大理藜芦中甾体生物碱的动态积累研究  

Study on the dynamic accumulations of steroidal alkaloid in Veratrum taliense

作　　者：朱金莲 王葳 杨礼攀 秦苗 陆云环 普春霞 ZHU Jinlian;WANG Wei;YANG Lipan;QIN Miao;LU Yunhuan;PU Chunxia(Chinese Medicine College,Yunnan University of Chinese Medicine,Kunming,Yunnan,650500 P.R.China)

机构地区：[1]云南中医药大学中药学院,云南昆明650500

出　　处：《华西药学杂志》2024年第6期692-696,共5页West China Journal of Pharmaceutical Sciences

基　　金：云南省科技厅科技计划项目(202101AZ070001-050);云南省院士专家工作站项目(202305AF150030)。

摘　　要：目的建立以介藜芦碱、藜芦胺和环巴胺为指标的大理藜芦质量控制方法;测定大理藜芦中代表性甾体生物碱的含量,研究其动态积累变化,为大理藜芦甾体生物碱的生物合成代谢调控研究奠定基础。方法采用Ecosil ODS-Extend 100A(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈(A)-0.03%三乙胺水溶液(B),进样量10μL,采用HPLC法测定大理藜芦中代表性甾体生物碱的含量,采用“中药色谱指纹图谱相似度评价系统(2012版)”进行指纹图谱分析;应用SPSS.25软件进行聚类分析。结果HPLC-PDA检测法精密度、重复性和稳定性的RSD均<3%,平均回收率>98.91%,RSD均<3%,3种甾体生物碱的线性范围125.0~997.6 mg·L^(-1)(r>0.9998)。大理藜芦甾体生物碱的动态积累具有一定的规律,地下部分甾体生物碱多样性及含量显著高于地上部分,乌龙坝居群甾体生物碱的含量明显高于梁王山居群。结论所用方法高效便捷,可用于定量检测大理藜芦中的介藜芦碱、藜芦胺、环巴胺。大理藜芦中的甾体生物碱在不同生长期具有不同的积累和转化规律,研究其动态积累,为生物合成代谢调控奠定基础,对大理藜芦的人工栽培及规范化种植也具有指导意义。OBJECTIVE To establish a quality control method of Veratrum taliense with the active indicators of jervine,veratramine and cyclopamine,and further,to determine the content of representative steroidal alkaloids in V.taliense and study its dynamic accumulations,building a foundation for the research on regulation of steroidal alkaloid biosynthesis and metabolism in V.taliense.METHODS The determination was performed on Ecosil ODS-Extend 100A(250 mm×4.6 mm,5μm)column with a mobile phase consisting of acetonitrile(A)-0.03%triethylamine(B)to establish an HPLC method(the sample size was 10μL)to determine the content of representative steroidal alkaloids in V.taliense.The similarity evaluation system of chromatographic fingerprints of TCM(2012)was used for fingerprint analysis,and SPSS.25 was used for cluster analysis.RESULTS The precision,repeatability,and stability RSD values of HPLC method were all less than 3%,with an average recovery rate of over 98.91%and RSD values of less than 3%.These three steroid alkaloids had a good linear relationship within the range of 125.0-997.6 mg·L^(-1),with correlation coefficient r^(2) was>0.9998.The dynamic accumulation of steroidal alkaloids in V.taliense was regular.The diversity and content of steroidal alkaloids in the underground part were significantly higher than those in the aerial plant,and the content of steroidal alkaloids in the Wulongba population was significantly higher than that in the Liangwangshan population.CONCLUSION The method is efficient and convenient,and can be used for the quantitative detection of jervine,veratramine and cyclopamine in Veratrum taliense.The steroidal alkaloids in V.taliense have different accumulation and transformation rules in different growth periods.Studying their dynamic accumulation establishes a basis for the regulation of steroidal alkaloid biosynthesis and metabolism in V.taliense,and also has guiding value for the artificial cultivation and standardized planting of V.taliense.

关 键 词：大理藜芦 甾体生物碱 介藜芦碱 藜芦胺 环巴胺 高效液相色谱法 动态积累 

分 类 号：R917[医药卫生—药物分析学]