中文摘要：基于免疫亲和柱(IAC)净化的提取方法已经用于检测药材中的黄曲霉毒素，但还不能在所有样品基质中得到令人满意的结果。难点在于药材的化学复杂性，及迫切需要弄清IAC净化的哪一个步骤限制了许多不同种类的药材中黄曲霉毒素的检测范围。本研究发现了两个主要因素：（1）高提取酸度和（2）高共萃取干扰化合物，严重降低了抗原抗体的识别，导致了严重的非特异性吸附。为此展开了一系列研究，优化提取效率。研究发现用0.1%磷酸盐缓冲液(pH 7.8, 2% Tween-20)以1：8稀释率稀释样品，能减轻高酸度的影响，并改善基体效应。经验证后，用该方法成功测定了实际样品中的黄曲霉毒素浓度。该方法可以用于从不同酸度的复合基质中提取黄曲霉毒素。
外文摘要：Although extraction methods based on immunoaffinity column (IAC) cleanup have been used to detect aflatoxins in medicinal herbs, they do not yield satisfactory results for all sample matrices. The difficulty arises from the chemical complexity of the herbs, and there is a pressing need to determine which steps in IAC cleanup limit the scope of aflatoxin detection in many different kinds of medicinal herbs. In this work, we found that there were two main factors that severely decreased antibody-antigen recognition and led to serious nonspecific adsorption: (1) high extract acidity and (2) high co-extraction of interfering compounds. We therefore carried out a systematic study to optimize extraction efficiency. We found that dilution of samples in 0.1 M phosphate buffer solution (pH 7.8, 2% Tween-20) at a 1:8 dilution ratio mitigated the effect of high acidity, decreased co-precipitation of compounds and nonspecific adsorption, and ameliorated the matrix effect. To validate this finding, and test if our method is widely applicable to in different kinds of herbal materials, we analyzed several representative complex sample matrices including fructus, cortex, and radix with varying extract pH values. The recovery efficiency was generally higher than 70%. We further validated our method by testing a certified reference material, and found that our approach accurately quantified aflatoxin concentration. After validation, we successfully used this method to determine the aflatoxin concentration of real samples. The approach described here could potentially be used to extract aflatoxin from other complex matrices with varying acidity.
外文关键词：Aflatoxins；Medicinal herb；Immunoaffinity column cleanup；Certified reference material
作者：Zhang, L；Dou, XW；Kong, WJ；Liu, CM；Han, X；Yang, MH
作者单位：Chinese Acad Med Sci
期刊名称：JOURNAL OF CHROMATOGRAPHY A
期刊影响因子：3.926
出版年份：2017
出版刊次：2
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