应用QuikPrep HSCCC系统实现共轭亚油酸及其异构体的分离

综述了高速逆流色谱(HSCCC)技术的原理,溶剂系统选择原则,影响因素以及应用范围,并应用该技术对共轭亚油酸(CLA)异构体进行分离。发现HSCCC可以对CLA实现高度的纯化。     

高速逆流色谱法分离纯化续随子种子中的七叶内酯

建立了高速逆流色谱(HSCCC)技术分离纯化续随子种子中七叶内酯的方法。将续随子种子的乙酸乙酯萃取物直接进行高速逆流色谱分离,考察了不同溶剂系统的分离效果。结果表明,最佳的溶剂系统为氯仿-甲醇-水(体积比为4:3:2),以其上相为固定相,下相为流动相。从200 mg续随子种子乙酸乙酯萃取物中分离得到80 mg七叶内酯,纯度为99.04%。HSCCC技术可高效分离纯化续随子种子中的七叶内酯,为得到高纯度的七叶内酯提供了制备技术。     

高速逆流色谱法快速分离制备枸杞中莨菪亭

建立了用高速逆流色谱(HSCCC)从枸杞中快速分离莨菪亭的方法。将枸杞的乙醇提取物经D-101大孔树脂初步纯化后直接进行高速逆流色谱分离,用薄层色谱-荧光法考察了莨菪亭在不同溶剂体系中的分配情况。结果表明,最佳的溶剂体系为氯仿-甲醇-水(10:7:3, v/v/v),取上相为固定相,下相为流动相,在主机转速为850 r/min、流速为1.5 mL/min、检测波长为365 nm的条件下,从200 mg样品中一次性分离制备可得到10.2 mg纯度达到98.3%的莨菪亭。制备所得的莨菪亭与对照品的高效液相色谱(HPLC)保留时间一致,且经核磁共振氢谱、碳谱鉴定结构;纯度经HPLC法测定。研究发现,氯仿-甲醇-水(10:7:3, v/v/v)体系可连续二次进样而样品的峰形未受明显的影响。实验结果表明用薄层色谱-荧光法可快速选定HSCCC溶剂体系,进而可快速、简便地制备高纯度的莨菪亭。     

半制备型高速逆流色谱分离制备铁棒锤根中的一种咪唑生物碱

采用高速逆流色谱(HSCCC)技术从铁棒锤根氯仿提取物中分离制备了一种高纯度咪唑类生物碱1H-imidazole-2-carboxylic acid,butyl ester(ICABE)。采用高效液相色谱(HPLC)测定目标化合物在两相溶剂中的分配系数,优化HSCCC分离ICABE的溶剂体系,确定了以正己烷-氯仿-乙醇-水(10∶1∶13∶2,v/v/v/v)为HSCCC的两相溶剂系统,以上相为固定相,下相为流动相,流动相流速为1.8 mL/min,主机转速850 r/min,检测波长为230 nm条件下进行分离制备,在350 min内从100 mg粗样品中一步分离得到7.5 mg ICABE,经HPLC检测其纯度达98%以上(峰面积归一化法),结构由UV、1H-NMR和13C-NMR得以鉴定。该方法简便、快速,所得产物纯度高,适合于铁棒锤中ICABE的制备分离。     

高速逆流色谱技术分离纯化天然产物中黄酮类化合物的研究进展

高速逆流色谱(HSCCC)技术是一种连续高效的新型液-液分配色谱技术,在中药、生化、食品、天然产物化学、环境分析等领域有着广泛的应用.本文综述了部分利用高速逆流色谱技术分离纯化已知和未知黄酮类化合物的文献报道,并介绍了几种新近发展的高速逆流色谱以及其在分离纯化黄酮类化合物和其它化合物中的应用.     

高速逆流色谱与质谱联用在中药分析中的应用

高速逆流色谱 ( High- speed countercurrent chromatography,HSCCC)是新型的液 -液分配色谱技术 ,它利用多层螺旋管同步行星式离心运动 ,在短时间内实现样品在互不相溶的两相溶剂系统中的高效分配 ,从而实现样品分离[1] . HSCCC分离纯化后 ,若用 MS离线检测 ,此时须挥发原 HSCCC的溶剂体系 ,才能实现样品转移 ,还可能会带来样品污染 .HSCCC与 MS的联用克服了这些缺陷 ,具有很强的实用价值 .丹参酮 A( Tashinone A)是一种结晶性菲醌类化合物 [2 ] ,在丹参乙醚提取物中含量丰富 .本文采用正己烷 -乙醚 -乙醇 -水体系 ,将 HSCC…     

流体动力学逆流色谱分离柱研究进展

近年来,以流体动力学为基础的逆流色谱仪在天然产物与生化药物分离中得到了广泛应用。柱设计是色谱体系的核心环节,分离柱管子的几何形状、安装方式,分离柱的位置、体积等因素都会对色谱行为和分离效率产生不同的影响。本文对基于流体动力学的现代逆流色谱分离柱的研制进展与应用情况进行综述,并对未来逆流色谱的改进进行了展望。     

逆流色谱仪器装置的研究进展

逆流色谱是一种快速、高效、无固定相载体的新型液-液分配色谱技术.综述了高速逆流色谱、双向逆流色谱、正交轴逆流色谱、pH-区带精制逆流色谱及螺线型圆盘柱式逆流色谱仪器装置的研究进展,介绍了高速逆流色谱-质谱联用技术、高速逆流色谱.蒸发光散射联用技术、高速逆流色谱.高效液相色谱联用技术及二维逆流色谱技术的发展状况.     

高速逆流色谱法制备罗汉果根中葫芦素类化合物

葫芦素作为四环三萜类化合物广泛存在于葫芦科植物中,但其含量较低、结构相似,采用常规的柱层析分离法较难得到大量、高纯度的单体化合物,导致其活性的研究与应用受到限制。研究采用高速逆流色谱法(HSCCC),建立了一种从罗汉果根提取物中制备葫芦素类化合物的方法。罗汉果根乙醇提取物经HPD-100大孔树脂、MCI、RP-C18柱层析分离后获得葫芦素粗品。通过基于薄层色谱的溶剂选择法(GUESS)对液-液萃取与高速逆流色谱的溶剂体系进行快速筛选,确定采用正己烷-乙酸乙酯-甲醇-水(3∶7∶3∶7, v/v/v/v)萃取葫芦素粗品,有机层萃取物再以正己烷-乙酸乙酯-甲醇-水(4∶6∶5∶5, v/v/v/v)进行HSCCC分离,以上相作为固定相,下相作为流动相,流速2.0 mL/min,主机转速860 r/min,进样量280 mg,一次分离即可获得5种葫芦素类化合物,经高效液相色谱检测其纯度分别为97.0%、95.4%、96.3%、91.6%和95.3%,并通过核磁共振技术、高分辨质谱技术及文献资料对比对各化合物的结构进行鉴定。经鉴定,5种化合物分别为葫芦素Q1、23,24-二氢葫芦素F-25-乙酸酯、葫芦素B、23,24-二氢葫芦素B和二氢异葫芦素B-25乙酸酯。该方法操作简单、快速,分离效果好,可作为规模化制备葫芦素类化合物的一种新方法。     

《制备色谱技术及应用》(第二版)

(ISBN 978-7-122-11958-2)该书从色谱科学的角度详细地阐述了制备色谱的原理、重要的实验技术、关键性色谱分离技巧及其应用。内容包括制备色谱的基础知识、制备薄层色谱、常压柱色谱、低压及中压柱色谱、高压制备液相色谱、高速逆流色谱、模拟移动床色谱、顶替色谱、制备气相色谱、电泳以及与制备色谱技术紧密相关的生物代谢产物的提取分离技术等。本书对制备色谱技术的系统介绍具有简明、系统、全面的特点。     

高速逆流色谱分离纯化EGCG3"Me的研究

首次采用高速逆流色谱法对经自制聚酰胺初步分离的表没食子儿茶素-3-(3″-O-甲基)没食子酸酯( EGCG3"Me)样品中的EGCG3"Me单体进行分离纯化.结果表明,选择水-甲醇-乙酸乙酯-正己烷(体积比5:2:9:1)为高速逆流色谱分离的两相溶剂系统,上相为固定相,下相为流动相,在主机转速700 r／min、流速2...     

An effective high-speed countercurrent chromatographic method for preparative isolation and purification of mollugin directly from the ethanol extract of the Chinese medicinal plant Rubia cordifolia

The medicinal plant Rubia cordifolia has been used widely in traditional Chinese medicine (TCM) for its antibacterial, antioxidant and anti-inflammatory activities. In this study, a preparative high-speed countercurrent chromatography (HSCCC) method for isolation and purification of the bioactive component mollugin directly from the ethanol extract of R. cordifolia was successfully established by using light petroleum (bp 60-90 degrees C)/ethanol/diethyl ether/water as the two-phase solvent system. The upper phase of light petroleum/ethanol/diethyl ether/water (5:4:3:1 v/v) was used as the stationary phase of HSCCC. Under the optimum conditions, 46 mg of mollugin at 98.5% purity, as determined by HPLC, could be yielded from 500 mg of the crude extract in a single HSCCC separation. The peak fraction of HSCCC was identified by 1H NMR and 13C NMR.     

高速逆流法对刺五加有效成分刺五加苷E的分离制备

 采用高速逆流法分离纯化刺五加粗品中的刺五加苷E。溶剂系统为氯仿 甲醇 异丙醇 水,体积比为5∶6∶1∶4,上相为固定相,下相为流动相,转速为800r/min,流速为2 0mL/min。所得刺五加苷E经高效液相分析,测定纯度为98%(峰面积百分比)。这为其他苷类等极性物质的分离纯化提供了依据。     

芦荟色酮的高速逆流色谱分离制备方法研究

 芦荟色酮是在芦荟叶中特有的一类具有抗炎和抑制酪氨酸酶等作用的活性物质。以芦荟全叶为原料,经过一系列的预处理手段,从脱色剂活性炭中获得芦荟色酮粗提物,再经过溶剂分配和富集后采用高速逆流色谱（HSCCC）对其中的色酮成分进行分离纯化。研究结果表明,采用氯仿-甲醇-水(体积比为4∶3∶2)混合溶液和二氯甲烷-甲醇-水(体积比为5∶4∶2)混合溶液作为溶剂分离系统,经过两步HSCCC可以分离纯化出色谱纯度在95％以上的芦荟色酮单体。经过紫外检测、快原子轰击质谱及核磁共振等方法的结构分析鉴定,证实分离所得物质为肉桂酰基-C-葡萄糖甙芦荟色酮。     

高速逆流色谱与硅胶柱色谱结合分离制备高纯度芦荟素异构体

芦荟素是芦荟叶中主要的蒽醌类成分,通常以芦荟素A和B两种非对映异构体形式并存,目前已成为许多芦荟产品质量控制指标之一。采用高速逆流色谱和硅胶柱色谱结合的方法,从老芦荟干粉中分离制备了高纯度的芦荟素A(纯度为98%)和芦荟素B(纯度为96%)样品,并采用快原子轰击质谱(FAB-MS)和核磁共振氢谱(1H NMR)以及GOESY(gradient-enhanced nuclear Overhauser effect spectroscopy)等方法对所得的两个芦荟素异构体的立体构象进行了确认。该法具有制备量大和分离效率高的特点。     

Separation of phenylsuccinic acid enantiomers using biphasic chiral recognition high‐speed countercurrent chromatography

High‐speed countercurrent chromatography (HSCCC) combined with biphasic chiral recognition was successfully applied to the resolution of phenylsuccinic acid enantiomers. d ‐Isobutyl tartrate and hydroxypropyl‐β‐cyclodextrin were employed as lipophilic and hydrophilic selectors dissolved in the organic stationary phase and aqueous mobile phase, respectively. The two‐phase solvent system was made up of n‐hexane/methyl tert‐butyl ether/water (0.5:1.5:2, v/v/v). Impacts of the type and concentration of chiral selectors, the pH value of the aqueous phase solution as well as the temperature on the separation efficiency were investigated. By means of preparative HSCCC, pure enantiomer was obtained by separating 810 mg of racemate with a purity >99.5% and a recovery rate between 82 and 85%. The experimental results indicate that biphasic recognition HSCCC provide a promising means for efficient separation of racemates.     

高速逆流色谱分离同分异构体

应用高速逆流色谱法对同分异构体的分离纯化进行研究。实验结果表明,以正己烷-乙酸乙酯-水(体积比为1∶10∶10)为两相溶剂系统,上相为固定相,下相为流动相,进行二次高速逆流色谱分离,可从茶多酚中分离出g级的儿茶素同分异构体——(-)-表没食子儿茶素没食子酸酯(EGCG)和(-)-没食子儿茶素没食子酸酯(GCG),其高效液相色谱纯度均在98%以上。选择四氯化碳-氯仿-甲醇-水(体积比为7∶3∶7∶3)为溶剂系统,下相为固定相,上相为流动相,经一次高速逆流色谱即可将药物中间体溴代苯胺同分异构体进行有效的分离。     

Rapid separation of cyanidin‐3‐glucoside and cyanidin‐3‐rutinoside from crude mulberry extract using high‐performance countercurrent chromatography and establishment of a volumetric scale‐up process

This study describes the rapid separation of mulberry anthocyanins; namely, cyanidin‐3‐glucoside and cyanidin‐3‐rutinoside, using high‐performance countercurrent chromatography, and the establishment of a volumetric scale‐up process from semi‐preparative to preparative‐scale. To optimize the separation parameters, biphasic solvent systems composed of tert‐butyl methyl ether/n‐butanol/acetonitrile/0.01% trifluoroacetic acid, flow rate, sample amount and rotational speed were evaluated for the semi‐preparative‐scale high‐performance countercurrent chromatography. The optimized semi‐preparative‐scale high‐performance countercurrent chromatography parameters (tert‐butyl methyl ether/n‐butanol/acetonitrile/0.01% trifluoroacetic acid, 1:3:1:5, v/v; flow rate, 4.0 mL/min; sample amount, 200–1000 mg; rotational speed, 1600 rpm) were transferred directly to a preparative‐scale (tert‐butyl methyl ether/n‐butanol/acetonitrile/0.01% trifluoroacetic acid, 1:3:1:5, v/v; flow rate, 28 mL/min; sample amount, 5.0–10.0 g; rotational speed, 1400 rpm) to achieve separation results identical to cyanidin‐3‐glucoside and cyanidin‐3‐rutinoside. The separation of mulberry anthocyanins using semi‐preparative high‐performance countercurrent chromatography and its volumetric scale‐up to preparative‐scale was addressed for the first time in this report.     

In vitro screening and isolation of human aromatase inhibitors from Cicer arietinum by a novel continuous online method combining chromatographic techniques

Ultrafiltration liquid chromatography with mass spectrometry can efficiently and rapidly screen and identify ligands from the seeds of Cicer arietinum for human aromatase. Using this method, we identified 11 major compounds, including organic acids, organic acid glycosides, flavone glycosides, isoflavones, and isoflavone glycosides, as potent human aromatase inhibitors. A continuous online method, including pressurized liquid extraction, countercurrent chromatography, and preparative liquid chromatography, was developed for scaling up the production of these compounds with high purity and efficiency. The bioactivity of the separated compounds was assessed by an in vitro enzyme inhibition assay. This novel approach using a combination of ultrafiltration liquid chromatography with mass spectrometry and pressurized liquid extraction with countercurrent chromatography and preparative liquid chromatography as well as an in vitro enzyme inhibition assay could be applied to efficiently screen and isolate human aromatase inhibitors from complex samples and to the large‐scale production of functional food and nutraceutical ingredients.     

虎杖中白藜芦醇和白藜芦醇甙的高速逆流色谱分离提纯及其分析

采用高速逆流色谱技术分离提纯中药虎杖中的活性多酚成分白藜芦醇和白藜芦醇甙。确定了样品前处理方法,选择了逆流色谱流动相体系,经高效液相色谱检验纯度,＾１ＨＮＭＲ及质谱判定结构,证实所得样品为白藜芦醇和白藜芦甙,得率分别为２．１８％和１．０７％。