Fig. 2: Targeted lipidomics for measuring eVDs and epoxy-eVDs. | Nature Communications

Fig. 2: Targeted lipidomics for measuring eVDs and epoxy-eVDs.

From: Anti-inflammatory dopamine- and serotonin-based endocannabinoid epoxides reciprocally regulate cannabinoid receptors and the TRPV1 channel

Fig. 2: Targeted lipidomics for measuring eVDs and epoxy-eVDs.

a Authentic standards 14’,15’-epoNADA and 14’,15’-epoNA5HT were synthesized. (i) m-CPBA, RT, 1 h, MeCN (ii) reverse-phase HPLC purify (iii) EDC, DMAP, DIPEA; ice bath 1 h; RT 8 h; 50:50 DCM:DMF (iv) reverse-phase HPLC purify. b Development of LC-MS/MS method for the separation of NADA, epo-NADA, NA5HT, epo-NA5HT and AEA (Method 1) and the different regioisomers of EET-EAs and epo-NADA and epo-NA5HT (Method 2). AEA m/z 348.3 → m/z 203.2; NADA m/z 440.2 → m/z 287.1; 14’,15’-epoNADA m/z 456.3 → m/z 137.1; 14’,15’-epoNA5HT m/z 479.3 → m/z 160.1; NA5HT m/z 463.3 → m/z 287.2; EET-EA m/z 264.2 → m/z 62.0. c NADA and NA5HT metabolism by BV2 microglial cells under lipopolysaccharide (LPS) stimulation in the presence of 1 µM t-AUCB (sEH inhibitor) and 10 µM NADA or NA5HT. The reversible CYP epoxygenase inhibitor SKF 525A was used to demonstrate CYP-mediated metabolism. Data represents the mean ± SEM of three experiments. Statistical significance based on a two-tailed t-test with equal variance. All data can be found in the Source Data file.

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