Get access

Development of a fast LC–MS/MS method for quantification of rilmenidine in human serum: elucidation of fragmentation pathways by HRMS

Authors

  • Lukáš Chytil,

    Corresponding author
    1. Institute of Forensic Medicine and Toxicology, 1st Faculty of Medicine, Charles University in Prague, 121 08 Prague 2, Czech Republic
    2. Institute of Pharmacology, 1st Faculty of Medicine, Charles University in Prague, 121 08 Prague 2, Czech Republic
    • Institute of Forensic Medicine and Toxicology, 1st Faculty of Medicine, Charles University in Prague, Ke Karlovu 2, 120 00 Praha 2, Czech Republic.
    Search for more papers by this author
  • Josef Cvačka,

    1. Institute of Organic Chemistry and Biochemistry Academy of Sciences of the Czech Republic, v.v.i., Flemingovo nám. 2, 166 10 Prague 6, Czech Republic
    Search for more papers by this author
  • Věra Marešová,

    1. Institute of Forensic Medicine and Toxicology, 1st Faculty of Medicine, Charles University in Prague, 121 08 Prague 2, Czech Republic
    Search for more papers by this author
  • Branislav Štrauch,

    1. Third Department of Internal Medicine, 1st Faculty of Medicine, Charles University in Prague, 121 08 Prague 2, Czech Republic
    Search for more papers by this author
  • Jiří Widimský Jr,

    1. Third Department of Internal Medicine, 1st Faculty of Medicine, Charles University in Prague, 121 08 Prague 2, Czech Republic
    Search for more papers by this author
  • Martin Štícha,

    1. Department of Organic Chemistry, Faculty of Science, Charles University in Prague, 128 43 Prague 2, Czech Republic
    Search for more papers by this author
  • Ondřej Slanař

    1. Institute of Pharmacology, 1st Faculty of Medicine, Charles University in Prague, 121 08 Prague 2, Czech Republic
    Search for more papers by this author

Abstract

Rilmenidine is an alpha 2 adrenoreceptor agonist used in the treatment of mild and moderate hypertension. In this study, a fast and accurate liquid chromatographic method with tandem mass spectrometric detection has been validated in order to assure quantification of rilmenidine in human serum. The fragmentation pathway of protonated rilmenidine was studied using high-resolution mass spectrometry (HRMS). This study compared selectivity, linearity, accuracy, precision, extraction efficiency, matrix effect and sensitivity using common liquid–liquid extraction (LLE) and solid-phase extraction (SPE) procedures. The limit of quantitation for both extraction techniques was 0.1 ng/ml. Several differences between the LLE and SPE have been observed in terms of linearity, accuracy, precision and matrix effect. Additionally, the advantages of SPE included less manual work load and increased recovery of rilmenidine in human serum to approximately 80% (LLE, 57%). The developed method involving SPE was found to be accurate (relative error (RE) < 5%), reproducible (relative standard deviation, RSD < 7%), robust and suitable for quantitative analysis of rilmenidine in serum samples obtained from patients under antihypertensive treatment. Copyright © 2010 John Wiley & Sons, Ltd.

Ancillary