pH区带逆流色谱法分离纯化丹参中的丹酚酸B和迷迭香酸

Separation and purification of salvianolic acid B and rosmarinic acid from Salvia miltiorrhiza Bge. by pH-zone-refining counter-current chromatography

采用pH区带精制逆流色谱技术(p H-ZRCCC)快速分离纯化丹参中的丹酚酸B和迷迭香酸。以乙酸乙酯-水(1∶1,V/V)为溶剂体系,上相加入三氟乙酸,使浓度达到10 mmol/L,作为固定相;下相加入氨水使浓度达到20 mmol/L,作为流动相;设置主机转速为850 r/min,流速为2.0 m L/min,检测波长为280 nm,对样品进行分离制备。从500 mg丹参粗提物中一次分离得到386 mg丹酚酸B以及25 mg迷迭香酸,经HPLC分析纯度分别为96.3%和98.1%,各化合物通过电喷雾电离(ESI)谱和1H-NMR进行了化学结构鉴定。研究结果表明,p H-ZRCCC是一种快速、高效分离纯化丹酚酸B和迷迭香酸的方法。

We rapidly separated and purified salvianolic acid B and rosmarinic acid from Salvia miltiorrhiza Bge. by pH-zone-refining counter-current chromatography （ pH-ZRCCC）. The extraction conditions included solvent system of ethyl acetate-water （1：1, V/V）, upper stationary phase of 10 mmol/L trifluoroacetic acid, lower mobile phase of 20 mmol/L NH3H2O, rotation speed of 850 r/min, flow speed of 2. 0 mL/min, and detection wavelength of 280 nm. Salvianolic acid B of 386 mg and rosmarinic acid of 25 mg were extracted from crude extract of 500 mg, whose purities were 96.3% and 98.1% through HPLC analysis. Their compound structures were identified by ESI-MS and 1^H-NMR. Experimental results show that pH-ZRCCC is a rapid and efficient method to separate and purify salvianolic acid B and rosmarinic acid.