title: Validation of a multi-analyte HPLC-DAD method for determination of uric acid, creatinine, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid and 2-methylhippuric acid in human urine.
abstract: During the last decades exposure sciences and epidemiological studies attracts more attention to unravel the mechanisms for the development of chronic diseases. According to this an existing HPLC-DAD method for determination of creatinine in urine samples was expended for seven analytes and validated. Creatinine, uric acid, homovanillic acid, niacinamide, hippuric acid, indole-3-acetic acid, and 2-methylhippuric acid were separated by gradient elution (formate buffer/methanol) using an Eclipse Plus C18 Rapid Resolution column (4.6mm×100mm). No interfering signals were detected in mobile phase. After injection of blank urine samples signals for the endogenous compounds but no interferences were detected. All analytes were linear in the selected calibration range and a non weighted calibration model was chosen. Bias, intra-day and inter-day precision for all analytes were below 20% for quality control (QC) low and below 10% for QC medium and high. The limits of quantification in mobile phase were in line with reported reference values but had to be adjusted in urine for homovanillic acid (45mg/L), niacinamide 58.5(mg/L), and indole-3-acetic acid (63mg/L). Comparison of creatinine data obtained by the existing method with those of the developed method showing differences from -120mg/L to +110mg/L with a mean of differences of 29.0mg/L for 50 authentic urine samples. Analyzing 50 authentic urine samples, uric acid, creatinine, hippuric acid, and 2-methylhippuric acid were detected in (nearly) all samples. However, homovanillic acid was detected in 40%, niacinamide in 4% and indole-3-acetic acid was never detected within the selected samples.
authors.affiliation: Institute of Forensic Medicine, University Hospital Jena, Jena, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Remane
authors.affiliation: Department of Metabolomics, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany.
authors.forename: Soeren
authors.initials: S
authors.lastname: Grunwald
authors.affiliation: Department of Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy, University of Leipzig, Leipzig, Germany.
authors.forename: Henrike
authors.initials: H
authors.lastname: Hoeke
authors.affiliation: Department of Proteomics, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Mueller
authors.affiliation: Department of Environmental Immunology, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Roeder
authors.affiliation: Department of Metabolomics, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany; Department of Proteomics, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany; Department of Biotechnology, Chemistry and Environmental Engineering Aalborg University, Aalborg, Denmark.
authors.forename: Martin
authors.initials: M
authors.lastname: von Bergen
authors.affiliation: Department of Metabolomics, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany. Electronic address: dirk.wissenbach@ufz.de.
authors.forename: Dirk K
authors.initials: DK
authors.lastname: Wissenbach
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D003404:Creatinine / Q000652:urine*; D006626:Hippurates / Q000652:urine*; D006719:Homovanillic Acid / Q000652:urine*; D006801:Humans; D007210:Indoleacetic Acids / Q000652:urine*; D009536:Niacinamide / Q000652:urine*; D014527:Uric Acid / Q000652:urine*
chemical_list: D006626:Hippurates; D007210:Indoleacetic Acids; D009536:Niacinamide; D014527:Uric Acid; C026113:2-methylhippuric acid; C030737:indoleacetic acid; D003404:Creatinine; C030514:hippuric acid; D006719:Homovanillic Acid
keywords: Creatinine; HPLC-DAD; Hippuric acid; Uric acid; Urine; Validation