Antibody-mediated extraction/negative-ion chemical ionization mass spectrometric measurement of thromboxane B2 and 2,3-dinor-thromboxane B2 in human and rat urine

Chiabrando, Chiara; Benigni, Ariela; Piccinell, A.; Carminati, Claudia Elisa; Cozzi, Emily; Fanelli, Roberto

doi:10.1016/0003-2697(87)90121-7

Cited by 51 publications

(6 citation statements)

References 13 publications

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“…Prostaglandins and thromboxanes were assayed in urine using immunoaffinity chromatography gas chromatography (GC)/electron capture mass spectrometry using a modification of the method of Chiabrando et al (1987). Briefly, urine samples (10 ml) were diluted 1:1 by volume with buffer at pH 8.0 and [2H4] 6-oxo-PGF1,,, [2H4] TXB2, [2H4] 2,3-dinor-6-oxo-PGF1ic and [2H4] 2,3-dinor-TXB2 (5 ng each) were added.…”

Section: Assaysmentioning

confidence: 99%

Differential effect of aspirin on thromboxane and prostaglandin biosynthesis in man.

Ritter

Cockcroft

Doktor

et al. 1989

Brit J Clinical Pharma

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1 Effects of a single intravenous dose of aspirin (600 mg) on bradykinin-stimulated prostaglandin (PG) and on thromboxane (TX) biosynthesis were determined in nine healthy male volunteers. Plasma concentrations of 6-oxo-PGF,1ll and 13,14-dihydro-15-oxo-PGF2a were measured in samples obtained during repeated 10 min intravenous infusions of bradykinin before and up to 6 h after the dose of aspirin. TXB2 was measured in serum from blood allowed to clot at 370 C. 2 Aspirin inhibited bradykinin stimulated PG and platelet TX biosynthesis 0.5 h after the dose. Serum TXB2 remained low, whereas PG synthesis recovered within 6 h. 3 Effects of intravenous sodium salicylate (600 mg) were studied identically in eight subjects. Prostanoid biosynthesis was not inhibited. 4 Biosynthesis of prostacyclin and TXA2 under basal conditions was studied in eight subjects by measuring 2,3-dinor-6-oxo-PGF1,, and 2,3-dinor-TXB2 in hourly urine samples obtained during and after intravenous infusion of aspirin and, on a separate occasion, of vehicle.5 Aspirin infusion reduced urinary excretion of both metabolites > 90%, but excretion of 2,3-dinor-6-oxo-PGF1a recovered more rapidly than did that of 2,3-dinor-TXB2. 6 We conclude that cyclo-oxygenase is rapidly synthesised in bradykinin-responsive tissues in vivo and that this reflects similarly rapid enzyme biosynthesis in tissues that produce PGI2 under basal conditions. Keywords thromboxane A2 prostacyclin prostaglandin metabolites bradykinin aspirin

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Section: Assaysmentioning

confidence: 99%

Differential effect of aspirin on thromboxane and prostaglandin biosynthesis in man.

Ritter

Cockcroft

Doktor

et al. 1989

Brit J Clinical Pharma

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“…Aliquots of IAC and C18 SPE extracts were dried and derivatized to ME-TMS or PFB-TMS and analyzed by GC-EI-MS or GC-NICI-MS, respectively. 2␣ -Because polyclonal antibodies raised against prostanoids coupled to the carrier protein via the carboxyl group often cross-react with the corresponding ␤-oxidation products (12,13,19), we tested three antibodies (A, B, and C) raised against 8-epi-PGF 2␣ (16) for their ability to extract any of its possible ␣-chain-shortened (C18 and C16) metabolites. Aliquots (10 ml) of a human urine pool were extracted using three different immunosorbents prepared with A, B, and C antibodies.…”

mentioning

confidence: 99%

Identification and Measurement of Endogenous β-Oxidation Metabolites of 8-epi-Prostaglandin F2α

Chiabrando¹,

Valagussa²,

Rivalta³

et al. 1999

Journal of Biological Chemistry

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F 2 -isoprostanes are prostaglandin-like compounds derived from nonenzymatic free radical-catalyzed peroxidation of arachidonic acid. 8-epi-Prostaglandin (PG) F 2␣ , a major component of the F 2 -isoprostane family, can be conveniently measured in urine to assess noninvasively lipid peroxidation in vivo. Measurement of major metabolites of endogenous 8-epi-PGF 2␣ , in addition to the parent compound, may be useful to better define its formation in vivo. 2,3-Dinor-5,6-dihydro-8-epi-PGF 2␣ is the only identified metabolite of 8-epi-PGF 2␣ in man, but its endogenous levels are unknown. In addition to this metabolite, we have identified another major endogenous metabolite, 2,3-dinor-8-epi-PGF 2␣ , in human and rat urine. The identity of these compounds, present at the pg/ml level in urine, was proven by a number of complementary approaches, based on: (a) immunoaffinity chromatography for selective extraction; (b) gas chromatography-mass spectrometry for structural analysis; (c) in vitro metabolism in isolated rat hepatocytes; and (d) chemical synthesis of the enantiomer of 2,3-dinor-5,6-dihydro-8-epi-PGF 2␣ as a reference standard. In humans, the urinary excretion rate of both dinor metabolites is comparable with that of 8-epi-PGF 2␣ . Both metabolites increase in parallel with the parent compound in cigarette smokers, and they are not reduced during cyclooxygenase inhibition. Another ␤-oxidation product, 2,3,4,5-tetranor-8-epi-PGF 2␣ , was identified as a major product of rat hepatocyte metabolism. In conclusion, at least two major ␤-oxidation products of 8-epi-PGF 2␣ are present in urine, which may be considered as additional analytical targets to evaluate 8-epi-PGF 2␣ formation and degradation in vivo.

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“…A second clean-up could involve a silica cartridge for removal of neutral lipids, then elution of eicosanoids in methanol. The most specific solid-phase extraction involves antibody binding of the analyte, with the antibody bound to a Sepharose matrix [7,8].…”

Section: Extractionmentioning

confidence: 99%

Analysis of eicosanoids and related lipid mediators using mass spectrometry

Maskrey

O'Donnell

2008

Biochemical Society Transactions

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Eicosanoids are oxidized arachidonate-derived lipids products generated by lipoxygenase, cyclo-oxygenase and cytochrome P450. Their bioactivity is mediated via receptor-dependent mechanisms and they are involved in a diverse array of processes in both health and disease. For many years, GC-MS was the method of choice for eicosanoid analysis, but, more recently, the availability of high-sensitivity electrospray LC (liquid chromatography)-MS/MS (tandem MS) has provided a new approach for quantification, while minimizing sample preparation. The present review summarizes the various methods available for routine quantification of eicosanoids, focusing ultimately on their analysis using a hybrid Q-Trap mass spectrometer.

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Antibody-mediated extraction/negative-ion chemical ionization mass spectrometric measurement of thromboxane B2 and 2,3-dinor-thromboxane B2 in human and rat urine

Cited by 51 publications

References 13 publications

Differential effect of aspirin on thromboxane and prostaglandin biosynthesis in man.

Differential effect of aspirin on thromboxane and prostaglandin biosynthesis in man.

Identification and Measurement of Endogenous β-Oxidation Metabolites of 8-epi-Prostaglandin F2α

Analysis of eicosanoids and related lipid mediators using mass spectrometry

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