Semi-micro high-performance liquid chromatographic analysis of tiropramide in human plasma using column-switching.

A rapid and sensitive column-switching semi-micro high-performance liquid chromatography method was developed for the direct analysis of tiropramide in human plasma. The plasma sample (100 microl) was directly injected onto Capcell Pak MF Ph-1 precolumn where deproteinization and analyte fractionation occurred. Tiropramide was then eluted into an enrichment column (Capcell Pak UG C(18)) using acetonitrile-potassium phosphate (pH 7.0, 50 mM) (12:88, v/v) and was analyzed on a semi-micro C(18) analytical column using acetonitrile-potassium phosphate (pH 7.0, 10 mM) (50:50, v/v). The method showed excellent sensitivity (limit of quantification 5 ng/ml), and good precision (C.V.<or=5.9%), and speed (total analysis time 20 min). The calibration curve was linear in the concentration range 5-200 ng/ml (coefficient of determination equal to 0.998). This method was successfully applied to the pharmacokinetic study of tiropramide in volunteers.