Application of ultra-performance LC-TOF MS metabolite profiling techniques to the analysis of medicinal Panax herbs
The morphological appearance and some ingredients of Panax ginseng, Panax notoginseng and Panax japonicus of the Panax genus are similar. However, their pharmacological activities are obviously different due to the significant differences in the types and quantity of saponins in each herb. In the present study, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOFMS) was used to profile the abundances of metabolites in the three medicinal Panax herbs. Multivariate statistical analysis technique, that is, principle component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were used to discriminate between the Panax samples. PCA of the analytical data showed a clear separation of compositions among the three medicinal herbs. The critical markers such as chikusetsusaponin IVa, ginsenoside R0, ginsenoside Rc, ginsenoside Rb1, ginsenoside Rb2 and ginsenoside Rg2 accountable for such variations were identified through the corresponding loading weights, and the tentative identification of biomarkers is completed by the accurate mass of TOFMS and high resolution and high retention time reproducibility performed by UPLC. The proposed analytical method coupled with multivariate statistical analysis is reliable to analyze a group of metabolites present in the herbal extracts and other natural products. This method can be further utilized to evaluate chemical components obtained from different plants and/or the plants of different geographical locations, thereby classifying the medicinal plant resources and potentially elucidating the mechanism of inherent phytochemical diversity.
KeywordsPanax Metabolite profiling UPLC-QTOFMS Saponin Multivariate statistical analysis
This study was financially supported by the International Collaboration Project (project number 2006DFA02700) of the Ministry Science and Technology of the People’s Republic of China. We also thank Yongbo Wang for assisting with sample collection.
- Akerele, O. (1992). WHO guidelines for the assessment of herbal medicines. Fitoterapia, 63, 99–104.Google Scholar
- Chan, E. C. Y., Yap, S. L., Lau, A. J., Leow, P. C., Toh, D. F., & Koh, H. L. (2007). Ultra-performance liquid chromatography/time-of-flight mass spectrometry based metabolomics of raw and steamed Panax notoginseng. Rapid Communications in Mass Spectrometry, 21, 519–528. doi: 10.1002/rcm.2864.PubMedCrossRefGoogle Scholar
- Cloarec, O., Dumas, M. E., Trygg, J., Craig, A., Barton, R. H., Lindon, J. C., et al. (2005). Evaluation of the orthogonal projection on latent structure model limitations caused by chemical shift variability and improved visualization of biomarker changes in 1H NMR spectroscopic metabonomic studies. Analytical Chemistry, 77, 517–526. doi: 10.1021/ac048803i.PubMedCrossRefGoogle Scholar
- Farag, M. A., Huhman, D. V., Lei, Z., & Sumner, L. W. (2007). Metabolic profiling and systematic identification of flavonoids and isoflavonoids in roots and cell suspension cultures of Medicago truncatula using HPLC-UV-ESI-MS and GC-MS. Phytochemistry, 68, 342–354. doi: 10.1016/j.phytochem.2006.10.023.PubMedCrossRefGoogle Scholar
- Guan, J., Lai, C. M., & Li, S. P. (2007). A rapid method for the simultaneous determination of 11 saponins in Panax notoginseng using ultra performance liquid chromatography. Journal of Pharmaceutical and Biomedical Analysis, 44, 996–1000. doi: 10.1016/j.jpba.2007.03.032.PubMedCrossRefGoogle Scholar
- Guo, Q., Yu, X. L., & Hu, X. X. (2004). Progress of chemical and pharmacological research of Panax japonicus. Research & Information on Traditional Chinese Medicine, 6, 17–19.Google Scholar
- Jiangsu New Medical College. (1985). Dictionary of Chinese Materia Medica. Shanghai Science and Technology Press, Shanghai, pp. 417–420.Google Scholar
- Long, M., Millar, D. J., Kimura, Y., Donovan, G., Rees, J., Fraser, P. D., et al. (2006). Metabolite profiling of carotenoid and phenolic pathways in mutant and transgenic lines of tomato: Identification of a high antioxidant fruit line. Phytochemistry, 67, 1750–1757. doi: 10.1016/j.phytochem.2006.02.022.PubMedCrossRefGoogle Scholar
- Mahady, G. B., Fong, H. H. S., & Farnsworth, N. R. (2001). Botanical Dietary Supplements: Quality, Safety, and Efficacy. Lisse, The Netherlands: Swets & Zeitlinger Publishers, B.V.Google Scholar
- National Committee of Pharmacopoeia. (2005). Pharmacopoeia of the People’ s Republic of China (Vol. I, pp. 10). Beijing: Chemical Industry Press.Google Scholar
- Shen, Y. F., Zhang, R., Moore, R. J., Kim, J., Metz, T. O., Hixson, K. K., et al. (2005). Automated 20 kpsi RPLC-MS and MS/MS with chromatographic peak capacities of 1,000–1,500 and capabilities in proteomics and metabolomics. Analytical Chemistry, 77, 3090–3100. doi: 10.1021/ac0483062.PubMedCrossRefGoogle Scholar
- Wagner, S., Scholz, K., Donegan, M., Burton, L., Wingate, J., & Völkel, W. (2006). Metabonomics and biomarker discovery: LC-MS metabolic profiling and constant neutral loss scanning combined with multivariate data analysis for mercapturic acid analysis. Analytical Chemistry, 78, 1296–1305. doi: 10.1021/ac051705s.PubMedCrossRefGoogle Scholar
- Wang, M., Lamers, R. J. A. N., Korthout, H. A. A. J., Van Nesselrooij, J. H. J., Witkamp, R. F., Van Der Heijden, R., et al. (2005). Metabolomics in the context of systems biology: Bridging Traditional Chinese Medicine and molecular pharmacology. Phytotherapy Research, 19, 173–182. doi: 10.1002/ptr.1624.PubMedCrossRefGoogle Scholar
- Wei, J. X., Chen, Y. G., & Du, Y. C. (1991). Introduction of separation method of dammarane-type saponins in Panax samples. Academic Journal of Kunming Medical College, 12, 56–64.Google Scholar
- Wilson, I. D., Nicholson, J. K., Castro-Perez, J., Granger, J. H., Johnson, K. A., Smith, B. W., et al. (2005). High resolution “ultra performance” liquid chromatography coupled to oa-TOF mass spectrometry as a tool for differential metabolic pathway profiling in functional genomic studies. Journal of Proteome Research, 4, 591–598. doi: 10.1021/pr049769r.PubMedCrossRefGoogle Scholar
- Xie, G. X., Plumb, R., Su, M. M., Xu, Z. H., Zhao, A. H., Qiu, M. F., et al. (2008). Ultra-performance LC/TOF MS analysis of medicinal Panax herbs for metabolomic research. Journal of Separation Science, 1015–1026. doi:10.1002/jssc.200700650.
- Zuo, R., Yuan, D. (2005). Research on chemical constituents and biological activities of Rhizoma of Panax japonicus. Lishizhen Medicine and Materia Medica Research, 16, 838–841.Google Scholar