A Simple and Rapid Ultra-performance Liquid Chromatography Method for Plasma Iohexol Determination
Keywords:
Glomerular filtration rate, Iohexol, Ultra-high-performance liquid chromatography, Method optimization, Method validationAbstract
Accurate measurement of iohexol in plasma is essential for determining glomerular filtration rate (GFR). In this study, we developed and validated a novel method for quantifying iohexol using ultra-performance liquid chromatography with ultraviolet detection (UPLC-UV). The method involved sample preparation and chromatographic separation for iohexol. Various parameters including the type of stationary phase, mobile phase composition, and pH of mobile phase were optimized to achieve optimal separation. The optimal chromatographic conditions are shown as follow; the mobile phase was 3.5% (v/v) of acetonitrile/water without adjusting pH, a flow rate was set at 0.2 mL/min, the analytical column was Poroshell 120 EC-C18 with a column temperature of 30 ำC. The developed method exhibited excellent linearity over a concentration range of 0 to 640 μg/mL (R2 = 0.9999). Precision analyses demonstrated %RSD values within acceptable limits (<2%) for within-run and between-run analyses. Recovery experiments yielded accurate results ranging from 97.70 ± 2.11% to 102.91 ± 1.47%. The method also achieved low limits of detection and quantitation (0.97 and 3.26 μg/mL, respectively) with a short analysis time per sample (6 minutes). Overall, the developed UPLC-UV assay provides a simple, rapid, and reliable method for iohexol determination in plasma, making it suitable for clinical applications in nephrology practice and research for direct GFR assessment.
References
Cusumano AM, Tzanno-Martins C, Rosa-Diez GJ. The glomerular filtration rate: from the diagnosis of kidney function to a public health tool. Front Med (Lausanne) 2021; 8: 769335.
Kaufman DP, Basit H, Knohl SJ. Physiology, glomerular filtration rate. StatPearls [serial on the Internet] 2024 Jan [cited 2024 March 4]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK500032/.
Rahn KH, Heidenreich S, Brückner D. How to assess glomerular function and damage in humans. J Hypertens 1999; 17: 309-17.
Vilhelmsdotter Allander S, Marké L, Wihlen B, Svensson M, Elinder CG, Larsson A. Regional variation in use of exogenous and endogenous glomerular filtration rate (GFR) markers in Sweden. Ups J Med Sci 2012; 117: 273-8.
Herget-Rosenthal S, Bökenkamp A, Hofmann W. How to estimate GFR-serum creatinine, serum cystatin C or equations? Clin Biochem 2007; 40: 153-61.
Levey AS, Titan SM, Powe NR, Coresh J, Inker LA. Kidney disease, race, and GFR estimation. Clin J Am Soc Nephrol 2020; 15: 1203-12.
Musso C, Alvarez J, Jauregui J, Macías J. Glomerular filtration rate equations: a comprehensive review. Int Uro Nephrol 2016; 48: 1105-10.
Lamb EJ, Levey AS, Stevens PE. The kidney disease improving global outcomes (KDIGO) guideline update for chronic kidney disease: evolution not revolution. Clin Chem 2013; 59: 462-5.
Ebert N, Bevc S, Bökenkamp A, et al. Assessment of kidney function: clinical indications for measured GFR. Clin Kidney J 2021; 14: 1861-70.
Caregaro L, Menon F, Angeli P, et al. Limitations of serum creatinine level and creatinine clearance as filtration markers in cirrhosis. Arch Intern Med 1994; 154: 201-5.
Sriperumbuduri S, Dent R, Malcolm J, et al. Accurate GFR in obesity-protocol for a systematic review. Syst Rev 2019; 8: 147.
Cvan Trobec K, Kerec Kos M, von Haehling S, et al. Iohexol clearance is superior to creatinine-based renal function estimating equations in detecting short-term renal function decline in chronic heart failure. Croat Med J 2015; 56: 531-41.
Levey AS, Coresh J, Tighiouart H, Greene T, Inker LA. Measured and estimated glomerular filtration rate: current status and future directions. Nat Rev Nephrol 2020; 16: 51-64.
Soman RS, Zahir H, Akhlaghi F. Development and validation of an HPLC-UV method for determination of iohexol in human plasma. J Chromatogr B 2005; 816: 339-43.
Slack A, Tredger M, Brown N, Corcoran B, Moore K. Application of an isocratic methanol-based HPLC method for the determination of iohexol concentrations and glomerular filtration rate in patients with cirrhosis. Ann Clin Biochem 2014; 51: 80-8.
Carrara F, Gaspari F. GFR measured by iohexol: the best choice from a laboratory perspective. J Lab Precis Med 2018; 3: 77.
Delanaye P, Ebert N, Melsom T, et al. Iohexol plasma clearance for measuring glomerular filtration rate in clinical practice and research: a review. Part 1: How to measure glomerular filtration rate with iohexol? Clin Kidney J 2016; 9: 682-99.
Pottel H, Schaeffner E, Ebert N, van der Giet M, Delanaye P. Iohexol plasma clearance for measuring glomerular filtration rate: effect of different ways to calculate the area under the curve. BMC Nephrol 2021; 22: 166.
Castagnet S, Blasco H, Vourc’h P, et al. Routine determination of GFR in renal transplant recipients by HPLC quantification of plasma iohexol concentrations and comparison with estimated GFR. J Clin Lab Anal 2012; 26: 376-83.
Ng DK, Schwartz GJ, Warady BA, Furth SL, Muñoz A. Relationships of measured iohexol GFR and estimated GFR with CKD-related biomarkers in children and adolescents. Am J Kid Dis 2017; 70: 397-405.
Schwertner HA, Weld KJ. Highperformance liquid-chromatographic analysis of plasma iohexol concentrations. J Chromatogr Sci 2015; 53: 1475-80.
Holleran JL, Parise RA, Guo J, et al. Quantitation of iohexol, a glomerular filtration marker, in human plasma by LC-MS/MS. J Pharm Biomed Anal 2020; 189: 113464.
Destefano JJ, Schuster SA, Lawhorn JM, Kirkland JJ. Performance characteristics of new superficially porous particles. J Chromatogr A 2012; 1258: 76-83.
Branch SK. Guidelines from the international conference on harmonization (ICH). J Pharm Biomed Anal 2005; 38: 798-805.
Shabir G. Step-by-step analytical methods validation and protocol in the quality system compliance industry. J Valid Technol 2004; 10: 314-24.
El Assri S, Sam H, El Assri A, et al. Iohexol assay for direct determination of glomerular filtration rate: optimization and development of an HPLC-UV method for measurement in serum and urine. Clin Chim Acta 2020; 508: 115-21.
Niculescu-Duvaz I, D’Mello L, Maan Z, et al. Development of an outpatient finger-prick glomerular filtration rate procedure suitable for epidemiological studies. Kidney Int 2006; 69: 1272-5.
Nilsson-Ehle P. Iohexol clearance for the determination of glomerular filtration rate: 15 years’ experience in clinical practice. eJIFCC 2001; 13: 48-52.
Farthing D, Sica DA, Fakhry I, et al. Simple HPLC–UV method for determination of iohexol, iothalamate, p-aminohippuric acid and n-acetyl-p-aminohippuric acid in human plasma and urine with ERPF, GFR and ERPF/GFR ratio determination using colorimetric analysis. J Chromatogr B 2005; 826: 267-72.
Cavalier E, Rozet E, Dubois N, et al. Performance of iohexol determination in serum and urine by HPLC: Validation, risk and uncertainty assessment. Clin Chim Acta 2008; 396: 80-5.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2024 Journal of the Medical Technologist Association of Thailand
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
