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钩苞大丁草的HPLC指纹图谱建立及含量测定
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| 篇名： | 钩苞大丁草的HPLC指纹图谱建立及含量测定 |
| TITLE： | Establishment of HPLC fingerprint and content determination of Gerbera delavayi |
| 摘要： | 目的 建立钩苞大丁草的指纹图谱及其11种成分的含量测定方法。方法采用高效液相色谱（HPLC）法，根据《中药色谱指纹图谱相似度评价系统（2012版）》建立13批（编号S1～S13）钩苞大丁草的指纹图谱并进行相似度评价，同时进行共有峰指认；采用SPSS25.0软件和SIMCA14.1软件进行分层聚类分析（HCA）、主成分分析（PCA）和正交偏最小二乘法-判别分析（OPLS-DA）；采用HPLC法测定样品中新绿原酸、绿原酸、隐绿原酸、3，8-二羟基-4-甲氧基-5-羧基-香豆精、咖啡酸、3-羟基-4-甲氧基-5-羧基-香豆精、木犀草素-7-O-β-D-葡萄糖苷、异绿原酸A、芹菜素-7-O-β-D-葡萄糖苷、异绿原酸C、花椒毒素11种成分的含量。结果13批钩苞大丁草的HPLC指纹图谱相似度为0.801～0.994；从中共标定了38个共有峰，并指认了其中13个共有峰。HCA、PCA结果均显示，S1～S5、S7聚为一类，S6聚为一类，S8聚为一类，S9、S11聚为一类，S10、S12～S13聚为一类；OPLS-DA结果显示，峰7（绿原酸）、峰21（异绿原酸A）、峰26（花椒毒素）、峰19（异绿原酸B）、峰33、峰13、峰23（异绿原酸C）、峰2（新绿原酸）、峰17（木犀草素-7-O-β-D-葡萄糖苷）的变量重要性投影值均大于1。上述11种成分在各自检测质量浓度范围内线性关系均良好（r均大于0.999）；精密度、重复性、稳定性试验的RSD均不大于2%（n均为6）；平均加样回收率为92.54%～105.55%，RSD为0.83%～1.93%（n＝6）；平均含量分别为0.744、5.014、0.646、0.431、0.069、0.582、0.979、2.754、0.157、1.284、2.943mg/g。结论本研究建立的HPLC指纹图谱和含量测定方法简单、准确、稳定，可为钩苞大丁草药材的质量控制提供依据。花椒毒素、绿原酸、异绿原酸A、木犀草素-7-O-β-D-葡萄糖苷、异绿原酸C、新绿原酸可作为钩苞大丁草药材的质量标志物。 |
| ABSTRACT： | OBJECTIVE To establish the fingerprint of Gerbera delavayi and the methods for the content determination of 11 components in G. delavayi. METHODS High-performance liquid chromatography（HPLC）was adopted to establish the fingerprints of 13 batches of G. delavayi（No. S1-S13）， and the similarities were evaluated according to Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition）， while the common peaks were identified. Hierarchical clustering analysis （HCA）， principal component analysis （PCA） and orthogonal partial least square-discriminant analysis （OPLS-DA） were carried out by using SPSS 25.0 software and SIMCA 14.1 software. The contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， 3，8-dihydroxy-4-methoxy-2-oxo-2H-1-benzopyran-5-carboxylic acid， caffeic acid， 3-hydroxy-4-methoxy-2- oxo-2H-1-benzopyran- 5-carboxylic acid， luteolin-7-O-β-D-glucoside， isochlorogenic acid A， apigenin-7-O-β-D-glucoside， isochlorogenic acid C and xanthotoxin were determined by HPLC. RESULTS The similarities in HPLC fingerprint of 13 batches of G. delavayi were 0.801-0.994； a total of 38 common peaks were identified and 13 common peaks were identified. The results of HCA showed that S1-S5 and S7 were clustered into one group， S6 into one category， S8 into one category， S9 and S11 into one category， S10， S12 and S13 into one category， and the results of PCA were consistent with them. The results of OPLS-DA showed that variable importance values for the projection of peak 7 （chlorogenic acid）， peak 21 （isochlorogenic acid A）， peak 26 （xanthotoxin）， peak 19 （isochlorogenic acid B）， peak 33， peak 13， peak 23 （isochlorogenic acid C）， peak 2 （new chlorogenic acid）， peak 17 （luteolin-7-O-β-D- glucoside） were greater than 1. The above 11 components had good linearity in their respective detection concentration ranges （r was greater than 0.999）. RSDs of precision， repeatability， and stability tests were not more than 2% （n＝6）. The average recovery rates were 92.54%-105.55%， and the RSDs were 0.83%-1.93% （n＝6）. The average contents of 11 components were 0.744， 5.014， 0.646， 0.431， 0.069， 0.582， 0.979， 2.754， 0.157， 1.284 and 2.943 mg/g， respectively. CONCLUSIONS The constructed HPLC fingerprint and content determination methods are simple， accurate and stable， which can provide reference for quality control of G. delavayi. Xanthotoxin， chlorogenic acid， isochlorogenic acid A， luteolin-7-O- β -D-glucoside， isochlorogenic acid C and new chlorogenic acid can be used as markers for G. delavayi. |
| 期刊： | 2025年第36卷第09期 |
| 作者： | 孙丽莎；蒋礼；李丽；⨩开；汪洋；潘洁；李月婷；李勇军 |
| AUTHORS： | SUN Lisha，JIANG Li，LI Li，TIAN Lin，WANG Yang，PAN Jie，LI Yueting，LI Yongjun |
| 关键字： | 钩苞大丁草；高效液相色谱法；指纹图谱；含量测定；化学模式识别 |
| KEYWORDS： | Gerbera delavayi； high-performance liquid chromatography method； fingerprint； content determination； chemical |
| 阅读数： | 7 次 |
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