Application Note 54
Targeted MRM Screening of U-13C Lipid Yeast Extracts for Robust Lipidomics Applications
Andrew J. Percy1, Nyasha Munjoma2, David Heywood2, Lee A. Gethings2, Krista Backiel1
1. Cambridge Isotope Laboratories, Inc. Tewksbury, MA USA
2. Waters Corporation, Manchester UK
Introduction
Fatty acids (FAs) and lipids are important biological compounds that are essential to the regulation and control of cellular functions (e.g., membrane structure, signal transduction, apoptosis) and metabolic pathways (e.g., fatty acid oxidation, ketone body synthesis, lipolysis).1 These biomolecules are also functionally important to an organism’s energy production and storage. Qualitative/quantitative analysis of these molecular species is a continual aim of lipidomics research. A driving factor is to better understand their underlying pathophysiology, as well as to identify new biomarkers (e.g., cardiovascular disease, diabetes, cancer) or diagnose existing ones toward improved precision / personalized medicine.2-4
Regardless of the research objective(s), a growing number of lipidomics methods utilize hyphenated analytical systems comprising a separation dimension (e.g., reversed-phase liquid chromatography, RPLC; hydrophilic interaction liquid chromatography, HILIC) and a mass spectrometer (operated in different acquisition modes e.g., multiple reaction monitoring or MRM).5 Critical to the qualitative/quantitative analysis is the incorporation of stable isotope-labeled standards. Among the benefits, their use helps determine an assay’s effectiveness (i.e., process efficiency), improve measurement precision, and enhance quantitative accuracy.
In this work, we qualified a condensed panel of stable isotope-labeled FAs / lipids in U-13C crude lipid yeast extracts (CIL catalog no. L-ISO1) on robust, high-throughput, analytical platforms (i.e., RPLC- and HILIC-MRM/MS). The verification results of this extract analyses are presented here along with a discussion of its potential use in different types of MS lipidomics applications.
Highlights
• Crude lipid yeast extracts (U-13C and unlabeled) comprise 100s of fatty acids and lipids
• Extracts afford broad application potential (e.g., quality control, credentialing, quantification)
• Targeted methods, results, and applications for the U-13C lipid yeast extract are described
• Methodologies are rapid (8-12 min) and simple to execute
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