Contents

Diagnostic-Fragment-Gated Reporter-Ion Normalization for Quantitative Mapping of Glycerophospholipid sn-Positional Isomers

Author(s): Louis E. Brus1
1Department of Chemistry, Columbia University, New York, New York 10027, USA
Louis E. Brus
Department of Chemistry, Columbia University, New York, New York 10027, USA

Abstract

Isomer-resolved lipidomics needs a direct analytical connection between structural assignment and relative abundance. Summed-composition names can merge distinct phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol arrangements into one lipid entry even when fatty-acyl placement on glycerol carries biochemical significance. This work establishes a diagnostic-fragment-gated version of Diagnostic Reporter-Ion Normalization (DRIN) for multistage mass spectrometry, in which quantitative ratios are accepted only when reporter ions arise from an assigned sn-positional fragment. The central question is whether reporter-ion measurements can be converted into calibrated and chemically interpretable lipid-isomer maps while keeping response linearity, matrix transfer, and plasma directionality as separate analytical claims. The measurement set comprises PC 18:1/16:0 calibration values, a two-concentration Escherichia coli lipid-extract comparison, and a human plasma phosphatidylcholine isomer profile. PC 18:1/16:0 produced near-proportional reporter response at both lipid and sn-isomer levels, with slopes of 1.0906 and 1.1274. In the bacterial extract comparison, 15 assigned PG and PE isomers recovered the intended two-fold relation with 87.0–97.5% accuracy. In plasma, PC 18:0/18:2, PC 16:0/18:2, PC 18:1/16:0, and PC 18:0/18:1 showed the strongest enrichment, whereas PC 14:0/20:4, PC 16:0/20:5, and PC 18:0/22:6 showed pronounced depletion. These findings answer the analytical question by showing that reporter-ion ratios become defensible isomer ratios only when diagnostic-fragment identity, calibration behavior, and matrix compatibility are evaluated together.

Keywords: analytical lipidomics; glycerophospholipid; sn-positional isomer; isobaric mass tag; diagnostic reporter ion; MS3; plasma phosphatidylcholine; Escherichia coli
Copyright © 2024 Louis E. Brus. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.