Fig. 2.

Quantitative analysis of neuronal NAPE by HPLC coupled to evaporative light-scattering detection. Representative chromatograms from (a) control neurons, (b) 1 μm ionomycin-stimulated neurons, and (c) control neurons to which synthetic NAPE (10 μg) was added before extraction, to verify its HPLC coelution with native NAPE. The arrows indicate the retention time of synthetic NAPE. Lipids were extracted and fractionated by column chromatography. The NAPE-containing fractions from two culture dishes were pooled and subjected to reversed-phase HPLC, as described in Materials and Methods. d, Response of the light-scattering detector as a function of injected synthetic NAPE (in 40 μl of chloroform). Photomultiplier voltage was set at 800 V, and nebulization temperature was set at 90°C.