RESEARCH PAPER Integrated comparative metabolite profiling via MS and NMR techniques for Senna drug quality control analysis Mohamed A. Farag & Andrea Porzel & Engy A. Mahrous & Mo’men M. El-Massry & Ludger A. Wessjohann Received: 24 September 2014 /Revised: 12 December 2014 /Accepted: 18 December 2014 # Springer-Verlag Berlin Heidelberg 2015 Abstract 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 chromato- graphy-mass spectrometry (UPLC-MS) analysis. Under opti- mized 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 anthra- quinone “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 composi- tional differences among Senna species. Keywords Senna alexandrina . S. bicapsularis . S. corymbosa . S.didymobotrya . S. alata . S. sophera . NMR . UPLC-MS . Sennosides . Principal component analysis . Laxative Abbreviations SLE Senna alexandrina leaf Egyptian SLI Senna alexandrina leaf Indian SPE Senna alexandrina pod Egyptian SPI Senna alexandrina pod Indian Introduction Plant-based medicines become increasingly popular all over the world. The authentication of herbal raw materials is im- portant to ensure their safety and efficacy. Some herbal drugs belonging to closely related species that differ in medicinal properties are difficult to identify owing to their close Electronic supplementary material The online version of this article (doi:10.1007/s00216-014-8432-1) contains supplementary material, which is available to authorized users. M. A. Farag (*) : E. A. Mahrous Pharmacognosy Department, College of Pharmacy, Cairo University, Kasr el Aini st., P.B. 11562, Cairo, Egypt e-mail: mfarag73@yahoo.com M. A. Farag e-mail: Mohamed.farag@pharma.cu.edu.eg A. Porzel : L. A. Wessjohann (*) Department of Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany e-mail: ludger.wessjohann@ipb-halle.de M. M. El-Massry National Organization for Drug Control and Research (NODCAR), Dokki, P.B. 12553, Cairo, Egypt Anal Bioanal Chem DOI 10.1007/s00216-014-8432-1