Leukotriene biosynthesis: direct chemical ionization mass spectrometry of underivatized arachidonic acid metabolites.
An improved direct chemical ionization (DCI) mass spectrometric technique, using a polyimide-coated fused silica fiber as an extended probe tip, was used to obtain molecular ions and diagnostic fragment ions of underivatized arachidonic acid, 5-hydroperoxyeicosatetraenoic acid, 15-hydroperoxyeicosatetraenoic acid, leukotriene B4 (LTB4) and, for the first time, of leukotriene A4 (LTA4)-free acid. In this technique, sample compounds are coated onto the fused silica fiber and vaporized in the plume of the reagent gas plasma of a chemical ionization source without external heating of the probe. Both ammonia and isobutane DCI spectra were obtained for each compound. A volatile alkaline eluent system was developed that allowed reversed-phase high-performance liquid chromatography of LTA4 to be followed rapidly by DCI mass spectrometry. With these techniques, the conversion of LTA4 to LTB4 during incubation with human liver microsomes was confirmed. Selected ion monitoring (SIM) of preselected ion fragments in the spectrum increases the selectivity of this technique and improves quantification in the range 100 ng to 10 pg.
Gut
J
J
Department of Anesthesia, Stanford University School of Medicine, California 94305.
Trudell
J R
JR
Jamieson
G C
GC
D001095:Arachidonic Acids / Q000032:analysis / Q000096:biosynthesis* / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D017572:Leukotriene A4; D007975:Leukotriene B4 / Q000096:biosynthesis*; D015289:Leukotrienes; D008054:Lipid Peroxides / Q000032:analysis / Q000378:metabolism; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000378:metabolism*
D001095:Arachidonic Acids; D017572:Leukotriene A4; D015289:Leukotrienes; D008054:Lipid Peroxides; D007975:Leukotriene B4; C025086:15-hydroperoxy-5,8,11,13-eicosatetraenoic acid; C025348:arachidonic acid 5-hydroperoxide