Aptamer-affinity column clean-up coupled with ultra high performance liquid chromatography and fluorescence detection for the rapid determination of ochratoxin A in ginger powder

被引:34
作者
Yang, Xihui [1 ,2 ]
Kong, Weijun [2 ]
Hu, Yichen [2 ]
Yang, Meihua [2 ]
Huang, Luqi [3 ]
Zhao, Ming [1 ]
Ouyang, Zhen [1 ]
机构
[1] Jiangsu Univ, Sch Pharm, Zhenjiang, Jiangsu, Peoples R China
[2] Chinese Acad Med Sci, Inst Med Plant Dev, Peking Union Med Coll, Key Lab Bioact Subst & Resources Utilizat Chinese, Beijing 100193, Peoples R China
[3] China Acad Chinese Med Sci, Natl Resource Ctr Chinese Mat Medica, Beijing, Peoples R China
基金
美国国家科学基金会;
关键词
Aptamer-affinity columns; Fluorescence detection; Ginger powder; Ochratoxin A; Ultra high performance liquid chromatography; ZINGIBER-OFFICINALE; EXTRACTION; MOLECULES; SELECTION;
D O I
10.1002/jssc.201301136
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Aptamers are single-stranded oligonucleotides with high affinity and specificity and are widely used in targets separation and enrichment. Here, an aptamer-affinity column (AAC) was firstly prepared in-house through a covalent immobilization strategy. Then, ochratoxin A (OTA) in ginger powder was absorbed and enriched using the new aptamer-based clean-up technology for the first time, and was further analyzed by ultra high performance liquid chromatography with fluorescence detection. After optimization, the average recoveries for blank samples spiked with OTA at 5, 15, and 45 g/kg ranged from 85.36 to 96.83%. Furthermore, the AAC exhibited a similar accuracy as an immunoaffinity column to clean up OTA in ginger powder. Above all, it exhibited better reusability, twice that of the immunoaffinity column, had lower toxicity and cost, and took less time. Of 25 contaminated ginger powder samples, OTA contamination levels ranged from 1.51 to 4.31 g/kg, which were lower than the European Union (EU) regulatory limits. All the positive samples were further confirmed by ultra-fast LC with MS/MS. In conclusion, the method of clean-up based on the AAC coupled to ultra-HPLC with fluorescence detection was rapid, specific, and sensitive for the quantitative analysis of OTA in a complex matrix.
引用
收藏
页码:853 / 860
页数:8
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