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Capillary electrophoresis with dual diode array detection and tandem mass spectrometry to access cardiovascular biomarkers candidates in human urine: trimethylamine-N-Oxide and L-carnitine
Zuzana Cieslarova ,
Mariana Magaldi ,
Lucélia Alcantara Barros ,
Claudimir Lucio do Lago ,
Daniel Rossado Oliveira ,
Francisco Antonio Helfenstein Fonseca ,
Maria Cristina Izar ,
Aline Soriano Lopes ,
Marina Franco Maggi Tavares ,
Aline Klassen
November 2018
November 2018
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Abstract
A capillary electrophoresis with diode array and tandem mass spectrometry detection
(CE-UV-MS/MS) method has been developed for the targeted assessment of cardiovascular
biomarkers candidates, trimethylamine-N-Oxide (TMAO) and l-carnitine, and creatinine
in human urine samples. The dual detection was applied due to the high concentration
of creatinine (monitored by UV detection at 200 nm) in relation to TMAO and l-carnitine
(quantified by selected reaction monitoring (SRM) mass spectrometry), in human urine.
All instrumental parameters, sheath liquid (SHL) and background electrolyte (BGE)
compositions were optimized with a pool of urine provided by adult healthy volunteers
and evaluated by signal-to-noise ratio (SNR) and peak shape of TMAO. The compositions
for the optimized BGE was formic acid at concentration of 0.10 mol L-1, and for SHL
was 70:30 MeOH:H2O containing 0.05% (v/v) formic acid, delivered at a flow rate of
5 μL min-1. Limits of detection for TMAO, l-carnitine and creatinine were 0.76, 0.54
and 303 μmol L-1, respectively. Limits of quantification were 2.5, 1.8 and 1000 μmol L-1,
respectively. Linearity was evaluated by ANOVA and presented R2 from 0.993 to 0.997.
Precision and accuracy were evaluated at three concentration levels. Coefficients
of variation (CV) from 1 to 21% were obtained for the intra-day precision evaluation
and from 2 to 16% for the inter-day precision evaluation. The recovery ranged from
75 to 116%. Quantitation of TMAO and l-carnitine in infarcted patients urine in comparison
to healthy individuals indicated a 2.2 fold increase of TMAO and a 7.0 fold increase
of l-carnitine. These results showed the potential applicability of the proposed method
for the evaluation of TMAO and l-carnitine in urine within a panel of candidate metabolites
in targeted metabolomics studies of cardiovascular diseases among other conditions.