A simple desalting method for direct MALDI mass spectrometry profiling of tissue lipids

Abstract

Direct MALDI-mass spectrometry (MALDI-MS) profiling of tissue lipids often observes isobaric phosphatidylcholine (PC) species caused by the endogenous alkali metal ions that bias the relative abundance of tissue lipids. Fresh rat brain cryosections were washed with 70% etha­nol (EtOH), water (H2O), or 150 mM ammonium acetate (NH4Ac), and the desalting effectiveness of each fluid was evaluated by MALDI-MS profiling of PC and sphingomyelin (SM) species in tissue and in the washing runoff. The results indicated that EtOH and H2O only partially desalted the tissue lipids, yet both substantially displaced the tissue lipids to the washing runoffs. On the other hand, NH4Ac effectively desalted the tissue lipids and produced a runoff containing no detectable PCs or SMs. NH4Ac wash also unveiled the underlying changes of PCs and SMs in the infarcted rat cortex previously masked by edema-caused increase of tissue sodium. The MS/MS of an isobaric PC in the infarcted cortex revealed the precursor change as the result of NH4Ac wash and confirmed the desalting effectiveness of such wash. Other than desalting, NH4Ac wash also removes contaminants in tissue, enhances the overall spectral quality, and benefits additionally in profiling of biological molecules in tissue.

Keywords

• desalting of tissue section
• direct tissue profiling
• matrix-assisted laser desorption/ionization