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

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

This quantity comprises 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 options, are provided. coaching and research of cytochrome P-450 metabolites of arachidonic acid are mentioned. The isolation, purification, and assay of enzymes very important within the biosynthesis and metabolism of those lipids are incorporated

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Falck, Biochem. Biophys. Res. Commun. 146, 638 (1987). 34 K. Yamashita, J. H. Capdevila, and I. A. Blair, unpublished observations. 35 B. W. Christman and I. A. Blair, Biomed. Environ. Mass Spectrom. 18, 258 (1989). 36 A. I. Mallet, Biomed. Environ. Mass Spectrom. 16, 207 (1988). /°P B 303I M"PFB ol100 ....... r.. 200 ..... m/z • o / ~o) o >~ -I "~ ....... 400 1325 o 1225 / 0 , ....... =: ........ t ......... 200 300 400 m/z FIG. 5. NCI mass spectra of (a) 14,15-EET PFB ester; (b) H6-14,15-EET PFB ester.

A. 85, 304 (1988). z3 D. J+ FitzGerald, J. Fragetta, and G. A. FitzGerald, J+ Clin. Invest. 82, 1708 (1988). 4 T 0/ ............... 100 { oPt=e M,-rFB 0/ ....... 100 I. . . 200 300 m/z , ......... 400 500 ......... ,, 07 . . . . . . . 200 ~ 0z 400 500 FIG. 3. NCI mass spectrum of (a) TMS ether, PFB ester derivative of 12(S)-HETE; (b) TMS ether, PFB ester derivative of Hs-12(S)-HETE. 4 pg/ml. 9 Prior to this study, literature values for normal concentrations of plasma 6-oxo-PGFi~ spanned a range of several orders of magnitude.

The lenses in the quadrupole assembly are tuned for optimal peak shape and peak heights across the mass range (100-633 D). The resolution is adjusted to give a 20% valley between the following pairs of ions: m/z 633/634, m/z 595/596, m/z 562/563 m/z 524/525, m/z 452/453, m/z 414/415, and m/z 281/282. The methane pressure is adjusted for optimal sensitivity. Tuning of the source and lenses is then repeated in order to ensure that all parameters are optimal for sensitivity and resolution. The instrument is then calibrated.

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