Development of quantitative analysis of plasma thromboxane B2 by gas chromatography-mass spectrometry.

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Abstract
To diagnose patients in thrombotic state, it is quite important to detect increased concentration of plasma thromboxane B2 (TXB2), a stable catabolite of TXA2. To determine plasma TXB2 levels with high sensitivity and selectivity, gas chromatography-mass spectrometry (GC/MS) was used. The trimethylsilyl (TMS) ether derivatives conventionally employed in GC/MS analysis of prostanoids were not suitable for quantitation of plasma prostanoids, because the mass spectra were deficient in ions with high intensity in the high mass range and TMS ether derivatives were sensitive to mousture. To solve these problems tert-butyldimethylsilyl (t-BDMS) ether derivatives were used, based on the observation that t-BDMS ether derivatives affored abundant ions at [M-57]+ and showed good hydrolytic stability. The reaction conditions of tert-butyldimethylsilylation were also examined to optimize the selected ion monitoring response. The t-BDMS ether derivatives of prostanoids were successfully analyzed with a short capillary column with a relatively large diameter, with maintaining good separation. In conjunction with the use of reversed-phase high performance liquid chromatography as purification procedure, a sensitive and reproducible stable isotope dilution assay of plasma TXB2 was developed.