Determination of Ivabradine in Rat Plasma by a Validated HPLC with Fluorescence Detection

OBJECTIVE To develop a HPLC-FLU method for the determination of ivabradine in plasma.METHODS The plasma samples with Varian C2 solid-extraction pretreatment were separated on a Kromasil  C18 column,using acetonitrile-10 mmol·L-1KH2PO4(containing 0.3% 10 mol·L-1 hydrochloride)(28∶72) as the mobile phase.The flow rate was 1.0 mL·min-1 at room temperature.Detection was carried out at λ1=328 nm and λ2=283 nm.RESULTS The method was linear for ivabradine in the range of 0.5-50 μg·L-1.The correlation coefficient was 0.999 6 and the LLOQ was 0.5 μg·L-1.The extraction recoveries were in the range of 78.74%-90.77%(n=5).The intra-day and inter-day precisions(RSD%) were 2.53-6.33(n=6) and 6.38-8.91(n=3).CONCLUSION The HPLC-FLU method was simple,sensitive and accurate.It was suitable for the determination of ivabradine in plasma.