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Efficient Method Development and Validation for the Determination of Cardiovascular Drugs in Human Plasma by SPE–UHPLC–PDA–FLD

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Abstract

This work reports the development and validation of an ultra-high-performance liquid chromatography method to separate 11 different cardiovascular drugs: acenocoumarol, amiloride, bisoprolol, fluvastatin, furosemide, glibenclamide, hydrochlorothiazide, rosiglitazone (internal standard), valsartan, verapamil and warfarin. Detection of the analytes was carried out by means of photodiode array and fluorescence detection. Initially, the most significant variables in liquid chromatography (pH of the mobile phase, nature of the organic modifier and stationary phase type) were studied simultaneously in a systematic way. Taking into account the resolution obtained for the different chromatographic separations, pH 4.5, methanol and C18 were chosen as mobile phase pH, organic modifier and stationary phase, respectively. The method was validated and applied to the analysis of real human plasma samples using a solid phase extraction sample treatment, and the robustness of the chromatographic separation was studied by experimental design. The reported method allows the separation of all the analytes and the IS in less than 6 min, and, furthermore, it simplifies the analysis of drugs used in combined cardiovascular therapy by applying a fast and simple methodology.

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Acknowledgements

Authors thank Ministry of Economy and Competitiveness (Project CTQ2013- 46179R), University of Basque Country (GIU 16/04 and UFI 11/23) for financial support and the pharmaceutical companies for the kind supply of studied drugs. M. E. Blanco also thanks the University of the Basque Country (UPV/EHU) for her postdoctoral grant.

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Authors and Affiliations

  1. Analytical Chemistry Department, Science and Technology Faculty, University of the Basque Country UPV/EHU, Sarriena s/n, 48940, Leioa, Biscay, Basque Country, Spain

    Oskar González Mendia, María Encarnación Blanco, Estitxu Rico, María Luz Alonso & Rosa María Alonso

  2. Painting Department, Fine Arts Faculty, University of the Basque Country UPV/EHU, 48940, Leioa, Biscay, Basque Country, Spain

    Miren Itxaso Maguregui

Authors
  1. Oskar González Mendia
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  2. María Encarnación Blanco
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  3. Estitxu Rico
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  4. María Luz Alonso
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  5. Miren Itxaso Maguregui
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  6. Rosa María Alonso
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Corresponding author

Correspondence to Oskar González Mendia.

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The authors declare that they have no conflict of interest.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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González Mendia, O., Blanco, M.E., Rico, E. et al. Efficient Method Development and Validation for the Determination of Cardiovascular Drugs in Human Plasma by SPE–UHPLC–PDA–FLD. Chromatographia 80, 605–615 (2017). https://doi.org/10.1007/s10337-017-3274-6

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  • DOI: https://doi.org/10.1007/s10337-017-3274-6

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