Performance of iohexol determination in serum and urine by HPLC: validation, risk and uncertainty assessment
Etienne Cavalier1, Eric Rozet, Nathalie Dubois
1Department of Clinical Chemistry, University of Liège, CHU Sart-Tilman, B-4000 Liège, Belgium. Etienne.cavalier@chu.ulg.ac.be
Background:
Determination of glomerular filtration rate plays an important role in nephrological practice. Iohexol is a reference marker for glomerular filtration rate determination. It is available and safe. The aim of this study was to develop a simple, efficient and easy to use analytical method for the quantification of iohexol in serum and urine by high performance liquid chromatography and to thoroughly validate this method.
Methods:
The HPLC method was inspired from the method published by Krutzen. The e.noval software V2.0 (Arlenda, Liège, Belgium) was used to compute all validation results.
Results:
The validation results indicate that the method will give accurate and reliable results for serum values ranging from 12.95 to 1295 microg/ml and for urine values ranging from 86.0 to 4144 microg/ml. In routine practice, iohexol concentrations found in plasma after injection range from 40 to 600 microg/ml. The expected urinary values are much wider. One should not hesitate to dilute urine samples to fit with the validated range over 5000 microg/ml.
Conclusion:
This is the first time that a reference method for the determination of GFR is validated with such a rigorous and thorough protocol. Contrary to other GFR markers, iohexol is now strongly validated from an analytical point of view.
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Key parameters for method validation include:
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