Preparative isolation of huperzines A and B from Huperzia serrata by displacement centrifugal partition chromatography.
The pH-zone refining centrifugal partition chromatography technique was used to separate the two acetylcholinesterase inhibitors huperzines A and B from a crude alkaloid extract of the club moss Huperzia serrata. Complete co-elution of huperzines A and B was initially observed with the well-known methyl tert-butyl ether-acetonitrile-water (4:1:5, v/v/v) solvent system with triethylamine (8mM) as the displacer and methane sulfonic acid (6mM) as the retainer. An efficient biphasic system was designed on the basis of solvent association that provided selectivity in the elution mode: n-heptane/ethyl acetate/n-propanol/water (5:15:35:45, v/v/v/v). Lowering the bridge solvent content (n-propanol) of this system increased the polarity difference between the two phases thus adapting it to the pH-zone refining mode. Thus, the purification of these compounds was achieved using the biphasic system n-heptane/ethyl acetate/n-propanol/water (10:30:15:45, v/v/v/v) with triethylamine (8mM) as the displacer and methane sulfonic acid (6mM) as the retainer.
Toribio
Alix
A
FRE CNRS 2715, IFR 53, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, BP 1039, 51687 Reims Cedex 2, France.
Delannay
Eldra
E
Richard
Bernard
B
Plé
Karen
K
Zèches-Hanrot
Monique
M
Nuzillard
Jean-Marc
JM
Renault
Jean-Hugues
JH
D000470:Alkaloids / Q000302:isolation & purification*; D002498:Centrifugation / Q000379:methods*; D003377:Countercurrent Distribution / Q000379:methods*; D032501:Huperzia / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D012717:Sesquiterpenes / Q000302:isolation & purification*
D000470:Alkaloids; D012717:Sesquiterpenes; C050426:huperzine A; C053016:huperzine B