ausblenden:
Schlagwörter:
Animals
Caenorhabditis elegans/*chemistry
Chromatography, High Pressure Liquid/methods
Complex Mixtures/*analysis
Folic Acid/*analogs & derivatives/*analysis/metabolism
Mass Spectrometry/methods
Metabolic Networks and Pathways
Sensitivity and Specificity
Tetrahydrofolate Dehydrogenase/genetics
Apci
C. elegans
Esi
dhfr-1
folic acid
Zusammenfassung:
Folic acid (FA) plays a vital role in central metabolism, including the one carbon cycle, nucleotide, and amino acid biosynthesis. The development of sensitive, accurate analytical methods to measure FA intermediates in tissues is critical to understand their biological roles in diverse physiological and pathological contexts. Here, we developed a highly sensitive method for the simultaneous quantification of FA intermediates in the nematode Caenorhabditis elegans as a model to dissect metabolic networks. The method was further validated by analyzing the worm folate pool upon RNAi knockdown of the dihydrofolate reductase gene dhfr-1. Comparative mass spectrometry behavior of the FA analogs using two different ion sources, electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI), revealed ESI-MS/MS to be more sensitive, but APCI-MS provided more detailed structure inferences, which can elucidate chemical investigation and synthesis of FA analogs. Finally, we report on the use of in vitro oxidation coupled with high-resolution mass spectrometry as a tool to discover new endogenous FA derivatives in the nematode.
