HPLC-ESI-MS测定中国健康人体内血浆中的双嘧达莫

HPLC-ESI-MS determination of dipyridamole in human plasma in healthy male Chinese volunteers

目的 建立一种简便、快速的高效液相色谱-质谱联用法测定(HPLC-ESI-MS)中国健康人体内血浆中双嘧达莫浓度的方法。方法 以乙腈沉淀处理血样。色谱柱为MACHEREY-MAGELNUCIEODUR 100-5 C_(18)柱(3μm,2.1mm×100mm),柱温50℃;流动相:乙腈-水(含30mmol·L^(-1)醋酸铵)=41.3:53.7(v/v);流速0.25mL·min^(-1)。质谱条件采用正离子检测的电喷雾电离方式(+)ESI其喷雾电压3.30kV,第一级锥孔电压59V;扫描方式为选择离子监测(SIR),用于定量分析监测的离子双嘧达莫为505.6 m/z,唑吡坦(内标)为308.3m/z。结果 双嘧达莫最低检测限为3.0μg·L^(-1),线性范围20～2500μg·L^(-1)(r=0.9910)。高、中、低浓度回收率分别为105.0％、105.6％、100.4％(n=5),日内RSD低于15.0％(n=5),日间RSD低于10.0％(n=5)。结论 该方法灵敏度高、快速、简便、无杂质干扰,适合于双嘧达莫血药浓度检测及药物动力学研究。

OBJECTIVE To develop a simple and rapid HPLC-ESI-MS method to quantify dipyridamole in buman plasma in healthy male Chinese volunteers. METHODS The blood plasma samples were extracted by cetonitrile. Chromatographic column was MACHEREY-MAGEL NUCLEODUR 100—5 C_(18) column (3μm, 2.1 mm×100 mm) and 50℃ of column temperature. The mobile phase was composed of acetonitrile and distilled water (contained 30 mmol·L^(-1) ammonium acetate) at the ratio of 41.3: 58.7 (v/v). The flow rate was 0.25 mL·min^(-1). The voltage of electrosprary ionization was 3.30 kV, cone voltage was 59 V and then SIR was used as a kind of scan model. The ion of dipyridamole was 505.6 m/z and zolpidem (an interal standard, IS) was 308.3 m/z. RESULTS The lower limit of detection was 3.0μg·L^(-1). The calibration curve was in good linearity over the range of 20～2500 μg·L^(-1) with the regression equation A/A_(IS)=0.000 572 7 C+0.012 47 (r^2=0.999 0). The methodology recoveries were much higher. The intra-day and inter-day RSD were lower than 15%. CONCLUSIONS The method is sensitive, rapid, free from impurity interference and convenient.