Separation of Alkaloids from Datura Metel. and Sophora Flavescens Ait by High-Speed Countercurrent Chromatography

Abstract

High-speed countercurrent chromatography was successfully applied to the separation of alkaloids from two medicinal herbs. Matrine and oxymatrine were isolated from the root of Sophora flavescens Ait with a two-phase solvent system composed of chloroform-0.07 M sodium phosphate (pH 6.4), while atropine and scopolamine were purified from the flowers of Datura mete L. with a similar solvent system by modifying the phosphate buffer pH at 6.5. Identification of each alkaloid was made by either TLC or paper partition chromatography using authentic pure compounds.     

高速逆流制备分离中药黄柏中的生物碱

 利用高速逆流对传统中药黄柏中的生物碱类活性成分进行了制备分离。两相溶剂分离系统采用氯仿 甲醇 0 5mol/LHCl(体积比为 2∶1∶1) ,并对制备物进行了薄层分析。证实了生物碱成分 ,检出了小檗碱。     

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。     

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。     

Application of pH-Zone-Refining Countercurrent Chromatography in the Preparative Separation of Alkaloids from Traditional Chinese Medicine

pH-Zone-refining countercurrent chromatography (CCC) is a recently developed new preparative separation method based on conventional CCC. The method uses a retainer base (or acid) in the stationary phase to retain the analyses in the column and an eluent acid (or base) to elute the analyses according to their pKa values and hydrophobicities. It produces a succession of highly concentrated rectangular peaks with minimum overlap similar to those observed in displacement chromatography. pH-zone-refining CCC has important advantages over the conventional CCC including an over 10-fold increase (up to 10 gram or more) in sample loading capacity, high concentration of fractions, and concentration of minor impurities. pH-zone-refining CCC has been successfully applied to the preparative separation of a variety of compounds including both acidic and basic derivatives of amino acid, hydroxyxanthene dyes, peptides, alkaloids, indole auxins, structural, geometrial and optical isomers.     

Separation of S-Triazine Herbicides by Countercurrent Chromatography

Abstract

Countercurrent chromatography (CCC) has been successfully applied for the separation of four chlorinated s-triazine derivatives, namely, simazine, atrazine, propazine, and trietazine, which are widely used as herbicides. Of several solvent systems investigated, n-hexane-ethyl acetate-methanol-water (8:2:5:5) gave the best range of the partition coefficient values because of the satisfactory solubility for all samples and therefore was used for the separation of s-triazines. Two types of countercurrent chromatographs, high speed CCC, and horizontal flow-through coil planet centrifuge, were used for s-triazines separation, and the identity of the separated fractions was unequivocally established by mass spectrometry. The potential practical application of CCC to herbicide analysis is discussed.     

The Application of High-Speed Countercurrent Chromatography on the Separation and Purification of Isoflavones from Radix Pueraria

High-speed countercurrent chromatography (HSCCC) is a special liquid-liquid partition chromatography without solid support matrix. It eliminates irreversible adsorption of sample in the conventional chromatographic method and has been successfully used for analysis and separation of various kinds of natural products.     

Evaluation of Analytical Countercurrent Chromatographs: High-Speed Countercurrent Chromatograph-4000 Vs. Analytical Toroidal Coil Centrifuge

Abstract

Performance of two countercurrent chromatographic models, high speed countercurrent chromatograph (HSCCC-4000) and analytical toroidal coil centrifuge (TCC), is evaluated in terms of theoretical plate number, resolution factor and separation time to assess their analytical capability. A series of experiments was conducted to investigate the effects of internal diameter and length of the coiled column, and flow rate of the mobile phase on the separation of indole auxins in two-phase solvent systems composed of n-hexane-ethyl acetate-methanol-water at different volume ratios. The three components of indole auxins were completely resolved in 16 min with a HSCCC system equipped with a multilayer coil of a 0.55 mm I.D. PTFE tube with theoretical plates ranging from 1290 to 829. Similar separation was achieved in 24 min with a TCC systems equipped with a 0.3 mm I.D. PTFE tube with theoretical plates ranging from 1811 to 969. It is concluded that both systems have comparable analytical capability at the present stage of development.     

中药粉防已中汉防已甲素和乙素的高效液相色谱分析

本文利用正相高效液相色谱（ＨＰＬＣ）系统建立了一个定性定量分析粉防已药材中的主成分汉防已甲素和乙素的方法，该方法准确、快速。回收率：甲素为９７．４％，乙素为９９．６％；变异系数：甲素为２．５０％，乙素为２．７７％；最低检出限：甲素为３．８７×１０－３ｍｍｏｌ／Ｌ，乙素为２．４８×１０－３ｍｍｏｌ／Ｌ。用上述方法分析实际样品，结果令人满意。     

Separation of Pesticides and Impurities by Countercurrent Chromatography

Abstract

Countercurrent chromatography (CCC) was chosen for study as an isolation technique for pesticides and the impurities found with them, because its high capacity makes possible comparatively large batches of purified materials and because its mild operating conditions minimize the danger of degradation during the separation. Technical grade fenthion, an organophosphorus insecticide, showed traces of six impurities. The principal one found in an analytical standard fenthion was identified as 3-methyl-4-(methylthio)anisole, well separated from the fenthion. The n-octyl ester of the herbicide 2,4,5-T was easily separated from the methyl ester impurity.     

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。     

逆流色谱分离镅(Ⅲ)和铕(Ⅲ)的研究

选择0.1 mol/L NaC lO4为流动相,以二氯苯基二硫代膦酸的二甲苯溶液为固定相,研究了萃取剂浓度、流动相pH及流动相流率对逆流色谱(CCC)分离Am3 和Eu3 效果的影响。结果表明,在实验条件下,两者的分离系数随着固定相中的萃取剂浓度增加、流动相的pH升高而增大;降低流动相流率后可明显改善分离效果。通过改变分离条件,Am3 和Eu3 可基本上实现相互分离,两者间的分离系数和分离效率分别达到2.87和0.74。     

Alkaloids of Stephania hernandifolia VI. Hernandifoline

Summary A new hasubanan alkaloid with the composition C₂₉H₃₃O₉N, mp 128–129°C₉ from ether), [] _D ²⁰ – 25° (c 2; ethanol) containing an ester group has been isolated from the herbStephania hernandifolia and has been called hernandifoline.All-Union Scientific-Research Institute for Medicinal Plants. Translated from Khimiya Prirodynkh Soedinenii, No. 2, pp. 158–164, March, 1971.     

Alkaloids of Stephania hernandifolia VIII. Methylhernandine

Summary A new hasubanan alkaloid with the composition C₂₀H₂₇O₆N, mp 152–153°C (ether-methanol) has been isolated from the herbStephania hernandifolia and has been called methylhernandine. The alkaloid has been shown to be identical with the methylamino alcohol obtained in the hydrolysis of hernandifoline.All-Union Scientific Research Institute of Medicinal Plants. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 455–457, July–August.     

Alkaloids of Stephania delovayi III. Stephodeline

Summary From the epigeal part ofStephania delovayi Diels (Menispermaceae) have been isolated cycleanine and a new amorphous alkaloid, stephodeline, with the composition C₂₁H₂₇O₅N, a structure for which is proposed.All-Union Scientific-Research Institute of Medicinal Plants. Translated from Khimiya Prirodnykh Soedinenii, No. 5, pp. 650–653, September–October, 1973.     

Alkaloids of Stephania delavayi IV. Isostephodeline

Summary The structure of the new alkaloid isostephodeline isolated from the roots ofStephania delavayi Diels has been established. The most probable relative configurations of isostephodeline and of the stephodeline isolated previously have been proposed.All-Union Scientific-Research Institute of Medicinal Plants. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 188–193, March–April, 1975.     

Alkaloids of Stephania hernandifolia VII. Hernandine

Summary The epigeal part ofStephania hernandifolia has given a new hasubanan alkaloid with the composition C₁₉H₂₅O₆N, mp 197–199°C (ethanol), [] _D ²⁰ -33° (c 1.8; ethanol), which has been called hernandine, and for which a partial structure has been proposed.All-Union Scientific-Research Institute for Medicinal Plants. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 180–184, March, 1971.