Determination of picogram amounts of lipoxin A4 and lipoxin B4 by high-performance liquid chromatography with electrochemical detection

Herrmann, Thomas; Steinhilber, Dieter; Knospe, Johanna; Roth, H. J.

doi:10.1016/s0378-4347(00)83914-6

Cited by 18 publications

(4 citation statements)

References 8 publications

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“…Detection limits of the LX and LT in this system were determined to be in the picogram range; these values were consistent with those reported by Herrmann et al (22). Mono-HETEs were resolved using an RP-HPLC system equipped with a Lambda Max UV detector, model 481, and the column (Beckman Ultrasphere-ODS, 5 um, 4.6 mm X 25 cm) was eluted at 1 ml/ min with MeOH/H20/acetic acid (75:25:0.01, vol/vol/vol).…”

Section: Methodssupporting

confidence: 92%

Human alveolar macrophages have 15-lipoxygenase and generate 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid and lipoxins.

Levy¹,

Romano²,

Chapman³

et al. 1993

J. Clin. Invest.

186

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Eicosanoids derived from lipoxygenase (LO)-catalyzed reactions play important roles in pulmonary inflammation. Here, we examined formation of LO-derived products by human alveolar macrophages (HAM). HAM converted 11-4Cl-arachidonic acid to a product carrying '4C-radiolabel that was identified as 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid (15-HETE) by physical methods. 15-LO mRNA was demonstrated in HAM by reverse transcription-polymerase chain reaction. Incubation of HAM for 3 d with interleukin 4 (IL-4) before exposure to j1-"4Clarachidonic acid led to both increased mRNA for 15-LO and a 4-fold increase in 15-HETE formation. In contrast, 5(S)-hydroxy-6-trans-8,11,14-cis-eicosatetraenoic acid generation was not significantly altered by prior exposure to IL-4. Additionally, lipoxins (LXA4 and LXB4) were detected from endogenous substrate, albeit in lower levels than leukotriene B4 (LTB4), in electrochemical detection/high performance liquid chromatography profiles from HAM incubated in the presence and absence of the chemotactic peptide (FMLP) or the calcium ionophore (A23187).Exposure of HAM to leukotriene A4 (LTA4) resulted in a 2-fold increase in LXA4 and 10-fold increase in LXB4. These results demonstrate the presence of 15-LO mRNA and enzyme activity in HAM and the production of LXA4 and LXB4 by these cells. Along with 5-LO-derived products, the biosynthesis of 15-LO-derived eicosanoids by HAM may also be relevant in modulating inflammatory responses in the lung. (J.

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Section: Methodssupporting

confidence: 92%

Human alveolar macrophages have 15-lipoxygenase and generate 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid and lipoxins.

Levy¹,

Romano²,

Chapman³

et al. 1993

J. Clin. Invest.

186

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“…Since these compounds do not contain aromatic or natural fluorescent systems, GC-MS and, likewise, HPLC combined with fluorescent detectors requires these compounds to be derivatized into a complex that fluoresces, which makes the analysis time-consuming and expensive [50,78]. Also, HPLC with electrochemical detection allows the determination of picogram amounts of lipoxin (LX) A4 (LXA4) and LXB4 in extracts of human polymorphonuclear granulocytes [79], and leukotriene (LT) B4 (LTB4) from human polymorphonuclear leukocytes [80]. However, this method is only suitable for the determination of electrochemically active substances, to which most oxylipins do not apply, in this case additional transformations of substances are required [81,82].…”

Section: High-performance Liquid Chromatographymentioning

confidence: 99%

Methods of the Analysis of Oxylipins in Biological Samples

et al. 2020

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Oxylipins are derivatives of polyunsaturated fatty acids and due to their important and diverse functions in the body, they have become a popular subject of studies. The main challenge for researchers is their low stability and often very low concentration in samples. Therefore, in recent years there have been developments in the extraction and analysis methods of oxylipins. New approaches in extraction methods were described in our previous review. In turn, the old analysis methods have been replaced by new approaches based on mass spectrometry (MS) coupled with liquid chromatography (LC) and gas chromatography (GC), and the best of these methods allow hundreds of oxylipins to be quantitatively identified. This review presents comparative and comprehensive information on the progress of various methods used by various authors to achieve the best results in the analysis of oxylipins in biological samples.

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“…A third RP-HPLC system was employed to detect picogram levels of 20-OH-LTB4 and 20-COOH-LTB4 as described by Herrmann et al (1987Herrmann et al ( ,1988. The column (Altex, Ultrasphere-ODs, 4.6 mm x 25 cm) was eluted with methanol/water (65:35, v/v) with TFA (1 mM) and the UV detector (Waters Assoc.…”

Section: Analysis Of Eicosanoidsmentioning

confidence: 99%

Lipoxin A₄ and lipoxin B₄ stimulate the release but not the oxygenation of arachidonic acid in human neutrophils: Dissociation between lipid remodeling and adhesion

Nigam

Fiore

Luscinskas

et al. 1990

Journal Cellular Physiology

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The profiles of actions of lipoxin A4 (LXA4) and lipoxin B4 (LXB4), two lipoxygenase-derived eicosanoids, were examined with human neutrophils. At nanomolar concentrations, LXA4 and LXB4 each stimulated the release of [1-14C]arachidonic acid from esterified sources in neutrophils. Lipoxin-induced release of [1-14C]arachidonic acid was both dose- and time-dependent and was comparable to that induced by the chemotactic peptide f-met-leu-phe. Time-course studies revealed that lipoxin A4 and lipoxin B4 each induced a biphasic release of [1-14C]arachidonic acid, which was evident within seconds (5-15 sec) in its initial phase and minutes (greater than 30 sec) in the second phase. In contrast, the all-trans isomers of LXA4 and LXB4 did not provoke [1-14C]AA release. Lipoxin-induced release of arachidonic acid was inhibited by prior treatment of the cells with pertussis toxin but not by its beta-oligomers, suggesting the involvement of guaninine nucleotide-binding regulatory proteins in this event. Dual radiolabeling of neutrophil phospholipid classes with [1-14C]arachidonic acid and [3H]palmitic acid showed that phosphatidylcholine was a major source of lipoxin-induced release of [1-14C]arachidonic acid. They also demonstrated that lipoxins rapidly stimulate both formation of phosphatidic acid as well as phospholipid remodeling. Although both LXA4 and LXB4 (10(-8)-10(-6) M) stimulated the release of [1-14C]arachidonic acid, neither compound evoked its oxygenation by either the 5- or 15-lipoxygenase pathways (including the formation of LTB4, 20-COOH-LTB4, 5-HETE, or 15-HETE). LXA4 and LXB4 (10(-7) M) each stimulated the elevation of cytosolic Ca2+ as monitored with Fura 2-loaded cells, albeit to a lesser extent than equimolar concentrations of FMLP. Neither lipoxin altered the binding of [3H]LTB4 to its receptor on neutrophils. In addition, they did not stimulate aggregation or induce adhesion of neutrophils to human endothelial cells. Results indicate that both LXA4 and LXB4 stimulate the rapid remodeling of neutrophil phospholipids to release arachidonic acid without provoking either aggregation or the formation of lipoxygenase-derived products within a similar temporal and dose range. Together they indicate that LXA4 and LXB4 display selective actions with human neutrophils and suggest that these eicosanoids possess unique profiles of action which may regulate neutrophil function during inflammation.

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Determination of picogram amounts of lipoxin A4 and lipoxin B4 by high-performance liquid chromatography with electrochemical detection

Cited by 18 publications

References 8 publications

Human alveolar macrophages have 15-lipoxygenase and generate 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid and lipoxins.

Human alveolar macrophages have 15-lipoxygenase and generate 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid and lipoxins.

Methods of the Analysis of Oxylipins in Biological Samples

Lipoxin A₄ and lipoxin B₄ stimulate the release but not the oxygenation of arachidonic acid in human neutrophils: Dissociation between lipid remodeling and adhesion

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Determination of picogram amounts of lipoxin A4 and lipoxin B4 by high-performance liquid chromatography with electrochemical detection

Cited by 18 publications

References 8 publications

Human alveolar macrophages have 15-lipoxygenase and generate 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid and lipoxins.

Human alveolar macrophages have 15-lipoxygenase and generate 15(S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid and lipoxins.

Methods of the Analysis of Oxylipins in Biological Samples

Lipoxin A4 and lipoxin B4 stimulate the release but not the oxygenation of arachidonic acid in human neutrophils: Dissociation between lipid remodeling and adhesion

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Lipoxin A₄ and lipoxin B₄ stimulate the release but not the oxygenation of arachidonic acid in human neutrophils: Dissociation between lipid remodeling and adhesion