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)


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 


  1. 1.
    Zappitelli M, Parvex P, Joseph L, Paradis G, Grey V, Lau S, Bell L (2006) Derivation and validation of cystatin C-based prediction equations for GFR in children. Am J Kidney Dis 48:221–230CrossRefPubMedGoogle Scholar
  2. 2.
    Schwartz GJ, Furth S, Cole SR, Warady B, Munoz A (2006) Glomerular filtration rate via plasma iohexol disappearance: pilot study for chronic kidney disease in children. Kidney Int 69:2070–2077CrossRefPubMedGoogle Scholar
  3. 3.
    Olsson B, Aulie A, Sveen K, Andrew E (1983) Human pharmacokinetics of iohexol. A new nonionic contrast medium. Invest Radiol 18:177–182CrossRefPubMedGoogle Scholar
  4. 4.
    Brown SC, O’Reilly PH (1991) Iohexol clearance for the determination of glomerular filtration rate in clinical practice: evidence for a new gold standard. J Urol 146:675–679PubMedGoogle Scholar
  5. 5.
    Gaspari F, Perico N, Ruggenenti P, Mosconi L, Amuchastegui CS, Guerini E, Daina E, Remuzzi G (1995) Plasma clearance of nonradioactive iohexol as a measure of glomerular filtration rate. J Am Soc Nephrol 6:257–263PubMedGoogle Scholar
  6. 6.
    Schwartz GJ, Work DF (2009) Measurement and estimation of GFR in children and adolescents. Clin J Am Soc Nephrol 4:1832–1843CrossRefPubMedGoogle Scholar

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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