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A method that involved the combination of pH-zone-refining counter-current chromatography and semipreparative reversed-phase liquid chromatography has been established for the preparative separation of alkaloids from Hypecoum leptocarpum. From 1.2 g of crude sample, 31 mg N-feruloyltyramine, 27 mg oxohydrastinine, 47 mg hydroprotopine, 25 mg leptopidine, and 18 mg hypecocarpine have been obtained. The structure of the new compound, hypecocarpine, is confirmed based on the analysis of spectroscopic data, including NMR, UV, and IR spectroscopy and positive electrospray ionization mass spectrometry. The known chemical structures were characterized on the basis of (1) H and (13) C NMR spectroscopy. The purities of the five alkaloids are all over 92.7% as determined by high-performance liquid chromatography. The alkaloids' cytotoxicity in breast cancer cells is assessed by using a Cell Counting Kit assay and their inhibitory effect on fatty acid synthase expression is assessed by a Western blot assay. These results suggest that leptopidine could suppress growth and induce cytotoxicity in breast cancer cells and that the cytotoxicity of leptopidine may be related to its inhibitory effect on fatty acid synthase expression.

High-speed counter-current chromatography (CCC) was firstly and successfully applied for the preparative separation and purification of alkaloids from crude extract of Hypecoum leptocarpum. After the measurement of partition coefficient of five target alkaloids in the two-phase solvent systems, the CCC was performed well with a two-phase solvent system composed of tetrachloromethane-chloroform-methanol-0.1 M HCl at a volume ratio of 1.5 : 2.5 : 3 : 2 (V/V/V/V). The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase. From 120 mg crude extract, 5 mg leptopidine, 32 mg oxohydrastinine, 27 mg (-)-N-methylanadine, 7 mg N-feruloyltyramine and 3 mg hypecoleptopine could be successfully separated. The amides alkaloid, N-feruloyltyramine, was firstly separated from H. leptocarpum. High-performance liquid chromatography analysis showed that the purity of each of the five target alkaloids was over 92%. Their chemical structures were confirmed by (1)H-NMR and (13)C-NMR data.

Anthocyanins are the main compounds in Nitraria tangutorun Bobr. The enrichment and purification of anthocyanins on macroporous resins were investigated. Regarding anthocyanin purification, static adsorption and desorption were studied. The optimal experimental conditions were the following: resin type: X-5; static adsorption time: 6h; desorption solution: ethanol-water-HCl (80:19:1, V/V/V; pH 1); desorption time: 40min. Furthermore, the in vitro and in vivo biological activities of the anthocyanins were evaluated. The anthocyanins showed ideal scavenging effects on free radicals in vitro, especially on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl free radical (OH). In the animal experiment, blood lipid metabolism of hyperlipidemia rats was regulated by anthocyanin contents. The superoxide dismutase (SOD) activity and the total antioxidant capacity (TAC) of hyperlipidemia rats were also improved by anthocyanins. These results showed that anthocyanins from N. tangutorun Bobr. fruits had potential biological activities in vivo as well as in vitro.