反应详情

In general, COX-2 enzymatic products can be detected or measured by a variety of methods. For example, reversed phase liquid chromatography-mass spectrometry is used to detect or measure COX-2 enzymatic produces. FIG. 12 is a selected-ion mass chromatogram of oxygenated 2-AG metabolites with (a) m/z=449.3 and (b) m/z=417.3 (Kozak et al. 2000, supra). For example, products are eluted with a 15-minute gradient of 20% to 100% acetonitrile in H2O (0.001% sodium acetate). LC/MS reveals the presence of five primary products in 2-AG/COX-2 reaction mixtures. Two closely eluting polar products each display m/z 449 consistent with the non-enzymatic PGH2-G isomerization products PGE2-G and PGD2-G (FIG. 12). Pentadeuterated standards of glyceryl esters of PGE2 and PGD2 are synthesized and coelute with these two polar products under multiple chromatographic conditions. An intermediate polarity product with m/z 377 is observed that is consistent with the PGH2-G degradation product 12-hydroxyheptadeca-5,8,10-trienoic acid (HHT-G, data not shown). Finally, two closely eluting non-polar 2-AG metabolites are detected with an m/z of 417, consistent with the glyceryl esters of HETEs (hydroxyeicosatetraenoic acids that are minor products of arachidonic acid oxygenation by COX-2) (FIG. 12).

1. customare eluted with a 15-minute
2. customreaction mixtures
3. washTwo closely eluting polar products each display m/z 449 consistent with the non-enzymatic PGH2-G
4. washFinally, two closely eluting non-polar 2-AG metabolites
5. customare detected with an m/z of 417, consistent with the glyceryl esters of HETEs (hydroxyeicosatetraenoic acids that are minor products of arachidonic acid oxygenation by COX-2) (FIG. 12)