摘要
目的 建立白前Cynanchum stauntonii色谱指纹图谱定性、多组分定量与化学计量学的整合分析方法,评价野生与栽培白前的质量差异。方法 采用infinity LabPoroshell 120 Aq-C_(18)(100 mm×4.6 mm,2.7μm)色谱柱,流动相为乙腈(B)-0.1%磷酸水溶液(A),梯度洗脱,检测波长为260 nm(0~14 min)、274 nm(14~52 min)、210 nm(52~70 min),体积流量为0.5m L/min,柱温为30℃,进样量为7μL。以对照药材为参照,建立了白前的超高效液相色谱法(UPLC)指纹图谱,结合聚类分析(cluster analysis,CA),主成分分析(principal component analysis,PCA)和正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)对20批白前进行质量评价,开发并验证了6种指标成分的含量测定分析方法。结果 指纹图谱及含量测定的方法学验证均良好,20批白前与对照药材的指纹图谱相似度均大于0.737,共标定14个共有峰,并指认了对羟基苯甲酸、香草酸、丁香酸、4-羟基苯乙酮、2,4-二羟基苯乙酮、anhydrohirundigenin 3-O-α-Ldiginopyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopy-ranosyl-(1→4)-β-D-thevetopyranoside(A-O-DCDT)6种指标成分;主成分得分结果与聚类分析结果一致,可将20批白前样品聚为2类,野生白前聚为一类,栽培白前聚为一类,OPLSDA筛选出引起野生与栽培白前质量差异的5个差异性标志物,分别为色谱峰7、8、11、12、14;同时,多成分定量测定结果表明,野生栽培白前中6个指标成分之间存在明显差异。结论 建立的20批白前UPLC指纹图谱、多指标成分含量测定和化学计量学相结合的质量评价方法,高效、准确、特征性强、重复性和稳定性好,可为白前质量评价体系的完善提供参考。
Objective To establish an integrated analysis method of chromatographic fingerprint,multi-component quantification and chemometrics,and to evaluate the quality difference between wild and cultivated Baiqian(Cynanchum stauntonii).Methods The infinity LabPoroshell 120 AQ-C_(18)(100 mm×4.6 mm,2.7μm)column was used for the analysis,with an acetonitrile(B)-0.1%phosphoric acid water solution(A)serving as the mobile phase.The detection wavelengths were 260 nm(0-14 min),274 nm(14-52 min)and 210 nm(52-70 min),flow rate of 0.5 mL/min,a column temperature of 30℃,and injection volume of 7μL.In this study,the ultra-high performance liquid chromatography(UPLC)fingerprint of C.stauntonii was established with reference to traditional Chinese medicinal materials,cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to evaluate the quality of 20 batches of C.stauntonii.A method for the determination of six index components of C.stauntonii was developed and validated.Results The methodological verification of fingerprint and content determination was good.The similarity of fingerprints between 20 batches of samples and the reference traditional Chinese medicinal materials of C.stauntonii was greater than 0.737.A total of 14 common peaks were calibrated,and six index components of p-hydroxybenzoic acid,vanillic acid,syringic acid,4-hydroxyacetophenone,2,4-dihydroxyacetophenone and A-O-DCDT were identified.The results of principal component score were consistent with the results of cluster analysis.The 20 batches of samples were clustered into two categories,wild C.stauntonii was clustered into one category,and cultivated C.stauntonii was clustered into another category.A total of five differential biomarkers causing differences in the quality of wild and cultivated C.stauntonii were screened using OPLS-DA,namely chromatographic peaks 7,8,11,12,and 14.Meanwhile,the results of multi-component quantitative analysis showed that there were significant differences in the six indicator components between wild and cultivated C.stauntonii.Conclusion The quality evaluation method,which combines UPLC fingerprinting,multi-component quantification,and chemometrics for 20 batches of C.stauntonii,is efficient,accurate,highly characteristic,reproducible,and stable.It can provide reference for the improvement of the quality evaluation system for C.stauntonii.
作者
刘世红
张红杨
冯书敏
黄必胜
陈诚
刘大会
雷咪
LIU Shihong;ZHANG Hongyang;FENG Shumin;HUANG Bisheng;CHEN Cheng;LIU Dahui;LEI Mi(Hubei Shizhen Laboratory/Hubei University of Chinese Medicine,Wuhan 430065,China;Xinxiangyuan Agricultural Science and Technology Development Co.,Ltd.,Wuhan 430065,China)
出处
《中草药》
CAS
CSCD
北大核心
2024年第22期7837-7846,共10页
Chinese Traditional and Herbal Drugs
基金
国家重点研发计划(2023YFC3503804)
中央本级重大增减支项目(2060302)。
关键词
白前
指纹图谱
多成分定量
化学计量学分析
质量评价
对羟基苯甲酸
香草酸
丁香酸
4-羟基苯乙酮
2
4-二羟基苯乙酮
Cynanchum stauntonii(Decne.)Schltr.ex H.Lév.
fingerprint
multi-component quantification
chemometric analysis
quality evaluation
p-hydroxybenzoic acid
vanillic acid
syringic acid
4-hydroxyacetophenone
2,4-dihydroxyacetophenone
