Assessment of Iohexol Serum Clearance by LC-MS/MS with Isotopically Labeled Internal Standard

Accurate measurement of the glomerular filtration rate (GFR) is essential for detecting renal insufficiency in living kidney donors. Iohexol is a “near-ideal” exogenous filtration marker for GFR measurement that has attracted increasing interest in clinical practice because it is non-toxic, non-radioactive, readily available, and easy to measure. In this chapter, we describe a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to measure iohexol in serum and to calculate GFR based on the rate of iohexol clearance. In this procedure, the contrast agent iohexol is administrated to the study subject in an outpatient setting, and three timed blood samples are collected. The serum proteins are precipitated, and the supernatant containing iohexol and the internal standard 2H5-iohexol is diluted prior to LC-MS/MS analysis. The LC-MS/MS method utilizes a Thermo Vanquish UHPLC coupled with TSQ Endura triple quadruple mass spectrometer, with a total run time of 2.5 min. The LC-MS/MS method has demonstrated good analytical performances, and the workflow can be used to reliably measure GFR in apparently healthy individuals without impaired renal function, such as living kidney donors.