Microbore high-performance liquid chromatographic determination of cisapride in rat serum samples using column...

For the determination of cisapride from serum samples, an automated microbore high-performance liquid chromatographic method with column switching has been developed. After serum samples (100 microl) were directly injected onto a Capcell Pak MF Ph-1 pre-column (10 x 4 mm I.D.), the deproteinization and concentration were carried out by acetonitrile-phosphate buffer (20 mM, pH 7.0) (2:8, v/v) at valve position A. At 2.6 min, the valve was switched to position B and the concentrated analytes were transferred from MF Ph-1 pre-column to a C18 intermediate column (35x2 mm I.D.) using washing solvent. By valve switching to position A at 4.3 min, the analytes were separated on a Capcell Pak C18 UG 120 column (250 x 1.5 mm I.D.) with acetonitrile-phosphate buffer (20 mM, pH 7.0) (5:5, v/v) at a flow-rate of 0.1 ml/min. Total analysis time per sample was 18 min. The linearity of response was good (r=0.999) over the concentration range of 5-200 ng/ml. The within-day and day-to-day precision (CV) and inaccuracy were less than 3.7% and 3.8%, respectively. The mean recovery was 96.5+/-2.4% with the detection limit of 2 ng/ml.