Abstract
Trifolium pratense L. is a Chinese herb that contains a large number of isoflavones. The common methods of screening and isolating isoflavones are typically labor-intensive and time-consuming. In this research, ultrafiltration liquid chromatography (UF-LC) was applied to screen α-glucosidase and xanthine inhibitors from T. pratense L., followed by high-speed countercurrent chromatography (HSCCC) to separate and isolate the active constituents. Four major compounds in T. pratense L. were identified as α-glucosidase and xanthine oxidase inhibitors through UF-LC. Five compounds, daidzein, genistein, irilone, prunetin, and maackiain, were isolated by HSCCC with purities of 95.65, 97.25, 96.35, 97.25, and 98.65 %, respectively, using the solvent system of n-hexane:ethyl acetate:ethanol:water [2.5:4:4:5 (v:v:v:v)]. The targeted compounds were identified by liquid chromatography coupled with mass spectrometry and NMR.
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Acknowledgments
This work was supported by the National Natural Science Foundations of China (Nos. 31170326, 31370374, 31500279, and 31400453), projects of Jilin Provincial Science and Technology Department (Nos. 20130521013JH, JH20140311032YY, and 20150520140JH), project of Jilin Provincial Education Department (No. [2013] 253) and National Natural Science Foundation of Changchun Normal University (No. [2013] 001).
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Wang, Y., Tang, Y., Liu, C. et al. Determination and isolation of potential α-glucosidase and xanthine oxidase inhibitors from Trifolium pratense L. by ultrafiltration liquid chromatography and high-speed countercurrent chromatography. Med Chem Res 25, 1020–1029 (2016). https://doi.org/10.1007/s00044-016-1548-4
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DOI: https://doi.org/10.1007/s00044-016-1548-4