Department of Bacteriology, University of Wisconsin-Madison, 1550 Linden Drive, Madison, WI 53706, USA.
Toxins (Basel). 2020 Jan 30;12(2):93. doi: 10.3390/toxins12020093.
Culture methods supplemented with high-performance liquid chromatography (HPLC) technique provide a rapid and simple tool for detecting levels of aflatoxins (AFs) produced by fungi. This study presents a robust method for simultaneous quantification of aflatoxin (AF) B1, B2, G1, and G2 levels in several fungal cultivation states: submerged shake culture, liquid slant culture, and solid-state culture. The recovery of the method was evaluated by spiking a mixture of AFs at several concentrations to the test medium. The applicability of the method was evaluated by using aflatoxigenic and non-aflatoxigenic . A HPLC coupled with the diode array (DAD) and fluorescence (FLD) detectors was used to determine the presence and amounts of AFs. Both detectors showed high sensitivity in detecting spiked AFs or AFs produced in situ by toxigenic fungi. Our methods showed 76%-88% recovery from medium spiked with 2.5, 10, 50, 100, and 500 ng/mL AFs. The limit of quantification (LOQ) for AFs were 2.5 to 5.0 ng/mL with DAD and 0.025 to 2.5 ng/mL with FLD. In this work, we described in detail a protocol, which can be considered the foremost and only verified method, to extract, detect, and quantify AFs employing both aflatoxigenic and non-toxigenic .
文化方法辅以高效液相色谱（HPLC）技术为检测真菌产生的黄曲霉毒素（AFs）水平提供了一种快速而简单的工具。本研究提出了一种同时定量检测几种真菌培养状态下黄曲霉毒素（AF）B1、B2、G1 和 G2 水平的可靠方法：浸没摇瓶培养、液体斜面培养和固态培养。该方法的回收率通过在测试培养基中添加几种浓度的 AF 混合物进行评估。该方法的适用性通过使用产毒和非产毒真菌进行评估。采用二极管阵列（DAD）和荧光（FLD）检测器的 HPLC 用于检测 AFs 的存在和数量。两种检测器都显示出对添加的 AFs 或产毒真菌原位产生的 AFs 的高灵敏度检测。我们的方法显示，从用 2.5、10、50、100 和 500 ng/mL AFs 污染的培养基中回收 76%-88%。DAD 的定量限（LOQ）为 2.5 至 5.0 ng/mL，FLD 的 LOQ 为 0.025 至 2.5 ng/mL。在这项工作中，我们详细描述了一种可以被认为是首要的和唯一经过验证的方法，该方法使用产毒和非产毒真菌提取、检测和定量 AFs。
