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Planta Med 2011; 77(5): 450-454
DOI: 10.1055/s-0030-1250459
Pharmacokinetic Investigations
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

Determination of Myrtucommulone from Myrtus communis in Human and Rat Plasma by Liquid Chromatography/Tandem Mass Spectrometry

Kathleen Gerbeth1 , Jürgen Meins1 , Oliver Werz2 , Manfred Schubert-Zsilavecz1 , 3 , Mona Abdel-Tawab1
  • 1Department for Research & Development, Central Laboratory of German Pharmacists, Eschborn, Germany
  • 2Department of Pharmaceutical Analytics, Pharmaceutical Institute, University of Tuebingen, Tuebingen, Germany
  • 3Institute of Pharmaceutical Chemistry, J. W. Goethe-University, ZAFES, Frankfurt, Germany
Further Information

Publication History

received May 27, 2010 revised August 25, 2010

accepted Sept. 25, 2010

Publication Date:
03 November 2010 (online)

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Abstract

Recent studies revealed that the non-prenylated acylphloroglucinol myrtucommulone (MC) from myrtle (Myrtus communis) potently suppresses the biosynthesis of eicosanoids by direct inhibition of cyclooxygenase-1, microsomal prostaglandin E2 synthase (mPGES)-1, and 5-lipoxygenase at IC50 values in the range of 1 to 29 µM. Moreover, MC showed potent efficacy in animal models of inflammation after intraperitoneal administration. Since the main prerequisite for therapeutic efficacy is sufficient bioavailability, it is important to evaluate whether the concentrations of MC achieved in plasma coincide with the pharmacological active concentrations determined in vitro. For that reason, a sensitive LC/MS/MS method has been developed and validated for the determination of MC in human plasma. This method is based on liquid-liquid extraction of plasma samples with 20 % ethyl acetate in tert-butyl methyl ether using the structurally related acylphloroglucinol hyperforin as the internal standard. Chromatographic separation was achieved on a Gemini C6 Phenyl column using a mixture of acetonitrile/water (85 : 15 v/v) containing 6 mM ammonium formate in a run time of 15 min at a flow rate of 1 mL/min, a column temperature of 40 °C, and an autosampler temperature of 5 °C. Mass spectrometric quantification was carried out in the negative ion mode using electrospray ionization (ESI) and multiple-reaction monitoring (MRM). The most intense [M–H]− MRM transition at m/z 667.4 → m/z 194.9 was used for quantification of MC and the transition at m/z 535.4 to m/z 383.2 was used to monitor hyperforin. The method was linear in the range of 1–100 ng/mL with r > 0.998, an intra- and inter-day RSD of 1.1–8.4 and 7.1–11.8 %, respectively, and a maximum R. E. of 13.8 % at the lowest concentration level. Moreover, cross validation revealed the suitability of the developed LC/MS method for application in rat studies.

Key words

myrtucommulone - myrtle - Myrtus communis – Myrtaceae - LC/MS - human/rat plasma

References

  • 1 Feißt C, Franke L, Appendino G, Werz O. Identification of molecular targets of the oligomeric nonprenylated acylphloroglucinols from Myrtus communis and their implication as anti-inflammatory compounds.  J Pharm Exp Ther. 2005;  315 389-396
    CrossrefPubMedGoogle Scholar
  • 2 Bonjar G H. Antibacterial screening of plants used in Iranian folkloric medicine.  Fitoterapia. 2004;  75 231-235
    CrossrefPubMedGoogle Scholar
  • 3 Elfellah M S, Akhter M H, Khan M T. Anti-hyperglycaemic effect of an extract of Myrtus communis in streptozotocin-induced diabetes in mice.  J Ethnopharmacol. 1984;  11 275-281
    CrossrefPubMedGoogle Scholar
  • 4 Sepici A, Gürbüz I, Cevik C, Yesilada E. Hypoglycaemic effects of myrtle oil in normal and alloxan-diabetic rabbits.  J Ethnopharmacol. 2004;  93 311-318
    CrossrefPubMedGoogle Scholar
  • 5 Levesque H, Lafont O. Aspirin throughout the ages: a historical review.  Rev Med Interne. 2000;  21 (Suppl. 1) 8s-17s
    CrossrefPubMedGoogle Scholar
  • 6 Rosa A, Deiana M, Casu V, Corona G, Appendino G, Bianchi F, Ballero M, Dessi M A. Antioxidant activity of oligomeric acylphloroglucinols from Myrtus communis L.  Free Radic Res. 2003;  37 1013-1019
    CrossrefPubMedGoogle Scholar
  • 7 Hayder N, Abdelwahed A, Kilani S, Ammar R B, Mahmoud A, Ghedira K, Chekir-Ghedira L. Anti-genotoxic and free-radical scavenging activities of extracts from (Tunisian) Myrtus communis.  Mutat Res. 2004;  564 89-95
    PubMedGoogle Scholar
  • 8 Romani A, Coinu R, Carta S, Pinelli P, Galvardi C, Vincieri F F, Franconi F. Evaluation of antioxidant effect of different extracts of Myrtus communis L.  Free Radic Res. 2004;  38 97-103
    CrossrefPubMedGoogle Scholar
  • 9 Appendino G, Bianchi F, Minassi A, Sterner O, Ballero M, Gibbons S. Oligomeric acylphloroglucinols from myrtle (Myrtus communis).  J Nat Prod. 2002;  65 334-338
    CrossrefPubMedGoogle Scholar
  • 10 Koeberle A, Pollastro F, Northoff H, Werz O. Myrtucommulone, a natural acylphloroglucinol, inhibits microsomal prostaglandin E(2) synthase-1.  Br J Pharmacol. 2009;  156 952-961
    CrossrefPubMedGoogle Scholar
  • 11 Rossi A, Paola R D, Mazzon E, Genovese T, Caminiti R, Bramanti P, Pergola C, Koeberle A, Werz O, Sautebin L, Cuzzocrea S. Myrtucommulone from Myrtus communis exhibits potent anti-inflammatory effectiveness in vivo.  J Pharm Exp Ther. 2009;  329 76-86
    CrossrefPubMedGoogle Scholar
  • 12 Gioti E M, Skalkos D C, Fiamegos Y C, Stalikas C D. Single-drop liquid phase microextraction for the determination of hypericin, pseudohypericin and hyperforin in biological fluids by high performance liquid chromatography.  J Chromatogr A. 2005;  1093 1-10
    CrossrefPubMedGoogle Scholar
  • 13 Rozio M, Fracasso C, Riva A, Morazzoni P, Caccia S. High-performance liquid chromatography measurement of hyperforin and its reduced derivatives in rodent plasma.  J Chromatogr B Anal Technol Biomed Life Sci. 2005;  816 21-27
    CrossrefPubMedGoogle Scholar
  • 14 Cui Y, Gurley B, Ang C Y, Leakey J. Determination of hyperforin in human plasma using solid-phase extraction and high-performance liquid chromatography with ultraviolet detection.  J Chromatogr B Anal Technol Biomed Life Sci. 2002;  780 129-135
    CrossrefPubMedGoogle Scholar
  • 15 Bauer S, Störmer E, Graubaum H J, Roots I. Determination of hyperforin, hypericin and pseudohypericin in human plasma using high-performance liquid chromatography analysis with fluorescence and ultraviolet detection.  J Chromatogr B Biomed Sci Appl. 2001;  765 29-35
    CrossrefPubMedGoogle Scholar
  • 16 Chi J D, Franklin M. Measurement of hyperforin a constituent of St.John's wort in plasma by high-performance liquid chromatography.  J Chromatogr B Biomed Sci Appl. 1999;  735 285-288
    CrossrefPubMedGoogle Scholar
  • 17 Paulke A, Schubert-Zsilavecz M, Wurglics M. Determination of St. John's wort flavonoid-metabolites in rat brain through high performance liquid chromatography coupled with fluorescence detection.  J Chromatogr B Anal Technol Biomed Life Sci. 2006;  832 109-113
    CrossrefPubMedGoogle Scholar
  • 18 Hollman P C H, van Trijp J M P, Buysman M N C P. Fluorescence detection of flavonols in HPLC by postcolumn chelation with aluminum.  Anal Chem. 1996;  68 3511-3515
    CrossrefPubMedGoogle Scholar
  • 19 Rückert U, Eggenreich K, Wintersteiger R, Wurglics M, Likussar W, Michelitsch A. Development of a high-performance liquid chromatographic method with electrochemical detection for the determination of hyperforin.  J Chromatogr A. 2004;  1041 181-185
    CrossrefPubMedGoogle Scholar
  • 20 Keller J-H, Karas M, Müller W E, Volmer D A, Eckert G P, Tawab M A, Blume H H, Dingermann T, Schubert-Zsilavecz M. Determination of hyperforin in mouse brain by high-performance liquid chromatography/tandem mass spectrometry.  Anal Chem. 2003;  75 6084-6088
    CrossrefPubMedGoogle Scholar
  • 21 Kim H, Roh H, Lee H J, Chung S Y, Choi S O, Lee K R, Han S B. Determination of phloroglucinol in human plasma by high-performance liquid chromatography-mass spectrometry.  J Chromatogr B Anal Technol Biomed Life Sci. 2003;  792 307-312
    CrossrefPubMedGoogle Scholar
  • 22 Riedel K D, Rieger K, Martin-Facklam M, Mikus G, Haefeli W E, Burhenne J. Simultaneous determination of hypericin and hyperforin in human plasma with liquid chromatography-tandem mass spectrometry.  J Chromatogr B Anal Technol Biomed Life Sci. 2004;  813 27-33
    CrossrefPubMedGoogle Scholar
  • 23 Müller H, Paul M, Hartmann D, Huch V, Blaesius D, Koeberle A, Werz O, Jauch J. Total synthesis of myrtucommulone A.  Angew Chem Int Ed. 2010;  49 2045-2049
    PubMedGoogle Scholar

Dr. Mona Abdel-Tawab

Department for Research & Development
Central Laboratory of German Pharmacists

Carl-Mannich-Str. 20

65760 Eschborn

Germany

Phone: +49 61 96 93 79 55

Fax: +49 61 96 93 78 10

Email: m.tawab@zentrallabor.com