Structural characterization of monohydroxyeicosatetraenoic acids and dihydroxy- and trihydroxyeicosatrienoic acids by ESI-FTICR

Cui, Lijie; Isbell, Marilyn A.; Chawengsub, Yuttana; Falck, John R.; Campbell, William B.; Nithipatikom, Kasem

doi:10.1016/j.jasms.2008.01.007

Cited by 16 publications

(12 citation statements)

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“…The identities of these metabolites were confirmed by LC-MS/MS. Two major products were detected with HPLC elution times of 13.1 and 15.5 min that comigrated with standards for 6-keto-PGF 1␣ and PGE 2 , respectively (Table 1) 5-HETE, respectively (12,42). Identical fragmentation patterns occurred with the respective HETE standards.…”

Section: Resultsmentioning

confidence: 83%

“…Desolvation and cone gas flow were 600 and 50 l/h, respectively. THETA fractions were analyzed using electrospray ionization with a Fourier transform ion cyclotron resonance mass spectrometer (ESI-FT/MS, IonSpec 7.0-tesla FTICR high-resolution mass spectrometer with a Waters Z spray ESI source) (12). Fractions were chromatographed using a Kromasil C-18 (5 m, 1 ϫ 150 mm) column.…”

Section: Animalsmentioning

confidence: 99%

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Role of arachidonic acid lipoxygenase metabolites in acetylcholine-induced relaxations of mouse arteries

Gauthier

Goldman

Aggarwal

et al. 2011

American Journal of Physiology-Heart and Circulatory Physiology

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Arachidonic acid (AA) metabolites function as EDHFs in arteries of many species. They mediate cyclooxygenase (COX)- and nitric oxide (NO)-independent relaxations to acetylcholine (ACh). However, the role of AA metabolites as relaxing factors in mouse arteries remains incompletely defined. ACh caused concentration-dependent relaxations of the mouse thoracic and abdominal aorta and carotid, femoral, and mesentery arteries (maximal relaxation: 57 ± 4%, 72 ± 4%, 82 ± 3%, 80 ± 3%, and 85 ± 3%, respectively). The NO synthase inhibitor nitro-L-arginine (L-NA; 30 μM) blocked relaxations in the thoracic aorta, and L-NA plus the COX inhibitor indomethacin (10 μM) inhibited relaxations in the abdominal aorta and carotid, femoral, and mesenteric arteries (maximal relaxation: 31 ± 10%, 33 ± 5%, 41 ± 8%, and 73 ± 3%, respectively). In mesenteric arteries, NO- and COX-independent relaxations to ACh were inhibited by the lipoxygenase (LO) inhibitors nordihydroguaiaretic acid (NDGA; 10 μM) and BW-755C (200 μM), the K(+) channel inhibitor apamin (1 μM), and 60 mM KCl and eliminated by endothelium removal. They were not altered by the cytochrome P-450 inhibitor N-methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide (20 μM) or the epoxyeicosatrienoic acid antagonist 14,15-epoxyeicosa-5(Z)-enoic acid (10 μM). AA relaxations were attenuated by NDGA or apamin and eliminated by 60 mM KCl. Reverse-phase HPLC analysis revealed arterial [(14)C]AA metabolites that comigrated with prostaglandins, trihydroxyeicosatrienoic acids (THETAs), hydroxyepoxyeicosatrienoic acids (HEETAs), and hydroxyeicosatetraenoic acids (HETEs). Epoxyeicosatrienoic acids were not observed. Mass spectrometry confirmed the identity of 6-keto-PGF(1α), PGE(2), 12-HETE, 15-HETE, HEETAs, 11,12,15-THETA, and 11,14,15-THETA. AA metabolism was blocked by NDGA and endothelium removal. 11(R),12(S),15(S)-THETA relaxations (maximal relaxation: 73 ± 3%) were endothelium independent and blocked by 60 mM KCl. Western immunoblot analysis and RT-PCR of the aorta and mesenteric arteries demonstrated protein and mRNA expression of leukocyte-type 12/15-LO. Thus, in mouse resistance arteries, 12/15-LO AA metabolites mediate endothelium-dependent relaxations to ACh and AA.

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Section: Resultsmentioning

confidence: 83%

Section: Animalsmentioning

confidence: 99%

Role of arachidonic acid lipoxygenase metabolites in acetylcholine-induced relaxations of mouse arteries

Gauthier

Goldman

Aggarwal

et al. 2011

American Journal of Physiology-Heart and Circulatory Physiology

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“…The unknown metabolites corresponding to THETAs/trioxilins from HPLC were identified by liquid chromatography–electrospray–Fourier transform ion cyclotron resonance mass spectrometry (LC‐ESI‐FTICR) (7.0 Tesla FTICR, IonSpec) as previously described (Cui et al . , Chawengsub et al . ).…”

Section: Methodsmentioning

confidence: 99%

Vascular hepoxilin and trioxilins mediate vasorelaxation through TP receptor inhibition in mouse arteries

et al. 2016

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Aim 12/15-lipoxygenase (12/15-LO) metabolizes arachidonic acid (AA) into several vasoactive eicosanoids. In mouse arteries, we previously characterized the enzyme’s 15-LO metabolites 12(S)-hydroxyeicosatetraenoic acid (HETE), 15-HETE, hydroxyepoxyeicosatrienoic acids (HEETAs) and 11,12,15-trihydroxyeicosatrienoic acids (11,12,15-THETAs) as endothelium-derived relaxing factors. However, the observed 12-LO metabolites remained uncharacterized. The purpose of this study was to determine the structure and biological functions of eicosanoids generated by the enzyme’s 12-LO activity. Methods Metabolites extracted from aortas of C57BL/6 male mice were separated using a series of reverse and normal phase chromatographic steps and identified as hepoxilin A3, trioxilin A3 and trioxilin C3 by mass-spectrometry. Activities of these natural compounds were tested on isometric tension and intracellular calcium release. The role of thromboxane (TP) receptor was determined in HEK293 cells overexpressing TPα receptor (TPα-HEK). Results All identified vascular 12-LO metabolites were biologically active. In mouse mesenteric arteries, trioxilin A3, C3 and hepoxilin A3 (3μM) relaxed arteries constricted with the thromboxane mimetic, U46619 (maximum relaxations of 78.9±3.2, 29.7±4.6, 82.2±5.0 and 88.0±2.4% respectively), but not phenylephrine-constricted arteries. In TPα-HEK cells, trioxilin A3, C3 and hepoxilin A3 (10μM) inhibited U46619 (10nM)-induced increases in intracellular calcium by 53.0±7.2%, 32.8±5.0% and 37.9±13.5%, respectively. In contrast, trioxilin B3 and hepoxilin B3 were not synthesized in arteries and exhibited little biological activity. Conclusion Trioxilin A3 and C3 and hepoxilin A3 are endogenous vascular relaxing factors. They are not endothelium-derived hyperpolarizing factors but mediate vascular relaxation by inhibiting TP agonist induced increases in intracellular calcium. Thus, they regulate vascular homeostasis by acting as endogenous TP antagonists.

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“…mentioned product ion spectra of 9-HETE and 12-HETE were similar and these two compounds must be separated chromatographically for their identification [23]. Yet, we were able to identify unique fragment m/z 135 (beak at C11–12 and loss of CO 2 ) [35] and m/z 151 for 12-HETE and 9-HETE, respectively, from the heatmap. The optimal transitions and the retention times for each of the 15 HETEs/EETs were listed in Table 1.…”

Section: Resultsmentioning

confidence: 99%

Comprehensive analysis of oxylipins in human plasma using reversed-phase liquid chromatography-triple quadrupole mass spectrometry with heatmap-assisted selection of transitions

Chen

Zhang

2018

Anal Bioanal Chem

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Oxylipins, a subclass of lipid mediators, are metabolites of various polyunsaturated fatty acids with crucial functions in regulation of systemic inflammation. Elucidation of their roles in pathological conditions requires accurate quantification of their levels in biological samples. We refined an ultra-performance liquid chromatography-multiple reaction monitoring-mass spectrometry (UPLC-MRM-MS)-based workflow for comprehensive and specific quantification of 131 endogenous oxylipins in human plasma, in which we optimized LC mobile phase additives, column and gradient conditions. We employed heatmap-assisted strategy to identify unique transitions to improve the assay selectivity and optimized solid phase extraction procedures to achieve better analyte recovery. The method was validated according to FDA guidelines. Overall, 94.4% and 95.7% of analytes at tested concentrations were within acceptable accuracy (80–120%) and precision (CV<15%), respectively. Good linearity for most analytes was obtained with R2 > 0.99. The method was also validated using a standard reference material – SRM 1950 frozen human plasma to demonstrate inter-lab compatibility.

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Structural characterization of monohydroxyeicosatetraenoic acids and dihydroxy- and trihydroxyeicosatrienoic acids by ESI-FTICR

Cited by 16 publications

References 44 publications

Role of arachidonic acid lipoxygenase metabolites in acetylcholine-induced relaxations of mouse arteries

Role of arachidonic acid lipoxygenase metabolites in acetylcholine-induced relaxations of mouse arteries

Vascular hepoxilin and trioxilins mediate vasorelaxation through TP receptor inhibition in mouse arteries

Comprehensive analysis of oxylipins in human plasma using reversed-phase liquid chromatography-triple quadrupole mass spectrometry with heatmap-assisted selection of transitions

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