Key result
Trisodium citrate anticoagulation, incubation for 1 hour at 37 °C, and freezing at -20 °C or -80 °C provided the optimal concentration range for measuring plasma thromboxane A2 metabolites.
Optimal pre-analytical conditions for measuring plasma thromboxane A2 metabolites include using trisodium citrate, 1-hour incubation at 37 °C, and freezing at -20 °C or -80 °C.
May enhance thromboxane metabolite assay reliability in research; leaves open clinical adoption or standardization.
AIMS: Platelet function testing is often affected by the existence of different pre-analytical variables that can cause platelet activation. The aim of this study was to assess the effect of such variables that are present when samples are taken (different anticoagulants, incubation temperature, and storage conditions) to select those which enable to reach optimal range of measured plasma concentrations of the two stable thromboxane A₂ metabolites, that is, thromboxane B₂ (TxB₂) and 11-dehydrothromboxane B₂ (11-dTxB₂). MATERIALS AND METHODS: For the purpose of this study, whole blood samples obtained from 20 volunteers were screened for TxB₂ and 11-dTxB₂ concentrations using commercial EIA kits (Cayman Chemicals™; Neogen™) in relation to the effect of different anticoagulants, using different incubation temperatures and storage conditions. RESULTS: Trisodium citrate has been shown not to be affecting the TxB₂ and 11-dTxB₂ concentrations compared with the values measured in the serum. Incubation of the samples for 1 h at 37 °C and freezing at -20 °C or -80 °C give the most suitable concentration range of both thromboxanes in the used EIA measurement. CONCLUSION: This study describes the setup of such pre-analytical phase conditions that enable the screening of platelet function in terms of the plasma concentrations of TxB₂ and 11-dTxB₂ in selected EIA measurement.
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Sadílková et al. (2012) studied this question. Pre-analytical variables (anticoagulants, incubation temperature, storage conditions) was evaluated on Plasma concentrations of TxB2 and 11-dTxB2. Trisodium citrate anticoagulation, incubation for 1 hour at 37 °C, and freezing at -20 °C or -80 °C provided the optimal concentration range for measuring plasma thromboxane A2 metabolites.
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