Fluorescamine-based fluorophore for spectrofluorimetric determination of heptaminol in human plasma; application to spiked human plasma

Spectrochim Acta A Mol Biomol Spectrosc. 2020 Feb 15:227:117711. doi: 10.1016/j.saa.2019.117711. Epub 2019 Oct 25.

Abstract

An innovative approach to determine heptaminol spectrofluorimetrically was developed, determining the optimum conditions needed, then validated for determination of heptaminol in its pure form, its tablets and in spiked human plasma. The presented method is based on the reaction between fluorescamine reagent with the primary amine group found in heptaminol, using a borate buffer at pH 9.0 that yields a highly fluorescent product, fluorescence was measured at 471 nm after excitation at 393 nm. The linearity of the constructed calibration curve was (75-850 ng/ml) with LOD and LOQ values 23.85 and 72.29 ng/ml respectively. The method was validated following the International Council for Harmonisation (ICH) guidelines indicating good accuracy and precision. Finally, the presented approach was adapted for in vitro study of heptaminol in spiked human plasma with a mean percentage recovery 100.52 ± 1.19% as well as in its tablets with a mean percentage recovery 99.47 ± 1.25%.

Keywords: Fluorescamine; Heptaminol; Spectrofluorimetric determination; Spiked plasma.

MeSH terms

  • Buffers
  • Fluorescamine / chemistry*
  • Fluorescent Dyes / chemistry*
  • Heptaminol / blood*
  • Heptaminol / chemistry
  • Humans
  • Hydrogen-Ion Concentration
  • Reproducibility of Results
  • Solvents / chemistry
  • Spectrometry, Fluorescence / methods*
  • Time Factors

Substances

  • Buffers
  • Fluorescent Dyes
  • Solvents
  • Fluorescamine
  • Heptaminol