Simultaneous determination of huperzine A and huperzine B in Seventeen species from Huperzioideae subfamily by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of huperzine A and huperzine B from seventeen species Huperzioideae subfamily (including 2 ineditus new species).METHODS The analysises of tartaricacid extracts of huperzine A and huperzine B were performed on a room temperature column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of methanol-spiritus mindereri flowing at 1.0 mL/min in a isocratic elution manner,and the detection wavelength was set at 310 nm.The contents were determined by external standards.RESULTS Huperzine A and huperzine B showed good linear relationships within 0.002 0-0.30 μg,0.001 8-0.27 μg (r=0.999 9),whose average recoveries were 103.86％,101.3％ with the RSDs of 1.85％,1.30％,respectively.Huperzine A and huperzine B were found in seventeen species,and there were significant differences in contents from species to species.The content of huperzine A in Phlegmariurus hamiltonii was the highest,which reached up to 0.22％,with higher levels of it in Phlegmariurus cryptomerianus,Phlegmariurus petiolatus,and Phlegmariurus phlegmaria;The content of huperzine B was the highest in P.phlegmaria.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of huperzine A and B from Huperzioideae subfamily.

Detection of Huperzine A and Huperzine B in fermentation broth of endophytic fungus Colletotrichum gloesporioides from Huperzia serrate by HPLC / 生物工程学报

In this study, we established a rapid and efficient HPLC method to determine the accumulation of Huperzine A and Huperzine B in the fermentation broth of endophytic fungus Colletotrichum gloesporioides from Huperzia serrate. The chloroform extracts of fermentation broth were dissolved in methanol and filtered before injection for HPLC analysis. The analysis was performed on an Agilent Eclipse plus-C18 column (250 mm×4.6 mm, 5 μm) by isocratic elution. The mobile phase was 0.015 mol/L ammonium acetate-methanol (70:30, V/V), the flow rate was 1 mL/min and the detection wavelength was set at 308 nm. Huperzine A and Huperzine B could be well separated within 25 min. Good linearity of Huperzine A was found in the range of 1.50-48.00 μg/mL (r=0.999 5), and that of huperzine B was in 0.25-7.50 μg/mL (r=0.999 7). The average recoveries of Huperzine A and Huperzine B were 106.83% and 108.06%, respectively (RSD=3.34%, 3.60%). The results demonstrate that this method can detect the content of huperzine A and huperzine B in fermentation broth simply, rapidly, accurately and in good reproducibility. Under the optimized conditions, the accumulated content of huperzine A and huperzine B were measured from the sixth to the fifteenth day. Huperzine A and Huperzine B reached the highest (12.417 0 μg/mL and 4.660 3 μg/mL, respectively) at the fourteenth and eighth days. The analysis methodology could contribute to the future study of huperzine A and huperzine B biosynthesis in C. gloeosporioides, consequently facilitate the development of new drug resources.