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

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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.

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Authors and Affiliations

  1. Department of Production Animal Medicine, Faculty of Veterinary Medicine, University of Helsinki, P.O. Box 57, 00014, Helsinki, Finland

    Merja Pöytäkangas, Ilkka Saastamoinen & Hannu Saloniemi

  2. Department of Equine and Small Animal Medicine, Faculty of Veterinary Medicine, University of Helsinki, P.O. Box 57, 00014, Helsinki, Finland

    Elise Saario-Paunio & Thomas Spillmann

  3. Finnish Food Safety Authority (Evira), Mustialankatu 3, 00790, Helsinki, Finland

    Tiina Putkonen

  4. Central Animal Laboratory, University of Turku, Turku, Finland

    Rafael Frias

Authors
  1. Merja Pöytäkangas
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  2. Elise Saario-Paunio
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  3. Tiina Putkonen
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  4. Ilkka Saastamoinen
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  5. Rafael Frias
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  6. Thomas Spillmann
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  7. Hannu Saloniemi
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Correspondence to Elise Saario-Paunio.

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Pöytäkangas, M., Saario-Paunio, E., Putkonen, T. et al. Rapid LC-UV Analysis of Iohexol in Canine Plasma for Glomerular Filtration Rate Determination. Chroma 71, 211–216 (2010). https://doi.org/10.1365/s10337-009-1445-9

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  • DOI: https://doi.org/10.1365/s10337-009-1445-9

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