Two-Step Separation of Nostotrebin 6 from Cultivated Soil Cyanobacterium (Nostoc sp.) by High Performance Countercurrent Chromatography

0440741 - MBÚ 2015 RIV CH eng J - Journal Article
Cheel, José - Kučerová, P. - Garrard, I. - Ignatova, S. - Hrouzek, Pavel - Kopecký, Jiří
Two-Step Separation of Nostotrebin 6 from Cultivated Soil Cyanobacterium (Nostoc sp.) by High Performance Countercurrent Chromatography.
Molecules. Roč. 19, č. 4 (2014), s. 8773-8787. E-ISSN 1420-3049
R&D Projects: GA MŠMT ED2.1.00/03.0110; GA MŠMT EE2.3.30.0059
Institutional support: RVO:61388971
Keywords : nostotrebin 6 * cyanobacterium * Nostoc * HPLC separation
Subject RIV: EE - Microbiology, Virology
Impact factor: 2.416, year: 2014

High performance countercurrent chromatography (HPCCC) was successfully applied for the separation of nostotrebin 6 from cultivated soil cyanobacteria in a two-step operation. A two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (4:5:4:5, v/v/v/v) was employed for the HPCCC separation. In the first-step operation, its neutral upper phase was used as stationary phase and its basic lower phase (1% NH3 in lower phase) was employed as mobile phase at a flow rate of 1 mL/min. In the second operation step, its neutral upper phase was used as stationary phase, whereas both its neutral lower phase and basic lower phase were employed as mobile phase with a linear gradient elution at a flow rate of 0.8 mL/min. The revolution speed and temperature of the separation column were 1,000 rpm and 30 degrees C, respectively. Using HPCCC followed by clean-up on Sephadex LH-20 gel, 4 mg of nostotrebin 6 with a purity of 99% as determined by HPLC/DAD-ESI-HRMS was obtained from 100 mg of crude extract. The chemical identity of the isolated compound was confirmed by comparing its spectroscopic data (UV, ESI-HRMS, ESI-HRMS2) with those of an authentic standard and data available in the literature.
Permanent Link: http://hdl.handle.net/11104/0243843