黄曲霉毒素
化学
轨道轨道
代谢物
色谱法
黄曲霉
检出限
乙酸乙酯
质谱法
致癌物
真菌毒素
甲酸
生物化学
食品科学
作者
Huali Xie,Xiupin Wang,Liangxiao Zhang,Tong Wang,Wen Zhang,Jun Jiang,Perng‐Kuang Chang,Zhi-Yuan Chen,Deepak Bhatnagar,Qi Zhang,Peiwu Li
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2018-11-16
卷期号:90 (24): 14331-14338
被引量:28
标识
DOI:10.1021/acs.analchem.8b03703
摘要
Aflatoxins, highly toxic and carcinogenic to humans, are synthesized via multiple intermediates by a complex pathway in several Aspergilli, including Aspergillus flavus. Few analytical methods are available for monitoring the changes in metabolite profiles of the aflatoxin biosynthesis pathway under different growth and environmental conditions. In the present study, we developed by a D-optimal mixture design a solvent system, methanol/dichloromethane/ethyl acetate/formic acid (0.36/0.31/0.32/0.01), that was suitable for extracting the pathway metabolites. The matrix effect from dilution of cell extracts was negligible. To facilitate the identification of these metabolites, we constructed a fragmentation ion library. We further employed liquid chromatography coupled with high-resolution mass spectroscopy (UHPLC-HRMS) for simultaneous quantification of the metabolites. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.002–0.016 and 0.008–0.05 μg/kg, respectively. The spiked recovery rates ranged from 81.3 to 100.3% with intraday and interday precision less than 7.6%. Using the method developed to investigate the time-course aflatoxin biosynthesis, we found that precursors, including several possible toxins (with a carcinogenic group similar to aflatoxin B1), occurred together with aflatoxin, and that production increased rapidly at the early growth stage, peaked on day four, and then decreased substantially. The maximum production of aflatoxin B1 and aflatoxin B2 occurred 1 day later. Moreover, the dominant branch pathway was the one for aflatoxin B1 formation. We revealed that the antiaflatoxigenicity mechanism of Leclercia adecarboxylata WT16 was associated with a factor upstream of the aflatoxin biosynthesis pathway. The design strategies can be applied to characterize or detect other secondary metabolites to provide a snapshot of the dynamic changes during their biosynthesis.
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