In-depth triacylglycerol profiling using MS3 Q-Trap mass spectrometry

Total triacylglycerol (TAG) level is a key clinical marker of metabolic and cardiovascular diseases. However, the roles of individual TAGs have not been thoroughly explored in part due to their extreme structural complexity. We present a targeted mass spectrometry-based method combining multiple reaction monitoring (MRM) and multiple stage mass spectrometry (MS3) for the comprehensive qualitative and semiquantitative profiling of TAGs. This method referred as TriP-MS3 -triacylglycerol profiling using MS3 -screens for more than 6,700 TAG species in a fully automated fashion. TriP-MS3 demonstrated excellent reproducibility (median interday CV similar to 0.15) and linearity (median R-2 = 0.978) and detected 285 individual TAG species in human plasma. The semiquantitative accuracy of the method was validated by comparison with a state-of-the-art reverse phase liquid chromatography (RPLC)-MS (R-2 = 0.83), which is the most commonly used approach for TAGs profiling. Finally, we demonstrate the utility and the versatility of the method by characterizing the effects of a fatty acid desaturase inhibitor on TAG profiles in vitro and by profiling TAGs in Caenorhabditis elegans. (C) 2021 The Authors. Published by Elsevier B.V.

Automated summary

TriP-MS3 is a mass spectrometry-based method that allows for comprehensive qualitative and semiquantitative profiling of over 6,700 TAG species. This method demonstrated excellent reproducibility and linearity, detecting 285 individual TAG species in human plasma. The accuracy of the method was validated by comparison with a state-of-the-art reverse phase liquid chromatography (RPLC)-MS.