Measurement of succinyl-carnitine and methylmalonyl-carnitine on dried blood spot by liquid chromatography-tandem mass spectrometry
| Autor: | Sara Boenzi, Cristiano Rizzo, Giancarlo la Marca, Eleonora Zelli, David W. Johnson, Rita Inglese, Carlo Dionisi-Vici, Tegra Barreiro Martinez, Maurizio Muraca |
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| Rok vydání: | 2014 |
| Předmět: |
Male
Adolescent Clinical Biochemistry Methylmalonic acidemia Biochemistry Limit of Detection Tandem Mass Spectrometry Liquid chromatography–mass spectrometry Carnitine medicine Humans Child Detection limit Chromatography Chemistry Biochemistry (medical) Selected reaction monitoring Infant Newborn Infant General Medicine medicine.disease Confidence interval Dried blood spot Methylmalonic aciduria Child Preschool Linear Models Female Dried Blood Spot Testing Chromatography Liquid medicine.drug |
| Zdroj: | Università degli studi di Firenze-IRIS |
| ISSN: | 0009-8981 |
| DOI: | 10.1016/j.cca.2013.11.016 |
| Popis: | Methylmalonic aciduria (MMA) is one of the most frequent organic acidurias, a class of diseases caused by enzymatic defects mainly involved in the catabolism of branched-chain amino acids. Recently, mild MMA and C4-dicarboxylyl-carnitine (C4DC-C) accumulation have been reported in patients carrying mutation in genes encoding the α-subunit (SUCLG1) and the β-subunit (SUCLA2) of the ADP-forming succinyl-CoA synthetase (SCS). We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify in dried blood spot the two isobaric compounds of C4DC-C, succinyl-carnitine and methylmalonyl-carnitine, to allow the differential diagnosis between classical MMA and SCS-related defects. This method, with an easy liquid-phase extraction and derivatization procedure, has been validated to demonstrate the specificity, linearity, recovery, lowest limit of quantification (LLOQ), accuracy and precision for quantitative determination of blood succinyl-carnitine and methylmalonyl-carnitine. The assay was linear over a concentration range of 0.025-10 μmol/L and achieved the LLOQ of 0.025 μmol/L for both metabolites. The average slope, intercept, and coefficient of linear regression (r(2)) were respectively: 0.3389 (95% confidence interval 0.2888-0.3889), 0.0113 (95% confidence interval -0.0157 to 0.0384), 0.9995 (95% confidence interval 0.9990-1.0000) for succinyl-carnitine and 0.5699 (95% confidence interval 0.5263-0.6134), 0.0319 (95% confidence interval -0.0038 to 0.0677), 0.9997 (95% confidence interval 0.9995-1.0000) for methylmalonyl-carnitine. Within-day and between-day coefficients of variation (CV) were 1.94% and 3.19% for succinyl-carnitine and 3.21%, and 2.56 for methylmalonyl-carnitine. This method is accurate and provides a new tool to differentiate patients with classical methylmalonic acidemia from those with SCS-related defects. |
| Databáze: | OpenAIRE |
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