Determination of Hypaphorine in Roots of Millettia Speciosa Champ.by UPLC
Objective:To optimize the extraction method of Hypaphorine in Millettia speciosa Champ,and to establish an ultra performance liquid chromatographic(UPLC)method for the determination of Hypaphorine in Millettia speciosa Champ,so as to compare the contents of Hypaphorine in different Millettia speciosa Champ.Methods:The extraction of Hypaphorine from Millettia speciosa Champ was optimized by single-factor and orthogonal experiments.Agilent XDB-C18column(1.8 μm,2.1×100 mm)and the mobile phase was eluted with a gradient of acetonitrile-water solution.The detection wavelength was set at 278 nm and the flow rate was 0.3 mL/min.The column temperature was maintained at 30 ℃.Results:The optimum conditions for the reflux extraction of Hypaphorine from Millettia speciosa Champ at 80 ℃ were determined as the material-liquid ratio of 1∶15 g/mL,the extraction time of 1.5 h,and the concentration of ethanol of 70％;the established method for the determination of Hypaphorine was rapid,accurate,stable and reliable.Conclusion:The content of Hypaphorine in 22 different Millettia speciosa Champ samples varied widely and was related to factors such as species,growing environment and growing years.
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