ONE STEP CHEMICAL DERIVATIZATION OF SECRETED CELL METABOLITES FOR SENSITIVE MASS SPECTROMETRY DETECTION

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Authors

PEČINKA Lukáš MORÁŇ Lukáš POROKH Volodymyr MARCHETTI-DESCHMANN Martina HAVEL Josef HAMPL Aleš VAŇHARA Petr

Year of publication 2022
Type Conference abstract
Citation
Description Metabolomics studies for biomarker discovery consist of a complete set of metabolites in complex biological systems (databases contain ~ 115 000 metabolites). Metabolome is much smaller than proteome (1 800 000 different proteins) making it relatively simple for data analysis. On the other hand, metabolites are chemically diverse compounds that occur at vide range of concentrations (ten orders of magnitude) which makes metabolomics analysis challenging. Various chemical derivatization procedures have been extensively studied to enhance ionization efficacy, retention time (chromatography), and shift molecular mass to a higher value (mass spectrometry). Functional groups including aldehyde, primary amines, and carboxylic acid have been targeted using different derivatization reagents. Around 80% of a total number of described metabolites can be derivatized and detected by mass spectrometry using i.e. Girard´s reagent T – GT (aldehyde groups), Coniferyl aldehyde – CA (primary amines), and 2-picolylamine – 2-PA (carboxylic acids). Metabolomics methods are highly used to analyze large-scale phenotypic changes in the cell from intracellular (cell lysate) or extracellular (cell secretome) biological matrices. Cell cultivation process is accompanied by secretion of a broad range of metabolites into the cultivation media (i.e., autocrine and paracrine factors such as immune receptors, cytokines, and hormones) which are characteristic for individual cell types. Herein, the method for derivatization of various classes of small molecules has been optimized and compared to the detection sensitivity of non-derivatized molecules. The secreted metabolites profile of human embryonic stem cells (hESCs) has been analyzed through multiple derivatized procedures coupled with metabolomics with the aim of a better understanding of hESCs therapeutical effects.
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