title: New solvent systems for gradient counter-current chromatography in separation of betanin and its derivatives from processed Beta vulgaris L. juice.
abstract: Betalains, natural plant pigments, are beneficial compounds due to their antioxidant and possible chemoprotective properties. A mixture of betalains: betanin/isobetanin, decarboxybetanins and neobetanin from processed red beet roots (Beta vulgaris L.) juice was separated in food-grade, gradient solvent systems using high-performance counter-current chromatography (HPCCC). The decarboxylated and dehydrogenated betanins were obtained by thermal degradation of betanin/isobetanin from processed B. vulgaris L. juice under mild conditions. Two solvent systems (differing in their composition by phosphoric acid and ethanol volume gradient) consisting of BuOH-EtOH-NaClsolution-H2O-H3PO4 (v/v/v/v/v, 1300:200-1000:1300:700:2.5-10) in the 'tail-to-head' mode were run. The flow rate of the mobile phase (organic phase) was 1.0 or 2.0 ml/min and the column rotation speed was 1,600 rpm (20°C). The retention of the solvent system stationary phase (aqueous phase) was ca. 80%. The system with the acid and ethanol volume gradient consisting of BuOH-EtOH-NaClsolution-H2O-H3PO4 (v/v/v/v/v, 1300:200-240:1300:700:2.5-4.5) pumped at 2.0 ml/min was the most effective for a separation of betanin/isobetanin, 17-decarboxy-betanin/-isobetanin, 2-decarboxy-betanin/-isobetanin, 2,17-bidecarboxy-betanin/-isobetanin pairs as well as neobetanin. The pigments were detected by LC-DAD and LC-MS. The results are crucial in the application of completely food-grade solvent systems in separation of food-grade compounds as well, and the systems can possibly be extended to other ionizable and polar compounds with potential health benefits. In particular, the method is applicable for the isolation and purification of betalains present in such rich sources as B. vulgaris L. roots as well as cacti fruits and Amaranthaceae flowering plants due to modification possibilities of the solvent systems polarity.
authors.affiliation: Department of Analytical Chemistry, Institute C-1, Faculty of Chemical Engineering and Technology, Cracow University of Technology, ul. Warszawska 24, Cracow 31-155, Poland. Electronic address: anetasporna@chemia.pk.edu.pl.
authors.forename: Aneta
authors.initials: A
authors.lastname: Spórna-Kucab
authors.affiliation: Brunel Institute for Bioengineering, Brunel University, Uxbridge, Middlesex, United Kingdom.
authors.forename: Ian
authors.initials: I
authors.lastname: Garrard
authors.affiliation: Brunel Institute for Bioengineering, Brunel University, Uxbridge, Middlesex, United Kingdom.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Ignatova
authors.affiliation: Department of Analytical Chemistry, Institute C-1, Faculty of Chemical Engineering and Technology, Cracow University of Technology, ul. Warszawska 24, Cracow 31-155, Poland.
authors.forename: Sławomir
authors.initials: S
authors.lastname: Wybraniec
mesh_terms: D027461:Beta vulgaris / Q000737:chemistry*; D050859:Betacyanins / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution; D010936:Plant Extracts / Q000737:chemistry; D012997:Solvents; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D050859:Betacyanins; D010936:Plant Extracts; D012997:Solvents; C020228:betanin
keywords: Beta vulgaris L.; Betalains; Betanin; Counter-current chromatography; Food-grade solvent systems