Rapid determination of creatinine in serum and urine by ion-pair high-performance liquid chromatography
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We report a simple and reliable high performance liquid chromatography method for measuring creatinine in serum and urine. The chromatographic run is performed on a C18 column after protein precipitation with acetone and addition of cimetidine as an internal standard. The separation is carried out in 20 min at a flow rate of 0.8 ml/min, with a mobile phase consisting of 100 mmol/l sodium dihydrogen phosphate solution, containing 30 mmol/l sodium lauryl sulfate pH 3.0 and acetonitrile (60:36, v/v). The absorbance is monitored at 200 nm. The relationship between creatinine concentration and the creatinine/internal standard peak area is linear up to 1,088 µmol/l. Within-run precision measured at three different creatinine concentrations ranges from 0.89% to 2.34% in serum and from 0.34% to 1.10% in urine. Between-run precision varies from 1.68% to 3.17% in serum and from 1.58% to 1.85% in urine over a wide range of concentrations. Analytical recovery is between 98.71% and 101.25% in serum and between 98.96% and 100.27% in urine. The detection limit is 3.24 µmol/l for a signal-to-noise ratio of 3. The method shows a good linearity with the reference isotope dilution gas chromatography-mass spectrometry procedure (r=0.999), without interferences, even in the presence of high bilirubin concentrations.
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- Rapid determination of creatinine in serum and urine by ion-pair high-performance liquid chromatography
International Journal of Clinical and Laboratory Research
Volume 29, Issue 3 , pp 103-109
- Cover Date
- Print ISSN
- Online ISSN
- Additional Links
- High-performance liquid chromatography
- Gas chromatography-mass spectrometry
- Author Affiliations
- 1. Department of Pediatrics, University Hospital of Padua, Via Giustiniani 3, I-35128, Padua, Italy
- 2. Department of Laboratory Medicine, University of Padua, Via Giustiniani 3, I-35128, Padua, Italy