Chromatographia

, Volume 71, Issue 3–4, pp 211–216 | Cite as

Rapid LC-UV Analysis of Iohexol in Canine Plasma for Glomerular Filtration Rate Determination

  • Merja Pöytäkangas
  • Elise Saario-Paunio
  • Tiina Putkonen
  • Ilkka Saastamoinen
  • Rafael Frias
  • Thomas Spillmann
  • Hannu Saloniemi
Original

Abstract

A rapid high-performance liquid chromatography UV method and a simple sample preparation for analyzing iohexol in canine plasma, for evaluating glomerular filtration rate (GFR) and intestinal permeability, were developed and validated. Trifluoroacetic acid (TFA) was used for protein precipitation and iohexol extraction from plasma, followed by vortex mixing and centrifugation. As an internal standard, 4-aminobenzoic acid (para-aminobenzoic acid, PABA) was added. The supernatant (5 μL) was injected into a Zorbax SB-C18 LC column maintained at 50 °C. The mobile phase of the LC method was a water–methanol gradient at pH 3.0 adjusted with TFA. Fast LC measurement was achieved by using a rapid-resolution LC technique. Total run time was 13 min, and UV wavelength was set at 246 nm. Precision of the method was 0.2–9.0%, depending on the iohexol concentration in plasma. Recovery of iohexol from plasma was over 90%, and recovery of the internal standard 99.1 ± 1.4%. The calibration curve was linear (r = 0.9997) over iohexol concentrations of 2.5–150 μg mL−1 (n = 5). This method is fast, simple, reliable and applicable in clinical settings.

Keywords

Rapid-resolution liquid chromatography Intestinal permeability Glomerular filtration rate Iohexol in canine plasma 

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Copyright information

© Vieweg+Teubner | GWV Fachverlage GmbH 2010

Authors and Affiliations

  • Merja Pöytäkangas
    • 1
  • Elise Saario-Paunio
    • 2
  • Tiina Putkonen
    • 3
  • Ilkka Saastamoinen
    • 1
  • Rafael Frias
    • 4
  • Thomas Spillmann
    • 2
  • Hannu Saloniemi
    • 1
  1. 1.Department of Production Animal Medicine, Faculty of Veterinary MedicineUniversity of HelsinkiHelsinkiFinland
  2. 2.Department of Equine and Small Animal Medicine, Faculty of Veterinary MedicineUniversity of HelsinkiHelsinkiFinland
  3. 3.Finnish Food Safety Authority (Evira)HelsinkiFinland
  4. 4.Central Animal LaboratoryUniversity of TurkuTurkuFinland

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