Produção, purificação e caracterização espectrométrica da micotoxina citreoviridina produzida pelo Penicillium citreonigrum em meio de cultura YES (Yeast Extract Sucrose)

TLDR: In this article, the authors established a protocol for the production and purification of citreoviridin in order to yield enough high purity material to conduct toxicological studies that would help to elucidate the events that occurred in Maranhão.

Abstract: Mycotoxins are secondary metabolites produced by fungi, which may develop in agricultural commodities in the field and/or during transport and storage. Citreoviridin, a mycotoxin produced mainly by Penicillium citreonigrum, was first isolated in 1947 in Japan during the investigation of the “yellow rice” disease, which caused acute cardiac beriberi. In May 2006, an outbreak of beriberi was reported in the State of Maranhão, Brazil, and rice samples collected in the region, was shown to be positive for P. citreonigrum and citreoviridin. The main objective of this work was to establish a protocol for the production and purification of citreoviridin in order to yield enough high purity material to conduct toxicological studies that would help to elucidate the events that occurred in Maranhão. Citreoviridin was produced by P. citreonigrum in 48 flasks of 500 mL YES liquid culture medium for 8 days at 25oC. The toxin was extracted with chloroform, purified by semi-preparative HPLC from the liquid medium and the mycelium of 48 culture flasks, and fully characterized by UV/VIS, FT-IR, H and C NMR, LC-MS/MS and LC-MSD TOF. Citreoviridin concentration was determined by HPLC/PDA against a commercial standard. In average, 29.7 and 85.7 mg of citreoviridin were recovered from the liquid medium and mycelium, respectively, yielding a total of 5.5 g citreoviridin from 15.3 g of crude extract. About 12 g of this extract was purified yielding 3.25 g of purified citreoviridin (27.1% yield), which identity was confirmed by the spectrometric analyses. Chromatographic and gravimetric analysis showed that the purified citreoviridin was 100% pure.