桑黄中黄嘌呤氧化酶抑制剂的筛选分离及质谱分析

Screening,separation and mass spectrometry analysis of xanthine oxidase inhibitors in Phellinus igniarius

以桑黄为研究对象,综合利用数学模型、色谱和质谱技术,建立了快速筛选分离桑黄中黄嘌呤氧化酶抑制剂的新方法。利用数学模型结合响应面优化桑黄中黄酮类活性成分的提取工艺,以黄嘌呤氧化酶为靶点,应用超滤-质谱技术快速筛选桑黄中黄嘌呤氧化酶的抑制剂;在活性指导下,利用高速逆流色谱技术分离纯化桑黄中活性成分,采用高效液相色谱-高分辨质谱结合核磁共振波谱技术对其进行结构鉴定。结果表明,在此条件下得到桑黄黄酮提取率为24.95 mg/g,优于响应面试验;从桑黄中筛选并分离出3个纯度均在90%以上的黄嘌呤氧化酶抑制剂,抑制效果强弱顺序为桑黄素D>桑黄素E>纤孔菌素A。本方法快速、灵敏,可为桑黄药理作用研究提供支撑,同时可为桑黄食品开发提供可行方法和理论支持。

A new method for the rapid screening and separation of xanthine oxidase inhibitors from Phellinus igniarius was developed using a combination of mathematical modeling,chromatography and mass spectrometry.The extraction process offlavonoid active ingredients in Phellinus igniarius was optimized by using mathematical modeling combined with response surface method,the ultrafiltration-mass spectrometry technique was applied to rapidly screen the inhibitors of xanthine oxidase in Phellinus igniarius;Under the guidance of activity,the active ingredients in Phellinus igniarius were separated and purified by high speed countercurrent chromatography(HSCCC),high performance liquid chromatography-high resolution mass spectrometry coupled with nuclear magnetic resonance spectroscopy was used for the structural identification of the active ingredients in Phellinus igniarius.The results showed that the extraction rate of Phellinus igniariusflavonoids was 24.95 mg/g,which was better than the response surface test;three xanthine oxidase inhibitors with the purity of above 90%were screened and isolated from Phellinus igniarius,and their inhibition effect was in the order of phelligridin D>phelligridin E>Fibrocidin A.This method is rapid,sensitive,and can provide support for the study of the pharmacological action of Phellinus igniarius,and also provide feasible method and theoretical support for the development of Phellinus igniarius food.