高速逆流色谱分离制备椭圆叶花锚中的2种口山酮苷元

椭圆叶花锚的主要活性成分为口山酮类化合物,这类化合物具有利胆、抗炎、抗菌及抗病毒活性。应用高速逆流色谱法建立了2种高纯度口山酮苷元的分离制备方法。对椭圆叶花锚氯仿萃取部位运用高速逆流色谱分离纯化,以正己烷-乙酸乙酯-甲醇-水(5:5:7:5, v/v/v/v)为两相溶剂系统,上相为固定相,下相为流动相。在主机转速800 r/min,流动相流速1.5 mL/min,检测波长254 nm条件下进行分离制备。所得产物经高效液相色谱分析检测,其化学结构由核磁共振氢谱(1H NMR)和核磁共振碳谱(13C NMR)鉴定。在此条件下,从100 mg粗样品中一步分离得到18 mg 1-羟基-2,3,5-三甲氧基口山酮,14 mg 1-羟基-2,3,4,5-四甲氧基口山酮。经高效液相色谱分析,其纯度均达98%以上。该方法简便、快速,所得产物纯度高,适合于椭圆叶花锚口山酮苷元的制备分离。     

高速逆流色谱分离纯化九里香中的黄酮类化合物

应用高速逆流色谱法分离纯化了九里香中的4种黄酮类化合物。以石油醚-乙酸乙酯-甲醇-水(5:5:4.8:5, v/v/v/v)作为两相溶剂系统,上相为固定相,下相为流动相,以主机转速800 r/min、流速2.0 mL/min、单次进样量200 mg的条件成功地从4.0 g九里香粗提物中分离纯化出54.31 mg 5,7,3′,4′,5′-五甲氧基黄酮(重结晶后)、107.68 mg 5-羟基-6,7,3′,4′-四甲氧基黄酮、215.54 mg 5-羟基-6,7,8,3′,4′-五甲氧基黄酮、84.36 mg 5-羟基-6,7,8,3′,4′,5′-六甲氧基黄酮,纯度均在95%以上。各化合物的结构均由质谱和核磁共振氢谱、碳谱鉴定。其中化合物5-羟基-6,7,3′,4′-四甲氧基黄酮为首次从九里香中分离得到。     

高良姜中二苯基庚烷类化合物的高速逆流色谱分离制备

应用高速逆流色谱法(HSCCC)分离纯化了高良姜中3种二苯基庚烷类化合物。以正己烷-乙酸乙酯-甲醇-水(2:3:1.75:1, v/v/v/v)为两相溶剂系统,下相为固定相,上相为流动相,在主机转速为858 r/min、流速1.5 mL/min的条件下,从122.20 mg高良姜石油醚萃取物中经一步HSCCC分离可制备得到5R-羟基-7-(4-羟基-3-甲氧基苯基)-1-苯基-3-庚酮(7.37 mg)、7-(4-羟基-3-甲氧基苯基)-1-苯基-4E-烯-3-庚酮(9.11 mg)和1,7-二苯基-4E-烯-3-庚酮(15.44 mg),经高效液相色谱分析,纯度均大于93%,各化合物的结构由质谱和核磁共振氢谱、碳谱鉴定确证。该方法简便、快速、高效,可用于高良姜中二苯基庚烷类化合物的快速分离制备。     

生附子中C19型二萜生物碱的高速逆流色谱分离及其结构鉴定

应用高速逆流色谱法分离制备了生附子中的3个C19型二萜生物碱类化合物。以正己烷-乙酸乙酯-甲醇-水(3:5:4:5, v/v/v/v)为两相溶剂系统,上相为固定相,下相为流动相,在主机转速850 r/min、流动相流速2.0 mL/min、检测波长235 nm条件下进行分离制备;一次性从90 mg附子总碱粗提物中分离制备得到15.3 mg北草乌碱,35.1 mg中乌头碱和22.7 mg次乌头碱,经高效液相色谱分析,测得它们的纯度分别为97.9%、96.2%和99.2%。并应用波谱(电喷雾离子质谱、核磁共振氢谱和核磁共振13C谱)解析法确定了它们的结构。利用该方法可以对生附子中的二萜类生物碱成分进行快速的分离和纯化     

夏天无生物碱的高速逆流色谱分离纯化

采用pH-区带精制逆流色谱与常规高速逆流色谱相结合的方法快速分离纯化夏天无总生物碱.利用pH-区带精制逆流色谱对夏天无总生物碱进行分离,以正己烷-乙酸乙酯-甲醇-水(5∶ 5∶ 2∶ 8, V/V, 上相加5 mmol/L三乙胺,下相加5 mmol/L HCl)为溶剂系统,上样量3.0 g,分离得到1个混合物和3个高纯度的生物碱单体:原阿片碱(375 mg)、苏元胡碱甲(362 mg)和比枯枯灵(246 mg), 其纯度分别为97.5%,95.6%和97.1%.所得混合物850 mg经常规高速逆流色谱二次分离,以正己烷-乙酸乙酯-甲醇-水(1∶ 0.8∶1.1∶ 1, V/V)为溶剂系统,得到比枯枯灵(105 mg)和四氢巴马亭(470 mg)单体,纯度分别为99.1%和99.7%.从该药材分得苏元胡碱甲,两种制备分离模式相结合,提高了夏天无生物碱的分离效率.     

高速逆流色谱分离纯化川西獐牙菜中3种(口山)酮苷元

建立了高速逆流色谱分离川西獐牙菜中3种(口山)酮苷元的方法.溶剂系统为V(正己烷):V(乙酸乙酯):V(甲醇):V(水)=5:5:6:4,上相为固定相,下相为流动相,转速为850 r/min,流速为2.5 mL/min,温度为25℃.从川西獐牙菜氯仿萃取部位的(口山)酮苷元混合物中制备得到18mg 1,8-二羟基-3,...     

含喹唑啉硫醚的杨梅素衍生物的设计、合成及生物活性研究

以杨梅苷为原料,通过活性拼接,设计并合成一系列含喹唑啉硫醚的杨梅素衍生物,其结构通过1H NMR、13C NMR、19F NMR和HRMS进行确证.生物活性测试结果表明,该类化合物对水稻白叶枯病菌(X.Oryzae)、柑橘溃疡病菌(X. Citri)和烟草青枯病菌(R. Solanacearum)表现出一定的抑制活性.其中, 5,7-二甲氧基-3-(3-((6-溴喹唑啉-4-基)硫基)丙氧基)-2-(3,4,5-三甲氧基苯基)-4H-色烯-4-酮(A15)对水稻白叶枯病菌的EC50值为13.9μg/mL,优于对照药叶枯唑(88.9μg/mL)和噻菌铜(68.1μg/mL);5,7-二甲氧基-3-(4-((6-氯喹唑啉-4-基)硫基)丁氧基)-2-(3,4,5-三甲氧基苯基)-4H-色烯-4-酮(A3)、5,7-二甲氧基-3-(3-((6-氯喹唑啉-4-基)硫基)丙氧基)-2-(3,4,5-三甲氧基苯基)-4H-色烯-4-酮(A14)、5,7-二甲氧基-3-(3-((6-溴喹唑啉-4-基)硫基)丙氧基)-2-(3,4,5-三甲氧基苯基)-4H-色烯-4-酮(A15)和5,7-二甲氧基-3-(3-((6-氟喹唑啉-4-基)硫基)丙氧基)-2-(3,4,5-三甲氧基苯基)-4H-色烯-4-酮(A16)对烟草青枯病菌的EC50值分别为1.1,14.0,11.9和7.5μg/mL,优于对照药叶枯唑(38.5μg/mL)和噻菌铜(184.8μg/mL).活体实验结果表明,化合物A15对水稻白叶枯病菌的具有良好治疗活性和保护活性.通过扫描电镜成像初步探讨了目标化合物A3对烟草青枯病菌和A15对水稻白叶枯病菌的抑菌作用机制.     

高海拔大马士革玫瑰的黄酮类化学成分

为研究高海拔种植大马士革玫瑰的化学成分,采用95%乙醇为溶剂进行连续回流提取,并采用硅胶、聚酰胺、C18及Sephadex LH-20凝胶等材料行分离纯化,最终得到了9个黄酮醇类化合物（1, 3, 5～11）和两个黄酮类化合物（2和4）,其结构经¹H NMR和¹³C NMR表征并结合理化方法鉴定为：5,7-二羟基-3,6,4'-二甲氧基黄酮醇（1）、 5,7-二羟基-6,4'-二甲氧基黄酮（2）、 5,7,4'-三羟基-3,6-甲氧基黄酮醇（3）、 5,7-二羟基-6,8,4'-三甲氧基黄酮（4）、 5,4'-二羟基-3,6,7-二甲氧基黄酮醇（5）、 5,7-二羟基-3,6,8,4'-三甲氧基黄酮醇（6）、 8-甲氧基山奈酚（7）、山奈酚（8）、槲皮素（9）、槲皮素 3-O-a-L-阿拉伯呋喃糖苷（10）、银锻苷（11）,其中化合物1～5为首次从蔷薇属植物中分离得到,化合物1～7、 10、 11为首次从大马士革玫瑰中分离得到。      

高速逆流色谱法分离制备鱼藤根中的2种鱼藤酮类化合物

建立了分离制备鱼藤根中2种鱼藤酮类化合物的高速逆流色谱法。以正己烷-乙酸乙酯-甲醇-水(体积比为7:0.25:5:3)为两相溶剂系统,上相为固定相,下相为流动相,在主机转速850 r/min、流速2.0 mL/min、检测波长254 nm条件下进行分离制备,从50 mg鱼藤根粗提物中得到了2种鱼藤酮类化合物,分别为6.4 mg纯度为96.60%的鱼藤酮和23.4 mg纯度为97.87%的鱼藤素。该方法为鱼藤酮类化合物的深入研究提供了物质基础。     

硅胶柱色谱结合高速逆流色谱法分离纯化荷花中黄酮类化合物

采用硅胶柱色谱结合高速逆流色谱法分离纯化了荷花中3种黄酮类化合物。荷花粗提物先经过硅胶柱色谱初步分离,得到黄酮含量高的组分,再经过高速逆流色谱分离,以乙酸乙酯-乙醇-水-乙酸(4:1:5:0.025, v/v/v/v)为两相溶剂系统,上相为固定相,下相为流动相,在主机转速800 r/min、流速2.0 mL/min、检测波长254 nm条件下,从150 mg样品中一次性分离制备得到6.1 mg槲皮素-3-O-β-D-葡萄糖醛酸苷(I), 14.8 mg杨梅素-3-O-β-D-葡萄糖苷(II)和20.2 mg紫云英苷(III),经高效液相色谱检测其纯度分别为97.0%、95.4%、96.3%,并通过质谱和核磁共振氢谱、碳谱鉴定各化合物的结构。该方法简便、快速、节省溶剂,可以对荷花中的黄酮类化合物进行快速有效的分离纯化,具有较好的实用价值,为荷花资源的进一步开发应用提供了参考依据。     

Preparative separation of flavonoid glycosides in leaves extract of Ampelopsis grossedentata using high-speed counter-current chromatography

Preparative separation of flavonoid glycosides in leaves extract of Ampelopsis grossedentata was conducted using high-speed counter-current chromatograph (HSCCC) with a solvent system composed of n-hexane-ethyl acetate-methanol-water (1:6:1.5:7.5, v/v). In a single operation, 28 mg of 5,7-dihydroxy-3',4'-trihydroxyflavone-3-O-6'-rhamnose and 18 mg of 5,7-dihydroxy-3',4'-dihydroxyflavone-3-O-6'-rhamnose was obtained from 150 mg of the extract. The chemical structure of the two compounds was elucidated by electrospray ionization (EIS) MS and NMR.     

Preparative isolation and purification of psoralen and isopsoralen from Psoralea corylifolia by high-speed counter-current chromatography

Psoralen and isopsoralen were separated from Psoralea corylifolia by high-speed counter-current chromatography (HSCCC). A two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (5:5:4.5:5.5, v/v) was used for HSCCC separation, and yielded, from 100 mg of crude extract, 39.6 mg of psoralen and 50.8 mg of isopsoralen each at over 99% purity as determined by high performance liquid chromatography (HPLC). The identification of psoralen and isopsoralen were performed with 1H NMR and 13C NMR.     

Preparative separation of rhein from Chinese traditional herb by repeated high-speed counter-current chromatography

High-speed counter-current chromatography (HSCCC) was repeatedly used for isolation and purification of rhein from Rheum officinale Baill (Dahuang) with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (3:7:5:5, v/v), which had been selected by analytical (HSCCC). Using two preparative units of the HSCCC centrifuge, about a 500 mg amount of the crude extract was separated, yielding 6.7 mg of rhein at a high purity of over 97%.     

Isolation and purification of oridonin from the whole plant of Isodon rubescens by high-speed counter-current chromatography

Semi-preparative high-speed counter-current chromatography (HSCCC) was successfully used for isolation and purification of oridonin from Isodon rubescens by using a two-phase-solvent system composed of n-hexane-ethyl acetate-methanol-water (2.8:5:2.8:5, v/v/v/v). The targeted compound isolated, collected and purified by HSCCC was analyzed by high performance liquid chromatography (HPLC). A total of 40.6 mg of oridonin with the purity of 73.5% was obtained in less than 100 min from 100 mg of crude Isodon rubescens extract. The chemical structure of the compound was identified by IR, 1H-NMR and 13C-NMR.     

Isolation of three sesquiterpene lactones from the roots of Cichorium glandulosum Boiss. et Huet. by high-speed counter-current chromatography

Because of the skeletal complexity and similarity of the polarity, little research was reported on the isolation of sesquiterpene lactones by high-speed counter-current chromatography (HSCCC). Herein, three sesquiterpene lactones were successfully purified from the ethyl acetate extract of the roots of the traditional Uyghur medicinal plant Cichorium glandulosum Boiss. et Huet. by HSCCC. The separation was performed in two steps with two solvent systems: n-hexane-ethyl acetate-methanol-water (1.5:5:2.75:5, v/v/v/v) and ethyl acetate-methanol-water (20:1:20, v/v/v). From 166 mg of the ethyl acetate extract, 19 mg of lactucopicrin was isolated with the first solvent system and 10 mg of 11beta,13-dihydrolactucin and 16 mg of lactucin were obtained with the second solvent system. All purified compounds were over 94% purity as determined by HPLC analysis, and these chemical structures were confirmed by (1)H NMR and (13)C NMR.     

Application of preparative high-speed counter-current chromatography for isolation and separation of schizandrin and gomisin A from Schisandra chinensis

Following an initial cleaning-up step on the D101 macroporous resin, a preparative high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (1:0.9:0.9:1, v/v) was used to isolate and separate schizandrin and gomisin A from Schisandra chinensis. A total of 107 mg schizandrin and 36 mg gomisin A with purities of 99.5% and 99.1% were obtained from 400 mg crude extract in one-step elution and less than 3 h, and the structure identification was performed by UV, IR, MS, 1H NMR and 13C NMR.     

SEPARATION OF THE MINOR FLAVONOLS FROM FLOS GOSSYPII BY HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY

An effective high-speed countercurrent chromatography (HSCCC) method was established for further separation and purification of four minor flavonols in addition to five major flavonols which were reported by our previous study from extracts of Flos Gossypii. HSCCC was performed with three two-phase solvent systems composed of n-hexane-ethyl acetate-methanol-water (7.5:15:6:7, v/v), (2.5:15:2:7, v/v) and (0:1:0:1, v/v). The separation was repeated 3 times, and 3.8 mg of 8-methoxyl-kaempferol-7-O-β-D-rhamnoside (HPLC purity 98.27%), 6.7 mg of astragalin (HPLC purity 94.18%), 3.3 mg of 4'-methoxyl-quercetin-7-O-β-D-glucoside (HPLC purity 94.30%) and 8.2 mg of hyperoside (HPLC purity 93.48%) were separated from 150 mg of the crude sample. The chemical structures of the flavonols were confirmed by MS, (1)H NMR and (13)C NMR. Meanwhile, the results indicated that the target compound with smaller K value (<0.5) can be separated by increasing column length of HSCCC. And four separation rules of flavonols according to the present study and references were summarized, which can be used as a useful guide for separation of flavonols by HSCCC.     

Flow rate gradient high-speed counter-current chromatography separation of five diterpenoids from Triperygium wilfordii and scale-up

In this paper, high-speed counter-current chromatography (HSCCC) instruments with different gravitational forces were applied for the separation of bioactive compounds from Triperygium wilfordii Hook.f. The critical parameters including sample concentration, sample volume and flow rate were first optimized on an analytical Mini-DE HSCCC system, and then scaled up to a preparative TBE 300A HSCCC system. Although this scale-up process was performed using different CCC instruments with different centrifuges and gravitational forces, the same resolutions were obtained and the elution time could be predictable. Five diterpenoid compounds and one unknown compound were separated from Triperygium wilfordii Hook.f. by HSCCC with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (HEMW) (3:2:3:2, v/v/v/v). This one-step flow gradient separation produced triptonide (25 mg), isoneotriptophenolide (77 mg), hypolide (83 mg), unknown compound (1 mg), triptophenolide (42 mg), triptonoterpene methyl ether VI (37 mg) from 320 mg crude extract with purities of 98.2%, 96.6%, 98.1%, 95.3%, 95.1%, and 96.5%, respectively. Their purities and structures were identified by high-performance liquid chromatography, mass spectrometry and NMR. This paper demonstrates that analytical CCC plays an important role in optimizing parameters and scale-up process when analytical CCC and preparative CCC are supplied by different manufacturers with different gravitational forces, and the scale-up process from analytical CCC to preparative CCC is still predictable.