UHPLC同时测定不同产地和不同采收期裸花紫珠及同属植物中9种活性成分的含量与质量评价  

Quality evaluation of Callicarpa nudiflora from different origins,picking stages and other plants of Callicarpa based on simultaneous determination of nine active components by UHPLC

作　　者：杨祖凡 杨扬[3] 杨义芳[3] 康兴东 李小峰 吴永忠 肖军平 王稳 贾强强 周斌[3] YANG Zu-fan;YANG Yang;YANG Yi-fang;KANG Xing-dong;LI Xiao-feng;WU Yong-zhong;XIAO Jun-ping;WANG Wen;JIA Qiang-qiang;ZHOU Bin(State Key Laboratory of Plateau Ecology and Agriculture,Qinghai University,Xining 810016,China;Medical College of Qinghai University,Xining 810001,China;Shanghai Institute of Pharmceutical Industry,Shanghai 201203,China;Jiangxi Puzheng Pharmaceutical Co.,Ltd.,Ji'an 331409,China)

机构地区：[1]青海大学三江源生态与高原农牧业国家重点实验室,青海西宁810016 [2]青海大学医学院,青海西宁810001 [3]上海医药工业研究院,上海201203 [4]江西普正药业有限公司,江西吉安331409

出　　处：《中草药》2023年第14期4631-4640,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(31460082);国家自然科学基金资助项目(R1402824);国家中医药管理局(ZYBZH-C-JX-40);青海省科技厅国际合作项目(2022-HZ-812)。

摘　　要：目的 利用UHPLC-UV建立裸花紫珠Callicarpanudiflora药材中9个活性成分同时定量的方法,并测定其在不同产地及不同采收期裸花紫珠中的含量,用于裸花紫珠药材的质量评价及产地、采收期的选择。方法 采用Phenomenex kintex XB-C_(18)色谱柱(150 mm×2.1 mm,1.7μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相进行梯度洗脱;体积流量0.3 mL/min;检测波长332nm和210nm;柱温为45℃;测定了83批裸花紫珠和7批同属不同种紫珠属样品中的9个活性成分含量。结合主成分分析及正交偏最小二乘法判别分析(orthogonal partial least squares discriminant analysis,OPLS-DA)对裸花紫珠进行全面分析。结果 建立了裸花紫珠中9个活性成分含量同时测定的方法。其中咖啡酸、木犀草苷、连翘酯苷B、毛蕊花糖苷、木犀草素-4’-O-β-D-吡喃葡萄糖苷、木犀草素-3’-O-β-D-吡喃葡萄糖苷、木犀草素、藿香黄酮醇和熊果酸分别在1.240~124.0、2.040~204.0、1.020~101.6、5.100~510.0、0.904 0~90.40、0.982 0~98.20、0.872 0~87.20、0.822 0~82.20、2.280~228.0μg/mL内线性关系良好,精密度、重复性、稳定性RSD值均小于3.00%,平均加样回收率在95.58%~101.73%,回收率RSD值均小于2.50%。主成分分析筛选出2个主成分,其累积方差贡献率为77.33%,表明这2个主成分可包含原有数据的大多数信息;OPLS-DA法标记出裸花紫珠药材中的4个差异性成分,分别为咖啡酸、连翘酯苷B、毛蕊花糖苷和熊果酸。结论 建立的多成分含量测定方法适用于裸花紫珠的质量评价,为裸花紫珠质量控制及相关经典名方开发提供参考。Objective An UHPLC-UV method was established for simultaneously determination of nine active components in Callicarpa nudiflora,and compare samples from different origins and picking stages.Methods Phenomenex Kintex XB-C_(18)column were performed on HPLC analysis,the mobile phase was acetonitrile(A)-0.1% phosphoric acid(B) for gradient elution.The flow rate was 0.3 mL/min,the wavelength was 280 nm and 332 nm and the column temperature was 45 ℃.The contents of nine active components in 83 batches of C.nudiflora and seven batches of different species of medicinal plants of Callicarpa were measured simultaneously,combined with principal component analysis and orthogonal partial least squares discriminant analysis(OPLS-DA),which were applied in the comprehensive analysis of C.nudiflora.Results A method for simultaneous quantification of nine active components in C.nudiflora was established.Caffeic acid,luteoloside,forsythoside B,acteoside,luteolin-4'-O-β-D-glucopyranoside,luteolin-3′-O-β-D-glucopyranoside,luteolin,pachypodol and ursolic acid had good linearity in the range of 1.240—124.0 μg/m L,2.040—204.0 μg/m L,1.020—101.6 μg/m L,5.100—510.0 μg/mL,0.9040—90.40 μg/mL,0.9820—98.20 μg/m L,0.8720—87.20 μg/m L,0.8220—82.20 μg/m L and 2.280—228.0 μg/m L,respectively.The RSD of precision,repeatability and stability was less than 3.00% and the average recovery value was between 95.58%—101.73%,with RSD less than 2.50%.Principal component analysis screened out two ingredients,and the cumulative variance contribution rate was 77.33%,indicating that the main ingredients contained the most information of the raw statistics.OPLS-DA screened out four differential components,including caffeic acid,forsythoside B,acteoside and ursolic acid.Conclusion The established method of multi-component quantitative analysis can be applied to the quality assessment of C.nudiflora,which also can provide reference for the quality control and the development of related prescriptions of C.nudiflora.

关 键 词：裸花紫珠 UHPLC-UV 主成分分析 多成分含量测定 苯丙烯酸类 黄酮类 咖啡酸 木犀草苷 连翘酯苷B 毛蕊花糖苷 木犀草素-4’-O-β-D-吡喃葡萄糖苷 木犀草素-3’-O-β-D-吡喃葡萄糖苷 木犀草素 藿香黄酮醇 熊果酸 

分 类 号：R286.2[医药卫生—中药学]