Multiplexed lipid metabolic tracing using click chemistry mass spectrometric reporter molecules

Abstract This protocol provides a method for sensitive multiplexed metabolic tracing. It is based on the combination of alkyne-labeled metabolic precursors and dedicated azido reporter molecules that are optimized for mass spectrometric detection after click reaction. After metabolic incorporation of alkyne fatty acids, sub-picomole to femtomole amounts of labeled material can be detected. Time resolution can be achieved by pulse-chase labeling.