高速逆流色谱法从小叶金钱草中分离制备三种黄酮苷类化合物

宋道光; 樊鑫宇; 徐顺连; 熊雯豆; 段向兰; 陈志

doi:10.13386/j.issn1002-0306.2019.13.005

高速逆流色谱法从小叶金钱草中分离制备三种黄酮苷类化合物

Preparation of Three Flavonoid Glycosides Compounds from Hydrocotyle sibthorpioides Lam. by High Speed Counter Current Chromatography

摘要

摘要: 应用高速逆流色谱法分离制备小叶金钱草中的三种大极性黄酮苷。小叶金钱草乙酸乙酯萃取物经硅胶柱色谱氯仿-甲醇(1:3)洗脱、葡聚糖凝胶70%甲醇纯化、高速逆流色谱正己烷-正丁醇-水-冰乙酸(1:1.7:1:0.1,V/V/V/V)两相溶剂系统分离,在主机转速800 r/min,流速2 mL/min,检测波长254 nm条件下进行分离制备,得到3个分离组分。对所得分离产物进行高效液相色谱分析及¹H、¹³C NMR鉴定,结果为杨梅素 3,3'-二-α-L-鼠李糖苷,木犀草素-7-O-β-D-葡萄糖醛酸苷和芹菜素-7-O-β-D-葡萄糖醛酸苷,其纯度达到90%以上。这三种化合物均为首次从小叶金钱草中分离得到,为今后小叶金钱草的化学成分研究提供依据。

Abstract: High-speed counter-current chromatography (HSCCC) was developed for separation of three high polarity flavonoid glycosides from EtOAc-soluble fractions of Hydrocotyle sibthorpioides Lam. The EtOAc-soluble fractions were subjected to silica gel column chromatography and further purification by Sephadex LH-20, eluted with gradient CHCl₃·MeOH (1:3) and MeOH·H₂O (7:3, V/V). After screening the solvent system, two phase solvent system consisting of n-hexane/n-BuOH/water/glacial acetic acid (1:1.7:1:0.1, V/V/V/V) were used to separation sample. In addition, the rotation speed, flow rate and detection wavelength were set at 2.0 mL/min, 800 r/min and 254 nm, respectively. The collected fractions were analyzed by HPLC and the purity were higher than 90%. Importantly, this three compounds were isolated from this plant for the first time. Their structures were identified by¹H NMR and¹³C NMR spectroscopy, named as myricetin 3, 3'-di-α-L-rhamnopyranoside, apigenin 7-O-β-D-glucuronide and luteolin 7-O-β-D-glucuronide. This work could provide supporting for study on the chemical constituent of Hydrocotyle sibthorpioides Lam.