Download PDF
Planta Med 2012; 78(5): 488-496
DOI: 10.1055/s-0031-1298170
Analytical Studies
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

Metabolome Classification of Commercial Hypericum perforatum (St. John's Wort) Preparations via UPLC-qTOF-MS and Chemometrics

Mohamed A. Farag1 , 2 , Ludger A. Wessjohann1
  • 1Department Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Halle (Saale), Germany
  • 2Pharmacognosy Department, College of Pharmacy, Cairo University, Cairo, Egypt
Further Information

Publication History

received October 8, 2011 revised Nov. 20, 2011

accepted Dec. 18, 2011

Publication Date:
23 January 2012 (online)

    Permissions and Reprints

Abstract

The growing interest in the efficacy of phytomedicines and herbal supplements but also the increase in legal requirements for safety and reliable contents of active principles drive the development of analytical methods for the quality control of complex, multicomponent mixtures as found in plant extracts of value for the pharmaceutical industry. Here, we describe an ultra-performance liquid chromatography method (UPLC) coupled with quadrupole time of flight mass spectrometry (qTOF-MS) measurements for the large scale analysis of H. perforatum plant material and its commercial preparations. Under optimized conditions, we were able to simultaneously quantify and identify 21 metabolites including 4 hyperforins, 3 catechins, 3 naphthodianthrones, 5 flavonoids, 3 fatty acids, and a phenolic acid. Principal component analysis (PCA) was used to ensure good analytical rigorousness and define both similarities and differences among Hypericum samples. A selection of batches from 9 commercially available H. perforatum products available on the German and Egyptian markets showed variable quality, particularly in hyperforins and fatty acid content. PCA analysis was able to discriminate between various preparations according to their global composition, including differentiation between various batches from the same supplier. To the best of our knowledge, this study provides the first approach utilizing UPLC-MS-based metabolic fingerprinting to reveal secondary metabolite compositional differences in Hypericum extract.

Key words

Hypericum perforatum L. - Hypericaceae - UPLC-MS - hyperforin - principal component analysis - quality control - St. John's wort

Supporting Information

References

  • 1 Kim H K, Choi Y H, Verpoorte R. NMR-based metabolomic analysis of plants.  Nat Protoc. 2010;  5 536-549
    Google Scholar
  • 2 Bedair M, Sumner L W. Current and emerging mass-spectrometry technologies for metabolomics.  Trac-Trend Anal Chem. 2008;  27 238-250
    Google Scholar
  • 3 Sumner L W, Mendes P, Dixon R A. Plant metabolomics: large-scale phytochemistry in the functional genomics era.  Phytochemistry. 2003;  62 817-836
    Google Scholar
  • 4 Wolfender J L, Glauser G, Grata E, Rudaz S. UPLC-TOF-MS for high resolution plant metabolite profiling and metabolomics.  Planta Med. 2008;  74 919-920
    Google Scholar
  • 5 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
    Google Scholar
  • 6 Hanhineva K, Kokko H, Siljanen H, Rogachev I, Aharoni A, Karenlampi S O. Stilbene synthase gene transfer caused alterations in the phenylpropanoid metabolism of transgenic strawberry (Fragaria xananassa).  J Exp Bot. 2009;  60 2093-2106
    Google Scholar
  • 7 Grata E, Guillarme D, Glauser G, Boccard J, Carrupt P A, Veuthey J L, Rudaz S, Wolfender J L. Metabolite profiling of plant extracts by ultra-high-pressure liquid chromatography at elevated temperature coupled to time-of-flight mass spectrometry.  J Chromatogr A. 2009;  1216 5660-5668
    Google Scholar
  • 8 Okada T, Nakamura Y, Kanaya S, Takano A, Malla K J, Nakane T, Kitayama M, Sekita S. Metabolome analysis of ephedra plants with different contents of ephedrine alkaloids by using UPLC-Q-TOF-MS.  Planta Med. 2009;  75 1356-1362
    Google Scholar
  • 9 Cao G, Zhang Y, Feng J A, Cai H, Zhang C R, Ding M J, Cong X D, Cai B C. A rapid and sensitive assay for determining the main components in processed fructus corni by UPLC-Q-TOF-MS.  Chromatographia. 2011;  73 135-141
    Google Scholar
  • 10 Dan M, Su M M, Gao X F, Zhao T, Zhao A H, Xie G X, Qiu Y P, Zhou M M, Liu Z, Jia W. Metabolite profiling of Panax notoginseng using UPLC-ESI-MS.  Phytochemistry. 2008;  69 2237-2244
    Google Scholar
  • 11 Xie G X, Ni Y, Su M M, Zhang Y Y, Zhao A H, Gao X F, Liu Z, Xiao P G, Jia W. Application of ultra-performance LC-TOF MS metabolite profiling techniques to the analysis of medicinal Panax herbs.  Metabolomics. 2008;  4 248-260
    Google Scholar
  • 12 Pongsuwan W, Bamba T, Harada K, Yonetani T, Kobayashi A, Fukusaki E. High-throughput technique for comprehensive analysis of Japanese green tea quality assessment using ultra-performance liquid chromatography with time-of-flight mass spectrometry (UPLC/TOF MS).  J Agric Food Chem. 2008;  56 10705-10708
    Google Scholar
  • 13 Barnes J, Anderson L A, Phillipson J D. St John's wort (Hypericum perforatum L.): a review of its chemistry, pharmacology and clinical properties.  J Pharm Pharmacol. 2001;  53 583-600
    Google Scholar
  • 14 Giese J. Tests for nutraceutical products.  Food Tech. 1999;  53 93
    Google Scholar
  • 15 Gobbi M, Mennini T. Is St John's wort a 'Prozac-like' herbal antidepressant?.  Trends Pharmacol Sci. 2001;  22 557-559
    Google Scholar
  • 16 Sarris J. Herbal medicines in the treatment of psychiatric disorders: a systematic review.  Phytother Res. 2007;  21 703-716
    Google Scholar
  • 17 Meruelo D, Lavie G, Lavie D. Therapeutic agents with dramatic antiretroviral activity and little toxicity at effective doses – aromatic polycyclic diones hypericin and pseudohypericin.  Proc Natl Acad Sci USA. 1988;  85 5230-5234
    Google Scholar
  • 18 Laakmann G, Dienel A, Kieser M. Clinical significance of hyperforin for the efficacy of Hypericum extracts on depressive disorders of different severities.  Phytomedicine. 1998;  5 435-442
    Google Scholar
  • 19 Laakmann G, Schule C, Baghai T, Kieser M. St. John's Wort in mild to moderate depression: the relevance of hyperforin for the clinical efficacy.  Pharmacopsychiatry. 1998;  31 54-59
    Google Scholar
  • 20 Medina M A, Martinez-Poveda B, Amores-Sanchez M I, Quesada A R. Hyperforin: more than an antidepressant bioactive compound?.  Life Sci. 2006;  79 105-111
    Google Scholar
  • 21 Butterweck V, Wall A, Lieflander-Wulf U, Winterhoff H, Nahrstedt A. Effects of the total extract and fractions of Hypericum perforatum in animal assays for antidepressant activity.  Pharmacopsychiatry. 1997;  30 117-124
    Google Scholar
  • 22 Kosuth J, Koperdakova J, Tolonen A, Hohtola A, Cellarova E. The content of hypericins and phloroglucinols in Hypericum perforatum L. seedlings at early stage of development.  Plant Sci. 2003;  165 515-521
    Google Scholar
  • 23 Kusari S, Lamshoft M, Zuhlke S, Spiteller M. An endophytic fungus from Hypericum perforatum that produces hypericin.  J Nat Prod. 2008;  71 159-162
    Google Scholar
  • 24 Tolonen A, Hohtola A, Jalonen J. Fast high-performance liquid chromatographic analysis of naphthodianthrones and phloroglucinols from Hypericum perforatum extracts.  Phytochem Anal. 2003;  14 306-309
    Google Scholar
  • 25 Lawaetz A, Schmidt B, Staerk D, Jaroszewski J, Bro R. Application of rotated PCA models to facilitate interpretation of metabolite profiles: commercial preparations of St. John's wort.  Planta Med. 2009;  75 271-279
    Google Scholar
  • 26 Rasmussen B, Cloarec O, Tang H R, Staerk D, Jaroszewski J W. Multivariate analysis of integrated and full-resolution H-1-NMR spectral data from complex pharmaceutical preparations: St. John's wort.  Planta Med. 2006;  72 556-563
    Google Scholar
  • 27 Schmidt B, Jaroszewski J W, Bro R, Witt M, Staerk D. Combining PARAFAC analysis of HPLC-PDA profiles and structural characterization using HPLC-PDA-SPE-NMR-MS experiments: commercial preparations of St. John's wort.  Anal Chem. 2008;  80 1978-1987
    Google Scholar
  • 28 Politi M, Zloh M, Pintado M E, Castro P M L, Heinrich M, Prieto J M. Direct metabolic fingerprinting of commercial herbal tinctures by nuclear magnetic resonance spectroscopy and mass spectrometry.  Phytochem Anal. 2009;  20 328-334
    Google Scholar
  • 29 Huck C, Abel G, Popp M, Bonn G. Comparative analysis of naphthodianthrone and phloroglucine derivatives in St. John's Wort extracts by near infrared spectroscopy, high-performance liquid chromatography and capillary electrophoresis.  Anal Chim Acta. 2006;  580 223-230
    Google Scholar
  • 30 Maggi F, Ferretti G, Pocceschi N, Menghini L, Ricciutelli M. Morphological, histochemical and phytochemical investigation on the genus Hypericum of the Central Italy.  Fitoterapia. 2004;  75 702-711
    Google Scholar
  • 31 Smith C A, Want E J, O'Maille G, Abagyan R, Siuzdak G. XCMS: processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification.  Anal Chem. 2006;  78 779-787
    Google Scholar
  • 32 Dunn W B, Broadhurst D, Brown M, Baker P N, Redman C W G, Kenny L C, Kell D B. Metabolic profiling of serum using ultra performance liquid chromatography and the LTQ-Orbitrap mass spectrometry system.  J Chromatogr B. 2008;  871 288-298
    Google Scholar
  • 33 van den Berg R A, Hoefsloot H C J, Westerhuis J A, Smilde A K, van der Werf M J. Centering, scaling, and transformations: improving the biological information content of metabolomics data.  BMC Genomics. 2006;  7 142
    Google Scholar
  • 34 Liu F, Pan C, Drumm P, Ang C Y W. Liquid chromatography-mass spectrometry studies of St. John's wort methanol extraction: active constituents and their transformation.  J Pharm Biomed Anal. 2005;  37 303-312
    Google Scholar
  • 35 Tatsis E C, Boeren S, Exarchou V, Troganis A N, Vervoort J, Gerothanassis I P. Identification of the major constituents of Hypericum perforatum by LC/SPE/NMR and/or LC/MS.  Phytochemistry. 2007;  68 383-393
    Google Scholar
  • 36 Lee J, Duke R K, Tran V H, Hook J M, Duke C C. Hyperforin and its analogues inhibit CYP3A4 enzyme activity.  Phytochemistry. 2006;  67 2550-2560
    Google Scholar
  • 37 Goodacre R, Shann B, Gilbert R J, Timmins E M, McGovern A C, Alsberg B K, Kell D B, Logan N A. Detection of the dipicolinic acid biomarker in Bacillus spores using Curie-point pyrolysis mass spectrometry and fourier transform infrared spectroscopy.  Anal Chem. 2000;  72 119-127
    Google Scholar
  • 38 van der Kooy F, Maltese F, Choi Y H, Kim H K, Verpoorte R. Quality control of herbal material and phytopharmaceuticals with MS and NMR based metabolic fingerprinting.  Planta Med. 2009;  75 763-775
    Google Scholar

Prof. Dr. Mohamed A. Farag

Pharmacognosy Department
College of Pharmacy
Cairo University

Kasr el Aini st.

Cairo 11562

Egypt

Phone: +011 20 22 36 22 45

Fax: +011 2 02 25 32 00 05

Email: mfarag73@yahoo.com

>