超滤配合LC-MS/MS法测定普伐他汀人血浆蛋白结合率

Determination of Protein Binding Rate of Pravastatin in Human Plasma by Ultrafiltration Coupled with LC-MS/MS

目的:测定普伐他汀人血浆蛋白结合率。方法:应用超滤法(A法)和溶剂萃取法(B法)处理样品,配合高效液相色谱串联质谱法分别检测普伐他汀游离型药物血浆样品与总的药物血浆样品,以瑞舒伐他汀为内标,采用正离子方式检测,多反应监测(MRM)模式扫描,检测离子通道为普伐他汀m/z448.2→327.7和瑞舒伐他汀m/z483.2→258.6。结果:普伐他汀血药浓度在0.25～50.00ng/ml范围内线性关系良好,定量限为0.25ng/ml;A、B法平均加样回收率分别是95.2%、87.2%、90.4%与87.5%、88.2%、90.7%。普伐他汀和瑞舒伐他汀的保留时间分别是1.0、1.3min;在1、2、4、6h时人血浆蛋白结合率分别为(50.5±5.7)%、(51.2±3.5)%、(49.8±6.8)%、(52.4±10.1)%。结论:本方法灵敏度高、分析时间短、专属性好、准确度高,可用于普伐他汀血浆蛋白结合率的测定。

OBJECTIVE： To determine the protein binding rate of pravastatin in human plasma. METHODS： After treated with ultrafiltration （method A） and solvent extraction method （method B）, LC-MS/MS were used to determine the plasma samples of free type and total type drug of pravastatin in plasma sample using rosuvastatin as internal standard. MS condition： in positive ion mode, MRM mode scanning, detection ion channels m/z 448.2→327.7 and m/z 483.2→258.6 for pravastatin sodium and rosuvas- tatin, respectively. RESULTS： The linear range of pravastatin was 0.25-50.00 ng/ml, and limit of quantification was 0.25 ng/ml; the average recoveries were 95.2%, 87.2% and 90.4% by method A and 87.5%, 88.2% and 90.7% by method B. The retention time of pravastatin and rosuvastatin were 1.0 min and 1.3 min, respectively. The protein binding rate of pravastatin in human plas- ma were （50.5 ± 5.7）%, （51.2 ± 3.5）%, （49.8 ± 6.8）% and （52.4 ±10.1）% at 1, 2, 4, 6 h. CONCLUSIONS： The method has high sensitivity, short analytic time, specificity and accuracy, and it can be used for the determination of plasma protein binding rate of pravastatin.