基于UPLC指纹图谱与多成分定量结合化学计量学的壮药茉莉花质量评价研究*

Study on quality evaluation of Zhuang medicinal jasmine based on UPLC fingerprint and multi-component quantitative combined with chemometric method

关键词:  茉莉花  指纹图谱  多成分定量  化学计量法  质量评价

Key words:  Jasmine  Fingerprint  Multiple component quantification  stoichiometry  Quality evaluation

摘要:

目的：建立壮药茉莉花的超高效液相色谱指纹图谱,并测定5种化学成分的含量,结合化学计量学更全面地评价茉莉花的质量。方法：采用Agilent ZORBAX RR StableBond C₁₈色谱柱(4.6×150mm,3.5μm),以乙腈-0.1%磷酸溶液为流动相,梯度洗脱；流速为0.3 mL?min_-1^,柱温30 ℃,检测波长为360 nm,建立茉莉花的UPLC指纹图谱,测定其中芦丁、金丝桃苷、槲皮素、山奈素和齐墩果酸共5个成分的含量,基于指纹图谱共有峰峰面积,采用聚类分析、主成分分析、正交偏最小二乘法-判别分析和统计学分析评价茉莉花的整体质量。结果：44批不同产地及批次茉莉花指纹图谱相似度均大于0.982,共标定22个共有峰,指认芦丁、金丝桃苷、槲皮素、山奈素与齐墩果酸5个共有峰,5个成分在各自检测浓度范围线性关系良好(R₂ ﹥0.9976),芦丁、金丝桃苷、槲皮素、山奈素与齐墩果酸的含量分别为0.430～1.319 mg ? g_-1^、0.156～0.572 mg ? g_-1^、 0.017～0.079mg ? g_-1^、0.002～0.005 mg ? g_-1^与1.083～2.326 mg ? g_-1^；聚类分析将44批茉莉花聚为3类,主成分分析共提取4个主成分,累计贡献率77.534%；主成分分析综合得分范围为－4.32～3.20,不同产地及批次茉莉花的质量存在较大差异；正交偏最小二乘法-判别分析(VIP﹥1)表明峰5、峰7(芦丁,VIP =1.236)、峰9(金丝桃苷,VIP =1.234)、峰6、峰12、峰10、峰4、峰15(槲皮素,VIP =1.180)、峰1、峰11、峰8、峰2、峰14和峰3共14个化合物可能是引起茉莉花质量差异的成分；Pearson(双侧)相关性分析显示芦丁、金丝桃苷、槲皮素、山奈素、齐墩果酸及5个成分总含量对指纹图谱综合得分的影响关系密切,可作为茉莉花质量的评价指标；以不用产地样品分组并进行方差分析,各组中黄酮类成分山奈素含量无显著性差异,芦丁、金丝桃苷和槲皮素3种黄酮类成分含量以广西组整体偏低,萜类成分齐墩果酸含量略有差异,以云南组的含量最高。结论：本文所建立的方法高效、简便、稳定,能较全面评价壮药茉莉花的整体质量,可为提升其质量标准提供科学依据。

Abstract:

Objective: To establish the ultra-high performance liquid chromatography (UPLC) fingerprint of Zhuang medicine jasmine, and determine the content of 5 chemical components, combined with stoichiometry to evaluate the quality of jasmine more comprehensively. Methods: The chromatography was performed on Agilent ZORBAX RR StableBond C18 column (4.6×150mm, 3.5μm) with acetonitrile- 0.1% phosphoric acid solution as mobile phase and gradient elution. The flow rate was 0.3mL?min_-1^{, the column temperature was 30} ℃, and the detection wavelength was 360 nm. The UPLC fingerprint of Jasmine was established, and the contents of rutin, hypericin, quercetin, saenetin and oleanolic acid were determined. Based on the fingerprint, the total peak-peak area was obtained. The whole quality of jasmine was evaluated by cluster analysis, principal component analysis, orthogonal partial least squares discriminant analysis and statistical analysis. Results: The similarity of fingerprints of 44 batches of jasmine from different origin and batches was greater than 0.982. A total of 22 common peaks were calibrated, which 5 common peaks of rutin, hypericin, quercetin, kaenetin and oleanolic acid were identified, the 5 components had a good linear relationship in their respective detection concentration ranges (R₂ ^> 0.9976). The contents of rutin, hypericin, quercetin and oleanic acid were respectively 0.430 ~ 1.319 mg ? g_-1^{, 0.156 ~ 0.572 mg ? g}_-1^{, 0.017 ~ 0.079mg ? g}_-1 ^{and 0.002 ~ 0.005 mg ? g}_-1 ^{and 1.083 ~ 2.326 mg ? g_-1}; 44 batches of jasmine flowers were grouped into 3 categories by cluster analysis, and 4 principal components were extracted by principal component analysis, with a cumulative contribution rate of 77.534%. The comprehensive scores of principal component analysis ranged from -4.32 to 3.20, which were great differences in the quality of jasmine from different origin and batches. OPLS-DA (VIP >1) showed that peak 5, peak 7 (rutin, VIP =1.236), peak 9 (hyperoside, VIP =1.234), peak 6, peak 12, peak 10, peak 4, peak 15 (quercetin; VIP =1.180), peak 1, peak 11, peak 8, peak 2, peak 14 and peak 3, a total of 14 compounds may be the components causing the quality difference of jasmine; Pearson (bilateral) correlation analysis showed that rutin, hypericin, quercetin, kaenetin, oleanolic acid and the total contents of the five components had a close relationship with the comprehensive score of fingerprint, and could be used as an evaluation index of jasmine quality. The contents of rutin, hyperoside and quercetin in Guangxi group were lower than those in other regions. The contents of oleanolic acid in Yunnan group were the highest. Conclusion: The established method is efficient, simple and stable, which can comprehensively evaluate the overall quality of Zhuang medicinal jasmine, and provide a scientific basis for improving its quality standards.