A novel reversed-phase HPLC method for the determination of urinary creatinine by pre-column derivatization with ethyl chloroformate: comparative studies with the standard Jaffé and isotope-dilution mass spectrometric assays

• Published in: Analytical and bioanalytical chemistry Vol. 406; no. 6; pp. 1807 - 1812
• Main Authors: Leung, Elvis M. K., Chan, Wan
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-02-2014; Springer; Springer Nature B.V
• Subjects: Analysis; Analytical Chemistry; Animals; Biochemistry; Biomarkers; Biomonitoring; Care and treatment; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chloroformate; Chromatography; Chromatography, High Pressure Liquid - methods; Colorectal cancer; Comparative studies; Creatinine; Creatinine - urine; Diagnosis; Food Science; Formic Acid Esters - chemistry; Fractionation; Glomerular filtration rate; High performance liquid chromatography; Hydrophobicity; Indicator Dilution Techniques; Laboratory Medicine; Limit of Detection; Liquid chromatography; Male; Mass spectrometry; Mass Spectrometry - methods; Metabolites; Methods; Monitoring/Environmental Analysis; Normalizing; Rats; Rats, Sprague-Dawley; Renal function; Scientific imaging; Spectrometers; Spectroscopy; Urine; Hong Kong China; China; Creatinine; Kidney function diagnosis; Urine normalization; High-performance liquid chromatography; Pre-column derivatization
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-013-7592-8

Creatinine is an important biomarker for renal function diagnosis and normalizing variations in urinary drug/metabolites concentration. Quantification of creatinine in biological fluids such as urine and plasma is important for clinical diagnosis as well as in biomonitoring programs and urinary metabolomics/metabonomics research. Current methods for creatinine determination either are nonselective or involve the use of expensive mass spectrometers. In this paper, a novel reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of creatinine of high hydrophilicity by pre-column derivatization with ethyl chloroformate is presented.
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-Ethyloxycarbonylation of creatinine significantly enhanced the hydrophobicity of creatinine, facilitating its chromatographic retention as well as quantification by HPLC. Factors governing the derivatization reaction were studied and optimized. The developed method was validated and applied for the determination of creatinine in rat urine samples. Comparative studies with isotope-dilution mass spectrometric method revealed that the two methods do not yield systematic differences in creatinine concentrations, indicating the HPLC method is suitable for the determination of creatinine in urine samples.
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