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.

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）.

高速逆流色谱分离纯化枸杞多糖在神经炎症中的作用

目的:构建高速逆流色谱技术分离纯化枸杞多糖的新方法,并对分离得到的枸杞多糖进行结构表征,探究其对神经炎症的影响。方法:通过分配系数K值和固定相保留率Sf,筛选并优化双水相体系偶联高速逆流色谱分离纯化枸杞多糖的条件,利用高效分子排阻色谱联用激光光散射仪和示差检测器串联检测法、1-苯基-3-甲基-5-吡唑啉酮柱前衍生结合高效液相色谱法、傅立叶变换红外光谱仪、热重分析仪、扫描电子显微镜对枸杞多糖进行初步结构表征,利用脂多糖诱导的小胶质细胞炎症模型研究枸杞多糖对神经炎症的作用。结果:确定最佳双水相体系为乙醇、饱和硫酸铵、水、异丙醇的质量比为1.4:1.0:2.58:0.3,高速逆流色谱最佳分离纯化条件为转速900 r/min,流动相流速0.8 mL/min。在上述条件下,分离得到3个枸杞多糖组分LBPs-1、LBPs-2和LBPs-3,多糖含量分别为78.35%±1.52%,69.03%±1.71%和62.11%±2.31%,分子量分别为7.40×10^(4) Da,2.73×10^(4) Da和1.47×10^(4) Da,单糖组成分别为甘露糖:鼠李糖:半乳糖醛酸:葡萄糖醛酸:葡萄糖:半乳糖:阿拉伯糖,其中摩尔比分别为1.9:4.9:1.3:8.1:7.6:32.6:41.8、2.1:4.3:1.5:8.3:5.1:28.6:37.9和5.9:4.6:1.1:0.7:13.8:66.8:5.3,同时热稳定性和微观形貌也有明显差异。生物活性研究结果表明,不同分子量的枸杞多糖可通过抑制炎症因子分泌,缓解脂多糖诱导的小胶质细胞炎症。结论:本研究构建了双水相体系偶联高速逆流色谱分离纯化枸杞多糖的新方法,为植物多糖高效分离纯化提供了技术支撑,为枸杞多糖在抑制神经炎症相关疾病中的应用提供了科学依据。

高速逆流色谱(HSCCC)在食品色素制备中的应用

综述了高速逆流色谱在食品色素制备中的应用情况,包括溶剂选择、色素种类等。

高速逆流色谱法(HSCCC)分离制备香水莲花中的柚皮素

采用高速逆流色谱法对香水莲花中的黄酮类化合物进行了分离,溶剂系统为正己烷-醋酸乙酯-甲醇-水(体积比1∶1.2∶2∶1),上层为固定相,下层为流动相,流速2.0 mL/min,转速830 r/min,检测波长280 nm,进样量300 mg,分离出80 mg化合物C,经HPLC测定纯度>98.6%,根据UV光谱、MS分析及1H-NMR、13C-NMR图谱分析结果,化合物C鉴定为柚皮素,该化合物为首次从该植物中分离得到,为香水莲花的开发应用提供了理论依据。

HSCCC-ELSD法分离纯化青葙子中的皂苷(英文)

连接有蒸发光散射检测器的高速逆流色谱仪首次成功的应用于制备和分离青葙子中的皂苷celosins A和B。二氯甲烷:正丁醇∶甲醇∶水(4∶0.3∶3∶2)+0.5%冰醋酸作为洗脱溶剂系统。从半制备型HSCCC收集到的组分进行HPLC分析,可以得到:celosin A纯度为98.9%,celosin B的纯度为98.1%。这是高速逆流色谱仪首次被用于纯化青葙子中的皂苷,两个化合物的结构通过碳谱和质谱来确定。

HSCCC法结合制备型HPLC技术靶向制备分离与鉴定大血藤中的多酚类化合物

目的:建立大血藤中多种多酚类化合物高效稳定的制备分离方法。方法:以正己烷-乙酸乙酯-甲醇-水(3∶7∶4∶6,v/v)为溶剂体系,利用高速逆流色谱对大血藤正丁醇萃取物进行纯化,再结合制备型液相色谱获得各单体化合物,根据UV、MS、1H-NMR波谱信息确定化学结构。结果:从大血藤中分离鉴定了8个多酚类成分,分别为:(1)没食子酸;(2)原儿茶酸;(3)绿原酸;(4)咖啡酸;(5)香草酸;(6)阿魏酸;(7)异丹叶大黄素;(8)赤松素。化合物(7)(8)为首次从该属植物中分得。结论:高速逆流色谱和制备型液相色谱联用技术可高效制备分离大血藤中多种多酚类化合物,为大血藤的药理研究及质量控制奠定了基础。

HSCCC分离纯化未成熟罗汉果皂苷类化合物

为建立高速逆流色谱分离未成熟罗汉果中皂苷类化合物的方法,该研究将罗汉果粗提物先经过大孔树脂富集皂苷类化合物,再采用高速逆流色谱分离罗汉果皂苷。结果表明:以氯仿-甲醇-正丁醇-水(5∶6∶1∶4,v/v/v/v)作为两相溶剂系统,上相为固定相,下相为流动相,在主机转速为860 r·min^(-1),流速为2.5 mL·min^(-1),检测波长为203 nm的条件下,一次性制备得到4个化合物,即11-O-罗汉果皂苷Ⅱ(Ⅰ)、罗汉果皂苷ⅡE(Ⅱ)、11-O-罗汉果皂苷Ⅲ(Ⅲ)和罗汉果皂苷Ⅲ(Ⅳ),经高效液相色谱检测纯度分别为95.5%、98.2%、80.1%和97.6%。该方法实现了未成熟罗汉果皂苷快速有效的分离,具有样品回收率高、损失少、避免样品失活等优点,提高了分离效率。该研究结果为更多的罗汉果皂苷化合物的分离纯化奠定了基础,补充与优化了罗汉果皂苷类化合物的分离方法。

应用高速逆流色谱(HSCCC)分离纯化山茱萸中的没食子酸

应用高速逆流色谱 ( HSCCC)分离山茱萸中的没食子酸并结合波谱技术进行结构鉴定。经过一次逆流色谱分离 ,可以得到纯度在 70 %以上的没食子酸 ,第二次分离即可使其纯度达到 97%以上。

猪苓中黄嘌呤氧化酶抑制剂的筛选分离及质谱分析

试验以猪苓为研究对象,综合利用遗传算法-BP神经网络数学模型、色谱和质谱等技术,建立快速筛选猪苓中黄嘌呤氧化酶抑制剂的新方法。首先,利用遗传算法-BP神经网络数学模型优化猪苓中麦角甾醇的提取工艺;其次,以黄嘌呤氧化酶为生物靶分子,应用超滤质谱技术,筛选猪苓中潜在的黄嘌呤氧化酶抑制剂,同时,应用分子对接技术验证超滤实验的准确性;最后,以活性筛选为导向,利用高速逆流色谱技术分离猪苓中黄嘌呤氧化酶抑制剂。结果显示:猪苓中麦角甾醇的最佳提取工艺为乙醇体积分数80%、液料比16 mL/g、提取时间1.4 h、提取次数3次,在此条件下,麦角甾醇得率为2.31%;从猪苓中筛选并分离出4个纯度均在90%以上的黄嘌呤氧化酶抑制剂,其抑制效果强弱顺序为猪苓酮A>猪苓酮B>麦角甾-4,6,8(14),22-四烯-3-酮>麦角甾-7,22-二烯-3-酮。该方法快速、灵敏,为猪苓抗痛风药理作用的进一步研究提供了一定的理论基础。

桑黄中黄嘌呤氧化酶抑制剂的筛选分离及质谱分析

以桑黄为研究对象,综合利用数学模型、色谱和质谱技术,建立了快速筛选分离桑黄中黄嘌呤氧化酶抑制剂的新方法。利用数学模型结合响应面优化桑黄中黄酮类活性成分的提取工艺,以黄嘌呤氧化酶为靶点,应用超滤-质谱技术快速筛选桑黄中黄嘌呤氧化酶的抑制剂;在活性指导下,利用高速逆流色谱技术分离纯化桑黄中活性成分,采用高效液相色谱-高分辨质谱结合核磁共振波谱技术对其进行结构鉴定。结果表明,在此条件下得到桑黄黄酮提取率为24.95 mg/g,优于响应面试验;从桑黄中筛选并分离出3个纯度均在90%以上的黄嘌呤氧化酶抑制剂,抑制效果强弱顺序为桑黄素D>桑黄素E>纤孔菌素A。本方法快速、灵敏,可为桑黄药理作用研究提供支撑,同时可为桑黄食品开发提供可行方法和理论支持。

活性引导结合高速逆流色谱分离蓝莓活性成分及其与α-葡萄糖苷酶的相互作用

基于活性引导结合高速逆流色谱(HSCCC)技术,分离蓝莓中具有抑制α-葡萄糖苷酶活性的组分。首先,经不同溶剂提取后,活性成分在水中得到富集;其次,通过HSCCC分离水提物,得到6种组分,其中F4对α-葡萄糖苷酶的抑制率显著高于其它几种;再次将F4采用Sephadex LH-20葡聚糖凝胶纯化,得到高纯的组分Ⅰ;最后通过紫外光谱、傅里叶红外光谱、高效液相色谱-质谱联用及核磁技术鉴定组分Ⅰ为矢车菊-3-O-葡萄糖苷(C3G),纯度为95.06%。此外,采用多光谱扫描和分子对接技术对C3G与α-葡萄糖苷酶的相互作用进行表征。研究发现C3G与α-葡萄糖苷酶通过氢键自发结合形成复合物。与C3G复合后,α-葡萄糖苷酶的二级结构发生不同程度的变化,其中α-螺旋和β-转角降低,β-折叠和不规则卷曲增加。分子对接模拟发现C3G与残基Leu 313、Ser 157、Tyr 158、Phe 314、Arg 315和Asp 307以氢键结合,并与周围4个疏水残基存在疏水作用,共同维持复合物结构。本研究结果对开发2型糖尿病功能性食品具有重要意义。

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。

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。