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 F1α (6-keto-PGF1α), 2,3-dinor-6-ketoprostaglandin F1α (2,3-dinor-6-keto-PGF1α), 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-PGF1α and anti-TXB2 antibodies (cross-reacting with 2,3-dinor-6-keto-PGF1α 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 PGF1α, 2,3-dinor-6-keto-PGF1α, 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-PGF1α 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-PGF1α or TXB2, since it allows profiling of a group of metabolites whose balance is important in several physiopathological conditions.
|Number of pages||11|
|Journal||Journal of Chromatography B: Biomedical Sciences and Applications|
|Publication status||Published - 1989|
ASJC Scopus subject areas
- Clinical Biochemistry
- Molecular Medicine