Preparative separation of quaternary ammonium alkaloids from Coptis chinensis Franch by pH-zone-refining counter-current chromatography

• Published in: Journal of Chromatography A Vol. 1370; pp. 156 - 161
• Main Authors: Sun, Changlei, Li, Jia, Wang, Xiao, Duan, Wenjuan, Zhang, Tianyou, Ito, Yoichiro
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 28.11.2014; Elsevier
• Subjects: Alkaloids; Alkaloids - isolation & purification; Biological and medical sciences; Chromatography; Chromatography, High Pressure Liquid - methods; Coptis - chemistry; Coptis chinensis Franch; Countercurrent Distribution - methods; General pharmacology; Hydrochloric acid; Hydrogen-Ion Concentration; Magnetic Resonance Spectroscopy; Medical sciences; pH-zone-refining counter-current chromatography; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; Plant Extracts - chemistry; Purity; Quaternary ammonium alkaloids; Quaternary Ammonium Compounds - isolation & purification; Separation; Solvents; Solvents - chemistry; Spectrometry; Spectrometry, Mass, Electrospray Ionization; Spectroscopy; Asia; China; Coptis chinensis Franch; Quaternary ammonium alkaloids; pH-zone-refining counter-current chromatography; Pentacyclic compound; Countercurrent chromatography; Organic cation; Purification; Iminium compounds; Angular nitrogen heterocycle; Pharmacognosy; Preparative chromatography; Ranunculaceae; Tetracyclic compound; Medicinal plant; Separation method; Alkaloid; Folk medicine; Dicotyledones; Angiospermae; Plant origin; pH; Isolation; Spermatophyta; Oxygen nitrogen heterocycle
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2014.10.043

•Highly polar compounds could be separated by pH-zone-refining CCC.•Five quaternary ammonium alkaloids with high purities are obtained in a single run.•Optimizing the concentration of the retainer and eluter is the key step.•A pre-purified sample is very important for an efficient separation. pH-zone-refining counter-current chromatography was successfully applied to the preparative separation of five quaternary ammonium alkaloids from the crude extract of Coptis chinensis Franch. The separation was performed with a two-phase solvent system composed of chloroform–methanol–water (4:3:3, v/v), where the upper aqueous stationary phase was added with 60mM of hydrochloric acid and the lower organic mobile phase with 5mM of triethylamine. From 1.0g of crude extract, 5.4mg of columbamine at 96.6% purity, 6.1mg of jateorhizine at 98.8% purity, 58.3mg of coptisine at 99.5% purity, 25.6mg of palmatine at 98.4% purity and 503.9mg of berberine at 99.5% purity were obtained. The purities of the isolated alkaloids were analyzed by HPLC and the chemical structures were identified by electrospray ionization-mass spectrometry and 1H NMR.