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免疫亲和萃取结合超高效液相色谱-串联质谱法测定中成药和中药材中的五种黄曲霉毒素

[Determination of five aflatoxins in Chinese patent medicines and medicinal herbs by immunoaffinity extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry].

作者信息

Han Shen, Liu Ying, Lu Meiling, Li Jianzhong, Wang Jinhua

机构信息

Technical Centre of Beijing Entry-Exit Inspection and Quarantine Bureau, Beijing 100026, China.

出版信息

Se Pu. 2011 Jul;29(7):613-7.

Abstract

A method for the determination of five aflatoxins (B1 , B2, G1 , G2, M1 ) in Chinese patent medicines and medicinal herbs by immunoaffinity extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed. The samples were extracted with 80% (v/v) methanol-water solution, followed by stepwise purification using an immunoaffinity column. The target compounds were then eluted with methanol. The extract was filtered then analyzed. With the gradient elution using a binary mobile phase containing of 0.1% formic acid-5 mmol/L ammonium acetate solution and methanol, the five aflatoxins were separated on an UHPLC BEH C18 column, followed by positive electrospray ionization and multi-reaction monitoring (MRM) provided by a triple-quadrupole tandem mass spectrometer. The limits of detection for the standard solution of aflatoxins ranged from 0.05-0.3 microg/L. The linear response was observed in the spiked concentration range of 0.5-100 microg/L with the correlation coefficients higher than 0.99. The spiked recoveries were within 62.3%-82.4% at the spiked levels of 1.0 microg/kg and 5.0 microg/kg for all the five aflatoxins with the relative standard deviations (RSDs) below 10% (n = 6). The developed method is sensitive, accurate, and reproducible with the reasonable recoveries, and can be applied to the determination of the 5 aflatoxins in the Chinese traditional patent medicines, medicinal herbs as well as other similar complex matrices.

摘要

建立了一种免疫亲和萃取结合超高效液相色谱 - 串联质谱法(UHPLC - MS/MS)测定中成药和中药材中5种黄曲霉毒素(B1、B2、G1、G2、M1)的方法。样品用80%(v/v)甲醇 - 水溶液提取,然后使用免疫亲和柱进行分步纯化。目标化合物随后用甲醇洗脱。提取物过滤后进行分析。采用含0.1%甲酸 - 5 mmol/L乙酸铵溶液和甲醇的二元流动相进行梯度洗脱,在UHPLC BEH C18柱上分离5种黄曲霉毒素,然后通过三重四极杆串联质谱仪进行正电喷雾电离和多反应监测(MRM)。黄曲霉毒素标准溶液的检测限为0.05 - 0.3 μg/L。在0.5 - 100 μg/L的加标浓度范围内观察到线性响应,相关系数高于0.99。对于所有5种黄曲霉毒素,在1.0 μg/kg和5.0 μg/kg的加标水平下,加标回收率在62.3% - 82.4%之间,相对标准偏差(RSD)低于10%(n = 6)。所建立的方法灵敏、准确、可重现,回收率合理,可应用于中成药、中药材以及其他类似复杂基质中5种黄曲霉毒素的测定。

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