Preparative purification of antiamyloidogenic stilbenoids from Vitis vinifera (Chardonnay) stems by centrifugal partition chromatography.

Five stilbenoids, E-resveratrol, E-piceatannol, (+) E-(epsilon)-viniferin, (+)-ampelopsin A and vitisin C were isolated from methyl tert-butyl ether (MtBE) stem extract of Vitis vinifera (Chardonnay cv). Their purification on a preparative scale was obtained by centrifugal partition chromatography (CPC) using quaternary Arizona solvent systems composed of n-heptane/ethyl acetate/methanol/water. We tested 23 Arizona solvent systems to partition the extract and found that systems K and M (Hept/EtOAc/MeOH/water, 1:2:1:2 and 5:6:5:6, respectively; v/v) were the best to separate the stilbenes mentioned above. This support-free liquid-liquid chromatographic procedure made it possible to isolate ampelopsin A from V. vinifera for the first time. The antiamyloidogenic activity of the isolated stilbenes was evaluated versus beta-amyloid fibrils. E-resveratrol and (+)-ampelopsin A were found to be the most active compounds with 63 and 46% inhibition at 10microM, respectively. These findings suggest that E-resveratrol and (+)-ampelopsin A may function as attractive new candidates for protecting against brain cell dysfunction in vivo in AD by inhibiting the aggregation of Abeta.

Fractionation of low-molecular-mass heparin by centrifugal partition chromatography in the ion-exchange displacement mode.

Centrifugal partition chromatography in the ion-exchange displacement mode allowed a preparative and efficient fractionation of low-molecular-mass heparins from enoxaparin sodium. Amberlite LA2--a lipophilic liquid secondary amine--was chosen as a weak anion exchanger. The biphasic system methyl isobutyl ketone-water was selected. Protonated LA2 (10%, v/v) was added to the organic stationary phase. Hydroxide (Na+, OH-) was chosen as a displacer in the aqueous mobile phase. The observed pH and concentration profiles are typical of displacement chromatography, as supported by numerical simulation. The Dubois test for the analysis of sugar content and an analysis of sulfur content (and consequently sulfatation rate) were carried out to monitor the effectiveness of the procedure. Moreover, the fractions were analyzed by high-performance size-exclusion chromatography and the 1H NMR spectra confirmed the fractionation of the sample of enoxaparin sodium.

Constituents of the stem bark of Ormosia wayana.

The isolation of lupeol, beta-sitosterol acetate and three quinolizidinic alkaloids, (+)-lupanine, (-)-sparteine and (-)-3 beta-hydroxy-13 alpha-tigloyloxy-lupanine (1), from the stem bark of Ormosia wayana is reported.

Isolation of indole alkaloids from Catharanthus roseus by centrifugal partition chromatography in the pH-zone refining mode.

Centrifugal partition chromatography (CPC) in the pH-zone refining mode allowed a preparative and efficient isolation of vindoline, vindolinine, catharanthine and vincaleukoblastine from a crude mixture of Catharanthus roseus alkaloids. The separation protocol was tested with a synthetic mixture of vindoline, catharanthine and vincaleukoblastine. The fraction content was analyzed and the results compared with theoretical chromatograms obtained by numerical simulation. The increase in injected sample mass results in an improvement of the purity of the isolated compounds. This observation, confirmed by theory, is of prime importance for the development of preparative pH-zone refining CPC as a preparative separation method.

Dammarane saponins from Zizyphus lotus.

Four dammarane-type saponins were isolated by means of centrifugal partition chromatography from the root bark of Zizyphus lotus. Their structures were elucidated using a combination of 1D and 2D 1H and 13C NMR spectra and mass spectroscopy. One of these glycosides is the known jujuboside A. The others are three new dammarane saponins, identified as 3-O-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl (1-->3)-[alpha-L-rhamnopyranosyl(1-->2)]-alpha-L-arabinopyranosyl jujubogenin = jujuboside C, 3-O-alpha-L-rhamnopyranosyl (1-->2)-[beta-D-glucopyranosyl(1-->3)]-beta-D-galactopyranosyl lotogenin = lotoside I, and 3-O-alpha-L-rhamnopyranosyl(1-->2)-[beta-D-glucopyranosyl (1-->3)]-beta-D-glucopyranosyl lotogenin = lotoside II. Lotogenin is a new dammarane derivative identified as (15R, 16R, 20R, 22R)-16 beta, 22-epoxydammar-24-ene-3 beta, 15 alpha, 16 alpha, 20 beta-tetraol.