Determination of endogenous tissue inflammation profiles by LC/MS/MS: COX- and LOX-derived bioactive lipids

LC/MS/MS method for analysis of E2 series prostaglandins and isoprostanes

“…However, because PGE 2 , PGD 2 , and 15-series isoPGE 2 have the same molecular weight and similar product ion spectrums, it is important to separate them in time before MS/MS detection. A number of HPLC conditions have been used to resolve PGE 2 /PGD 2 -like molecules ( 13,20,29,(31)(32)(33)(34)(35)(36). However, to the best of our knowledge, none of the reported conditions allowed for PGE 2 , PGD 2 , and major isoPGE 2 separation in a single run without derivatization.…”

“…Liquid chromatography with tandem mass spectrometer detection (LC/MS/MS) methods are considered to be the most specifi c and less laborious for isoPG measurement as compared with enzyme immunoassay and radioimmunoassay methods, high performance liquid chromatography (HPLC) with ultraviolet or fl uorescence detection, and gas chromatography with MS or fl ame ionization detection because of a better isomer separation and no derivatization or multi-step purifi cation requirements ( 29,30 ). However, the LC/MS/MS methods currently used do not allow for simultaneous separation of the major brain PGE 2 /D 2 and isoPGE 2 such as PGE 2 , entPGE 2 , PGD 2 , 8-isoPGE 2 , and 11 ␤ -PGE 2 without derivatization and multiple HPLC separations ( 13,20,29,(31)(32)(33)(34)(35)(36).…”

Characterization of an arachidonic acid-deficient (Fads1 knockout) mouse model

“…Eicosanoids were extracted from colonic mucosa, small intestine mucosa, and lung using a previously described method ( 19,20 ). Briefl y, snap-frozen tissues were ground to a fi ne powder and homogenized with an Ultrasonic Processor (Misonix).…”

“…Protein concentration was determined by the method of Bradford according to the manufacturer's instructions (Pierce). Liquid chromatography/tandem mass spectroscopic analyses were performed using a QuattroUltima mass spectrometer (Waters) equipped with an Agilent 1100 binary pump high-performance liquid chromatography system (Agilent) using a modifi ed version of the method of Yang et al ( 19 ). Eicosanoids of interest were chromatographically separated using a Luna 3 m phenyl-hexyl 4.6 × 100 mm analytic column (Phenomenex).…”

An improved LC–MS/MS method for the quantification of prostaglandins E2 and D2 production in biological fluids

“…To overcome the limitations of GC-MS and immunoassays for prostaglandin measurements, LC-MS [15][16][17] and LC-MS-MS [18][19][20][21][22][23][24][25][26][27] based methods have evolved as powerful tools for measuring prostaglandins in biological samples because of their high sensitivity, high selectivity and simplicity of sample preparation. Although five of these LC-MS or LC-MS-MS methods have measured PGD 2 [15][16][17][18][19], none have controlled for the inherent chemical instability of PGD 2 which is critical for determining accurate levels and for comparison with the values of other, more stable eicosanoids.…”