thromboxane-b3 and Leukemia--Erythroblastic--Acute

We devised a simple and effective purification for the microdetermination of thromboxane B3 (TXB3), a hydrolysis product of TXA3- [18O2]TXB3 was synthesized by the repeated base-catalyzed hydrolysis of methyl ester derivatives in [18O]water, to obtain an internal standard (IS) for the gas chromatography/selected ion monitoring (GC/SIM) of TXB3. The methyl ester (ME)-methoxime (MO)-dimethylisopropylsilyl (DMIPS) ether derivative was prepared, then GC/SIM was carried out by monitoring the ion at m/z 668 for TXB3 and that at m/z 672 for IS. A good linear response over the range of 10 pg approximately 10 ng was demonstrated. We were able to detect the levels of TXB3 in the medium of human erythroleukemia (HEL) cell cultured with eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA). This method can be used to determine 3-series thromboxane in biological samples.

Topics: Calibration; Chromatography, Gas; Culture Media, Conditioned; Docosahexaenoic Acids; Eicosapentaenoic Acid; Humans; Isotope Labeling; Leukemia, Erythroblastic, Acute; Microchemistry; Oxygen Isotopes; Reproducibility of Results; Thromboxane B2; Thromboxanes; Tumor Cells, Cultured