Aflatoxins M1 and M2: Preparation and purification

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Abstract
A crude product containing aflatoxins M1 and M2, as well as large quantities of aflatoxins B1 and B2, obtained by fermentation of rice withAspergillus flavus NRRL 3251 was chromatographed on a silicic acid column. Almost all the B1 and B2 were separated from M1 and M2. Aflatoxins M1 and M2 were eluted together with ethanol‐chloroform (5∶95 v/v). The combined M1 and M2 fraction was placed on a Merck silica gel (0.05–0.2 mm) column to be washed with hexane‐chloroform (1∶1 v/v) and chloroform to remove traces of B1 and B2 and eluted with ethanol‐chloroform (1.5∶98.5 v/v) to obtain aflatoxin M1 and mixtures of M1 and M2. Rechromatography of M1 on another silica gel column gave pure M1 which was crystallized from acetonitrile. Aflatoxin M2 was prepared by hydrogenation of M1 and crystallized from acetonitrile.