Quantitative determination of exo- and endo-iohexol in canine and feline samples using high performance liquid chromatography with ultraviolet detection

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• 作者：S. De Baere^a ; ^{Siegrid.debaere@ugent.be} ; P. Smets^b ; ^{Pascale.smets@ugent.be} ; N. Finch^c ; ^{Natalie.finch@bristol.ac.uk} ; R. Heiene^d ; ^{Reidun.heiene@nvh.no} ; P. De Backer^a ; ^{Patrick.debacker@ugent.be} ; S. Daminet^b ; ^{Sylvie.daminet@ugent.be} ; S. Croubels^a ; ^{siska.croubels@ugent.be}
• 关键词：CRS ; Chemical reference substance ; CS ; Cushing's syndrome ; DTPA ; Diethylenetriaminepentaacetic acid ; EDTA ; Ethylenediaminetetraacetic acid ; g ; Goodness-of-fit coefficient ; GFR ; Glomerular filtration rate ; HPLC ; High-performance liquid chromatography ; IS
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：2012
• 出版时间：5 March, 2012
• 年：2012
• 卷：61
• 期：Complete
• 页码：50-56
• 全文大小：578 K

文摘

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 ¦ÌL of trifluoroacetic acid to 100 ¦ÌL of serum/plasma. Following vortexing and centrifugation, an aliquot of the supernatant was injected onto a polymeric PLRP-S column (250 mm ¡Á 4.6 mm i.d., dp: 8 ¦Ìm, 100 ?), maintained at 30 ¡ãC. The mobile phase consisted of water (A) and methanol (B) and a gradient elution (flow-rate: 1.0 mL min^?, 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 ¦Ìg mL^?) and endo-iohexol (0.62-93.0 ¦Ìg mL^?). 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 ¦Ìg mL^? for exo-iohexol and 0.62 and 0.20 ¦Ìg mL^? 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.