Development and Validation of Targeted Metabolomics Methods Using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) for the Quantification of 235 Plasma Metabolites

Kangkang Xu; Franz Berthiller; Barbara U. Metzler-Zebeli; Heidi E. Schwartz-Zimmermann

doi:10.3390/molecules30030706

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University of Veterinary Medicine Vienna
PHAIDRA
Detail o:3928

Title (eng)

Development and Validation of Targeted Metabolomics Methods Using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) for the Quantification of 235 Plasma Metabolites

Author

Kangkang Xu

Franz Berthiller

Barbara U. Metzler-Zebeli

University of Veterinary Medicine Vienna

Heidi E. Schwartz-Zimmermann

Abstract (eng)

Plasma contains metabolites with diverse physicochemical properties, ranging from highly polar to highly apolar, and concentrations spanning at least nine orders of magnitude. Plasma metabolome analysis is valuable for monitoring health and evaluating medical interventions but is challenging due to the metabolome's diversity and complexity. This study aims to develop and validate targeted LC-MS/MS methods for quantifying 235 mammalian metabolites from 17 compound classes in porcine plasma without prior derivatization. Utilizing reversed-phase and hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry, each analyte is identified and quantified using two selected reaction monitoring (SRM) transitions. Fast polarity switching and scheduled SRM enhance the metabolome coverage and throughput, enabling the analysis of one sample in about 40 min. A simple "dilute and shoot" sample preparation protocol was employed, with samples injected at two dilution levels to align metabolite concentrations within calibration curve ranges. Validation in porcine plasma included assessments of carryover, linearity, detection and quantification limits, repeatability and recovery. The method was further applied to plasma samples from various animal species, demonstrating its applicability to human and animal studies. This study establishes two robust LC-MS/MS methods for comprehensive porcine plasma metabolome quantification, advancing large-scale targeted metabolomics in biomedical research.

Keywords (eng)

Tandem Mass Spectrometry MethodsAnimalsMetabolomics MethodsSwineMetabolomeChromatography, Liquid MethodsPlasma Chemistry MetabolismReproducibility of ResultsLiquid Chromatography-Mass Spectrometry

Type (eng)

journal article

Language

[eng]

Persistent identifier

https://phaidra.vetmeduni.ac.at/o:3928

DOI

10.3390/molecules30030706

Is in series

Title (eng)

Molecules

Volume