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.
Keywords: counter-current chromatography; large scale; liquid–liquid chromatography; mycotoxin purification.