title: Solvent system selectivities in countercurrent chromatography using Salicornia gaudichaudiana metabolites as practical example with off-line electrospray mass-spectrometry injection profiling.
abstract: For the development of an efficient two-stage isolation process for high-speed countercurrent chromatography (HSCCC) with focus on principal metabolites from the ethyl acetate extract of the halophyte plant Salicornia gaudichaudiana, separation selectivities of two different biphasic solvent systems with similar polarities were evaluated using the elution and extrusion approach. Efficiency in isolation of target compounds is determined by the solvent system selectivity and their chronological use in multiple separation steps. The system n-hexane-ethyl acetate-methanol-water (0.5:6:0.5:6, v/v/v/v) resulted in a comprehensive separation of polyphenolic glycosides. The system n-hexane-n-butanol-water (1:1:2, v/v/v) was less universal but was highly efficient in the fractionation of positional isomers such as di-substituted cinnamic acid quinic acid derivatives. Multiple metabolite detection performed on recovered HSCCC tube fractions was done with rapid mass-spectrometry profiling by sequential off-line injections to electrospray mass-spectrometry (ESI-MS/MS). Selective ion traces of metabolites delivered reconstituted preparative HSCCC runs. Molecular weight distribution of target compounds in single HSCCC tube fractions and MS/MS fragment data were available. Chromatographic areas with strong co-elution effects and fractions of pure recoverable compounds were visualized. In total 11 metabolites have been identified and monitored. Result of this approach was a fast isolation protocol for S. gaudichaudiana metabolites using two solvent systems in a strategic sequence. The process could easily be scaled-up to larger lab-scale or industrial recovery.
authors: Universidade Federal do Rio de Janeiro, Instituto de Pesquisas de Produtos Naturais, Centro de Ciências da Saúde, Avenida Carlos Chagas Filho, Rio de Janeiro, RJ 21941-590, Brazil. Electronic address: fncosta@nppn.ufrj.br.
authors: Fernanda das Neves
authors: Fd
authors: Costa
authors: Institute of Food Chemistry, Technische Universität Braunschweig, Schleinitzstrasse 20, 38106 Braunschweig, Germany.
authors: Gerold
authors: G
authors: Jerz
authors: Universidade Federal do Rio de Janeiro, Instituto de Pesquisas de Produtos Naturais, Centro de Ciências da Saúde, Avenida Carlos Chagas Filho, Rio de Janeiro, RJ 21941-590, Brazil.
authors: Fabiana de Souza
authors: Fde S
authors: Figueiredo
authors: Institute of Food Chemistry, Technische Universität Braunschweig, Schleinitzstrasse 20, 38106 Braunschweig, Germany.
authors: Peter
authors: P
authors: Winterhalter
authors: Universidade Federal do Rio de Janeiro, Instituto de Pesquisas de Produtos Naturais, Centro de Ciências da Saúde, Avenida Carlos Chagas Filho, Rio de Janeiro, RJ 21941-590, Brazil.
authors: Gilda Guimarães
authors: GG
authors: Leitão
mesh_terms: D005591:Chemical Fractionation; D002623:Chemistry Techniques, Analytical; D019663:Chenopodiaceae / Q000737:chemistry*; D003377:Countercurrent Distribution; D008970:Molecular Weight; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013057:Spectrum Analysis
chemical_list: D012997:Solvents
keywords: Dicaffeoyl-quinic acid derivatives; Flavonoid-glycosides; High-speed countercurrent chromatography; Off-line ESI-MS/MS injection metabolite profiling; Salicornia gaudichaudiana; Solvent system selectivity