Preparative isolation and purification of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high-speed counter-current chromatography

An efficient method for the isolation and purification of 12,13-dihydroxyeuparin from Radix Eupatorii Chinensis by high speed counter-current chromatography (HSCCC) was established in this paper. The ether extracts of Radix Eupatorii Chinensis were purified by HSCCC with a solvent system of hexyl hydride-ethyl acetate-methanol-water (1:2:1:2, v/v/v/v). The upper phase was used as the stationary phase and the lower phase as the mobile phase. About 8.4 mg of 12, 13-dihydroxyeuparin was obtained from 200 mg of ether extracts from Radix Eupatorii Chinensis in one-step HSCCC separation, with the purity of 96.71%, as determined by HPLC. After methanol- water recrystallization, the purity of 12,13-dihydroxyeuparin reached 99.83%. Such a simple and effective method was fairly useful to prepare pure compound as reference substances for related study on Radix Eupatorii Chinensis.

Preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus by high-speed counter-current chromatography

A high-speed counter-current chromatography （HSCCC） method was successfully developed for the preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus in one step. The purity of deoxyschizandrin was 98.5%, and the structure was identified by MS, UV and NMR. This method was simple, fast, convenient and appropriate to prepare pure compound as reference substances for related research on Schisandrae Sphenantherae Fmctus.

An Economical Method for Preparative Purification of Five Alkaloids from Coptis Chinensis Franch by High-Speed Counter-Current Chromatography Using Singled Prepared Solvent System by GC

Coptis chinensis Franch, a widely used Traditional Chinese Medicine, shows various kinds of bioactivity. The major active components of the herb are considered to be alkaloids. Thus, preparative separation of these alkaloids is critical important for further pharmacology and mechanism studies. In the paper, five alkaloids from C. chinensis were purified by HSCCC using the solvent system composed of chloro-form-metha- nol-water (2:1:1, v/v/v) single prepared. The content of each solvent in solvent system were determined by gas chromatography (GC), then according the ratios of solvents in each phase to prepare the mobile and stationary phase respectively. And a comparative study was carried out between together preparation and single preparation of the solvent system. The purities and recoveries of all the products were over 98.5% and 92%. However, 134 mL chloroform, 336 mL methanol and 452 mL water were saved when the two phase were singled by GC. Our research showed an economical method for separating alkaloids from C. chinensis by HSCCC using the solvent system single prepared by GC.

Separation and Purification of Acetophenones from Cynanchum bengei Decne Root Bark by Combination of Silica Gel and High-speed Counter-current Chromatography

[Objectives] To develop a method for separation and purification of acetophenones from Cynanchum bengei Decne root bark by combination of silica gel and high-speed counter-current chromatography( HSCCC). [Methods]The crude extract of Cynanchum bengei Decne root bark was separated by silica gel column chromatography,and parts A and B containing acetophenones were obtained. Then,parts A and B were separated by HSCCC with a two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-water( 4∶ 6∶ 4. 5∶ 5. 5 and4∶ 6 ∶ 3 ∶ 7, V/V), respectively. [Results] From 260 mg of part A, four compounds with p-dihydroxybenzene 3. 9 mg(Ⅰ),4-hydroxyacetophenone 17. 1 mg( Ⅱ),2,5-di-hydroxyacetophenone 13. 3 mg(Ⅲ) and 2,4-dihydroxyaceto-phenone 21. 0 mg(Ⅳ) were obtained. And from 300 mg of part B,136 mg of Radix Cynanchi Bungei benzophenone(Ⅴ) was obtained. The purity of compounds determined by HPLC was 97. 0%,96. 6%,99. 2%,99. 7%,99. 5%,respectively. [Conclusions] The established method is simple and efficient. It can be used for separation of acetophenones from Cynanchum bengei Decne root bark and has better practical value,which could provide a reference basis for development and utilization of Cynanchum bengei Decne root bark.

Isolation and Purification of Isoaloeresin D and Aloin from Aloe vera by High-speed Counter-current Chromatography

Objective To develop an efficient method to isolate and purify the main components isoaloeresin D and aloin from Aloe vera for its industrial production.Methods High-speed counter-current chromatography was used to isolate isoaloeresin D and aloin in a one-step separation from dried crude extract of A.vera.The biphasic solvent system composed of hexane-ethyl acetate-acetone-water(0.2:5:1.5:5) was used at a flow rate of 1.0 mL/min,while the lipophilic phase was selected as the mobile phase and the apparatus was rotated at 840 r/min.The effluent was detected at 254 nm.Results Isoaloeresin D(53.1 mg) and aloin(106.9 mg) were separated from the crude extract(384.7 mg) with the purities of 98.6% and 99.5%,respectively.Conclusion HSCCC is a powerful technique for isolation and separation of chemical composition from aloe.

Separation of Five Quinolone Alkaloids from Fruits of Evodia rutaecarpa by High-speed Counter-current Chromatography

Objective To develop a suitable method for the large-scale separation of quinolone alkaloids from the fruits of Evodia rutaecarpa by high-speed counter-current chromatography （HSCCC）. Methods Two solvent systems were developed for the separation method. The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase at 35 oCwith the flow rate of 2 mL/min and rotation speed of 855 r/min. Results Using the described method, 1-methyl-2-undecyl- 4（1H）-quinoione （1）, evocarpine （2）, 1-methy-2-[（6Z,9Z}]-6,9-pentade-cadienyl-4- （1H）-quinolone （3）, dihydroevocarpine （4）, and the mixture （5） of 1-methyl-2-[（z^- 10-pentadecenyl]-4（1H）-quinolone （Va） and 1 -methyl-2-[（Z^-6-pentadecenyl]- 4（1H）-quinolone （Vb） could be isolated from a petroleum ether extract. They were identified by 1H-NMR, ~3C-NMR, and MS/MS, and the purities were 94.3%, 95.2%, 96.8%, 98.3%, and 96.8%, respectively. Conclusion Five quinolone alkaloids from the fruits of E. rutaecarpa could be systematically isolated and purified using HSCCC. The presented method is simple and efficient with good potentials on the preparation of reference substances, especially on the quality control of Chinese materia medica.

Preparative Separation of Four Alkaloids from Gelsemium elegans by High-speed Counter-current Chromatography

Objective To develop an efficient preparative method for the separation of Gelsemium alkaloids from Gelsemium elegans. Methods High-speed counter-current chromatography （HSCCC） with several two-phase solvent systems was investigated for the separation of Gelsemium alkaloids. The purity and structure identification of the purified compounds were performed with HPLC and NMR spectra, respectively. Results In a single operation, 206.6 mg of crude alkaloid sample was separated to yield 28.7 mg of koumine, 24.9 mg of gelsemine, 26.9 mg of humantenine, and 7.2 mg of gelsevirine, with the purities of 97.8%, 95.4%, 97.4%, and 93.5%, respectively. Conclusion A preparative HSCCC method is successfully established for the separation of four Gelsemium alkaloids from G. elegans with a modified two-phase solvent system com posed of n-hexane-ethyl acetate-ethanol-O. 5% triethylamine-H2O （3：5：3：4）.

高速逆流色谱分离与鉴定鹿药中黄酮类化合物

采用高速逆流色谱(HSCCC)与其它色谱联用的方法分离纯化鹿药中的化学成分,得到5个黄酮类化合物:5,7,3′,4′-四羟基-3-甲氧基-8-甲基黄酮(1)、8-甲基木犀草素(2)、3′-甲氧基木犀草素(3)、木犀草素(4)和槲皮素(5),它们均为首次自该种及该属植物中分离得到。HSCCC以V(氯仿):V(甲醇):V(水)=4:3:2混合液为溶剂,上相为固定相,下相为流动相,分离纯化得到3个分别含化合物1,4和5的主要部分,经HPLC检验纯度分别为98.3%,96.7%和95.3%;还有1个含有化合物2和3的较纯部分,通过SephadexLH-20柱色谱,以V(甲醇):V(水)=1:1混合液洗脱将二者分离。通过理化性质及紫外、红外、质谱、核磁等波谱分析确定化合物结构。

白花败酱草中异牡荆苷和异荭草苷的高速逆流色谱分离和制备

应用高速逆流色谱分离制备白花败酱草中的异荭草苷和异牡荆苷,以乙酸乙酯∶乙醇∶水(4∶1∶5)为两相溶剂系统,上相为固定相,下相为流动相,流速2.0 mL/m in,主机800 r/m in,检测波长254 nm。以此分离条件经一步洗脱,从300 mg白花败酱草粗提物中制备得到异荭草苷24.1 mg和异牡荆苷49.8 mg,产物纯度经HPLC检测高达98.0%,结构经UV、IR、MS1、H NMR和13C NMR鉴定,二者均为首次从败酱属植物中分离得到。

高速逆流色谱制备分离紫甘薯花色苷

采用高速逆流色谱法分离纯化紫甘薯花色苷。以正丁醇-乙酸乙酯-0.5%乙酸(3∶1∶4,V/V)为溶剂体系,上相为固定相,下相为流动相,流速2 mL/min,进样量300 mg,分离得到两种花色苷的混合物;混合物再以0.2%三氟乙酸-正丁醇-甲基叔丁基醚-乙腈(6∶5∶2∶1,V/V)为溶剂体系,上相为固定相,下相为流动相,流速1.5 mL/min,进样量100 mg,分离得到纯度分别为98.5%和96.7%的两个花色苷单体组分。通过紫外-可见光谱、质谱、核磁等技术进行结构鉴定,确定组分1为3-O-{6-O-(E)-咖啡酰-2-O-[6-O-(E)-对羟基苯甲酰-β-D-吡喃葡糖基]-β-D-吡喃葡糖苷}-[5-O-(β-D-吡喃葡糖苷)]芍药素,组分2为3-O-{6-O-(E)-咖啡酰-2-O-[6-O-(E)-阿魏酰-β-D-吡喃葡糖基]-β-D-吡喃葡糖苷}-[5-O-(β-D-吡喃葡糖苷)]芍药素。实验从450 g紫甘薯中获得63 mg组分1和48 mg组分2,为紫甘薯花色苷分离提供了高效、稳定、可靠的方法。

高速逆流色谱法分离茶叶中的生物碱

以氯仿－甲醇－磷酸二氢钠缓冲液（２３ｍｍｏｌ／Ｌ，ｐＨ５．６）（体积比４∶３∶２）为溶剂系统，上相为固定相，下相为移动相，从茶叶的总生物碱中分离出三个成分，其中一个是咖啡碱，一个是茶碱，另一个待定。将分离结果与ＴＬＣ分离结果相比较，证实了高速逆流色谱法的有效及实用性。

罗布麻中黄酮的超声波强化提取及高速逆流色谱分离纯化

本研究采用超声波强化提取罗布麻中总黄酮,选择乙醇浓度、超声功率、超声时间、料液比为因素进行正交试验优选出超声提取的最佳工艺条件:60%浓度乙醇,料液比为1:15,在300W的超声波下超声提取10min。此条件下,提取率达90%以上。将提取后的总黄酮应用高速逆流色谱进行分离纯化,两相溶剂系统为乙酸乙酯:乙醇:水:乙酸(4:1:5:0.25,V/V),进样量为150mg,得45mg纯度为98.6%的罗布麻甲素。

苦参生物碱的高速逆流色谱法制备研究——色谱参数和仪器参数的最佳化

应用新型的液液分配技术——高速逆流色谱法 (High Speed Counter- current Chromatography,HSCCC)对苦参生物碱类成分的分离制备进行探讨。选用氯仿 -磷酸 -磷酸钠缓冲液 (p H6 .2 0 )溶剂系统 ,经正交试验确定了HSCCC的最佳运行参数 ,将苦参粗提物分离得到 6个固态收集物 ,经 TL C4种不同展开系统证实其中有一个为单一组分。实验表明 ,HSCCC法是一种高效、简便的分离制备中草药有效成分纯品的新方法。

高速逆流色谱法分离纯化黄芪中的芒柄花素和毛蕊异黄酮

采用高速逆流色谱法(HSCCC),以正己烷氯仿甲醇水组成二相系统作为固定相与流动相,对黄芪的乙酸乙酯粗提物进行了分离纯化。结果发现:以正己烷氯仿甲醇水(体积比为1.5∶3∶3∶2)组成的系统可以从黄芪的乙酸乙酯粗提物中分离出毛蕊异黄酮,纯度可达95%以上,并可以初步纯化芒柄花素;接着用正己烷氯仿甲醇水(体积比为4∶4∶5∶4)组成的系统进一步纯化芒柄花素,其纯度达95%以上。利用该方法,可以对中药黄芪中的异黄酮进行快速的分离和纯化。

高速逆流色谱分离制备陈皮中的黄酮类化合物

应用高速逆流色谱法分离制备了陈皮中3种黄酮类化合物。以石油醚-乙酸乙酯-甲醇-水(体积比为2∶4∶3∶3)为两相溶剂系统,在主机转速850r/min、流动相流速1.7mL/min、检测波长280nm条件下进行分离制备,6h内从4.0g陈皮粗提物中一步分离制备得到橙皮苷10.1mg、桔皮素49.8mg和5-羟基-6,7,8,3′,4′-五甲氧基黄酮50.6mg,纯度均达97.0%以上,各化合物结构经质谱和核磁共振氢谱、碳谱鉴定。利用该方法可以对陈皮中的黄酮类化合物进行快速的分离和纯化。

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800r/min、流速2.0mL/min、单次进样量200mg的条件成功地从4.0g九里香粗提物中分离纯化出54.31mg5,7,3′,4′,5′-五甲氧基黄酮(重结晶后)、107.68mg5-羟基-6,7,3′,4′-四甲氧基黄酮、215.54mg5-羟基-6,7,8,3′,4′-五甲氧基黄酮、84.36mg5-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。

高速逆流色谱结合大孔树脂从龙胆中快速分离高纯度龙胆苦苷

目的:建立龙胆中高纯度龙胆苦苷的快速分离制备方法。方法:药材醇提取液经D101大孔吸附树脂柱的层析物直接进行高速逆流色谱(high-speed counter-current chromatography,HSCCC)分离纯化,以醋酸乙酯-正丁醇-水(2∶1∶3)为溶剂系统,下相为流动相,流速1.5 mL.min-1,检测波长254 nm,分离温度30℃,所得产物经高效液相色谱与质谱分析检测,并与标准品对照。结果:在此条件下,经一步分离从300 mg粗提物中得到136 mg纯度为99.6%的龙胆苦苷。结论:该法简便、快速、所得产物纯度高,适合于龙胆苦苷标准品的制备。

高速逆流色谱法分离纯化银杏叶中白果内酯和银杏内酯A、B、C

目的从银杏叶中分离纯化白果内酯和银杏内酯A、B、C。方法银杏叶提取液经醋酸乙酯萃取、D-101大孔吸附树脂柱和Al2O3柱纯化后得到总内酯提取物,提取物再经两次高速逆流色谱分离制备4种内酯单体。结果25%乙醇热提取、醋酸乙酯萃取、D-101柱、pH4.0的Al2O3柱对总内酯的提纯最终使总内酯质量分数达到44.98%。经高速逆流色谱制备得到的白果内酯和银杏内酯A、B、C最高质量分数分别为98.3%、98.9%、98.8%、98.4%。结论本法简便快速、回收率高。

高速逆流色谱分离制备紫锥菊中的菊苣酸

建立了高速逆流色谱分离制备紫锥菊有效成分菊苣酸的新方法。溶剂系统为V(正己烷):V(乙酸乙酯):V(甲醇):V(0.5%乙酸)=1:4:2:5.5,上相为固定相,下相为流动相。从200mg紫锥菊粗提物一次分离得到纯度为96.8%的菊苣酸33.6mg,并用LC-MS-MS鉴定了化学结构。

大麻药中两个新异戊烯基黄酮的高速逆流色谱分离制备

应用高速逆流色谱分离制备大麻药的化学成分,以石油醚∶乙酸乙酯∶乙醇∶水(1∶1.2∶1.2∶1,V/V)为两相溶剂系统,上相为固定相,下相为流动相,流速2.0 mL/min,主机转速800 r/min,分离温度30℃,检测波长280 nm。以此分离条件经一步洗脱从400 mg大麻药粗提物中制备得到95.3 mg和84.5 mg的两个单体化合物,经理化反应和光谱分析分别鉴定为(2S)-5,2′,6′-三羟基-8-异戊烯基-6,7-(3-异戊烯基-2,2-二甲基-1-酮-环己二烯)-二氢黄酮(1)和(2S)-5,2′,6′-三羟基-8-异戊烯基-6,7-(3-异戊烯基-2,2-二甲基吡喃)-3′,4′-(2,2-二甲基-1-酮-环己二烯)-二氢黄酮(2)。所得的化合物1和2均为新化合物。