HPLC测定不同产地叶下珠中6种成分含量及指纹图谱研究  

HPLC fingerprint studies and determination of six components in Phyllanthus urinaria L.collected from different habitats

作　　者：王艳红[1] 潘新波[1] 赵丽娟 赵唯年 丁丽[1] 贾忠 孟敏[3] Wang Yanhong;Pan Xinbo;Zhao Lijuan;Zhao Weinian;Ding Li;Jia Zhong;Meng Min(Institute of Liver Disease,the Second People's Hospital of Lanzhou City,Lanzhou 730046,China;Veterinary College,Gansu Agricultural University,Lanzhou 730070,China;Department of Pharmacy,Gansu Provincal Hospital,Lanzhou 730000,China)

机构地区：[1]兰州市第二人民医院肝病研究所,兰州730046 [2]甘肃农业大学兽医学院,兰州730070 [3]甘肃省人民医院药剂科,兰州730000

出　　处：《国际中医中药杂志》2022年第12期1408-1414,共7页International Journal of Traditional Chinese Medicine

基　　金：甘肃省自然科学基金项目(20JR10RA370);兰州市科技计划指导性项目(2022-ZD-18);兰州市人才创新创业项目(2019-RC-3、2021-RC-121)。

摘　　要：目的建立HPLC法同时测定叶下珠中没食子酸、短叶苏木酚、柯里拉京、老鹳草素、鞣花酸和芦丁6种成分含量的方法及指纹图谱分析方法。方法叶下珠以50%甲醇超声提取,采用Phenonmenex Luna C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,流速1.0 ml/min,柱温25℃,进样量10μl,检测波长270 nm。建立不同产地叶下珠HPLC指纹图谱,采用主成分分析法(PCA)和正交偏最小二乘判别(OPLS-DA)分析不同产地叶下珠化学成分的差异性。结果没食子酸、短叶苏木酚、柯里拉京、老鹳草素、鞣花酸和芦丁分别在0.0428~0.6416、0.0334~0.5014、0.1422~2.1331、0.3831~5.7465、0.0631~0.9462、0.0192~0.2878μg范围内与峰面积线性关系良好,平均回收率分别为103.65%、96.39%、101.85%、95.04%、98.79%、98.33%。不同产地叶下珠HPLC指纹图谱共有14个特征峰,鉴别指认其中6种化合物。PCA分析结果显示不同产地叶下珠所含化学成分具有差异性,OPLS-DA筛选出老鹳草素、鞣花酸和柯里拉京3个化合物为不同产地叶下珠的差异性成分。结论该方法高效、准确、灵敏度高,可用于叶下珠药材中上述6种成分含量的测定,建立的不同产地叶下珠HPLC指纹图谱结合6个特征性指标成分含量测定方法可用于叶下珠的质量控制评价。Objective To establish a method of measuring the contents of gallic acid,brevifolin,corilagin,geraniin,ellagic acid and rutin in Phyllanthus urinaria L.simultaneously with fingerprint study for analysis.Methods Phyllanthus urinaria L.was extracted by ultrasound with 50%methanol.Chromatographic separation was performed on a Phenonmenex Luna C18(4.6 mm×250 mm,5μm).The mobile phase consisted of acetonitrile(A)and 0.1%phosphoric acid aqueous solution(B)with gradient elution.The flow rate was 1.0 ml/min.The column temperature was 25℃,and the injection volume was 10μl.The detection wavelength was 270 nm.HPLC fingerprintsof Phyllanthus urinaria L.from different habitats was established.PCA and OPLS-DA were used to analyze the differences in chemical components of different habitats.Results Gallic acid,brevifolin,corilagin,geraniin,ellagic acid and rutin showed good linearity at 0.0428-0.6416,0.0334-0.5014,0.1422-2.1331,0.3831-5.7465,0.0631-0.9462 and 0.0192-0.2878μg,respectively.The average recovery rate of them was 103.65%,96.39%,101.85%,95.04%,98.79%and 98.33%,respectively.The HPLC fingerprints of different habitats contained 14 characteristic common peaks,and six compounds characteristic peaks were identified.PCA analysis showed that the chemical components of Phyllanthus urinaria L.from different habitats were different.Geraniin,ellagic acid and corilagin were screened by OPLS-DA.Conclusions The method is efficient,accurate and sensitive,which can be used to measure the six components in Phyllanthus urinaria L..The established HPLC fingerprint of different habitats combined with the measrurement method of six components can be used for the quality control and evaluation of Phyllanthus urinaria L..

关 键 词：叶下珠 高效液相色谱法 中药产地 含量测定(中药) 指纹图谱 

分 类 号：R284.1[医药卫生—中药学] O657.72[医药卫生—中医学]