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Quantification of Iohexol in Serum by High-Performance Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

  • Faye B. Vicente
  • Gina Vespa
  • Alan Miller
  • Shannon HaymondEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1383)

Abstract

Iohexol is a nonradioactive contrast medium, and its clearance from serum or urine is used to measure glomerular filtration rate (GFR). GFR is the most useful indicator of kidney function and progression of kidney disease. GFR determination using iohexol clearance is increasingly being applied in clinical practice, given its advantages over and correlation with inulin. We describe a high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) method for iohexol clearance, requiring only 50 μL of serum. The sample preparation involves protein precipitation with LC/MS-grade methanol, containing ioversol as the internal standard. Samples are centrifuged and supernatant is dried under nitrogen gas at room temperature. Samples are reconstituted with mobile phase (ammonium acetate—formic acid—water). Iohexol is separated using an HPLC gradient method on a C-8 analytical column. MS/MS detection is in the multiple-reaction monitoring (MRM) mode and the transitions monitored are m/z 822.0 to m/z 804.0 and m/z 807.0 to m/z 588.0 for iohexol and ioversol, respectively.

Key words

Iohexol Glomerular filtration rate Mass spectrometry Liquid chromatography Serum Quantification 

References

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Faye B. Vicente
    • 1
  • Gina Vespa
    • 1
  • Alan Miller
    • 1
  • Shannon Haymond
    • 1
    Email author
  1. 1.Department of PathologyNorthwestern University Feinberg School of MedicineChicagoUSA

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