Novel HILIC-ESI-MS method for urinary profiling of MSUD and methylmalonic aciduria biomarkers

Mathew, Elizabeth Mary and Lewis, Leslie Edward Simon and Rao, Pragna and Nalini, K and Kamath, Asha and Moorkoth, Sudheer (2019) Novel HILIC-ESI-MS method for urinary profiling of MSUD and methylmalonic aciduria biomarkers. Journal of Chromatographic Science, 57 (8). pp. 1-9. ISSN 0021-9665

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Abstract

Methyl malonic acid and branched-chain keto acids are important biomarkers for the diagnosis of cobalamin deficiencies and maple syrup urine disease. We report the development and validation of a HILIC-ESI-MS2 method for the quantification of these organic acids from neonatal urine. The samples were 100 times diluted and analyzed on a ZIC- ILIC column with 25-mM formic acid in water: 25-mM formic acid in acetonitrile (45:55) at a flow rate of 0.8 mL/min with a runtime of only 6 minutes. The method demonstrated a lower limit of detection of 10 ng/mL, Limit of Quantification (LOQ) of 50 ng/mL, linearity of r2 ≥ 0.990 and recoveries of 87–105% for all analytes.The intraday and interday precision CV’s were <10% and 12%, respectively. Extensive stability studies demonstrated the analytes to be stable in stock and in matrix with a percent change within ±15%. The Bland–Altman analysis of the developed method with the gold standard GCMS method demonstrated a bias of 0.44, 0.11, 0.009 and –0.19 for methyl malonic acid, 3-methyl-2-oxovaleric acid, 2- ydroxy-3methylbutyric acid and 4-methyl-2-oxovaleric acid, respectively, proving the methods are comparable. The newly developed method involves no derivatization and has a simple sample preparation and a low runtime, enabling it to be easily automated with a high sample throughput in a cost-effective manner

Item Type: Article
Subjects: Medicine > KMC Manipal > Biochemistry
Departments at MU > Statistics
Medicine > KMC Manipal > Paediatrics
Pharmacy > MCOPS Manipal > Pharmaceutical Quality Assurance
Depositing User: KMC Library
Date Deposited: 06 Aug 2019 09:29
Last Modified: 06 Aug 2019 09:29
URI: http://eprints.manipal.edu/id/eprint/154286

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