Simple high-performance liquid chromatographic method to analyze serum creatinine has several advantages over the Jaffe picric acid reaction as demonstrated with a cimetidine dose response in rhesus monkeys

A simple method for creatinine determination was developed using high-perfo rmance liquid chromatography (HPLC) to more accurately monitor serum creati nine levels in experimental animal models when compared to the Jaffe method . The new HPLC procedure will replace the traditional Jaffe method for rhes us monkey kidney function studies. We developed an isocratic method using a polymeric, hydrophilic, silica-based strong cation-exchange bed with a 5.0 mmol/l lithium acetate matrix, pH 4.9, which isolates creatinine with no d etectable impurities as determined by three-dimensional ultraviolet-visible spectral analysis. Sample preparation includes deproteination with acetoni trile, evaporation, and resolubilization in mobile phase followed by quanti tation with UV detection at 234 nm. Extraction efficiency across the measur ed range was 96 +/-2%. From numerous extracted rhesus monkey creatinine cur ves (n=38) a slope of 251 100 +/- 756 (95% CI) and an intercept of 675.6 +/ - 712.7 (95% CI) was calculated. Extraction efficiency and peak purity test s with human plasma were cross-compared with rhesus monkey serum producing equivalent results. An average of 120 samples can be run daily. (C) 2001 El sevier Science B.V. All rights reserved.