高效液相色谱法同时测定不同采收期枸骨叶中4种三萜类成分含量  

Simultaneous Determination of Four Triterpenoids in The Leaves of Ilex cornuta at Different Harvest Time by HPLC

作　　者：王存琴 汪雷 陈颖 臧莉 黄超 张子祥 姚倩 张克仲 WANG Cunqin;WANG Lei;CHEN Ying;ZANG Li;HUANG Chao;ZHANG Zixiang;YAO Qian;ZHANG Kezhong(College of Pharmacy,Wannan Medical College,Wuhu 241002 Anhui,China)

机构地区：[1]皖南医学院药学院,安徽芜湖241002

出　　处：《中药新药与临床药理》2020年第9期1092-1096,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：安徽省重点研究与开发计划项目(202004a07020042);皖南医学院校级重点科研基金项目(WK2019Z03);国家自然科学基金项目(81402818);安徽省高校优秀青年人才支持计划重点项目(gxyqZD2016177)。

摘　　要：目的建立高效液相色谱(HPLC)法同时测定枸骨叶中长梗冬青苷、27-p-Coumaroyloxy ursolic acid、3β-Hydroxyurs-11-en-28,13β-olide和乌发醇的定量分析方法,并观察各化合物在不同采收时期的含量差异。方法采用HPLC法对枸骨叶中4种三萜类成分进行含量测定。色谱柱:YMC-ODS C18(4.6 mm×250 mm,5μm);乙腈-0.4%磷酸水溶液为流动相,梯度洗脱;检测波长:205 nm;柱温:30℃;流速:1.0 mL·min^-1。结果枸骨叶中4种三萜类化合物均有良好的分离度,长梗冬青苷、27-p-Coumaroyloxy ursolic acid、3β-Hydroxyurs-11-en-28,13β-olide和乌发醇分别在54.4~1 740μg·mL^-1(r=0.999 7)、47.2~755.2μg·mL^-1(r=0.999 9)、45.2~867.84μg·mL^-1(r=0.999 7)、31.2~998.4μg·mL^-1(r=0.999 8)范围内呈现较好的线性关系,平均加样回收率在99.85%~101.74%之间,RSD均小于3%(n=6)。结论该方法简单准确,具有良好的重复性和稳定性,适用于测定枸骨叶中4种三萜类成分的含量,可为枸骨叶的质量控制提供科学依据。不同采收时期枸骨叶中4种三萜类成分含量存在一定差异。Objective A method was established for the simultaneous determination of 4 triterpenoids in the leaves of Ilex cornuta by HPLC,including pedunculoside,27-p-Coumaroyloxy ursolic acid,3β-Hydroxyurs-11-en-28,13β-olide and Uvaol.The differences and dynamic accumulation of 4 triterpenoids in the leaves of Ilex cornuta during different harvesting periods were studied.Methods The analysis was performed on a YMC-ODS C18 column(4.6 mm×250 mm,5 μm) with gradient elution of acetonitrile-0.4% phosphoric acid solution system at a flow rate of 1.0 mL·min^-1,and the column temperature was 30℃.The detection wavelength was 205 nm.Results The four triterpenoids had good resolutions and exhibited good linear relationships in the ranges of 54.4-1 740 μg·mL^-1(r=0.999 7),47.2-755.2 μg·mL^-1(r=0.999 9),45.2-867.84 μg·mL^-1(r=0.999 7),31.2-998.4 μg·mL^-1(r=0.999 8),respectively.The average recovery rates were 99.85%-101.74%,and the RSDs were less than 3%(n=6).Conclusion The method is simple,accurate,reproducible and stable.It is suitable for the determination of four triterpenoids in leaves of Ilex cornuta and can provide scientific basis for the quality control of leaves of Ilex cornuta.There were some differences in the contents of four triterpenoids in the leaves of Ilex cornuta during different harvesting periods.

关 键 词：枸骨 不同采收期 三萜类成分 高效液相色谱法 含量测定 

分 类 号：R284.1[医药卫生—中药学]