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Preparative isolation and purification of urolithins from the intestinal metabolites of pomegranate ellagitannins by high-speed counter-current chromatography
- Zhao, Wenhua, Wang, Yuji, Hao, Weijia, Yang, Hua, Song, Xueying, Zhao, Ming, Peng, Shiqi
- Journal of Chromatography B 2015 v.990 pp. 111-117
- X-radiation, colorectal neoplasms, countercurrent chromatography, high performance liquid chromatography, human cell lines, hydrogen peroxide, malondialdehyde, metabolites, nuclear magnetic resonance spectroscopy, oxidative stress, pomegranates, solvents, superoxide dismutase
- Urolithins were separated from the intestinal metabolites of pomegranate ellagitannins by high-speed counter current chromatography in two steps using two solvent systems composed of n-hexane-ethyl acetate-methanol-acetic acid-water (2.5:2:0.25:5, v/v/v/v/v) and n-hexane-ethyl acetate-methanol-acetic acid-water (2.5:0. 8:0.25:5, v/v/v/v/v) for the first time. Each injection of 100mg extract yielded 21mg of pure urolithin A and 10mg of pure urolithin B. High-performance liquid chromatography analyses revealed that the purity of urolithin A and urolihtin B was over 98.5%. The structures of urolithin A and urolitihn B were identified by high resolution-MS, NMR and single crystal x-ray analysis. Urolithins reduced the oxidative stress status in colon cancer by decreasing the intracellular ROS and malondialdehyde levels, and increasing SOD activity in H2O2 treated Caco-2 cells.