Arachidonate Related Lipid Mediators by John N. Abelson, Melvin I. Simon, Robert C. Murphy, Frank A.

By John N. Abelson, Melvin I. Simon, Robert C. Murphy, Frank A. Fitzpatrick

This quantity contains details on arachidonic acid metabolites and platelet activating issue. Assays for the quantitative dimension of prostaglandins, leukotrienes, and platelet-activating issue, together with enzyme immunoassays and mass spectrophotometric thoughts, are provided. instruction and research of cytochrome P-450 metabolites of arachidonic acid are mentioned. The isolation, purification, and assay of enzymes vital within the biosynthesis and metabolism of those lipids are integrated

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Limbird, J. Biol. Chem. 261, 8660-8666 (1986). [2] NEGATIVE-ION MASS SPECTROMETRY OF LIPIDS 19 leased arachidonate was converted to PGD2, PGE2, and PGF2~. Quantification of the PGs by capillary column G C - N C I - M S allowed the size of the initial pool of released arachidonate to be quantified in the presence of a relatively high background of TxB2. Interest in the analysis of trace quantities of the four regioisomeric EETs (cytochrome P-450 metabolites of arachidonic acid) stimulated the development of N C I - M S methods for these compounds) 2'33 The NCI characteristics of the EET-PFB esters were a little different from the other eicosanoid PFB derivatives.

The residue is dissolved in decane (10 ~1) ready for GC-MS analysis. Gas Chromatography-Mass Spectrometry GC-MS is carried out on a Nermag R1010C quadrupole mass spectrometer (Delsi Instruments, TX) interfaced with a Varian Vista gas chromatograph (Varian, Palo Alto, CA). Data reduction and analysis are carried out using Nermag SIDAR software. ~m coating thickness, Supelco, Bellefonte, PA) is passed through the transfer line from the gas chromatograph until the end is just inside the ion source.

Biol. Chem. 261, 3501 (1986). 13 D. F. Tuber, M. A. Phillips, and W. A. Hubbard, Prostaglandins 22, 349 (1981). [2] NEGATIVE-ION MASS SPECTROMETRY OF LIPIDS i 0/ta) 1303 15 I 0 | ......... i......... , ......... , ......... ,~........ I tO0 0 .... tO0 150 , .... i .... 150 200 , .... m/z ~ ......... 200 m/z 250 ~ ......... 250 300 t 300 :]50 ....... 350 FIo. 1. NCI mass spectra of (a) arachidonic acid PFB ester, (b) C-20-2H3-1abeled arachidonic acid PFB ester. 5% of protiem impurity at rn/z 303 and the [M - PFB] anion at m/z 306 was the base peak in the NCI mass spectrum (Fig.

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