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

Molecules

José Cheel²  Petra Kuპrová²  Ian Garrard¹  Svetlana Ignatova¹  Pavel Hrouzek² 

[1] Brunel Institute for Bioengineering, Brunel University, Kingston Lane, Uxbridge, Middlesex UB8 3PH, UK; E-Mails:;Department of Phototrophic Microorganisms-ALGATECH, Institute of Microbiology, Academy of Sciences of the Czech Republic, Opatovický mlýn, Třebon 379 81, Czech Republic; E-Mails:

关键词: cyanobacteria;    nostotrebin-6;    countercurrent chromatography;    polyphenolic compound;   

DOI  :  10.3390/molecules19078773

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.