Simultaneous determination of leukotrienes B4 and E4 in whole blood and of leukotriene E4 in urine of rabbit by reversed-phase high-performance liquid chromatography

Antonio Celardo, Giuseppe Dell'Elba, Zeinab Mohamed Eltantawy, Virgilio Evangelista, Chiara Cerletti

Research output: Contribution to journalArticle

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Abstract

Sulfidopeptide leukotrienes E4 (LTE4) and B4 (LTB4) were simultaneously extracted from rabbit whole blood with acetonitrile. LTC4 and LTD4 were converted to LTE4 by γ-glutamyl transpeptidase and leucine-amino peptidase before extraction. LTE4 was extracted from urine with C18 Sep-Pak cartridges. The compounds were resolved and quantitated by reversed-phase high-performance liquid chromatography (HPLC) with a diode-array detector; in selected cases the collected fractions were assayed for LTB4 and LTE4 by specific enzyme immunoassay (EIA). The correlation factor of the measured increase in LTE4 concentrations and addition of incremental amounts of LTC4 to blood was r = 0.998; slope of 1.05 ± 0.01 (mean ± S.D.). Concentrations of LTE4 measured by HPLC correlated with those obtained with EIA (r = 0.996; slope = 0.98 ± 0.03 and r = 0.991; slope = 0.97 ± 0.04 in blood and urine, respectively). For blood LTB4 the correlation of HPLC versus EIA was r = 0.990; slope = 1.12 ± 0.04. The method described is accurate and reproducible, allowing measurement of both LTB4 and LTE4 in whole blood after a single extraction procedure. Simultaneous measurement of these metabolites after specific stimulation or in pathological conditions is recommended for in vivo investigations of LTs production.

Original languageEnglish
Pages (from-to)261-269
Number of pages9
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume658
Issue number2
DOIs
Publication statusPublished - Aug 19 1994

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Leukotriene E4
Leukotriene B4
High performance liquid chromatography
Blood
Leukotriene C4
Enzymes
Leukotriene D4
gamma-Glutamyltransferase
Metabolites
Leucine
Diodes
Peptide Hydrolases

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Simultaneous determination of leukotrienes B4 and E4 in whole blood and of leukotriene E4 in urine of rabbit by reversed-phase high-performance liquid chromatography. / Celardo, Antonio; Dell'Elba, Giuseppe; Eltantawy, Zeinab Mohamed; Evangelista, Virgilio; Cerletti, Chiara.

In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 658, No. 2, 19.08.1994, p. 261-269.

Research output: Contribution to journalArticle

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abstract = "Sulfidopeptide leukotrienes E4 (LTE4) and B4 (LTB4) were simultaneously extracted from rabbit whole blood with acetonitrile. LTC4 and LTD4 were converted to LTE4 by γ-glutamyl transpeptidase and leucine-amino peptidase before extraction. LTE4 was extracted from urine with C18 Sep-Pak cartridges. The compounds were resolved and quantitated by reversed-phase high-performance liquid chromatography (HPLC) with a diode-array detector; in selected cases the collected fractions were assayed for LTB4 and LTE4 by specific enzyme immunoassay (EIA). The correlation factor of the measured increase in LTE4 concentrations and addition of incremental amounts of LTC4 to blood was r = 0.998; slope of 1.05 ± 0.01 (mean ± S.D.). Concentrations of LTE4 measured by HPLC correlated with those obtained with EIA (r = 0.996; slope = 0.98 ± 0.03 and r = 0.991; slope = 0.97 ± 0.04 in blood and urine, respectively). For blood LTB4 the correlation of HPLC versus EIA was r = 0.990; slope = 1.12 ± 0.04. The method described is accurate and reproducible, allowing measurement of both LTB4 and LTE4 in whole blood after a single extraction procedure. Simultaneous measurement of these metabolites after specific stimulation or in pathological conditions is recommended for in vivo investigations of LTs production.",
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