天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (9): 1469-1479.doi: 10.16333/j.1001-6880.2023.9.001

• 研究论文 •    下一篇

茯苓皮与白茯苓中三萜类成分的HPLC指纹图谱与化学模式识别研究

占慧慧1,2,3,丁婵1,2,3,彭思源1,2,3, 刘媛1,2,3,孟俊华1,2,3,肖作为1*,崔培梧1,2,3   

  1. 1湖南中医药大学药学院;2国家中医药管理局中药药性与药效三级科研实验室;3湖南中医药大学菌物药研究室,长沙 410208
  • 出版日期:2023-09-22 发布日期:2023-09-21
  • 基金资助:
    国家自然科学基金(81973211/C0033375);湖南省中医药科研计划(D2022139(A2022005-17));湖南省大学生创新创业训练计划(湘教通[2022]174号-2918);湖南中医药大学“十四五”重点学科-生物工程学科(校行发规字[2023]2号)

Study on HPLC fingerprint and chemical pattern recognition of triterpenoids in Poriae Cutis and White Poria

ZHAN Hui-hui1,2,3,DING Chan1,2,3,PENG Si-yuan1,2,3,LIU Yuan1,2,3,MENG Jun-hua1,2,3,XIAO Zuo-wei1*,CUI Pei-wu1,2,3*   

  1. 1College of Pharmacy,Hunan University of Chinese Medicine;2Research Lab of TCM Property & Efficacy,Tier 3,National Administration of TCM;3Mycomedicine Research Lab,Hunan University of Chinese Medicine,Changsha 410208,China
  • Online:2023-09-22 Published:2023-09-21

摘要: 建立茯苓皮与白茯苓中三萜类成分的HPLC指纹图谱,并对其进行化学模式识别研究,为菌物药茯苓的质量控制及标准制定提供依据。采用HPLC法,Agilent 5 TC-C18 (2)色谱柱(250 mm × 4.6 mm,5 μm),以乙腈-0.3%磷酸水溶液为流动相梯度洗脱,流速1.0 mL/min,柱温25 ℃,变波长方式检测,采集波长为242、203 nm,进样量10 μL。建立了茯苓皮与白茯苓中三萜类成分指纹图谱,应用聚类分析(CA)、主成分分析(PCA)手段结合相似度评价对实验数据进行识别,以分析不同茯苓样品的相似性和差异性,找出可反映茯苓皮与白茯苓三萜成分特征的关键色谱峰。茯苓皮与白茯苓三萜成分指纹图谱的共有模式下确认了6个共有峰,分别为2号峰(茯苓新酸B)、3号峰(去氢土莫酸)、4号峰(茯苓新酸A)、9号峰(去氢茯苓酸)、10号峰(茯苓酸)、12号峰(松苓新酸);不同来源的茯苓样品相同药用部位间相似度均在0.90以上;依据CA和PCA分析结果将茯苓样品药用部位分为两大类。PCA分析结果显示包括代表去氢土莫酸、去氢茯苓酸、茯苓酸成分在内的7个共有色谱峰(3、5和6、8~11号峰)在评价茯苓皮及白茯苓质量时必不可少,其余包括代表茯苓新酸B、茯苓新酸A、松苓新酸成分在内的8个色谱峰(1和2、4、7、12~15号峰)可作为评价茯苓皮质量的特征鉴别峰;白茯苓中15个共有色谱峰间的差异比茯苓皮小。本研究建立的茯苓三萜HPLC指纹图谱和化学模式识别可为不同茯苓药材质量控制与评价提供参考。

关键词: 茯苓, 三萜, 指纹图谱, 聚类分析, 主成分分析, 质量控制

Abstract:

In this study,HPLC analysis combined with chemical pattern recognition was employed to establish the HPLC fingerprint related to triterpenoids in Poriae Cutis and White Poria,which can provide a reference for quality control and standard establishment of decoction pieces derived from mycomedicine Poria cocos.HPLC method was adopted and performed on an Agilent 5 TC-C18 (2) column (250 mm × 4.6 mm,5 μm).The mobile phase was consisted of acetonitrile and 0.3% phosphoric acid solution,and gradient elution procedure was employed with the flow rate setting at 1.0 mL/min.The column temperature was set at 25 °C,the detection wavelength was set at 242 and 203 nm,and the injection volume was 10 μL,respectively.To establish the HPLC fingerprint and chemical pattern recognition of Poriae Cutis and White Poria,cluster analysis (CA),principal component analysis (PCA) and similarity evaluation were adopted to process the experimental data,the similarity and difference of Poriae Cutis and White Poria samples were analyzed subsequently.Finally,the key chromatographic peaks representing triterpenoids were pointed out from the established HPLC fingerprint.Among the common chemical components showed in the chromatograms,six peaks related to poricoic acid B (peak 2),dehydrotumulosic acid (peak 3),poricoic acid A (peak 4),dehydropachymic acid (peak 9),pachymic acid (peak 10),and dehydrotrametenolic acid (peak 10) were indentified,respectively.The similarity of tested decoction pieces from the same part of P. cocos sclerotium were all above 0.90.According to CA and PCA data,all the decoction pieces analyzed in this study can be divided into two groups representing different medicinal parts of P. cocos sclerotium.Seven common chromatographic peaks including peak 3 (dehydrotumulosic acid),peak 5,peak 6,peak 8,peak 9 (dehydropachymic acid),peak 10 (pachymic acid) and peak 11 were essential for the quality evaluation of Poriae Cutis and White Poria,the other eight chromatographic peaks including peak 1,peak 2 (poricoic acid B),peak 4 (poricoic acid A),peak 7,peak 12 (dehydrotrametenolic acid) and peaks 13 to 15 could be used as the characteristic identification peaks for evaluating the quality of Poriae Cutis according to the loading scatter analysis of PCA data.The characteristic difference deduced from the 15 common chromatographic peaks of White Poria samples was smaller than that of Poriae Cutis samples.The established HPLC fingerprint of Poriae Cutis and White Poria combined with chemical pattern recognition can provide a scientific reference for the quality control and evaluation of samples or products derived from P. cocos .

Key words: Poria cocos, triterpenoids, fingerprint, cluster analysis, PCA, quality control

中图分类号:  R284.1