目的 探究胶体金免疫层析技术用于定量检测中药材中黄曲霉毒素B1的可行性。方法 采用不同水平加标的方式,利用阴性样本考察基质干扰情况;通过考察方法精密度、灵敏度、线性、测定重复性及加标回收率等,对快检方法性能进行评价;采用快检方法完成样品测定,并结合高效液相色谱法对快检结果进行验证。结果 对阴性样本基质分别加入高浓度和低浓度黄曲霉毒素B1对照品,测定后发现陈皮和决明子等样本存在基质干扰,且低浓度加标时,干扰更大,经研究采用高稀释倍数可降低其基质干扰。快检方法精密度RSD(n=10)为4.6%,重复性RSD(n=6)为4.1%,各样本加标回收率在72.8%～112.8%之间,整体来看方法性能良好。采用两种方法检测僵蚕、全蝎、水蛭等19种药材共计43批次,测定结果表明,胶体金免疫层析快检方法与高效液相色谱方法检测结果符合率为83.7%,两种测定方法结果一致性较高。结论 本实验结果表明,胶体金免疫层析快检方法可用于部分中药材中黄曲霉毒素B1的定量检测,同时为相关快检检测标准方法的建立和提高提供了技术支持。
Abstract
OBJECTIVE To explore the feasibility of using colloidal gold immunochromatography for quantitative detection of aflatoxin B1 in traditional Chinese medicine. METHODS Negative samples were used to investigate matrix interference by different levels of spikes.The rapid inspection performance was evaluated by examining the precision, sensitivity, linearity, repeatability and recovery rate. The sample was determined by rapid test method and verified by HPLC. RESULTS High-concentration and low-concentration aflatoxin B1 reference materials were added to the negative sample matrix. After the measurement, it was found that there were matrix interferences in the samples such as tangerine peel and cassia seed, and the interference was greater when the concentration was increased. So high dilution factor was used to reduce the interference. The precision RSD of the rapid test method was 4.6% (n=10), the reproducibility RSD was 4.1% (n=6), and the recoveries of different samples were between 72.8% and 112.8%. The overall performance of the method was good. A total of 43 batches of 19 kinds of medicinal materials such as silkworm, cockroach and leeches were detected by two methods. The coincidence rate between the fast test and the HPLC test was 83.7%. Therefore, the results obtained by the two detection methods were considered to be approximate. CONCLUSION Colloidal gold immunochromatographic rapid test method can be used for the quantitative detection of aflatoxin B1 in some traditional Chinese medicines, and provides technical support for the establishment and improvement of relevant rapid detection standard methods.
关键词
中药材 /
黄曲霉毒素B1 /
快检 /
胶体金 /
高效液相色谱法
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Key words
traditional Chinese medicine /
aflatoxin B1 /
rapid determine /
colloidal gold method /
HPLC
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中图分类号：
R917
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参考文献
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脚注
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基金
国家十二五“重大新药创制”课题“中药质量安全检测和风险控制技术平台”资助(2014ZX09304307-002)
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