HPLC-荧光检测法测定大鼠血浆伊伐布雷定浓度

 目的 建立测定血浆中伊伐布雷定浓度的高效液相-荧光(FLU)测定方法，用于临床血浆伊伐布雷定浓度的测定。方法 血浆样品经Varian C₂柱固相萃取后采用HPLC-FLU法在Kromasil C₁₈柱分离，流动相采用乙腈-10 mmol·L - 1磷酸二氢钾（含0.3%10 mmol·L-1盐酸）（28∶72），流速为1 mL ·min- 1，柱温为25 ℃，荧光检测波长：λ₁=328 nm，λ₂=283 nm。结果 内源性杂质不干扰测定，伊伐布雷定在0.5～50 μg·L-1内线性良好（r=0.999 6），最低定量限为0.5 μg·L-1。萃取回收率为78.74%～90.77%（n=5），批内和批间精密度（RSD%）分别为2.53～6.33（n=6）和6.38～8.91（n=3）。结论 建立的固相萃取-HPLC-FLU方法简便、灵敏、准确、所需血浆量少，可以用于临床血药浓度测定。

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.