High-performance liquid chromatographic determination of taurine in human plasma using pre-column extraction and derivatization

Plasma samples (100 mu l) were treated with 150 mu l of acetonitrile and centrifuged at 5800g for 10 min and 50 mu l of 10 mM berate buffer (pH 9.2) were added to the supernatant solution. This was followed by the addition of a 50 mu l aliquot of 5 mM fluorescamine in acetonitrile and immediate vortex mixing. A 20 mu l sample was injected on to a reversed-phase HPLC system using a Bondclone C-18 10 mu m analytical column (300 mm x 3.9 mm). The mobile phase was tetrahydrofuran-acetonitrile-phosphate buffer (15 mM, pH 3.5) (4:24:72, v/v/v). The taurine derivative was detected by measuring the UV absorbance of 385 nm. Platelet-poor plasma samples were spiked with known amounts of taurine and inter- and intra-assay calibration curves were obtained. The method was applied to the determination of taurine in platelet-rich plasma.