View More View Less
  • 1 Universität Stuttgart Institute of Biology, Department of Zoology Pfaffenwaldring 57 70569 Stuttgart Germany
  • 2 University of Hohenheim Institute of Food Chemistry Garbenstrasse 28 70599 Stuttgart Germany
Restricted access

Rapid analysis by coupling HPTLC with bioluminescence and mass spectrometry enables very fast response to bioactive substances in unknown samples. In this study marine sponges were screened for new bioactive compounds. After chromatographic separation of twelve methanolic marine sponge extracts the HPTLC plates were coated with bioluminescent bacteria ( Vibrio fischeri ) by a simple dipping procedure. If separated compounds were bioactive they inhibited or enhanced the bacterial luminescence and could be identified as dark zones on the luminescent background. This micro-biological detection revealed new compounds compared with physical (absorbance or fluorescence measurement) or chemical (microchemical derivatization) detection techniques. Effect-directed analysis turned out to be superior to target analysis in the search for natural products with a distinct effect. For identification of unknown bioactive zones the HPTLC system was coupled to a high-resolution mass spectrometer to obtain the exact masses of the unknowns. Thus, a Vibrio fischeri -bioactive zone was identified as avarone, a bioactive metabolite so far only known to be synthesized by the sponge Dysidea avara . This methodology proved very effective not only for detection but also for identification of unknown bioactive metabolites in sponges.

  • G. Eberz, H.G. Rast, K. Burger, W. Kreiss and C. Weisemann , Chromatographia 43 (1996) 5–9.

    Weisemann C. , '' (1996 ) 43 Chromatographia : 5 -9.

  • M. Vuillaume and M. Barbier , Comparative Biochemistry and Physiology, Part B: Biochemistry & Molecular Biology 80B (1985) 547–550.

    Barbier M. , '' (1985 ) 80B Comparative Biochemistry and Physiology, Part B: Biochemistry & Molecular Biology : 547 -550.

  • G.A. Goodrich and H.R. Burreli , Anal. Biochem. 127 (1982) 395–401.

    Burreli H.R. , '' (1982 ) 127 Anal. Biochem. : 395 -401.

  • W. Kreiss, G. Eberz, H.G. Rast and C. Weisemann , Patent EP 1359226, 2003.

  • J.E. Becvar and L.E. Becvar , Patent US 6017722, 1991.

  • S.M. Verbitsky, J.D. McChesney, G.T. Gourdin and L.M. Ikenouye , Patent US 2007184514, 2007.

  • J.E. Becvar and L.E. Becvar , Patent US 6017722, 1991.

  • C. Weisemann, W. Kreiss, H.G. Rast and G. Eberz , Patent DE 4230264, 1992.

  • S.A. Jassim, A. Ellison, S.P. Denyer and G.S. Stewart , J. Biolumin. Chemilumin. 5 (1990) 115–122.

    Stewart G.S. , '' (1990 ) 5 J. Biolumin. Chemilumin. : 115 -122.

  • E.A. Meighen and P.V. Dunlap , Adv. Microb. Physiol. 34 (1993) 1–67.

    Dunlap P.V. , '' (1993 ) 34 Adv. Microb. Physiol. : 1 -67.

  • K.L.E. Kaiser and V.S. Palabrica , Water Poll. Res. J. Canada 26 (1991) 361–431.

    Palabrica V.S. , '' (1991 ) 26 Water Poll. Res. J. Canada : 361 -431.

  • C. Buch, J. Sigh, J. Nielsen, J.L. Larsen and L. Gram , Systematic Applied Microbiol. 26 (2003) 338–349.

    Gram L. , '' (2003 ) 26 Systematic Applied Microbiol. : 338 -349.

  • L. Ravn, A.B. Christensen, S. Molin, M. Givskov and L. Gram , J. Microbiol. Methods 44 (2001) 239–251.

    Gram L. , '' (2001 ) 44 J. Microbiol. Methods : 239 -251.

  • International Organization for Standardization (ISO) , ISO 11348 (2007).

  • G. Morlock and W. Schwack , LC-GC July (2008) 366–371.

  • W. Weber, W. Seitz, A. Aichinger and R. Albert , CBS CAMAG Bibliogr. Serv. 94 (2005) 2–4.

    Albert R. , '' (2005 ) 94 CBS CAMAG Bibliogr. Serv. : 2 -4.

  • S.M. Verbitsky, S. Hickey, G.T. Gourdin and L.M. Ikenouye , CAMAG Bibliogr. Serv. CBS 99 (2007) 11–13.

    Ikenouye L.M. , '' (2007 ) 99 CAMAG Bibliogr. Serv. CBS : 11 -13.

  • G. Morlock, W. Schwack , J. Planar Chromatogr. 20 (2008) 399–406.

    Schwack W. , '' (2008 ) 20 J. Planar Chromatogr. : 399 -406.

  • S.M. Verbitski, G.T. Gourdin, L.M. Ikenouye and J.D. McChesney , American Biotechnology Laboratory 24 (2006) 40–42.

    McChesney J.D. , '' (2006 ) 24 American Biotechnology Laboratory : 40 -42.

  • S.M. Verbitski, G.T. Gourdin, L.M. Ikenouye and J.D. McChesney , Abstracts of Papers, 230th ACS National Meeting, Washington, DC, United States, 2005.

  • U. Baumann, C. Brunner, E. Pletscher and N. Tobler , Umweltwissenschaften und Schadstoff-Forschung 15 (2003) 163–167.

    Tobler N. , '' (2003 ) 15 Umweltwissenschaften und Schadstoff-Forschung : 163 -167.

  • T. Reemtsma, A. Putschew and M. Jekel , Vom Wasser 92 (1999) 243–255.

    Jekel M. , '' (1999 ) 92 Vom Wasser : 243 -255.

  • S.M. Verbitski, G.T. Gourdin, L.M. Ikenouye, J.D. McChesney and J. Hildreth , J. AOAC Int. 91 (2008) 268–275.

    Hildreth J. , '' (2008 ) 91 J. AOAC Int. : 268 -275.

  • L. Minale, R. Riccio and G. Sodano , Tetrahedron Lett. 38 (1974) 3401–3404.

    Sodano G. , '' (1974 ) 38 Tetrahedron Lett. : 3401 -3404.

  • US Patent 4946869, 2008.

  • US Patent 5001162, 2008.

  • G. Morlock and Y. Ueda , J. Chromatogr. A 1143 (2007) 243–251.

    Ueda Y. , '' (2007 ) 1143 J. Chromatogr. A : 243 -251.

  • K. Dreisewerd, J. Muething, A. Rohlfing, I. Meisen, Z. Vukelic, J. Peter-Katalinic, F. Hillenkamp and S. Berkenkamp , Anal. Chem. 77 (2005) 4098–4107.

    Berkenkamp S. , '' (2005 ) 77 Anal. Chem. : 4098 -4107.

  • B. Fuchs, J. Schiller, R. Suess, M. Schuerenberg and D. Suckau , Anal. Bioanal. Chem. 389 (2007) 827–834.

    Suckau D. , '' (2007 ) 389 Anal. Bioanal. Chem. : 827 -834.

  • G.J. Van Berke and V. Kertesz , Anal. Chem. 78 (2006) 4938–4944.

    Kertesz V. , '' (2006 ) 78 Anal. Chem. : 4938 -4944.

  • A. Alpmann and G. Morlock , Anal. Bioanal. Chem. 386 (2006) 1543–1551.

    Morlock G. , '' (2006 ) 386 Anal. Bioanal. Chem. : 1543 -1551.

  • U. Jautz and G. Morlock , J. Chromatogr. A 1128 (2006) 244–250.

    Morlock G. , '' (2006 ) 1128 J. Chromatogr. A : 244 -250.

  • V. Kertesz, M.J. Ford and G.J. Van Berkel , Anal. Chem. 77 (2005) 7183–7189.

    Berkel G.J. , '' (2005 ) 77 Anal. Chem. : 7183 -7189.

  • H. Luftmann, M. Aranda and G. Morlock , Rapid Commun. Mass Spectrom. 21 (2007) 3772–3776.

    Morlock G. , '' (2007 ) 21 Rapid Commun. Mass Spectrom. : 3772 -3776.

  • C. Thoms , Alkaloide in Schwämmen der Gattung Aplysina-chemische Ökologie und assoziierte Bakterien, PhD thesis, Heinrich-Heine-Universität, Düsseldorf (2004).

    Thoms C. , '', in Alkaloide in Schwämmen der Gattung Aplysina-chemische Ökologie und assoziierte Bakterien , (2004 ) -.

    • Search Google Scholar
  • W.E.G. Müller, R.K. Zahn, M.J. Gasic, N. Dogovic, A. Maidhof, C. Becker, B. Diehl-Seifert and E. Eich , Comp. Biochem. Physiol. C 80 (1985) 47–52.

    Eich E. , '' (1985 ) 80 Comp. Biochem. Physiol. C : 47 -52.

  • A. Putz, A. Klöppel, M. Pfannkuchen, F. Brümmer and P. Proksch , J. Chem. Ecology (2008) submitted.

  • E. Dytkiewitz and G. Morlock , J. AOAC Int. 91 (2008) 1237–1244.

    Morlock G. , '' (2008 ) 91 J. AOAC Int. : 1237 -1244.

Monthly Content Usage

Abstract Views Full Text Views PDF Downloads
Jun 2020 0 6 6
Jul 2020 6 0 0
Aug 2020 9 0 0
Sep 2020 1 0 0
Oct 2020 6 0 0
Nov 2020 0 0 0
Dec 2020 0 0 0