title: Development of an efficient method for the preparative isolation and purification of chlorophyll a from a marine dinoflagellate Amphidinium carterae by high-speed counter-current chromatography coupled with reversed-phase high-performance liquid chromatography.
abstract: In our research into chlorophylls of marine dinoflagellates, chlorophyll a was separated rapidly from the hexane extract of Amphidinium carterae in three steps. The first step was silica gel column chromatography, where elution was performed with 0-50% ethyl acetate in n-hexane. The second was high-speed counter-current chromatography using a two-phase solvent system consisting of n-hexane-ethyl acetate-methanol-water (5:5:5:1, v/v), and the third step was preparative reversed-phase high-performance liquid chromatography using a solvent system of acetone-water (89:11, v/v). HPLC analysis showed that the purity of chlorophyll a from the second step was over 83%, and after the third it was over 99%. Thirty milligrams of chlorophyll a was isolated from a crude sample of 250 mg of chlorophylls, and its structure was identified by analyzing its MS, 1H NMR and 13C NMR spectra.
authors.affiliation: Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, People's Republic of China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Long
authors.forename: Yang
authors.initials: Y
authors.lastname: Song
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.forename: Li
authors.initials: L
authors.lastname: Lei
authors.forename: Kai
authors.initials: K
authors.lastname: Huang
authors.forename: Benwen
authors.initials: B
authors.lastname: Long
mesh_terms: D000818:Animals; D002734:Chlorophyll / Q000302:isolation & purification*; D002853:Chromatography, Liquid / Q000379:methods*; D004141:Dinoflagellida
chemical_list: D002734:Chlorophyll