[关键词]
[摘要]
目的 建立UPLC多指标成分定量测定与指纹图谱结合化学模式识别法对不同基原十大功劳木Mahoniae Caulis药材进行质量评价,为十大功劳木质评质控体系的构建及其基原鉴别提供参考和依据。方法 采用UPLC法,色谱柱为Agilent Zorbax SB-Aq(150 mm×2.1 mm,1.8μm),流动相为乙腈-0.1%磷酸水溶液,梯度洗脱;体积流量0.3 mL/min,指纹图谱和含量测定检测波长分别为220 nm和345 nm,柱温15 ℃,进样量为2 μL。通过建立不同基原十大功劳木指纹图谱,同时测定十大功劳木中的4个指标成分,结合聚类分析(hierarchical cluster analysis,HCA)、主成分分析(principal component analysis,PCA)、正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)和方差分析对不同基原十大功劳木作差异性鉴别研究。结果 建立了4种基原十大功劳木药材UPLC指纹图谱,同一基原的十大功劳木相似度均大于0.968,4种基原十大功劳木相似度为0.864~0.964。其中宽苞十大功劳木M. eurybracteata、小果十大功劳木M. bodinieri、安平十大功劳木M. ganpinensie和长柱十大功劳木M. duclouxiana分别确定了17、15、14、9个共有峰,并指认了4种基原十大功劳木共有峰中的4个成分,分别是非洲防己碱、盐酸药根碱、盐酸巴马汀和盐酸小檗碱;运用HCA、PCA和OPLS-DA能将4种基原明显区分,OPLS-DA筛选得到7个差异性标志物。定量测定结果得出不同基原的4指标成分差异明显,安平十大功劳木基原质量较好。结论 建立的指纹图谱和定量测定方法合理、可靠,能准确有效地鉴别十大功劳木不同基原药材,为十大功劳木药材的质量控制和基原鉴别提供科学依据和参考。
[Key word]
[Abstract]
Objective To evaluate the quality of Mahoniae Caulis from different species by UPLC multi-component determination and fingerprint combine with chemical pattern recognition method, and provide a reference and basis for the construction of the quality assessment of Shidagonglaomu (Mahoniae Caulis) and the identification of species. Methods The analysis was performed on an UPLC with Agilent Zorbax SB-Aq column (150 mm×2.1 mm, 1.8 μm), the mobile phase consisted of acetonitrile-0.1% phosphoric acid solution for gradient elution, the volume flow rate was 0.3 mL/min, the detection wavelength of fingerprint and content determination were 220 nm and 345 nm, column temperature was 15 ℃, and injection volume of 2 μL. The fingerprint of Mahoniae Caulis from different species were established by UPLC method, and the content of four active ingredients in Mahoniae Caulis were simultaneously determined. The differences of different Mahoniae Caulis species were studied by hierarchical cluster analysis (HCA), principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA) and analysis of variance.Results The UPLC fingerprints of Mahoniae Caulis from four species were established. The similarity of Mahoniae Caulis from the same species was greater than 0.968,and similarity of four Mahoniae Caulis species ranged from 0.864 to 0.964. A total of 17, 15, 14 and 9 common peaks were identified for Mahonia eurybracteata, M. bodinieri, M. ganpinensie, M. duclouxiana, respectively. And four of them were identified by comparing with reference substance, which were columbamine, jatrorrhizine hydrochloride, palmatine hydrochloride, and berberine hydrochloride. The results of HCA, PCA and OPLS-DA could accurately distinguish four species of Mahoniae Caulis. There were obvious differences in chemical components among the different species and seven inter-species significant differential components were found by OPLS-DA. The content determination results showed that there were obvious differences in the four index components of Mahoniae Caulis from different species, and M. ganpinensie had better quality. Conclusion The established fingerprint and multi-component content determination are reasonable and reliable, which can significantly distinguish different Mahoniae Caulis medicinal plants, and provide scientific basis and reference for the species identification and quality evaluation of Mahoniae Caulis medicinal materials from different species.
[中图分类号]
R286.2
[基金项目]
中药配方颗粒关键技术研究与应用合同编号(黔科合支撑[2022]186)