| Betulin,a kind of lupanoid triterpenoids,has strong biological activities in anti-HIV,anti-tumor.Recently,it has been attracted more and more attention.In this paper,we are investigating the extraction,identification and its teratogenic effect on zebrafish of betulin from the bark of Betula platyphylla suk.The findings will provide a basic scientific foundation for the development of betulinol and its derivatives.The major research results are as follows:1.The main parameters affecting betulin extraction were optimized.Ultrasound-assisted organic solvent extraction strategywas used to extract betulinol from the bark of birch corticoids.Through single factor design and response surface optimization,the optimum extraction conditions were: ethanol concentration 65%,solid-liquid ratio 1:25,extraction temperature 45 oC,extraction time 30 min.Under the optimum conditions,betulin extraction rate was 92.67%.The betulin extract was preliminarily purified by AB-8 macroporous resin and quantitatively and qualitatively determined by ultraviolet-visible spectrophotometry.2.Betulin was preliminarily determined in this paper.Betulin sample was firstly detected by HPLC-MS according to the retention time and molecular weight,which were compared with the betulin standard.From the MS profiles,the extract was determined to be betulin,and the molecular weight was 443.6.3.The teratogenicity of betulin sample was preliminarily demonstrated through zebrafish experiments.Three different concentrations(0.1-2.5 mg/mL)of betulin sample and drug control group were set,and changes of zebrafish phenotype were observed for continuous three days to evaluate betulin safety.The results show that Zebrafish embryos(6hpf)and juveniles(3dpf)are not damaged under microscope(4×100)observation,indicating betulin sample shows no teratogenic and lethal effects on zebrafish embryos and juveniles when the concentration of betulin ranges from 0.1mL to 2.5 mg/mL. |
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