Development of a candidate reference method for the simultaneous quantification of betaine, choline and trimethylamine N-oxide in serum samples by two-dimensional liquid chromatography and isotope dilution tandem mass spectrometry
Daniela Pineda-Cevallos, María Castañón Apilánez, Elena López-Cancio, Belén Prieto García, J. Ignacio García Alonso, Pablo Rodríguez-González
TL;DR
A new method was developed to measure betaine, choline, and trimethylamine N-oxide in serum, which could help predict cardiovascular risks.
Contribution
The novel method combines two-dimensional liquid chromatography and isotope dilution tandem mass spectrometry for simultaneous quantification of three metabolites in serum.
Findings
The method was validated according to CLSI guidelines for accuracy and reliability.
The technique was applied to 74 serum samples from ischemic stroke patients to study these metabolites as potential biomarkers.
The approach enables rapid and precise separation of the three analytes using a two-dimensional chromatography setup.
Abstract
Serum choline, betaine and trimethylamine N-oxide levels are associated with the risk of cardiovascular events. However, no reference procedure for the determination of these compounds in serum has been developed so far. This work describes the combination of IDMS and two-dimensional liquid chromatography operating in multiple heart-cutting mode for the simultaneous quantification of the three compounds in human serum by isotope dilution tandem mass spectrometry. A reversed-phase separation is proposed as the first dimension and coupled with cation exchange chromatography in the second dimension. The online isolation of the single fraction in which the three analytes co-elute from the first dimension enables a rapid chromatographic separation in the second dimension through cation exchange. The method was validated according to the Clinical and Laboratory Standards Institute (CLSI)…
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Taxonomy
TopicsMetabolomics and Mass Spectrometry Studies · Analytical Chemistry and Chromatography · Electrochemical sensors and biosensors