Separation And Purification Of Alkaloids From Lycoris Radiate, Coptis Chinensis, Litsea Cubeba, And Berberis Pliretii By PH-Zone-Refining Countercurrent Chromatography

Objective: This current article describes the application of pH-zone-refining countercurrent chromatography in the separation and purification of alkaloids from TCM. A method used for the separation of alkaloids from Lycoris radiate, Coptis chinensis Franch, Litsea cubeba(Lour) Pers., and Berberis pliretii Schneid by pH-zone-refining countercurrent chromatography has been established, which could provide scientific basis for the purification and separation of alkaloids from TCM rapidly and efficiently. At the same time, it also provides technical support for the rapid preparation of standard substance from TCM and the research and development of new drugs.Method: pH-Zone-refining countercurrent chromatography and preparative high performance liquid chromatography are used for the separation and purification alkaloids from Lycoris radiate. And pH-zone-refining countercurrent chromatography is employed for preparative separation of quaternary ammonium alkaloids from Coptis chinensis Franch. In the separation of aporphine alkaloids from Litsea cubeba(Lour) Pers., the combination of pH-zone-refining countercurrent chromatography and high-speed countercurrent chromatography are used. pH-Zone-refining countercurrent chromatography is also applied to separate the alkaloids from Berberis pliretii Schneid. The analysis of these separated compounds is carried out by high performance liquid chromatography and their chemical structures are identified by UV, MS, and NMR data.Result: Seven alkaloids were obtained from Lycoris radiate, which were identified as ungiminorine, lycorine, hemanthamine, 11-hydroxyvittatine, 9-norpluviine, galantamine, and lycoramine. The purities were all over 95%. Five quaternary ammonium alkaloids including columbamine, jateorhizine, coptisine, palmatine, and berberine with high purity all being over 96% were obtained from Coptis chinensis Franch. From Litsea cubeba(Lour) Pers., six aporphine alkaloids were obtained which including laurolitsine, boldine, laurotetanine, reticuline, isoboldine, and norisocorydine. The purity of each separated alkaloid was over 93%. Jateorhizine, palmatine, and berberine were obtained with the purity of all being over 93% from Berberis pliretii Schneid by pH-zone-refining countercurrent chromatography.Conclusion: pH-Zone-refining countercurrent chromatography could be used to separate and purify compounds with the quantity of several grams by selecting different solvent systems and concentration of retainer and eluter and combining with other chromatographic techniques. Without solid supports, pH-zone-refining countercurrent chromatography could avoid sample loss, inactivation, and degeneration caused by irreversible adsorption. The column volume of semi-preparative apparatus of high-speed countercurrent chromatography is a few hundred milliliters. It is a technique that is solvent-saving, simple, efficient, and fit for the separation of compounds without chromophores. Therefore, alkaloids were deemed as good candidates for the separation by pH-zone-refining countercurrent chromatography from natural products.