高速逆流色谱用于天然产物分离和指纹图谱构建

利用国产高速逆流色谱分离纯化雪莲黄酮类成分和丹参醌类成分。雪莲分离选用氯仿 甲醇 水 (10∶7∶3)体系 ,固定相保留率 72 % ,仪器参数 80 0r min 2mL min ,采用一步洗脱法 ,7h内得到 14个组分 ;丹参分离选用正己烷 乙醇 水 (10∶5 5∶4 5 )体系 ,固定相保留率达到 78 8% ,采用分步洗脱 ,3个产地丹参在 13h内各分离得到 12个洗脱组分。HSCCC洗脱图谱可以表现出不同产地丹参的差别 ,并且各对应洗脱峰保留时间的相对标准偏差 <3% ,因此提出将HSCCC作为构建中药指纹图谱的方法之一 ,其可行性需要通过与常规的指纹图谱构建方法比较之后做出评价。

Application of high-speed counter-current chromatography on separation of natural products and development of fingerprint

High-speed counter-current chromatography (HSCCC) is a liquid-liquid partition system, which is practical in separation and purification of Chinese traditional medicines. In this study, separation of Chinese traditional medicinal plant -Saussurea involucrate Kar. Et Kin .and Salvia miltiorrhiza Bunge was performed on HSCCC. HSCCC (TBE-300) is from Shenzhen Tauto Biotech Co. Ltd., China, with three preparative coils connected in series (diameter of tube=2.6 mm, total volume=300 mL) and a 20 mL sample loop. Saussurea involucrate Kar. Et Kin. from Xinjiang province was separated by HSCCC in a one-step elution. HSCCC system was performed at a speed of 800 r/min and at a flow-rate of 1.7 mL/min. 14 peak fractions were eluted within 7hours. Salvia miltiorrhiza Bunge from 3 different growth locations were separated by HSCCC in a step-wise elution program:0-470 min, with solvent system A; then with solvent system B. HSCCC system was performed at a speed of 900rpm and at a flow-rate of 2 mL/min. All 12 peak fractions could be eluted within 13 h. Based on the above results, HSCCC was forwarded as a method to develop the fingerprint of Chinese traditional medicines. There would be a further study on the feasibility of HSCCC in fingerprinting.