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Title: Two-step separation of nostotrebin 6 from cultivated soil cyanobacterium (Nostoc sp.) by high performance countercurrent chromatography
Authors: Cheel, J
Kučerová, P
Garrard, I
Ignatova, S
Hrouzek, P
Kopecký, J
Keywords: Countercurrent chromatography;Cyanobacteria;Nostotrebin-6;Polyphenolic compound
Issue Date: 2014
Publisher: MDPI AG
Citation: Molecules, 19(7): 8773 - 8787, (2014)
Abstract: 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 °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.
ISSN: 1420-3049
Appears in Collections:Brunel Institute for Bioengineering (BIB)

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