制备型高效液相色谱法从槐角转化液中纯化5,7,8,4’-四羟基异黄酮  

Purification of 5,7,8,4’-tetrahydroxyisoflavone from Fermentation of Fructus Sophorae by pre-HPLC

作　　者：郑丹 彭雯鑫 毛慧淇 沈红兰 周学 张玉千 ZHENG Dan;PENG Wenxin;MAO Huiqi;SHEN Honglan;ZHOU Xue;ZHANG Yuqian(School of Biotechnology and Chemical Engineering,Taizhou College,Nanjing Normal University,Taizhou 225300,China)

机构地区：[1]南京师范大学泰州学院化学与生物工程学院,江苏泰州225300

出　　处：《生物化工》2022年第6期81-84,91,共5页Biological Chemical Engineering

基　　金：江苏省大学生创新创业训练计划项目(202113843010Y);江苏省自然科学基金面上项目(BK20211128);南京师范大学泰州学院横向项目(202018)。

摘　　要：5,7,8,4’-四羟基异黄酮是一种重要的酪氨酸酶抑制剂。本研究利用制备型高效液相色谱(pre-HPLC)从槐角的微生物转化液中分离纯化5,7,8,4’-四羟基异黄酮。考察流动相体系、进样量、流速和检测波长等因素对纯化效果的影响,确定较优的参数,并对纯化之后的产物进行结构鉴定。结果表明,55%B(甲醇∶乙腈=12∶30,V∶V)线性洗脱、流速为7 mL/min、进样量为2 mL条件下,单个样品运行时间为20 min,产物纯度可以达到96%。通过化学反应、质谱分析、红外光谱分析、核磁共振分析,确定物质为5,7,8,4’-四羟基异黄酮。利用该方法可以快速、高效地制备高纯度的5,7,8,4’-四羟基异黄酮。5,7,8,4’-tetrahydroxyisoflavone is an important tyrosinase inhibitor.In this study,5,7,8,4’-tetrahydroxyisoflavone are isolated and purified by preparative high performance liquid chromatography(pre-HPLC)from microbial transformation liquid of fructus sophorae.The influence of mobile phase system,injection amount,flow rate and detection wavelength on the purification effect is investigated,the optimal parameters are determined,and the structure of the purified product is identified.The results show that 55%B(methanol:acetonitrile=12:30,V:V)linear elution with a flow rate of 7 mL/min and an injection volume of 2 mL can achieve 96%product purity with a single sample run time of 20 min.By chemical reaction,mass spectrometry,infrared spectroscopy,and nuclear magnetic resonance analysis,the substance is determined to be 5,7,8,4’-tetrahydroxyisoflavone.This method has the characteristics of fast,high efficiency and high purity,and can be used to prepare 5,7,8,4’-tetrahydroxyisoflavone with high purity.

关 键 词：制备型高效液相色谱法 5 7 8 4’-四羟基异黄酮 槐角 棘孢曲霉 

分 类 号：R284.2[医药卫生—中药学]