Quantitative profiling of 6-ketoprostaglandin F1α, 2,3-dinor-6-ketoprostaglandin F1α, thromboxane B2 and 2,3-dinor-thromboxane B2 in human and rat urine by immunoaffinity extraction with gas chromatography-mass spectrometry

Chiara Chiabrando, Vittorio Pinciroli, Andrea Campoleoni, Ariela Benigni, Antonella Piccinelli, Roberto Fanelli

Research output: Contribution to journalArticlepeer-review

Abstract

A rapid and simple method based on immunoaffinity extraction, stable isotope dilution and gas chromatography-mass spectrometry has been developed for profiling urinary metabolites of prostacyclin and thromboxane. 6-Ketoprostaglandin F (6-keto-PGF), 2,3-dinor-6-ketoprostaglandin F (2,3-dinor-6-keto-PGF), thromboxane B2 (TXB2) and 2,3-dinor-thromboxane B2 (2,3-dinor-TXB2) were quantitatively extracted from human or rat urine spiked with deuterated internal standards using mixed-bed columns containing immobilized anti-6-keto-PGF and anti-TXB2 antibodies (cross-reacting with 2,3-dinor-6-keto-PGF and 2,3-dinor-TXB2, respectively). The extract was directly derivatized to form pentafluorobenzyl ester, methyloxime, trimethylsilyl ether derivatives. Quantitation was performed by stable isotope dilution assay and high-resolution gas chromatography-negative ion chemical ionization mass spectrometry, by monitoring the carboxylate anions (M - 181) of the derivatized metabolites. The method was applied to evaluate the urinary excretion of 6-keto PGF, 2,3-dinor-6-keto-PGF, TXB2 and 2,3-dinor-TXB2 in humans and rats. Results were in accordance with previously reported data obtained by other methods. Novel data on the urinary excretion of 2,3-dinor-6-keto-PGF in rats under basal conditions are presented. This sensitive and selective method represents a significant advance in terms of rapidity and simplicity over other immunoaffinity-gas chromatography-mass spectrometry methods for measuring single prostanoids, such as 6-keto-PGF or TXB2, since it allows profiling of a group of metabolites whose balance is important in several physiopathological conditions.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume495
Issue numberC
DOIs
Publication statusPublished - 1989

ASJC Scopus subject areas

  • Chemistry(all)
  • Clinical Biochemistry
  • Molecular Medicine

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