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Development of an Oxylipin Library Using Liquid Chromatography-Ion Mobility Quadrupole Time-of-Flight: Application to
Maria Moran-Garrido1, Ameer Y Taha2,3,4, Ángel Gaudioso5
1Centro de Metabolómica y Bioanálisis (CEMBIO), Facultad de Farmacia, Universidad San Pablo-CEU, CEU Universities, Urbanización Montepríncipe, 28660 Boadilla del Monte, Madrid, Spain.
Analytical Chemistry
|February 10, 2025
Summary
This study introduces a comprehensive oxylipin library using LC-IM-QTOF-MS to resolve challenging isomeric separations. Ion mobility significantly enhances the identification of bioactive lipid mediators in complex biological samples.
Area of Science:
- Lipidomics
- Analytical Chemistry
- Biochemistry
Background:
- Oxylipins are critical bioactive lipid mediators derived from polyunsaturated fatty acids (PUFAs).
- Analyzing and identifying oxylipins is difficult due to their numerous isomeric forms.
- Ion mobility coupled with LC-MS offers a powerful approach for separating isomeric compounds.
Purpose of the Study:
- To develop an extensive oxylipin library for improved analysis and identification.
- To evaluate the efficacy of ion mobility in separating oxylipin isomers.
- To establish a reliable method for oxylipin quantification in biological samples.
Main Methods:
- Development of an oxylipin library with retention time (RT), m/z, and collision cross-section (CCS) values for 74 standards.
- Utilized liquid chromatography-ion mobility-quadrupole time-of-flight mass spectrometry (LC-IM-QTOF-MS) in positive and negative ionization modes.
- Investigated various adducts and calculated CCS differences (ΔCCS%) for over 1000 isomeric pairs.
Main Results:
- The library demonstrated the efficacy of ion mobility in separating 15 oxylipin isomer categories.
- Positive ionization mode and sodium adducts significantly improved isomer separation, with 274 pairs baseline separated.
- Application to mouse brain samples successfully identified 19 oxylipins, resolving coeluting isomers using drift time separation.
Conclusions:
- The developed LC-IM-QTOF-MS method and oxylipin library provide high accuracy and reproducibility for oxylipin analysis.
- Ion mobility is crucial for resolving complex oxylipin isomer mixtures.
- This approach enables confident identification and quantification of oxylipins in biological systems.
