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DOI: 10.1055/s-0029-1185577
© Georg Thieme Verlag KG Stuttgart · New York
Metabolome Analysis of Ephedra Plants with Different Contents of Ephedrine Alkaloids by Using UPLC‐Q‐TOF‐MS
Publikationsverlauf
received October 15, 2008
revised February 24, 2009
accepted March 9, 2009
Publikationsdatum:
20. April 2009 (online)
Abstract
Metabolome analysis of four varieties of Ephedra plants, which contain different amounts of ephedrine alkaloids, was demonstrated in this study. The metabolites were comprehensively analyzed by using ultra performance liquid chromatography (UPLC) coupled with quadrupole time-of-flight mass spectrometry (Q‐TOF‐MS) and the ephedrine alkaloids were also profiled. Subsequently, multivariate analyses of principal component analysis (PCA) and batch-learning self-organizing mapping (BL‐SOM) analysis were applied to the raw data of the total ion chromatogram (TIC). PCA was performed to visualize the fingerprints characteristic for each Ephedra variant and the independent metabolome clusters were formed. The metabolite fingerprints were also visualized by BL‐SOM analysis and were displayed as a lattice of colored individual cells which was characteristic for each Ephedra variant. BL‐SOM analysis was also used for identification of chemical marker peaks because the information assigned to a cell represented either increases or decreases in peak intensities. Using this analysis, ephedrine alkaloids were successfully selected from the TICs as chemical markers for each Ephedra variant and this result suggested that BL‐SOM analysis was an effective method for the selection of marker metabolites. We report our study here as a practical case of metabolomic study on medicinal resources.
Key words
Ephedra - Ephedraceae - metabolomics - UPLC‐Q‐TOF‐MS - multivariate analysis - ephedrine alkaloids
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- Supporting Information .
References
- 1 Pichersky E, Gang D R. Genetics and biochemistry of secondary metabolites in plants: an evolutionary perspective. Trends Plant Sci. 2000; 5 439-445
- 2 Abourashed E A, El-Alfy A T, Khan I A, Walker L. Ephedra in perspective – a current review. Phytother Res. 2003; 17 703-712
- 3 Caveney S, Charlet D A, Freitag H, Maier-Stolte M, Starratt A N. New observations on the secondary chemistry of world Ephedra (Ephedraceae). Am J Bot. 2001; 88 1199-1208
- 4 Leung A Y, Foster S. Encyclopedia of common natural ingredients, 2nd edition. New York; John Wiley & Sons 1996
- 5 Pullela S V, Takamatsu S, Khan S, Khan I A. Isolation of lignans and biological activity studies of Ephedra viridis. Planta Med. 2005; 71 789-791
- 6 Kasahara Y, Hikino H, Tsurufuji S, Watanabe M, Ohuchi K. Anti-inflammatory actions of ephedrines in acute inflammations. Planta Med. 1985; 51 325-331
- 7 Konno C, Mizuno T, Hikino H. Isolation and hypoglycemic activity of ephedrans A, B, C, D and E, glycans of Ephedra distachya herbs. Planta Med. 1985; 51 162-163
- 8 Bino R J, Hall R D, Fiehn O, Kopka J, Saito K, Draper J, Nikolau B J, Mendes P, Roessner-Tunali U, Beale M H, Trethewey R N, Lange B M, Wurtele E S, Sumner L W. Potential of metabolomics as a functional genomics tool. Trends Plant Sci. 2004; 9 418-425
- 9 Rhee S Y, Dickerson J, Xu D. Bioinformatics and its applications in plant biology. Annu Rev Plant Biol. 2006; 57 335-360
- 10 Schauer N, Fernie A R. Plant metabolomics: towards biological function and mechanism. Trends Plant Sci. 2006; 11 508-516
- 11 Kim H K, Choi Y H, Erkelens C, Lefeber A W, Verpoorte R. Metabolic fingerprinting of Ephedra species using 1H‐NMR spectroscopy and principal component analysis. Chem Pharm Bull. 2005; 53 105-109
- 12 Kanaya S, Kinouchi M, Abe T, Kudo Y, Yamada Y, Nishi T, Mori H, Ikemura T. Analysis of codon usage diversity of bacterial genes with a self-organizing map (SOM): characterization of horizontally transferred genes with emphasis on the E. coli O157 genome. Gene. 2001; 276 89-99
- 13 Xie G, Plumb R, Su M, Xu Z, Zhao A, Qiu M, Long X, Liu Z, Jia W. Ultra-performance LC/TOF MS analysis of medicinal Panax herbs for metabolomic research. J Sep Sci. 2008; 31 1015-1026
- 14 Grata E, Boccard J, Guillarme D, Glauser G, Carrupt P A, Farmer E E, Wolfender J L, Rudaz S. UPLC‐TOF‐MS for plant metabolomics: a sequential approach for wound marker analysis in Arabidopsis thaliana. J Chromatogr B. 2008; 871 261-270
- 15 Malla K J, Mitsuhashi A, Miyajima T, Watanabe T, Kitajima J, Arai Y, Takano A. Studies on medicinal plant resources of the Himalayas (5), variation in alkaloid content in cultivated Nepalese Ephedra (1). Natural Medicines. 2005; 59 237-240
- 16 Malla K J, Sugi N, Watanabe T, Kondo S, Kitajima J, Arai Y, Takano A. Studies on medicinal plant resources of the Himalayas (6), – cultivation of Nepalese Ephedra collected at upper Mustang. Natural Medicines. 2005; 59 245-248
- 17 Mikage M, Kondo N. Micro-botanical studies of Ephedra plants from the Himalayan region. Part I. Anatomical studies on the herbal stems and botanical origin of Tibetan crude drugs “Tshe” and “Balu”. J Jpn Bot. 1996; 71 323-332
- 18 Sagara K, Oshima T, Misaki T. A simultaneous determination of norephedrine, pseudoephedrine, ephedrine and methylephedrine in Ephedrae Herba and oriental pharmaceutical preparations by ion-pair high-performance liquid chromatography. Chem Pharm Bull. 1983; 31 2359-2365
- 19 Gay M L, White K D, Obermeyer W R, Betz J M, Musser S M. Determination of ephedrine-type alkaloids in dietary supplements by LC/MS using a stable-isotope labeled internal standard. J AOAC Int. 2001; 84 761-769
- 20 Hirai M Y, Klein M, Fujikawa Y, Yano M, Goodenowe D B, Yamazaki Y, Kanaya S, Nakamura Y, Kitayama M, Suzuki H, Sakurai N, Shibata D, Tokuhisa J, Reichelt M, Gershenzon J, Papenbrock J, Saito K. Elucidation of gene-to-gene and metabolite-to-gene networks in Arabidopsis by integration of metabolomics and transcriptomics. J Biol Chem. 2005; 280 25590-25595
- 21 Hirai M Y, Yano M, Goodenowe D B, Kanaya S, Kimura T, Awazuhara M, Arita M, Fujiwara T, Saito K. Integration of transcriptomics and metabolomics for understanding of global responses to nutritional stresses in Arabidopsis thaliana. Proc Natl Acad Sci USA. 2004; 101 10205-10210
- 22 Oikawa A, Nakamura Y, Ogura T, Kimura A, Suzuki H, Sakurai N, Shinbo Y, Shibata D, Kanaya S, Ohta D. Clarification of pathway-specific inhibition by fourier transform ion cyclotron resonance/mass spectrometry-based metabolic phenotyping studies. Plant Physiol. 2006; 142 398-413
- 23 Sander L C, Sharpless K E, Satterfield M B, Ihara T, Phinney K W, Yen J H, Wise S A, Gay M L, Lam J W, McCooeye M, Gardner G, Fraser C, Sturgeon R, Roman M. Determination of ephedrine alkaloids in dietary supplement standard reference materials. Anal Chem. 2005; 77 3101-3112
- 24 Nakamura Y, Kimura A, Saga H, Oikawa A, Shinbo Y, Kai K, Sakurai N, Suzuki H, Kitayama M, Shibata D, Kanaya S, Ohta D. Differential metabolomics unraveling light/dark regulation of metabolic activities in Arabidopsis cell culture. Planta. 2007; 227 57-66
- 25 Moco S, Bino R J, Vorst O, Verhoeven H A, de Groot J, van Beek T A, Vervoort J, de Vos C H. A liquid chromatography-mass spectrometry-based metabolome database for tomato. Plant Physiol. 2006; 141 1205-1218
- 26 Kim J K, Bamba T, Harada K, Fukusaki E, Kobayashi A. Time-course metabolic profiling in Arabidopsis thaliana cell cultures after salt stress treatment. J Exp Bot. 2007; 58 415-424
Dr. Taketo Okada
Faculty of Pharmaceutical Sciences at Kagawa Campus,
Tokushima Bunri University
Shido 1314–1
Sanuki-city
Kagawa 769-2193
Japan
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