Farag Mohamed A, Porzel Andrea, Mahrous Engy A, El-Massry Mo'men M, Wessjohann Ludger A
Pharmacognosy Department, College of Pharmacy, Cairo University, Kasr el Aini st., P.B. 11562, Cairo, Egypt,
Anal Bioanal Chem. 2015 Mar;407(7):1937-49. doi: 10.1007/s00216-014-8432-1. Epub 2015 Jan 11.
Senna alexandrina Mill (Cassia acutifolia and Cassia angustifolia) are used for the laxative medicine Senna. Leaves and pods from two geographically different sources were distinguished from each other via proton nuclear magnetic resonance (1H-NMR) and ultra performance liquid chromatography-mass spectrometry (UPLC-MS) analysis. Under optimized conditions, we were able to simultaneously quantify and identify 107 metabolites including 21 anthraquinones, 24 bianthrones (including sennosides), 5 acetophenones, 25 C/O-flavonoid conjugates, 5 xanthones, 3 naphthalenes, 2 further phenolics, and 9 fatty acids. Principal component analysis (PCA) and hierarchical clustering analysis (HCA) were used to define both similarities and differences among samples. For sample classification based on tissue type (leaf and pod), both UPLC-MS and NMR were found to be more effective in separation than on geographical origin. Results reveal that the amounts of the major classes of bioactives in Senna, i.e., flavonoids and sennosides, varied significantly among organs. Leaves contained more flavonoids and ω-3 fatty acids but fewer benzophenone derivatives than pods. In contrast, pods were more enriched in bianthrones (sennosides). PCA analysis was found to be equally effective in predicting the origin of the commercial Senna preparation using NMR and/or UPLC-MS datasets. Furthermore, a selection of six so far uninvestigated Senna species were analyzed by UPLC-MS. Results revealed that the Senna alata leaf in terms of secondary metabolite composition is the most closely related species to S. alexandrina Mill, showing the highest levels of the anthraquinone "rhein" and flavonoid conjugates. To the best of our knowledge, this study provides the first approach utilizing both UPLC-MS and NMR to reveal secondary metabolite compositional differences among Senna species.
亚历山大番泻叶（Cassia acutifolia和Cassia angustifolia）被用于制作泻药番泻叶。通过质子核磁共振（1H-NMR）和超高效液相色谱-质谱联用（UPLC-MS）分析，区分了来自两个地理区域不同来源的叶片和荚果。在优化条件下，我们能够同时定量和鉴定107种代谢物，包括21种蒽醌、24种双蒽酮（包括番泻苷）、5种苯乙酮、25种C/O-黄酮共轭物、5种呫吨酮、3种萘、2种其他酚类和9种脂肪酸。主成分分析（PCA）和层次聚类分析（HCA）用于确定样品之间的异同。对于基于组织类型（叶片和荚果）的样品分类，发现UPLC-MS和NMR在分离方面比基于地理来源更有效。结果表明，番泻叶中主要生物活性成分类别，即黄酮类化合物和番泻苷，在不同器官中的含量差异显著。叶片中黄酮类化合物和ω-3脂肪酸含量较多，但二苯甲酮衍生物比荚果少。相比之下，荚果中双蒽酮（番泻苷）含量更高。发现PCA分析在使用NMR和/或UPLC-MS数据集预测市售番泻叶制剂的来源方面同样有效。此外，通过UPLC-MS对六种尚未研究的番泻叶物种进行了分析。结果表明，就次生代谢物组成而言，翅荚决明叶是与亚历山大番泻叶最密切相关的物种，显示出最高水平的蒽醌“大黄酸”和黄酮共轭物。据我们所知，本研究首次采用UPLC-MS和NMR相结合的方法揭示番泻叶物种之间次生代谢物组成的差异。
