title: Schinus terebinthifolius scale-up countercurrent chromatography (Part I): High performance countercurrent chromatography fractionation of triterpene acids with off-line detection using atmospheric pressure chemical ionization mass spectrometry.
abstract: 'Countercurrent chromatography' (CCC) is an ideal technique for the recovery, purification and isolation of bioactive natural products, due to the liquid nature of the stationary phase, process predictability and the possibility of scale-up from analytical to preparative scale. In this work, a method developed for the fractionation of Schinus terebinthifolius Raddi berries dichloromethane extract was thoroughly optimized to achieve maximal throughput with minimal solvent and time consumption per gram of processed crude extract, using analytical, semi-preparative and preparative 'high performance countercurrent chromatography' (HPCCC) instruments. The method using the biphasic solvent system composed of n-heptane-ethyl acetate-methanol-water (6:1:6:1, v/v/v/v) was volumetrically scaled up to increase sample throughput up to 120 times, while maintaining separation efficiency and time. As a fast and specific detection alternative, the fractions collected from the CCC-separations were injected to an 'atmospheric pressure chemical ionization mass-spectrometer' (APCI-MS/MS) and reconstituted molecular weight MS-chromatograms of the APCI-ionizable compounds from S. terebinthifolius were obtained. This procedure led to the direct isolation of tirucallane type triterpenes such as masticadienonic and 3β-masticadienolic acids. Also oleanonic and moronic acids have been identified for the first time in the species. In summary, this approach can be used for other CCC scale-up processes, enabling MS-target-guided isolation procedures.
authors.affiliation: Institute of Food Chemistry, Technische Universität Braunschweig, Schleinitzstrasse 20, 38106 Braunschweig, Germany. Electronic address: m.neves-vieira@tu-braunschweig.de.
authors.forename: Mariana Neves
authors.initials: MN
authors.lastname: Vieira
authors.affiliation: Institute of Food Chemistry, Technische Universität Braunschweig, Schleinitzstrasse 20, 38106 Braunschweig, Germany; Institute of Natural Products Research, Federal University of Rio de Janeiro, CCS, Bl. H, Ilha do Fundão 21941-590, RJ, Brazil.
authors.forename: Fernanda das Neves
authors.initials: Fd
authors.lastname: Costa
authors.affiliation: Institute of Natural Products Research, Federal University of Rio de Janeiro, CCS, Bl. H, Ilha do Fundão 21941-590, RJ, Brazil.
authors.forename: Gilda Guimarães
authors.initials: GG
authors.lastname: Leitão
authors.affiliation: Advanced, Bioprocessing Centre, Institute of Environment, Health & Societies, Brunel University, Uxbridge, Middlesex UB8 3PH, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Garrard
authors.affiliation: Advanced, Bioprocessing Centre, Institute of Environment, Health & Societies, Brunel University, Uxbridge, Middlesex UB8 3PH, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Hewitson
authors.affiliation: Advanced, Bioprocessing Centre, Institute of Environment, Health & Societies, Brunel University, Uxbridge, Middlesex UB8 3PH, UK.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Ignatova
authors.affiliation: Institute of Food Chemistry, Technische Universität Braunschweig, Schleinitzstrasse 20, 38106 Braunschweig, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Winterhalter
authors.affiliation: Institute of Food Chemistry, Technische Universität Braunschweig, Schleinitzstrasse 20, 38106 Braunschweig, Germany.
authors.forename: Gerold
authors.initials: G
authors.lastname: Jerz
mesh_terms: D027926:Anacardiaceae / Q000737:chemistry*; D001274:Atmospheric Pressure; D005591:Chemical Fractionation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification*; D012997:Solvents / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D014315:Triterpenes / Q000302:isolation & purification
chemical_list: D010936:Plant Extracts; D012997:Solvents; D014315:Triterpenes; C459621:masticadienolic acid; C514393:tirucallane
keywords: CCC off-line APCI-MS/MS profiling; High performance countercurrent chromatography; Method optimization; Scale-up; Schinus terebinthifolius; Triterpene acids