The Purification Step is not Crucial in EIA Measurements of Thromboxane B 2 and 11-Dehydrothromboxane B2 in Human Plasma
Sadilkova, L.; Paluch, Z.; Mottlova, J.; Bednar, F.; Alusik, S.
Clinical Laboratory 58(1-2): 177-183
2012
ISSN/ISBN: 1433-6510 PMID: 22372363 Document Number: 15539
Background: Thromboxane B 2 (TxB 2 ) and particularly 11-dehydrothromboxane B 2 (11-dTxB 2 ) are widely used as prognostic risk markers of platelet activation in cardiovascular diseases. The main errors in TxB 2 and 11-dTxB 2 determination include either low concentrations of circulating TxB 2 (1 - 2 pg/mL) and 11-dTxB 2 (0.9 - 4.3 pg/mL) or rather high transiency (mean TxB 2 half-life is approximately 5 minutes) as well as an incorrect preanalytical phase set up. The aim of this study was to investigate the impact of a widely used purification step on the results of enzyme immunosorbent assay (EIA) - based measurement of the two selected thromboxanes. Methods: For the purpose of this study, 20 plasma samples (10 healthy donors, 10 patients under treatment with acetylsalicylic acid) were screened for TxB 2 and 11-dTxB 2 concentrations using commercial competitive EIA kits (Cayman Chemicals TM , Tallinn, Estonia; Neogen TM , Lexington, KY, USA) with or without the introduction of the purification procedure. Results: The purification step does not significantly affect the results of EIA measurements of the two of TxA 2 metabolites (TxB 2 , 11-dTxB2) in human plasma. The levels of TxB 2 and 11-dTxB 2 determined in the plasma samples were not significantly changed (p 2 and 11-dTxB 2 EIA measurement for routine basic screening of platelet function.
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