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HPLC micro-fractionation coupled to a cell-based assay for automated on-line primary screening of calcium antagonistic components in plant extracts

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Abstract

High performance liquid chromatography (HPLC) micro-fractionation was successfully coupled to an automated 45Ca2+ uptake assay using GH4C1 cells for the separation of natural product extracts and for the primary detection of their calcium antagonistic components. The reliability of the procedure was first established with a reference solution consisting of pure compounds with a known effect on the Ca2+ uptake. No loss of activity was observed to occur after HPLC micro-fractionation. Extracts of Peucedanum palustre and Pinus sylvestris, showing high and no inhibition of Ca2+ uptake as total extracts, respectively, were analysed and the inhibitory activity of the P. palustre extract could be traced to two components, identified as columbianadin and isoimperatorin. As expected, no significant inhibition was observed with the micro-fractionated P. sylvestris samples. In summary, the procedure was found to be applicable for primary detection of calcium antagonistic components in complex matrices and to significantly reduce the time previously needed for bioactivity-guided isolation.

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Acknowledgements

This work was supported by the National Technology Agency in Finland (grants no. 40045/02 and 40083/03), and by a personal grant (for PT) from the Finnish Cultural Foundation. The authors would like to express their gratitude to Prof. Kid Törnquist from Åbo Akademi University (Finland) for providing the GH4C1 cell line.

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Authors and Affiliations

  1. Viikki Drug Discovery Technology Center (DDTC), Faculty of Pharmacy, University of Helsinki, P.O. Box 56, 00014, Finland

    Päivi Tammela & Pia Vuorela

  2. Division of Pharmacognosy, Faculty of Pharmacy, University of Helsinki, P.O. Box 56, 00014, Finland

    Tero Wennberg & Heikki Vuorela

Authors
  1. Päivi Tammela
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  2. Tero Wennberg
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  3. Heikki Vuorela
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  4. Pia Vuorela
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Correspondence to Pia Vuorela.

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Tammela, P., Wennberg, T., Vuorela, H. et al. HPLC micro-fractionation coupled to a cell-based assay for automated on-line primary screening of calcium antagonistic components in plant extracts. Anal Bioanal Chem 380, 614–618 (2004). https://doi.org/10.1007/s00216-004-2795-7

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  • DOI: https://doi.org/10.1007/s00216-004-2795-7

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