基于高效液相色谱法结合化学计量学分析宣木瓜和资丘木瓜三萜酸成分的差异  

Analysis of the Differences of Triterpenoid Acids Between “Xuanmugua”and“Ziqiumugua”Based on HPLC and Chemometric Methods

作　　者：胡丹 张亚莉 喻格 李娜 李道文 张玲 HU Dan;ZHANG Yali;YU Ge;LI Na;LI Daowen;ZHANG Ling(School of Pharmacy,Anhui University of Traditional Chinese Medicine,Hefei 230012 Anhui,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012

出　　处：《中药新药与临床药理》2022年第12期1694-1700,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：安徽省高校自然科学研究重点项目(KJ2020A0442);安徽中医药大学校级科研项目(2020zryb08);安徽中医药大学校级探索性项目(2021hxts23,2021zxts16)。

摘　　要：目的 建立高效液相色谱法(HPLC)同时测定木瓜中山楂酸、科罗索酸、白桦脂酸、齐墩果酸、熊果酸和乙酰熊果酸6种三萜酸的含量,并结合化学计量学方法分析宣木瓜和资丘木瓜三萜酸成分的差异。方法 采用Shim-pack CLC-ODS(M)色谱柱(4.6 mm×250 mm,5μm);测定波长为210 nm;流动相为甲醇-水-冰乙酸-三乙胺(255∶45∶0.15∶0.05);流速:0.9 mL·min^(-1);柱温:25℃。运用聚类分析、偏最小二乘判别分析(PLS-DA)和主成分分析(PCA)探究宣木瓜和资丘木瓜三萜酸成分的差异性。结果 木瓜中6种三萜酸均能实现较好分离,在考察的浓度范围内线性关系良好(r≥0.999 9),加样回收率为90%~100%,回收率的RSD均小于3%。聚类分析发现,组间距为15时能将宣木瓜和资丘木瓜聚为两类,PLS-DA表明齐墩果酸、熊果酸、山楂酸、白桦脂酸是宣木瓜和资丘木瓜的主要差异性成分,PCA发现宣木瓜综合得分普遍高于资丘木瓜。结论 宣木瓜和资丘木瓜中三萜酸含量存在明显差异,所建立的HPLC结合化学计量学方法可有效区分两个主产地的木瓜。Objective To establish an HPLC method to simultaneously determine the content of 6 kinds of triterpenoid acids(including maslinic acid, corosolic acid, betulinic acid, oleanolic acid, ursolic acid and acetylursolic acid)in Chaenomelis Fructus. The differences of the content of triterpene acids between Xuanmugua(Chaenomelis Fructus from Xuancheng city, Anhui province) and Ziqiumugua(Chaenomelis Fructus from Changyang county, Hubei province) were analyzed by chemometric methods. Methods High performance liquid chromatography(HPLC)was performed on a Shim-pack CLC-ODS(M)column(4.6 mm×250 mm,5 μm). The detection wavelength was 210 nm, while the mobile phase was methanol-water-glacial acetic acid-triethylamine(255∶45∶0.15∶0.05)and the flow rate of mobile phase was 0.9 mL·min^(-1). The column temperature was 25 ℃.Cluster analysis,partial least squares discriminant analysis(PLS-DA)and principal component analysis(PCA)were used to explore the differences of the content of 6 kinds of triterpene acids between “Xuanmugua” and“Ziqiumugua”. Results The 6 kinds of triterpenoid acids in Chaenomelis Fructus could be well separated and the linear relationships were good(r≥0.999 9)within the investigated concentration range. The average recoveries ranged from 90% to 100%(RSD<3%). Cluster analysis suggested that when group interval was 15, 22 batches of Chaenomelis Fructus could be clustered into two clades. PLS-DA showed that oleanolic acid,ursolic acid,maslinic acid and betulinic acid were the main different components of Chaenomelis Fructus in two producing areas. The results of PCA also showed that the comprehensive score of“Xuanmugua”was generally higher than that of“Ziqiumugua”. Conclusion There are obvious differences in the content of triterpenoids acids between“Xuanmugua”and“Ziqiumugua”. The established HPLC combining with chemometric methods can be used to distinguish effectively Chaenomelis Fructus from the two main producing areas.

关 键 词：木瓜 宣木瓜 资丘木瓜 三萜酸 高效液相色谱法 聚类分析 偏最小二乘判别分析 主成分分析 

分 类 号：R284.1[医药卫生—中药学]