title: Quantitative determination of exo- and endo-iohexol in canine and feline samples using high performance liquid chromatography with ultraviolet detection.
abstract: A sensitive and specific high performance liquid chromatography-ultraviolet detection (HPLC-UV) method for quantitative determination of exo- and endo-iohexol in cat and dog serum/plasma is presented. Sample preparation consisted of a protein precipitation step performed by adding 15 μL of trifluoroacetic acid to 100 μL of serum/plasma. Following vortexing and centrifugation, an aliquot of the supernatant was injected onto a polymeric PLRP-S column (250 mm × 4.6 mm i.d., dp: 8 μm, 100 Å), maintained at 30 °C. The mobile phase consisted of water (A) and methanol (B) and a gradient elution (flow-rate: 1.0 mL min(-1), total run-time: 21 min). The UV detector was set at a wavelength of 254 nm. Matrix-matched calibration graphs were prepared for both exo- (0.44-657 μg mL(-1)) and endo-iohexol (0.62-93.0 μg mL(-1)). Correlation and goodness-of-fit coefficients were between 0.9985-0.9999 and 4.44-9.87%, respectively. Limits of quantification and detection were 0.44 and 0.15 μg mL(-1) for exo-iohexol and 0.62 and 0.20 μg mL(-1) for endo-iohexol, respectively. Results for within-day and between-day precision and accuracy fell within the ranges specified. The reported method is simple and cost-effective. It has been successfully used for the analysis of exo- and endo-iohexol in serum/plasma samples of cats and dogs as part of pharmacokinetic studies with iohexol in order to determine plasma clearance of exo- and endo-iohexol. This indicates the usefulness of the developed method for application in the field of veterinary clinical practice and research.
authors.affiliation: Dept. of Pharmacology, Toxicology and Biochemistry, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium. Siegrid.debaere@ugent.be
authors.forename: S
authors.initials: S
authors.lastname: De Baere
authors.forename: P
authors.initials: P
authors.lastname: Smets
authors.forename: N
authors.initials: N
authors.lastname: Finch
authors.forename: R
authors.initials: R
authors.lastname: Heiene
authors.forename: P
authors.initials: P
authors.lastname: De Backer
authors.forename: S
authors.initials: S
authors.lastname: Daminet
authors.forename: S
authors.initials: S
authors.lastname: Croubels
mesh_terms: D000818:Animals; D002415:Cats; D011232:Chemical Precipitation; D002626:Chemistry, Pharmaceutical / Q000379:methods* / Q000592:standards; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D004285:Dogs; D007472:Iohexol / Q000737:chemistry* / Q000378:metabolism; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods / Q000592:standards
chemical_list: D007472:Iohexol