| HPLC法检测蛇足石杉内生真菌胶胞炭疽发酵液中石杉碱甲和石杉碱乙的含量 |
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| 引用本文： | 胡莉琴,康信聪,沈鹏原,陈田,张家银,刘东波. HPLC法检测蛇足石杉内生真菌胶胞炭疽发酵液中石杉碱甲和石杉碱乙的含量[J]. 生物工程学报, 2018, 34(5): 777-784 |
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| 作者姓名： | 胡莉琴 康信聪 沈鹏原 陈田 张家银 刘东波 |
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| 作者单位： | 湖南农业大学园艺园林学院;国家中医药管理局亚健康干预技术实验室;湖南省作物种质创新与资源利用重点实验室;湖南省植物功能成分利用协同创新中心 |
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| 基金项目： | 国家自然科学基金 (No. 81773850) 资助。 |
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| 摘 要： | 建立了高效液相色谱(HPLC)测定蛇足石杉(Huperzia serrate)内生真菌胶胞炭疽发酵液中石杉碱甲(Huperzina A)和石杉碱乙(Huperzine B)含量的方法,并以此方法检测胶胞炭疽发酵液中石杉碱甲和石杉碱乙含量的积累。内生真菌发酵液经氯仿萃取、甲醇溶解、过滤后进行高效液相检测分析,选用Agilent Eclipse plus-C18色谱柱(250 mm×4.6 mm,5μm),以0.015 mol/L乙酸铵(p H 6.8)和甲醇溶液(70∶30)为流动相进行等度洗脱,流速1 mL/min,检测波长为308 nm,连续检测内生真菌胶胞炭疽发酵液中第6–15天石杉碱甲和石杉碱乙的含量积累。结果表明,发酵提取液中的石杉碱甲和石杉碱乙可在25 min内进行很好的分离和分析,石杉碱甲在1.50-48.00μg/mL范围内线性关系良好(相关系数r为0.999 5),石杉碱乙在0.25-7.50μg/mL范围内线性关系良好(相关系数r为0.999 7),石杉碱甲和石杉碱乙的平均加标回收率分别为106.83%、108.06%,相对标准偏差(RSD)分别为3.34%、3.60%。该方法简便、快速、精密度高、结果准确,适用于内生真菌发酵液中石杉碱甲和石杉碱乙含量检测。在发酵过程中,内生真菌发酵液中石杉碱甲和石杉碱乙的含量呈现先增后减,随后有所增加继而又减少的趋势。石杉碱甲和石杉碱乙的含量分别在内生真菌发酵第14天、第8天达到最高,分别为12.417 0μg/mL、4.660 3μg/mL。该方法学的建立为内生真菌胶胞炭疽合成石杉碱甲与石杉碱乙的机制研究提供了检测手段,从而有利于药物新资源的开发。
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| 关 键 词： | 高效液相色谱，次生代谢产物，千层塔，内生真菌，石杉碱甲，石杉碱乙 |
| 收稿时间： | 2017-10-02 |
Detection of Huperzine A and Huperzine B in fermentation broth of endophytic fungus Colletotrichum gloesporioides from Huperzia serrate by HPLC |
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| Liqin Hu,Xincong Kang,Pengyuan Shen,Tian Chen,Jiayin Zhang and Dongbo Liu. Detection of Huperzine A and Huperzine B in fermentation broth of endophytic fungus Colletotrichum gloesporioides from Huperzia serrate by HPLC[J]. Chinese journal of biotechnology, 2018, 34(5): 777-784 |
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| Authors: | Liqin Hu Xincong Kang Pengyuan Shen Tian Chen Jiayin Zhang Dongbo Liu |
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| Affiliation: | 1 Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, Hunan, China; 2 State Key Laboratory of Subhealth Intervention Technology, Changsha 410128, Hunan, China,1 Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, Hunan, China; 2 State Key Laboratory of Subhealth Intervention Technology, Changsha 410128, Hunan, China,1 Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, Hunan, China; 2 State Key Laboratory of Subhealth Intervention Technology, Changsha 410128, Hunan, China,1 Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, Hunan, China; 2 State Key Laboratory of Subhealth Intervention Technology, Changsha 410128, Hunan, China,2 State Key Laboratory of Subhealth Intervention Technology, Changsha 410128, Hunan, China; 3 Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128, Hunan, China and 1 Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, Hunan, China; 2 State Key Laboratory of Subhealth Intervention Technology, Changsha 410128, Hunan, China; 3 Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128, Hunan, China; 4 Hunan Co-innovation Center for Utilization of Botanical Functional Ingredients, Changsha 410128, Hunan, China |
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| Abstract: | |
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| Keywords: | HPLC secondary metabolites Huperzia serrate endophytic fungi Huperzine A Huperzine B |
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