Evidente A, Amalfitano C, Pengue R, Altomare C
Dipartimento di Scienze Chimico-Agrarie, Università di Napoli Federico II, Portici, Italy.
Nat Toxins. 1999;7(4):133-7. doi: 10.1002/(sici)1522-7189(199907/08)7:4<133::aid-nt60>3.0.co;2-i.
A simple, very sensitive and rapid HPLC method was developed for the simultaneous quantitative analysis of both fusapyrone (FP) and deoxyfusapyrone (DFP), the two antifungal 3-substituted-4-hydroxy-6-alkyl-2-pyrones isolated from rice culture of Fusarium semitectum, in crude extracts. Such method was optimized on C-18 reverse phase column, using the isolated metabolites as standards, with a sequence of linear elution steps with a MeOH-H(2)O mixture and using an ultraviolet detector fixed at 285 nm, where both alpha-pyrones showed a characteristic absorption maximum. This method was used to quantify the bioactive metabolites in crude organic extracts from two F. semitectum strains. The recovery of FP and DFP was measured in a crude extract from a poor metabolite producer F. semitectum strain. The recovery values ranged from 84% to 99% for FP and from 99% to 101% for DFP, indicating that the method was close to quantitative recovery. Furthermore, an efficient medium pressure column chromatography and TLC combined method was developed for the isolation and purification of FP and DFP from fungal culture extracts.
建立了一种简单、灵敏且快速的高效液相色谱法，用于同时定量分析从半裸镰刀菌水稻培养物中分离得到的两种抗真菌3-取代-4-羟基-6-烷基-2-吡喃酮——镰孢吡喃酮（FP）和脱氧镰孢吡喃酮（DFP）在粗提物中的含量。该方法在C-18反相柱上进行优化，以分离得到的代谢产物为标准品，采用甲醇-水混合物进行线性洗脱，使用固定在285nm的紫外检测器，在此波长下两种α-吡喃酮均显示出特征吸收最大值。该方法用于定量两种半裸镰刀菌菌株粗有机提取物中的生物活性代谢产物。在代谢产物产量较低的半裸镰刀菌菌株的粗提物中测定了FP和DFP的回收率。FP的回收率在84%至99%之间，DFP的回收率在99%至101%之间，表明该方法接近定量回收。此外，还开发了一种高效中压柱色谱和薄层色谱相结合的方法，用于从真菌培养提取物中分离和纯化FP和DFP。
