RESUMO
A sensitive and selective method for the separation and quantification of the three organic acids 3-hydroxy-3-methylglutaric acid, 3-methylglutaric acid, and glutaric acid in human urine samples by CE with mass spectrometry detection has been developed. This methodology is faster, simpler and less time-consuming, than other methodologies previously described, and requires of reduced amounts of reagents as well. Samples are first filtered and then diluted in water. For the electrophoretic separation, a 20mM ammonium acetate and 10% methanol solution at pH 9.1 was selected as the running electrolyte. With 5-s hydrodynamic injection, detection limits ranging from 15.5 to 39.3µM and linear responses ranging from the LOQ calculated for each analyte to more than 400µM were obtained for the analysis of the different organic acids in less than 13min. Remarkable selectivity is achieved by mass spectrometry detection using 0.25% of formic acid in 50% v/v 2-propanol-water solution as sheath liquid, and enough sensitivity without interferences from the matrices was obtained as well. This methodology has revealed as an efficient approach to help the 3-hydroxy-3-methylglutaric aciduria diagnoses in order to discard or confirm the occurrence of the disease as of the presence or absence of the expected increased levels of these analytes in samples of potential patients.
