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Measurement of Enzymatic and Nonenzymatic Polyunsaturated Fatty Acid Oxidation Products in Plasma and Urine of Macular Degeneration Using LC‐QTOF‐MS / MS
Author(s) -
Leung Ho Hang,
Leung Kin Sum,
Durand Thierry,
Galano JeanMarie,
Lee Jetty ChungYung
Publication year - 2020
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1002/lipd.12264
Subject(s) - isoprostanes , chemistry , polyunsaturated fatty acid , repeatability , enzyme , urine , chromatography , neurodegeneration , biochemistry , fatty acid , lipid peroxidation , medicine , disease
Oxidized polyunsaturated fatty acids (PUFA) are associated to pathogenesis of diseases including cardiovascular and neurodegeneration. The novel products are not only biomarkers but also lipid mediators in gene regulation and signaling pathways. Herein, simultaneous quantitation of 28 products derived from nonenzymatic and enzymatic oxidation of PUFA i.e. 5‐, 15‐F 2t ‐isoprostanes, 7‐, 17‐F 2t ‐dihomo‐isoprostanes, 7‐, 17‐F 2t ‐dihomo‐isofurans, 5‐, 8‐, 18‐F 3t ‐isoprostanes, 4‐, 10‐, 13‐, 14‐, 20‐F 4t ‐neuroprostanes, 5‐, 8‐, 9‐, 11‐,12‐, 15‐, 20‐HETE, 4‐, 7‐, 11‐, 14‐, 17‐HDHA, RvE1, and NPD1 using LC‐(ESI)‐QTOF‐MS/MS was developed. These products were measurable in a single sample and the analytical time was relative short (~15 min). Furthermore, we showed that the use of internal standards is a requisite to normalize matrix effects and preparation loss for the quantitation. Validation assays indicated the method to be robust for plasma and mid‐stream urine sample analysis in particular from those of age‐related macular degeneration subjects, where the accuracy of quantitation displayed good repeatability.