Michel Boutin1, Nancy Presse2,3,4, Tristan Martineau1, Audrey Perreault2, Pierrette Gaudreau5,6, Christiane Auray-Blais1. 1. Division of Medical Genetics, Department of Pediatrics, Centre de Recherche-CHUS, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke J1H 5N4, QC, Canada. 2. Research Center on Aging, Centre Intégré Universitaire de santé et de Services Sociaux de l'Estrie-CHUS, Sherbrooke J1H 4C4, QC, Canada. 3. Department of Community Health Sciences, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke J1H 5N4, QC, Canada. 4. Centre de recherche, Institut Universitaire de Gériatrie de Montréal, Montréal H3W 1W5, QC, Canada. 5. Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal H2X 0A9, QC, Canada. 6. Département de Médecine, Faculté de Médecine, Université de Montréal, Montréal H2X 0A9, QC, Canada.
Abstract
Aim: Vitamin B12 deficiency is characterized metabolically by increased serum and urine methylmalonic acid (MMA). Urinary MMA/creatinine ratio is suggested for screening for metabolic vitamin B12 deficiency in older populations. Results: A UPLC-MS/MS method for the analysis of urinary MMA and creatinine was developed/validated. A good separation of MMA from succinic acid, its structural isomer, was achieved. Intra- and interday accuracy biases and precision coefficients were all ≤6.3% for MMA and creatinine. Urine and serum samples of 34 individuals of the NuAge Biobank were analyzed for technical comparisons showing that urinary MMA/creatinine ratios by UPLC-MS/MS strongly correlated with GC-MS values, and with serum MMA values. Conclusion: The UPLC-MS/MS method developed is rapid/reliable for the analysis of urinary MMA/creatinine ratios.
Aim: Vitamin B12 deficiency is characterized metabolically by increased serum and urine methylmalonic acid (MMA). Urinary MMA/creatinine ratio is suggested for screening for metabolic vitamin B12 deficiency in older populations. Results: A UPLC-MS/MS method for the analysis of urinary MMA and creatinine was developed/validated. A good separation of MMA from succinic acid, its structural isomer, was achieved. Intra- and interday accuracy biases and precision coefficients were all ≤6.3% for MMA and creatinine. Urine and serum samples of 34 individuals of the NuAge Biobank were analyzed for technical comparisons showing that urinary MMA/creatinine ratios by UPLC-MS/MS strongly correlated with GC-MS values, and with serum MMA values. Conclusion: The UPLC-MS/MS method developed is rapid/reliable for the analysis of urinary MMA/creatinine ratios.
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Keywords:
MMA; UPLC–MS/MS; creatinine; liquid chromatography; mass spectrometry; method development and validation; methylmalonic acid; urine; vitamin B12