TY - JOUR
T1 - 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
AU - Remane, Daniela
AU - Grunwald, Soeren
AU - Hoeke, Henrike
AU - Mueller, Andrea
AU - Roeder, Stefan
AU - von Bergen, Martin
AU - Wissenbach, Dirk K.
PY - 2015/8/15
Y1 - 2015/8/15
N2 - 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 (4.6. mm. ×. 100. mm).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 (45. mg/L), niacinamide 58.5(mg/L), and indole-3-acetic acid (63. mg/L). Comparison of creatinine data obtained by the existing method with those of the developed method showing differences from -120. mg/L to +110. mg/L with a mean of differences of 29.0. mg/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.
AB - 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 (4.6. mm. ×. 100. mm).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 (45. mg/L), niacinamide 58.5(mg/L), and indole-3-acetic acid (63. mg/L). Comparison of creatinine data obtained by the existing method with those of the developed method showing differences from -120. mg/L to +110. mg/L with a mean of differences of 29.0. mg/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.
KW - Creatinine
KW - Hippuric acid
KW - HPLC-DAD
KW - Uric acid
KW - Urine
KW - Validation
UR - http://www.scopus.com/inward/record.url?scp=84935138100&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2015.06.021
DO - 10.1016/j.jchromb.2015.06.021
M3 - Journal article
AN - SCOPUS:84935138100
SN - 1570-0232
VL - 998-999
SP - 40
EP - 44
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
ER -