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The Purification Step is not Crucial in EIA Measurements of Thromboxane B2 and 11-Dehydrothromboxane B2 in Human Plasma by Lenka Sadilkova, Zoltan Paluch, Jirina Mottlova, Frantisek Bednar, Stefan Alusik

Background: Thromboxane B2 (TxB2) and particularly 11-dehydrothromboxane B2 (11-dTxB2) are widely used as prognostic risk markers of platelet activation in cardiovascular diseases.
The main errors in TxB2 and 11-dTxB2 determination include either low concentrations of circulating TxB2 (1 - 2 pg/mL) and 11-dTxB2 (0.9 - 4.3 pg/mL) or rather high transiency (mean TxB2 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 TxB2 and 11-dTxB2 concentrations using commercial competitive EIA kits (Cayman ChemicalsTM , Tallinn, Estonia; NeogenTM, 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 TxA2 metabolites (TxB2, 11-dTxB2) in human plasma. The levels of TxB2 and 11-dTxB2 determined in the plasma samples were not significantly changed (p<0.05) when the purification step was omitted compared to the purified samples.
Conclusions: This study establishes a protocol allowing for reliable and reproducible plasma TxB2 and 11-dTxB2 EIA measurement for routine basic screening of platelet function.

DOI: Clin. Lab. 2012;58:177-183