Jinzhou 121001, China;
2.
Beijing Research Center for Agricultural Standards and Testing, Agricultural Product Quality and Safety Risk Assessment Laboratory of the Department of Agriculture (Beijing), Beijing Municipal Key Laboratory of Agriculture Environment Monitoring, Beijing 100097, China;
3.
Capital Normal University, Beijing 100097, China
Received Date: 11 January 2020
Revised Date: 06 March 2020
Fund Project:
This work was supported by the Science and Technology Innovation Special Construction Funded Program of Beijing Academy of Agriculture and Forestry Sciences (No.KJCX20170401) and the Beijing Agricultural Forestry Academy Youth Foundation (No. QNJJ201903).

Abstract: An aptamer affinity column (AAC) was applied to specific identification and enrichment of aflatoxin B₁ (AFB₁). Aptamer was used as a novel recognition element for the high affinity and strong specificity of target. By optimizing the loading, washing and eluting conditions of AFB₁ in the sample, AFB₁ aptamer affinity column exhibited a good sample capacity (334.6±18.2) ng. The content of AFB₁ lotus seeds was detected by photo chemical reactor for enhanced high-performance liquid chromatography combined with photo chemical reactor for enhanced detection and fluorescence detection (HPLC-PCD-FLD) and AAC clean-up, with LOD of 0.05 ng/mL and excellent recoveries of 91.8%-108.6%. Meanwhile, the AAC could be reused at least twenty times. This study provides a new method for the enrichment and purification of trace toxins in complex matrix such as food, agricultural products and traditional Chinese medicines, and has a good application prospect.

Key words:

Aptamer affinity column
/ Aflatoxin B₁
/ Solid-phase extraction
/ N-Hydroxy succinimide-activated sepharose
/ Lotus seed

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核酸适配体亲和柱净化-高效液相色谱法测定莲子中黄曲霉毒素B1

English

Determination of Aflatoxin B1 in Lotus Seed by High Performance Liquid Chromatography with Aptamer Affinity Column for Purification and Enrichment

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核酸适配体亲和柱净化-高效液相色谱法测定莲子中黄曲霉毒素B₁

Determination of Aflatoxin B₁ in Lotus Seed by High Performance Liquid Chromatography with Aptamer Affinity Column for Purification and Enrichment