Study On The Chromatographic Separation Method Of Alkaloids In The Huperzia Serrata

In this thesis, aimed to solve the problem of separation for alkaloids, the extract of Huperzia serrata is chosen as object, and studied from the following three aspects: quantitative analysis of Huperzine A, building various modes of2D liquid chromatography methods and preparation of pure compounds from the complex extract.Firstly, a high performance liquid chromatography method was established to determine the Huperzine A in the extract of Huperzia serrata. After extracted by methanol/water/formic acid (10/90/0.2, v/v/v), the sample was filtered for HPLC analysis. The separation was performed on an XCharge C18column by gradient elution with water (containing0.1%trifluoroacetic acid,v/v) and acetonitrile (containing0.09%trifluoroacetic acid,v/v) as the mobile phases. Rapid separation was achieved within10min at a flow rate of2mL/min with ultraviolet absorption detection at a wavelength of310nm. Through the investigation of standard curves, precision of instrument, stability of method and recovery, the results demonstrate that the created method was simple, rapid and accurate with good reproducibility and could be used to evaluate the quality of Huperzia serrata.Secondly, based on the different separation mechanism of RPLC, IEX and Mixed-mode, three kinds of two-dimensional liquid chromatography systems was built, including XCharge C18/XAqua C3、XCharge C18/XCharge SCX and XCharge C18/C18WCX for the separation of Huperzia serrata. And a simple evaluation system was applied to observe the orthogonality. The results showed that every system was high orthogonal, which satisfied the efficient analysis for the complex components of Huperzia serrata extract.Lastly, the purification preparation of Huperzia serrata extract was perfectly completed using one of2D liquid chromatography systems. By means of MS and1H NMR, the structures of Huperzine A and B had been confirmed.