Coupled with evaporative light scattering detection, a high-speed counter-current chromatography (HSCCC) method was developed for preparative isolation and purification of three glycine-conjugated cholic acids, glycochenodeoxycholic acid (GCDCA), glycohyodeoxycholic acid (GHDCA), and glycohyocholic acid (GHCA) from Pulvis Fellis Suis (Pig gallbladder bile) for the first time. The separation was performed with a two-phase solvent system consisted of chloroform-methanol-water-acetic acid (65:30:10:1.5, v/v/v/v) by eluting the lower phase in the head-to-tail elution mode. The revolution speed of the separation column, flow rate of the mobile phase, and separation temperature were 800 rpm, 2 mL/min and 25°C, respectively. In a single operation, 33 mg of GCDCA, 38 mg of GHDCA, and 23 mg of GHCA were obtained from 200 mg of crude extract with the purity of 95.65%, 96.72%, and 96.63%, respectively, in one-step separation. The HSCCC fractions were analyzed by high-performance liquid chromatography (HPLC) and the structures of the three glycine-conjugated cholic acids were identified by ESI-MS, 1H NMR, and 13C NMR.

中文翻译：

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通过高速逆流色谱结合 ELSD 检测从 Pulvis Fellis SUIS 中分离和纯化三种甘氨酸共轭胆酸

结合蒸发光散射检测，开发了一种高速逆流色谱 (HSCCC) 方法，用于制备性分离和纯化三种甘氨酸结合胆酸、甘氨鹅脱氧胆酸 (GCDCA)、甘草脱氧胆酸 (GHDCA) 和甘草胆酸 ( GHCA）首次来自 Pulvis Fellis Suis（猪胆囊胆汁）。使用由氯仿-甲醇-水-乙酸 (65:30:10:1.5, v/v/v/v) 组成的两相溶剂系统进行分离洗脱模式。分离柱转速、流动相流速、分离温度分别为800rpm、2mL/min、25℃。在一次手术中，33 mg GCDCA，38 mg GHDCA，从200mg粗提物中一步分离得到GHCA和23mg，纯度分别为95.65%、96.72%和96.63%。HSCCC 馏分通过高效液相色谱 (HPLC) 进行分析，三种甘氨酸共轭胆酸的结构通过 ESI-MS、1H NMR 和 13C NMR 进行鉴定。