Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Közép fasor 52, H-6726 Szeged, Hungary.
Toxins (Basel). 2023 Feb 25;15(3):178. doi: 10.3390/toxins15030178.
Aflatoxins (AFs) are a group of secondary metabolites that cause several diseases in both animals and humans. Since the discovery of this group of toxins, several effects were revealed, such as hepatic changes, carcinoma, failure, and cancer of the liver. In the European Union, there are concentration limits for this group of mycotoxins in food and feed products; thus, these substances are required in their pure forms to prepare reference standards or certified reference materials. In our present work, a liquid-liquid chromatographic method utilizing a toluene/acetic acid/water ternary system was improved. In order to enhance the purification and gain a higher amount of pure AFs in one separation run, a scale-up of the previous separation was carried out. In several scale-up steps-including the determination of the maximum concentration and volume to load on a 250 mL rotor via a loop and via a pump as well, and the quadruplication of the entire separation procedure to a 1000 mL rotor-an efficient scale-up was achieved. Utilizing a 250 mL rotor in an 8-hour workday, altogether approximately 2.2 g of total AFs could be purified with 8.2 liters of solvent, while on a 1000 mL column, approximately 7.8 g AFs could be prepared, utilizing around 31 liters of solvents.
黄曲霉毒素（AFs）是一组次级代谢物，可导致动物和人类的多种疾病。自发现该毒素以来，人们揭示了其多种作用，如肝变化、癌、衰竭和肝癌。在欧盟，食品和饲料产品中对该组霉菌毒素有浓度限制；因此，需要这些物质的纯物质来制备参考标准或认证参考材料。在我们目前的工作中，改进了一种利用甲苯/乙酸/水三元体系的液-液色谱方法。为了提高纯化效果并在一次分离运行中获得更多的纯 AFs，对先前的分离进行了放大。在几个放大步骤中，包括通过环路和泵确定在 250 mL 转子上的最大浓度和加载体积，以及将整个分离过程扩大四倍到 1000 mL 转子上，实现了有效的放大。在 8 小时工作日内使用 250 mL 转子，总共可以使用 8.2 升溶剂来纯化约 2.2 克总 AFs，而在 1000 mL 柱上，大约可以使用 31 升溶剂来制备约 7.8 克 AFs。
