高效液相色谱法测定人血浆中咪唑安定的质量浓度

HPLC for determination of plasma midazolam concentration

目的利用高效液相色谱测定人血浆中咪唑安定的质量浓度。方法以安定为内标,Hypersil ODS C18为色谱柱,以甲醇-乙腈-磷酸盐缓冲液为流动相;柱温：35℃;流速：1.0 mL.min-1;检测波长：220 nm。结果咪唑安定质量浓度在10~1 000μg.L-1范围内与峰面积比呈良好的线性关系,标准曲线方程为y=0.012 x＋0.434（r=0.9964,P〈0.001）,最低检测质量浓度为1μg.L-1。日内RSD≤4.83%,日间RSD≤5.79%;低、中、高质量浓度的提取回收率分别为79.43%、83.98%和85.75%,方法回收率分别为96.56%、104.87%和102.98%。结论利用高效液相色谱法测定人血浆中咪唑安定的质量浓度方法可靠、专属性理想、稳定性好,适合临床进行咪唑安定的药动学研究。

Objective To develop a HPLC assay for determining human plasma level of midazolam.Methods Diazepam was used as internal standard.The analytical column was packed with Hypersil ODS C18,and temperature was set at 35℃.The mobile phase consisted of methanol-acetonitrile-phosphate buffer,pumped at 1.0 mL·min-1,and measured at 220 nm.Results The linear range of midazolam was 10~1000 μg·L-1,and the minimal limit of detection was 1 μg·L-1.The regression equation was expressed as y=0.012x＋0.434（r=0.9964,P〈0.001）.The intraday and interday RSD was ≤4.83% and ≤5.79%,respectively.The extraction and assay recovery of low,middle and high concentrations was 79.43%,83.98%,85.75% and 96.56%,104.87%,102.98%,respectively.Conclusion HPLC assay is reliable,accurate and sensitive for determination of midazolam in human plasma,and is suitable for pharmacokinetics of midazolam.