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Figure 1 HPLC – Chromatogram at 270 nm of A. fumigatus IP 2279-94, growth at 25°C in MEM and UV spectra of metabolites detected in this culture: fumigaclavine A [A], gliotoxin [B], fumigaclavine C [C], fumitremorgin C [D], verruculogen [E], and fumagillin [F]. The first fractionation was performed as follows: four 10-minute fractions (F1-4) and one 6-minute fraction (F5). F3 fraction was secondarily split into two-minute fractions (F'1-5). Pink line: gradient line.
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Figure 2 Effect of the five fractions obtained by HPLC-DAD of the organic phase of the 37°C A. fumigatus culture filtrate at 1/2 h. Only F3 (20–30 min) showed a significant increase in transepithelial potential differences (Vt) and a decrease, albeit not significant, in transepithelial resistance (Rt). *: significant difference compared with HNEC culture medium with DMSO as controls (p
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Figure 3 Effect of serial dilutions of standard fumagillin at 1/2 h. The first significant effects observed, i.e. a decrease in transepithelial potential differences (Vt) and an increase in transepithelial resistance (Rt), were the opposite of those observed with the whole organic phase. *: significant difference compared with HNEC culture medium with DMSO as controls (p
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Figure 4 Effect of serial dilutions of standard verruculogen at 1/2 h. The first significant effects observed, i.e. an increase of transepithelial potential differences (Vt) and a decrease of transepithelial resistance (Rt), were similar to those observed with the whole organic phase. *:significant difference compared with HNEC culture medium with DMSO as controls (p
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Figure 5 Positive mode electrospray ionization MS analysis of the F'3 fraction. Despite the fact that the molecular ion at m/z 512 (M+ H)+ is not observed, the presence of ions at both m/z 494 (M- H2O)+ and m/z 534 (M + Na)+ enables this compound to be identified as verruculogen [16].
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