高效液相色谱法测定咪达唑仑在小鼠体内的分布

Determination of Midazolam in Mouse Plasma Serum Concentration Distribution by Reversed-Phase HPLC

目的采用HPLC测定咪达唑仑在小鼠体内的分布情况。方法采用Kromasil C18色谱柱(4.6 mm×150 mm,5μm),甲醇-水(含0.1%二乙胺,0.05%磷酸溶液,60∶40)为流动相,流速1.0 mL.min-1,检测波长225 nm。以三唑仑为内标物,按内标法定量。结果咪达唑仑在5～400 ng.mL-1内线性关系良好(r=0.999 3),咪达唑仑最低检测质量浓度为2.5 ng.mL-1,平均回收率为(96.95±1.8)%,日内及日间的RSD均小于2%。结论本方法具有快速、简便、灵敏、准确、经济等特点,适用于测定咪达唑仑在小鼠体内的分布,为该药物的临床检测提供依据。

OBJECTIVE Determination by HPLC of midazolam in mouse distribution, and the method was applied to detect midazolam in mouse distribution. METHODS The analytical column was packed with Kromasil C18 （4. 6 mm× 150 mm, 5 ×μm） the mobile phase was methanol-water 60： 40, containing 0. 1% diethyl amine, 0. 05 % phosphoric acid , the flow rate was 1 mL . min- 1, and detection wave length was set at 225 nm, with the triazolanl as the internal standard, according to the internal standard to quantitate. RESULTS The linear ranges were 5 - 400 ng .mL- 1 for midazolam in human plasima. ： The correlation coefficients were 0. 999 3. The midazolam low limit of detection was 2.5 ng .mL-1. The method recoveries were （96. 95 ± 1.8）%. Withinday and betweenday RSD were less than 2%. CONCLUSION The method is rapid, simple, sensitive, accurate and.economic, and applied to provide basis for clinical testing for midazolam in mouse distribution.