半制备HPLC分离纯化海星皂苷novaeguinoside A  

Isolation and Purification of novaeguinoside A from Starfish by Semi-preparative HPLC

作　　者：康俊霞[1,2] 康永锋[2] 韩华[1] 

机构地区：[1]浙江大学动物科学学院,浙江杭州310058 [2]上海海洋大学食品学院,上海201306

出　　处：《实验室研究与探索》2013年第10期17-20,共4页Research and Exploration In Laboratory

基　　金：浙江省教育厅科研项目(Y200803020)

摘　　要：采用半制备反相高效液相色谱法从镶边海星(Craspidaster sp.)中分离纯化皂苷,并建立纯化条件以获得单体化合物,为半制备反相高效液相色谱纯化海星皂苷提供实验参考依据。对得到的海星粗总皂苷通过正相、反相硅胶及葡聚糖凝胶LH-20柱层析进行分离,然后确定分离制备条件,用半制备高效液相色谱对薄层层析板显示的同一斑点进行纯化获得单体化合物。通过检测获得制备单体化合物的色谱条件为:Zorbax 300 SB-C 18柱(5μm;250 mm×9.4 mm i.d.),RID示差检测器,柱温30℃,流动相MeOH-H_2O(47:53),流速1.5 mL/min,进样量110μL(样品浓度18 mg/mL),在保留时间32.96 min处收集到36 mg的白色粉末状单体化合物。经核磁共振谱等广泛一维和二维谱图鉴定其结构为novaeguinoside A。Asterosaponins were isolated and purified from starfish（ Craspidaster sp. ）by reversed-phase semi-preparative HPLC, and the optimum purificated condition was developed to search for active compounds. Asterosaponins in the n- BuOH extracted from starfish were isolated and purified by different chromatographic methods including silica gel CC, ODS CC and Sephadex LH-20 CC. Then the fraction which displayed a single spot detected by TLC was purified by reversed-phase semi-preparative HPLC. The chromatographic conditions for preparation compounds were investigated and determined to be Zorba 300 SB-C 18 column （5μm; 250 mm × 9.4 mm i. d. ） , refractive index detector, column temperature：30℃ , mobile phase：47% MeOH, flow rate： 1.5 mL/min） and the injection amount was 110 μL with sample concentration ：18 mg/mL. Finally the compound a-3, a colorless crystalline powder, was obtained 36 mg at ta 32.96 min and its structure was elucidated as novaeguinoside A by analysis of its ESI-MS and 1D and 2D NMR spectroscopic data. This is the first time to isolated and purified Asterosaponins from starfish Craspidaster sp. which was collected in the South China Sea, China by reversed-phase semi-preparative HPLC. This is the first time to isolate novaeguinoside A from the specie starfish.

关 键 词：海星 皂苷 高效液相色谱 

分 类 号：R965[医药卫生—药理学]