title: Countercurrent chromatography separation of saponins by skeleton type from Ampelozizyphus amazonicus for off-line ultra-high-performance liquid chromatography/high resolution accurate mass spectrometry analysis and characterisation.
abstract: Ampelozizyphus amazonicus Ducke (Rhamnaceae), a medicinal plant used to prevent malaria, is a climbing shrub, native to the Amazonian region, with jujubogenin glycoside saponins as main compounds. The crude extract of this plant is too complex for any kind of structural identification, and HPLC separation was not sufficient to resolve this issue. Therefore, the aim of this work was to obtain saponin enriched fractions from the bark ethanol extract by countercurrent chromatography (CCC) for further isolation and identification/characterisation of the major saponins by HPLC and MS. The butanol extract was fractionated by CCC with hexane - ethyl acetate - butanol - ethanol - water (1:6:1:1:6; v/v) solvent system yielding 4 group fractions. The collected fractions were analysed by UHPLC-HRMS (ultra-high-performance liquid chromatography/high resolution accurate mass spectrometry) and MSn. Group 1 presented mainly oleane type saponins, and group 3 showed mainly jujubogenin glycosides, keto-dammarane type triterpene saponins and saponins with C31skeleton. Thus, CCC separated saponins from the butanol-rich extract by skeleton type. A further purification of group 3 by CCC (ethyl acetate - ethanol - water (1:0.2:1; v/v)) and HPLC-RI was performed in order to obtain these unusual aglycones in pure form.
authors.affiliation: Universidade Federal do Rio de Janeiro, Instituto de Pesquisas de Produtos Naturais, CCS, bloco H, Ilha do Fundão, 21941-590, RJ, Brazil.
authors.forename: Fabiana
authors.initials: F
authors.lastname: de Souza Figueiredo
authors.affiliation: Università di Salerno, Dipartimento di Farmacia, Via Giovanni Paolo II 132, 84084, Fisciano, Italy.
authors.forename: Rita
authors.initials: R
authors.lastname: Celano
authors.affiliation: Universidade Federal do Rio de Janeiro, Departamento de Produtos Naturais e Alimentos, Faculdade de Farmácia, CCS, bloco A2, Ilha do Fundão, 21941-590, RJ, Brazil.
authors.forename: Danila
authors.initials: D
authors.lastname: de Sousa Silva
authors.affiliation: Universidade Federal do Rio de Janeiro, Instituto de Pesquisas de Produtos Naturais, CCS, bloco H, Ilha do Fundão, 21941-590, RJ, Brazil.
authors.forename: Fernanda
authors.initials: F
authors.lastname: das Neves Costa
authors.affiliation: Advanced Bioprocessing Centre, Institute of Environment, Health & Societies, CEDPS, Brunel University London, Middlesex, UB8 3PH, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Hewitson
authors.affiliation: Advanced Bioprocessing Centre, Institute of Environment, Health & Societies, CEDPS, Brunel University London, Middlesex, UB8 3PH, UK.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Ignatova
authors.affiliation: Università di Salerno, Dipartimento di Farmacia, Via Giovanni Paolo II 132, 84084, Fisciano, Italy.
authors.forename: Anna Lisa
authors.initials: AL
authors.lastname: Piccinelli
authors.affiliation: Università di Salerno, Dipartimento di Farmacia, Via Giovanni Paolo II 132, 84084, Fisciano, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Rastrelli
authors.affiliation: Universidade Federal do Rio de Janeiro, Departamento de Produtos Naturais e Alimentos, Faculdade de Farmácia, CCS, bloco A2, Ilha do Fundão, 21941-590, RJ, Brazil.
authors.forename: Suzana
authors.initials: S
authors.lastname: Guimarães Leitão
authors.affiliation: Universidade Federal do Rio de Janeiro, Instituto de Pesquisas de Produtos Naturais, CCS, bloco H, Ilha do Fundão, 21941-590, RJ, Brazil. Electronic address: ggleitao@yahoo.com.br.
authors.forename: Gilda
authors.initials: G
authors.lastname: Guimarães Leitão
mesh_terms: D000440:Butanols / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003377:Countercurrent Distribution / Q000379:methods*; D006027:Glycosides / Q000737:chemistry; D006586:Hexanes / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D010936:Plant Extracts / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry; D027823:Rhamnaceae / Q000737:chemistry*; D012503:Saponins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D012997:Solvents / Q000737:chemistry; D014315:Triterpenes / Q000737:chemistry / Q000302:isolation & purification; D000097669:Dammaranes
chemical_list: D000440:Butanols; D006027:Glycosides; D006586:Hexanes; D010936:Plant Extracts; D012503:Saponins; D012997:Solvents; D014315:Triterpenes; D000097669:Dammaranes; C400159:jujubogenin; C026385:n-hexane
keywords: Ampelozizyphus amazonicus; Countercurrent chromatography; HPLC–MS; Rhamnaceae; Saponin skeleton type; Triterpene saponins