Isolation And Purification Of Effective Compounds From Rheum Officinale Etc Traditional Chinese Medicinal Herbs By Adsorption Chromatography On 12% Cross-linked Agarose Gel Media

Gel media based on cross-linked dextran and cross-linked agarose are widely used for size exclusion chromatography of large biomolecules, which was developed in the 1960's. Superose 12, the gel that is prepared by spherical agarose with cross-linking two times. The high cross-linked structure increased its rigidity and stability observably. So it can be used in high velocity. And now Superose 12 has been widely used in the separation and purification of high molecular weight compounds. It was also used in the separation and purification of active components from natural products.It has become an urgent task to investigate Chinese traditional medicinal herbs with modern technological instrument and establish integrate quality appraisal system. And it is also the road that must be taken in the development of the Chinese native medicine enterprise. So carrying on the research of the new method of traditional Chinese medicine plays an important role in promoting the development of modern traditional Chinese pharmacology and the Chinese medicine modernization. Successful application of adsorption chromatography on cross-linked 12% agarose has been reported for the purification of flavones and polyphenols.In the present paper, under the optimized experiment conditions, active components from common traditional Chinese herbs (Rheum officinale Baill., Glycine max (L.)Merr Polygonum cuspidatum Sieb.et Zucc, etc.) were separated and purified. The purity of the obtained compounds after Superose 12 separation was above 95% determined by high performance liquid chromatography (HPLC) analysis and their chemical structures were identified by 1H-NMR, 13C-NMR and standard samples. And also discussed the separated mechanism of hydroxyanthraquinone, flavonoid and lignans compounds on cross-linked 12% agarose.1. Isolation and purification of hydroxyanthraquinone compounds from the roots of Rheum officinale Baill. by Superose 12A chromatographic method for isolation and purification of chemical constituents from the well-known traditional Chinese drugDa-huang (roots of Rheum officinale Baill.) was established by using 12% cross-linked agarose gel, Superose 12, as the separation media. 55% methanol (including 0.01% HAC) was used as the eluent for separation of cinnamic acid, rhein, aloe-emodin, chrysophanol, physcion and emodin from Da-huang crude extract. As a result, cinnamic acid and five kinds of hydroxyanthraquinones including rhein, aloe-emodin, chrysophanol, physcion and emodin were obtained in one-stepp seperation with high purity.The retention behavior of hydroxyanthraquinones on Superose 12 was studied according to the chemical structure and the retention time. The results of the present study clearly demonstrate that Superose 12 can be used for the separation of hydroxyanthraquinones and that the retention of hydroxyanthraquinones on Superose 12 is based on a mixture of hydrogen bonding and hydrophobic interactions between the hydroxyl groups of hydroxyanthraquinones and the residues of the cross-linking reagents used in the manufacturing process of Superose 12.2. Isolation and purification of Sobyean isoflavones from sobyean by Superose 12 Six soy isoflavones including daidzin, genistin, glycitin, daidzein, glycitein and genistein with the purity of 99.0%, 99.0%, 90.5%, 99.0%, 94.5% and 99.0%, were isolated and purified by adsorption chromatography Superose 12 from soybean in one-step separation. When 40% and 60% methanol were used as the mobile phase in gradient mode at a flow rate of 0.5 ml/min, good separation results could be obtained.The retention behavior of soy isoflavones on Superose 12 was studied according to the chemical structure and the retention time. The results of the present study clearly demonstrate that Superose 12 can be used for the separation of soy isoflavones and that the retention of soy isoflavones on Superose 12 is based on a mixture of hydrogen bonding and hydrophobic interactions between the hydroxyl groups of soy isoflavones and the residues of the cross-linking reagents used in the manufacturing process of Superose 12.3. Isolation and purification of four compounds from the Chinese medicinal herb Polygonum cuspidatum Sieb.et Zucc by adsorption chromatography on cross-linked 12% agaroseThe four compounds of the well-known traditional Chinese drug Polygonum cuspidatum Sieb. et Zucc can be separated by adsorption chromatography on the cross-linked 12% agarose gel Superose 12. Sixty five percent methanol was used as the eluent for separation of piceid, resveratrol, anthraglycoside B and emodin from Hu-zhang crude extract. The purity of each compound is over 95% as determined by HPLC.The retention behavior of the compounds from Polygonum cuspidatum Sieb.et Zucc on Superose 12 was studied according to the chemical structure and the retention time. The results of the present study clearly demonstrate that Superose 12 can be used for the separation of the compounds from Polygonum cuspidatum Sieb.et Zucc and that the retention of these compounds on Superose 12 is based on a mixture of hydrogen bonding and hydrophobic interactions between the hydroxyl groups of the compounds and the residues of the cross-linking reagents used in the manufacturing process of Superose 12.4. Isolation and purification of four compounds from the Chinese medicinal herb Rhizoma Anemarrhenae by adsorption chromatography on 12% cross-linked agarose Four bioactive compounds including two lignans compounds cishinkiresinol, (-)-4′-O-methylnyasol and two flavonoid compounds mangiferin, neomangiferin were separated from the methanol extract of Rhizoma Anemarrhenae by adsorption chromatography. Sixty percent methanol was used as the mobile phase. The purities of the four compounds reached 96.1%, 98.0%, 97.2% and 98.1%.The retention behavior of the compounds from Rhizoma Anemarrhenae on Superose 12 was studied according to the chemical structure and the retention time. The results of the present study clearly demonstrate that Superose 12 can be used for the separation of the compounds from Rhizoma Anemarrhenae and that the retention of these compounds on Superose 12 is based on a mixture of hydrogen bonding and hydrophobic interactions between the hydroxyl groups of the compounds and the residues of the cross-linking reagents used in the manufacturing process of Superose 12.5. One step isolation and purification of four compounds from the Chinese medicinal Radix Glycyrrhiza by adsorption chromatography on 12% cross-linked agarose Adsorption chromatography (Superose 12) was successfully used for the isolation and purification of four compounds from Radix Glycyrrhiza, the Chinese herbal drug. Sixty percent methanol was used as the mobile phase. Four kinds of compounds including licuraside, licorice-glycoside A, isoliquiritigenin and glycycoumarin were obtained after one step separation, with the purity of 91.3%, 95.1%, 91.8% and 96.7%, respectively, as determined by high performance liquid chromatography (HPLC).The retention behavior of the compounds from Radix Glycyrrhiza on Superose 12 was studied according to the chemical structure and the retention time. The results of the present study clearly demonstrate that Superose 12 can be used for the separation of the compounds from Radix Glycyrrhiza and that the retention of these compounds on Superose 12 is based on a mixture of hydrogen bonding and hydrophobic interactions between the hydroxyl groups of the compounds and the residues of the cross-linking reagents used in the manufacturing process of Superose 12.6. Isolation and purification of three compounds from the leaves of Forsythia suspensa by adsorption chromatography on 12% cross-linked agarose Superose 12, adsorption chromatography was successfully applied to the separation and purification of forsythiaside, phillyrin and rutin from the leaves of Forsythia suspensa.The optimum solvent system composed of methanol-water (30:70, v/v) led to the successful preparation of forsythiaside, phillyrin and rutin with high purity (99.2%, 96.7% and 99.2%).The retention behavior of the compounds from the leaves of Forsythia suspensa on Superose 12 was studied according to the chemical structure and the retention time. The results of the present study clearly demonstrate that Superose 12 can be used for the separation of the compounds from the leaves of Forsythia suspensa and that the retention of these compounds on Superose 12 is based on a mixture of hydrogen bonding and hydrophobic interactions between the hydroxyl groups of the compounds and the residues of the cross-linking reagents used in the manufacturing process of Superose 12.