Effects of intraperitoneally administered chromanols on pulmonary levels of lipid mediators in ovalbumine sensitized mice

Female mice were pretreated i.p. with α-T-13'-COOH (α-13′-carboxychromanol) and α-AC (α-amplexichromanol) (10 mg kg-1) or vehicle (DMSO 2%, 0.5 ml) 30 min before each OVA (ovalbumine) challenge. Animals were sacrificed at day 14 to evaluate in the lung homogenate COX products and 12/15-LOX-derived products formed from arachidonic acid (12-HETE and 15-HETE), eicosapentaenoic acid (12-HEPE and 15-HEPE) and docosahexaenoic acid (17-HDHA, 14-HDHA) analyzed by UPLC-MS/MS.

Raw analyst files (.wiff and .wiff.scan) of the UPLC-MS/MS results were uploaded, together with an excel file for the sample list.

The methods and results were published in Cerqua et al., Pharmacol. Res.,  2022 Jul;181:106250.doi: 10.1016/j.phrs.2022.106250 

Non-esterified fatty acids and lipid mediators (LM) were extracted from plasma or lung homogenates using reversed phase cartridges (Sep-Pak® Vac 6cc 500 mg/6 ml C18; Waters). Internal standards added: d4-LTB4, d4-prostaglandin (PG)E2, d8–5S-hydroxyeicosatetraenoic acid (HETE), d5-lipoxin A4, d5-resolvin D2, (200 nM, each, Cayman Chemicals), and d8-arachidonic acid (AA, 10 μM, Cayman Chemicals). Lipid mediators were separated on an Acquity UPLC BEH C18 column (2.1 × 100 mm, Waters) using an Acquity UPLC system (Waters) and detected using a QTRAP 5500 mass spectrometer (SCIEX), equipped with an electrospray ionization source. Diagnostic ion fragments were determined by scheduled multiple reaction monitoring in the negative ion mode for peak identification.