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Development and validation of a HILIC‐MS/MS multitargeted method for metabolomics applications
Author(s) -
Virgiliou Christina,
Sampsonidis Ioannis,
Gika Helen G.,
Raikos Nikolaos,
Theodoridis Georgios A.
Publication year - 2015
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201500208
Subject(s) - hydrophilic interaction chromatography , chromatography , bioanalysis , chemistry , metabolomics , metabolite , analyte , triple quadrupole mass spectrometer , elution , metabolome , mass spectrometry , protein precipitation , high performance liquid chromatography , tandem mass spectrometry , selected reaction monitoring , biochemistry
The paper reports the development of a multianalyte method and its application in metabolic profiling of biological fluids. The initial aim of the method was the quantification of metabolites existing in cell culture medium used in in‐vitro fertilization (IVF) and in other biological fluids related to embryo growth. Since most of these analytes are polar primary metabolites a hydrophilic interaction liquid chromatography system was selected. The analytical system comprised Ultra‐HPLC with detection on a triple quadrupole mass spectrometer operating in both positive and negative modes. Mobile phase and gradient elution conditions were studied with the aim to achieve the highest coverage of metabolic space in a single injection namely the largest number of analytes that could be detected and quantified. The developed method provides absolute quantitation of ca. 100 metabolites belonging to key metabolite classes such as sugars, aminoacids, nucleotides, organic acids, and amines. Following validation, the method was applied for the metabolic profiling of hundreds of samples of spent culture medium originating from human IVF procedures and several hundreds of biological samples such as amniotic fluid, human urine and blood serum from pregnant women. The bioanalytical end‐point was to provide assistance in the process of embryo transfer and improving IVF success rates but also to provide insight in complications related to the subsequent embryo growth during pregnancy.