Electron capture gas chromatographic detection of thromboxane B2

An electron capture gas chromatographic method is described for the detection of thromboxane B2. Thromboxane B2 is esterified with diazomethane, followed by treatment with pentafluorobenzylhydroxylamine hydrochloride and silylation with BSA. In pyridine, the free aldehyde form of the acetal ring is favored allowing rapid formation of a novel thromboxane B2 pentafluorobenzyloxime. The method has been applied to detect thromboxane B2 formation during aggregation of washed platelets. It must be emphasized that by ordinary analytical standards, the derivatization reproducibility from 50-375 nanograms is poor (+/- 11% - +/- 42%); however, the improved selectivity of the method and its ability to detect nanogram levels of thromboxane B2 make it a useful complement to commonly employed bioassay techniques.