High-Performance Liquid-Chromatographic Analysis of Plasma Iohexol Concentrations.
In this study, a high-performance liquid-chromatographic (HPLC) method using photodiode array detection and isocratic conditions was developed for the analysis of plasma iohexol concentrations. Plasma proteins were precipitated with 1:1 volume of plasma and acetonitrile-ethanol-water (60:38.4:1.6, v/v/v). Iohexol concentrations in the supernatant phase were analyzed on a Waters Symmetry C-18 reversed-phase column under isocratic conditions at 245 nm. The extraction recoveries of iohexol from plasma were >95% and the plasma iohexol calibration curves were linear (R(2) ≥ 0.9998) from 10 to 1500 µg/mL. The within-day coefficients of variation (CVs) at plasma iohexol concentrations of 100, 375, 750 and 1500 µg/mL were 5.1, 3.5, 1.3 and 2.5%, respectively; the between-day CVs at 100, 375, 750 and 1500 µg/mL were 8.6, 4.2, 4.0 and 3.7%, respectively. The day-to-day accuracies of the method at plasma iohexol concentrations of 50, 100, 375, 750 and 1500 µg/mL were 89.0, 99.4, 108.4, 103.6 and 101.2%, respectively (n = 5). The lower limit of plasma iohexol quantitation was 10 µg/mL and no interferences >9 µg/mL were found in over 75 pre-dose porcine plasma samples. The applicability of the method was demonstrated by determining the glomerular filtration rates of iohexol in the porcine (Sus scrofa) model.
Clinical Research Division, Wilford Hall Ambulatory Surgical Center, JBSA Lackland, 2200 Bergquist Drive, San Antonio, TX 78236, USA harvey.schwertner@us.af.mil.
Harvey A
HA
Schwertner
Department of Nephrology, JBSA Lackland, San Antonio, TX 78236, USA.
Kyle J
KJ
Weld
D000818:Animals; D001774:Blood Chemical Analysis; D001798:Blood Proteins / Q000302:isolation & purification; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007472:Iohexol / Q000032:analysis*; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine
D001798:Blood Proteins; D007472:Iohexol