霉菌毒素对作物如中药(TCMs)的污染是全世界公认的问题。因此,在生产和加工过程中测定TCM中的霉菌毒素是非常必要的,这意味着快速,需要灵敏和准确的分析方法。在这项工作中,提出了一种基于96孔微量滴定板的可视化蛋白质芯片新方法。结合比色法,五种霉菌毒素(曲霉毒素A,玉米赤霉烯酮,脱氧雪腐镰刀菌烯醇,可以在30分钟内同时检测到90个样品(TCM)中的黄曲霉毒素B1和伏马菌素B1)。五种真菌毒素的检出限为0.25μg/kg,0.33μg/kg,11.84μg/kg,0.06μg/kg,和3.58μg/kg,能充分满足《中国药典》(2020年版)对霉菌毒素的规定要求。在反复的条件下,在实际样品上进行了实验,验证了该方法的可行性。结果表明,所有分析物的回收率在70%~120%之间,相对标准偏差小于15%。与LC-MS/MS相比,这种方法大大减少了所需的时间,与基于荧光的筛选测定相比,比色技术提供了更直接的结果。该方法显示出用于质量控制的TCM的快速和灵敏的现场检测的巨大潜力。
The pollution of mycotoxins to crops such as traditional Chinese medicines (TCMs) is an established problem throughout the world. Thus, mycotoxin determination in TCMs during production and processing is significantly necessary, which means rapid, sensitive and accurate analytical methods are needed. In this work, a new method of visual protein microarray based on a 96-well microtiter plate was proposed. Combined with a colorimetric method, five mycotoxins (ochratoxin A, zearalenone, deoxynivalenol, aflatoxin B1 and fumonisin B1) in 90 samples (TCMs) could be detected simultaneously within 30 minutes. The detection limits for the five mycotoxins are 0.25 μg/kg, 0.33 μg/kg, 11.84 μg/kg, 0.06 μg/kg, and 3.58 μg/kg, which can satisfy specified requirements of mycotoxins in the Chinese Pharmacopoeia (2020 edition) adequately. Under repeated conditions, experiments were carried out on actual samples to verify the feasibility of the method. The results showed that the recoveries of all analytes were between 70 % and 120 %, and the relative standard deviations were less than 15 %. In comparison to LC-MS/MS, this method significantly reduces the required time, and the colorimetric technique offers more direct results compared to fluorescence-based screening assays. This method exhibits substantial potential for the rapid and sensitive on-site detection of TCMs for quality control.