Preparative LC isolation and purification of fumonisin B1 from rice culture

Procedures are presented for the production and rapid isolation of fumonisin B 1 (FB 1 ) from fumonisin B 2 (FB 2 ) and fumonisin B 3 (FB 3 ), and for the purification of the mycotoxin FB 1 from cultures of Fusarium moniliforme MRC 826 grown on rice. The toxin was extracted from rice culture with acetonitrile :water (1 :1 ; 5mL/g of culture material), filtered, and reduced in volume on a rotary evaporator to remove the acetonitrile. A three-step liquid chromatographic (LC) method was developed for the isolation of FB 1 , FB 2 , and FB 3 , and the purification of FB 1 . Preparative reversed-phase LC was used to isolate and partially purify the FB 1 . In the first step, the extract was applied to a Waters Bondapak PrepPak 500 C 18 reversed-phase cartridge and eluted with a methanol :water gradient. Fractions containing partially purified FB 1 were collected, reduced in volume, and subjected to a second preparative LC procedure. The extract was applied to two Bondapak PrepPak cyano cartridges, and isocratic elution with water :0.5% pyridine was used in the purification step. Recovery of the total FB 1 was 97% from the first isolation step and was 93% from the second purification step. Analytical high-performance liquid chromatography and fast-atom bombardment/mass spectrometry were used in determining the purity of the FB 1 . Recovery of the high purity FB 1 (95% or greater) was 77% or 2811 mg/3647 mg of starting material.