Collisionally induced dissociation of epoxyeicosatrienoic acids and epoxyeicosatrienoic acid-phospholipid molecular species.
Four isomers of epoxyeicosatrienoic acid (EET) can be formed by cytochrome P-450 oxidation of arachidonic acid: 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acid. The collision-induced dissociation of the [M-H]- anion at m/z 319 from each of these isomers, using negative-ion fast atom bombardment ionization and a triple quadrupole mass spectrometer, resulted in a series of common ions as well as ions characteristic of each isomer. The common ions were m/z 301 [M-H2O]- and 257 [M-(H2O + CO2)]-. Unique ions resulted from cleavages alpha to the epoxide moiety to form either conjugated carbanions or aldehydes. Mechanisms involving charge site transfer are suggested for the origin of these ions. A distonic ion series that may involve a charge-remote fragmentation mechanism was also observed. The epoxyeicosatrienoic acids were also incorporated into cellular phospholipids following incubation of the free acid with murine mast cells in culture. Negative fast atom bombardment mass spectrometry of purified glycerophosphoethanolamine-EET species and glycerophosphocholine-EET species yielded abundant [M-H]- and [M-CH3]- ions, respectively. The collision-induced dissociation of these specific high-mass ions revealed fragment ions characteristic of the epoxyeicosatrienoic acids incorporated (m/z 319, 301, and 257) and the same unique ions as those seen with each isomeric epoxyeicosatrienoic acid. With this direct method of analysis, phospholipids containing the four positional isomers of EET, including the highly labile (5,6-EET), could be identified as unique molecular species in mast cells incubated with EET.
Bernstrom
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K
National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206.
Kayganich
K
K
Murphy
R C
RC
D015117:5,8,11,14-Eicosatetraynoic Acid / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D015126:8,11,14-Eicosatrienoic Acid / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001853:Bone Marrow / Q000737:chemistry; D002461:Cell Line, Transformed / Q000737:chemistry; D004852:Epoxy Compounds / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D008407:Mast Cells / Q000737:chemistry; D051379:Mice; D010715:Phosphatidylglycerols / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D016339:Spectrometry, Mass, Fast Atom Bombardment / Q000379:methods*
D004852:Epoxy Compounds; D010715:Phosphatidylglycerols; D010743:Phospholipids; D015117:5,8,11,14-Eicosatetraynoic Acid; D015126:8,11,14-Eicosatrienoic Acid