title: HPLC estimation of iothalamate to measure glomerular filtration rate in humans.
abstract: Glomerular filtration rate (GFR) is usually determined by estimation of iothalamate (IOT) clearance. We have developed and validated an accurate and robust method for the analysis of IOT in human plasma and urine. The mobile phase consisted of methanol and 50 mM sodium phosphate (10:90; v/v). Flow rate was 1.2 mL/min on a C18 reverse phase column, Synergi-hydro (250 × 4.6 mm) 4 µm 80 Å, with an ultraviolet detector set to 254 nm. Acetonitrile was used for the deproteination and extraction of IOT from human plasma and urine. Precision and accuracy were within 15% for IOT in both plasma and urine. The recoveries of IOT in urine and plasma ranged between 93.14% and 114.74 and 96.04-118.38%, respectively. The linear range for urine and plasma assays were 25-1500 and 1-150 µg/mL respectively. The lower limits of detection were 0.5 µg/mL for both urine and plasma, with no interference from plasma and urine matices. This method has been fully validated according to FDA guidelines and the new HPLC assay has been applied to a new formulation of IOT (Conray™ 43), to calculate GFR in healthy volunteers. The new method is simple, less expensive and it would be instrumental in future clinical and pharmacokinetic studies of iothalamate in kidney patients.
authors.affiliation: Department of Chemistry, College of Science, United Arab Emirates University, Al Ain, UAE.
authors.forename: Iltaf
authors.initials: I
authors.lastname: Shah
authors.affiliation: School of Life Sciences, Pharmacy and Chemistry, Kingston University, Penrhyn Road, Kingston-upon-Thames, Surrey, KT1 2EE UK.
authors.forename: James
authors.initials: J
authors.lastname: Barker
authors.affiliation: School of Life Sciences, Pharmacy and Chemistry, Kingston University, Penrhyn Road, Kingston-upon-Thames, Surrey, KT1 2EE UK.
authors.forename: Declan P
authors.initials: DP
authors.lastname: Naughton
authors.affiliation: School of Life Sciences, Pharmacy and Chemistry, Kingston University, Penrhyn Road, Kingston-upon-Thames, Surrey, KT1 2EE UK.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Barton
authors.affiliation: Department of Chemistry, College of Science, United Arab Emirates University, Al Ain, UAE.
authors.forename: Syed Salman
authors.initials: SS
authors.lastname: Ashraf
keywords: GFR; Glomerular filtration rate; HPLC; Human urine and plasma; Iothalamate; Iothalamic acid