反相高效液相色谱-紫外法测定血浆中咪达唑仑浓度

Determination of the Concentration of Midazolam in Plasma by RP-HPLC-UV Detection

目的 :建立血浆中咪达唑仑含量的反相高效液相色谱 -紫外检测方法。方法 :采用HypersilODS反相色谱柱 (250mm×4 0mm ,5μm ) ,甲醇 -乙腈 -0 02mol/L磷酸钾缓冲液 ( pH7 4) (65∶25∶10 ,V/V )为流动相 ,样品经碱化提取分离后流动相溶解 ,在221nm波长处检测。结果 :本法测定的标准曲线在50～1600ng/ml范围内呈现良好线性关系 (r=0.9999) ,最低检测浓度为2ng/ml ,绝对回收率在90 8 %～95 4 %之间 ,方法回收率在99 3 %～101 3 %之间 ,日内、日间变异系数分别为1 94 %～5 16 %、3 00 %～6 39 %。结论 :用高效液相色谱 -紫外法检测咪达唑仑的血药浓度 ,方法简便、稳定、灵敏度高 。

OBJECTIVE:To build up a method of determining the concentration of midazolam in plasma by RPHPLC-UV detection.METHODS:The separation was carried out by a reversed-phase Hypersil ODS column(250mm×4.0mm,5μm) with a mobile phase consisting of methanol-acetonitrile-0.02mol/L potassium phosphate buffer(pH 7.4) (65∶25∶10,V/V).Mida_zolam was extracted from alkalinizing plasma and soluted in the mobile phase then detected at 221nm.RESULTS:The calibration curve had the fine linearity in the concentration range 50～1 600ng/ml(r=0.9 999).The detection limit was 2ng/ml.The absolute recovery was 90.8%～95.4%,the relative recovery was 99.3%～101.3%.The within-day and between-day precision(CV%) was 1.94%～5.16%,3.00%～6.39% respectively.CONCLUSION:The method is simple,stable and highly sensitive and could meet with the research of clinical pharmacokinetics.