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Clinical Chemistry 39: 2470-2477, 1993;
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Clinical Chemistry, Vol 39, 2470-2477, Copyright © 1993 by American Association for Clinical Chemistry

Rapid, direct enzyme immunoassay of 11-keto-thromboxane B2 in urine, validated by immunoaffinity/gas chromatography-mass spectrometry

R Djurup, C Chiabrando, A Jorres, R Fanelli, M Foegh, HU Soerensen and PN Joergensen
Novo Nordisk, Immunochemical Department, Bagsvaerd, Denmark.

We have developed a direct enzyme immunoassay (EIA) for quantifying immunoreactive 11-keto-thromboxane B2 (iKTXB) in unprocessed human urine. Cross-reactivity with other thromboxane metabolites and prostanoids was negligible. Analytical recovery of 11-keto-TXB2 in urine specimens was 97.4% to 99.8%. Total imprecision for two clinical specimens was 8.5% and 12.2%. Intake of acetylsalicylic acid decreased the measured concentration of iKTXB. Cardiopulmonary bypass, a procedure known to activate platelets, increased the mean excretion rate of iKTXB 10-fold. Simultaneous gas chromatography-mass spectrometry analysis of 11-keto-TXB2 and 11-keto-2,3-dinor TXB2 in urine specimens (n = 17) from healthy subjects indicated that urinary iKTXB concentrations measured by EIA represented a sum of the two 11- keto metabolites. We conclude that the direct EIA is sufficiently sensitive, rapid, simple, and specific to allow screening for alterations in thromboxane biosynthesis in patients.


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