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ARS Home » Plains Area » Grand Forks, North Dakota » Grand Forks Human Nutrition Research Center » Dietary Prevention of Obesity-related Disease Research » Research » Publications at this Location » Publication #364241

Title: Simplified mass spectrometric analysis of ceramides using a common collision energy

item Picklo, Matthew
item Hanson, Benjamin
item Bukowski, Michael

Submitted to: Lipids Journal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/17/2019
Publication Date: 7/24/2019
Publication URL:
Citation: Picklo, M.J., Hanson, B.K., Bukowski, M.R. 2019. Simplified mass spectrometric analysis of ceramides using a common collision energy. Lipids Journal.

Interpretive Summary: Ceramides are fatty acid metabolites associated with diseases such as insulin resistance and cancer. Accurate measurement of ceramides requires an advanced method using mass spectrometry. In this work, we developed a simplified mass spectrometry method for the analysis of ceramides in plasma and liver. Use of this simplified method reduces sample processing time and reduces the use of purified ceramides standards, thus creating a savings in human resources and funds.

Technical Abstract: Ceramides (CER) are biologically active sphingolipid precursors that are mechanistically linked to several pathogenic states including cancer, insulin resistance, and neurodegeneration. CER are commonly quantified through mass spectrometry-based methods founded upon a precursor ion scan (PIS) in positive mode to produce a characteristic m/z 264 ion. The ionization efficiency (IE) CER species decreases with an increase in CER mass, thus quantitation of CER typically involves application mass dependent-response factors (RF) for each CER species. In this work, we observed that the RF were systematically dependent on the number of fatty acid acyl carbons and the collision energy (CE) used to generate the m/z 264 ion. Using these complimentary trends, we determined an “isosbestic” CE where the RF for all CER species were equivalent, allowing for CER quantitation without post-collection correction factors. A comparison of this common CE/common RF method to the multiple RF method demonstrated good agreement between the two methods. Use of the common CE/common RF method will reduce data processing and the reduce the use of multiple CER species standards.