Wild edible macro fungi Floccularia luteovirens proved to be a valuable source for the identification of novel lead molecules with therapeutic potential. Nevertheless, the chemical constituents of Floccularia luteovirens are rarely reported due to absence of efficient purification methods. In this study, a hydrophilic interaction chromatography directed by on-line HPLC-DPPH assay has been developed and successfully applied for the isolation of free radical inhibitor from the methanolic extract of Floccularia luteovirens. Using a hydrophilic interaction chromatographic column coupled with the HPLC-DPPH assay for screening the potential radical scavengers, the mid-pressure hydrophilic interaction chromatography (HILIC) proved to be more efficient in the pretreatment stage, yielding the fraction rich in free radical scavengers in good yield (5.9% recovery from 130.0 g of fresh F. luteovirens). From highly potent fraction, the target compound was isolated using the Click XION preparative chromatography with 17.2% recovery. The isolated compound was L-(+)-ergothioneine, where the purity (>95%) and antioxidant activity of were confirmed by chromatography and HPLC-DPPH assay, while the structure of this compound was elucidated from HR ESI-MS and NMR data. This method proved to be very efficient for the recognition and isolation of highly polar free radical inhibitors from fungi extracts, and is also applicable for the purification of highly polar compounds from other sources.

中文翻译：

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使用在线HPLC-DPPH分析指导的亲水相互作用色谱法，从黄粉虫中制备高极性自由基抑制剂。

野生食用大型真菌Floccularia luteovirens被证明是鉴定具有治疗潜力的新型先导分子的宝贵来源。然而，由于缺乏有效的纯化方法，鲜有黄腐絮状菌的化学成分鲜有报道。在这项研究中，开发了一种通过在线HPLC-DPPH测定指导的亲水相互作用色谱，并成功地用于从黄全絮甲醇提取物中分离自由基抑制剂。使用亲水相互作用色谱柱和HPLC-DPPH分析法筛选潜在的自由基清除剂，中压亲水相互作用色谱（HILIC）在预处理阶段被证明是更有效的，从而产生了富含自由基清除剂的馏分产量（5。从130.0 g新鲜的F. luteovirens中回收9％。使用Click XION制备型色谱从高效馏分中分离出目标化合物，回收率为17.2％。分离出的化合物为L-（+）-麦角硫氨酸，其纯度（> 95％）和抗氧化活性通过色谱和HPLC-DPPH分析得以证实，而该化合物的结构则通过HR ESI-MS和NMR数据得以阐明。事实证明，该方法对于从真菌提取物中识别和分离高极性自由基抑制剂非常有效，也可用于从其他来源纯化高极性化合物。分离的化合物为L-（+）-麦角硫氨酸，其纯度（> 95％）和抗氧化活性通过色谱和HPLC-DPPH测定得以证实，而该化合物的结构则通过HR ESI-MS和NMR数据得以阐明。事实证明，该方法对于从真菌提取物中识别和分离高极性自由基抑制剂非常有效，也可用于从其他来源纯化高极性化合物。分离的化合物为L-（+）-麦角硫氨酸，其纯度（> 95％）和抗氧化活性通过色谱和HPLC-DPPH测定得以证实，而该化合物的结构则通过HR ESI-MS和NMR数据得以阐明。事实证明，该方法对于从真菌提取物中识别和分离高极性自由基抑制剂非常有效，也可用于从其他来源纯化高极性化合物。