Development and validation of an HPLC-UV method for iodixanol quantification in human plasma

Area of Science:

• Analytical Chemistry
• Clinical Chemistry
• Pharmacokinetics

Background:

• Iodixanol is an iso-osmolar contrast agent used in medical imaging.
• Accurate quantification of iodixanol is crucial for its use as an exogenous marker for Glomerular Filtration Rate (GFR) measurement.
• Existing methods may require optimization for large-scale clinical applications.

Purpose of the Study:

• To develop and validate a High-Performance Liquid Chromatography with Ultraviolet detection (HPLC-UV) method for quantifying iodixanol in human plasma.
• To establish a reliable analytical technique for assessing iodixanol levels in clinical settings.
• To support the use of iodixanol as a marker for GFR estimation.

Main Methods:

• Plasma samples were processed by adding iohexol as an internal standard and perchloric acid, followed by neutralization.
• Extraction and quantification were performed using a C18 analytical column with a gradient elution method.
• HPLC-UV analysis was conducted with a mobile phase of sodium formate buffer and acetonitrile.

Main Results:

• The HPLC-UV method demonstrated high recovery (>95%) for both iodixanol and iohexol.
• The assay achieved a lower limit of quantification of 10 microg/ml and linearity up to 750 microg/ml.
• Intra- and inter-day accuracy exceeded 95%, with precision below 10% coefficient of variation.

Conclusions:

• A simple, accurate, precise, and robust HPLC-UV method for iodixanol quantification in human plasma was successfully developed and validated.
• This validated method is suitable for large-scale clinical studies requiring iodixanol measurement.
• The findings support the utility of iodixanol as an exogenous marker for GFR assessment.