Quantitative Determination of Several Prostaglandins and Thromboxane B2 in the Isolated Perfused Rat Brain
A GC-MS assay was developed to investigate the time course of the tissue concentrations of PGE2, PGF2α, PGD2, 6-keto-PGF1α as well as TXB2 in the isolated rat brain, perfused with a synthetic fluoro-carbon perfusion medium. After perfusion, the brains were frozen in isopentane at −56°, freeze-dried at −40° and pulverized (liq. N2). The powder (~100 mg) was spiked with 4 deuterated analytes and extracted with methanol/water. The supernatants were applied to a BondElut RP-18 cartridge. The eluate was dried down, the residue dissolved in ether/p.e.⌽ and the solution applied to a silica gel column. The eluate was dried down and the residue subjected to methoximation, PFB esterification and silylation. The derivatized COP’s were separated and quantitated by NCI GC-MS.
KeywordsPerfusion Medium Ischaemic Brain Absolute Detection Limit Microwave Fixation Ischaemic Brain Damage
petroleum ether (light petroleum)
negative-ion chemical ionization
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