Thin layer chromatographic identification of thromboxane B2 in serum, and platelet rich plasma stimulated by arachidonic acid and collagen.

Abstract

The thromboxane B2(TxB2) level in platelet rich plasma (PRP), after arachidonic acid and stimulus, and its presence in serum, may help determine abnormalities in the platelet prostaglandin pathway} For example, if no TxB2 is produced in response to any aggregating agent, this could be due to cyclo-oxygenase deficiency, thromboxane synthetase deficiency or more commonly aspirin ingestion. Similarly, TxB2 is absent from serum2 h after the ingestionof aspirin. T~ is usually measured by radioimmunoassay (RIA) and by high performance liquid chromatography (HPLC). RIA is expensive and not well suited for small batch analysis. The lack of ultraviolet absorption of Tx~ means that a derivitization stage is necessary prior to analysis by HPLC.2 The difficulties encountered in the determination of TxB2, usually mean that its estimation is rarely undertaken during platelet studies. Thin layer chromatography (TLC) can be performed at reasonable cost, with minimal equipment, and has previously been used to separate TxB2 from other prostaglandins.! Since TxB2is synthesized in large amounts in response to arachidonic acid and collagen and the serum concentration of TxB2 is of the same order as that in PRP stimulated with arachidonic acid, the simplicity and general availability of TLC would make it a useful method in the identification of TxB2 during the study of arachidonic acid metabolism in platelets.