Abstract
ABSTRACTComprehensive metabolomic and lipidomic mass spectrometry methods are in increasing demand; for instance, in research related to nutrition and aging. The nematode Caenorhabditis elegans is a key model organism in these fields, owing to the large repository of available C. elegans mutants and their convenient natural lifespan. Here, we describe a robust and sensitive analytical method for the semi-quantitative analysis of >100 polar (metabolomics) and >1000 apolar (lipidomics) metabolites in C. elegans, using a single-sample preparation. Our method is capable of reliably detecting a wide variety of biologically relevant metabolic aberrations in, for example, glycolysis and the tricarboxylic acid cycle, pyrimidine metabolism and complex lipid biosynthesis. In conclusion, we provide a powerful analytical tool that maximizes metabolic data yield from a single sample.This article has an associated First Person interview with the joint first authors of the paper.
Highlights
Considerable advances in high-performance liquid chromatography (HPLC), mass spectrometry (MS), nuclear magnetic resonance (NMR) make it possible to reliably detect tens of thousands of compounds[1]
We demonstrate that metabolomic and lipidomic analysis can be performed on a single sample using a single extraction protocol, reducing sample preparation and throughput time without compromising metabolite identification
Applying these internal standards for the data normalization led to even better linearity of for pyruvate, cytidine monophosphate (CMP), adenosine triphosphate (ATP), and NAD+ (Figure 1J-M, Table S2)
Summary
Considerable advances in high-performance liquid chromatography (HPLC), mass spectrometry (MS), nuclear magnetic resonance (NMR) make it possible to reliably detect tens of thousands of compounds[1]. Metabolomic analysis has seen a surge in popularity over the last decades and the importance and intricacies of metabolism in health and disease are becoming increasingly evident[2]. This has prompted increased demand for reliable and robust metabolomic methods for polar and apolar metabolite analyses in model organisms and human tissues[3]. Metabolic network models for C. elegans were recently constructed[10,11] and a curated consensus is currently being assembled in a European-led consortium[12] The success of such endeavors relies heavily on accurate and robust metabolomics methods[13]
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