title: Measurements of leukotrienes in human plasma by solid phase extraction and high performance liquid chromatography.
abstract: Leukotrienes (LTs) are biologically active compounds derived from lipoxygenase catalyzed metabolism of arachidonic acid in mammalian tissues. The present report describes a simple method for extraction and isolation of dihydroxy-LTs; LTB4, LTB5 and the peptido-LTs; LTC4, LTD4 and LTE4 from human plasma, using a pretreatment cartridge which utilizes both hydrophobic and ion-exchange interactions. 5 ml acidified plasma or acetate buffer containing commercially available LT standards were passed through the cartridges under suction, and the absorbed LTs were subsequently eluted in a stepwise manner with acetate buffer containing increasing amounts of methanol. The eluted LTs were analyzed by reversed-phase high performance liquid chromatography (HPLC) on octadecylsilyl (ODS)-silica, using a Waters HPLC unit. Both with plasma and acetate buffer the present methodology resulted in good separation of the LTs with a total run-time of less than 32 min. Recovery of dihydroxy-LTs was approximately 80% (range 73-82%) both when the standards were dissolved in plasma and in acetate buffer. Recovery of the peptido-LTs was, however somewhat lower (47-50%). It should be noted that the present method has the advantage that exposure to chemicals of high toxicity is avoided.
authors.affiliation: Department of Medical Physiology, Institute of Medical Biology, University of Tromoø, Norway.
authors.forename: T
authors.initials: T
authors.lastname: Henden
authors.forename: H
authors.initials: H
authors.lastname: Strand
authors.forename: E
authors.initials: E
authors.lastname: Børde
authors.forename: A G
authors.initials: AG
authors.lastname: Semb
authors.forename: T S
authors.initials: TS
authors.lastname: Larsen
mesh_terms: D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006801:Humans; D015289:Leukotrienes / Q000097:blood* / Q000302:isolation & purification
chemical_list: D015289:Leukotrienes