液相色谱-质谱联用法测定人血浆中达那唑的浓度

 目的建立测定人血浆中达那唑的液相色谱-质谱联用法(LC-MS)。方法取血浆,加入内标物炔诺酮,经正己烷液-液萃取后,在60℃水浴中氮气吹干,残渣用流动相溶解,进行LC-MS测定。色谱柱为Agilent Zorbax Eclipse XDB-C₁₈柱(4.6mm×150mm,5μm);以甲醇和0.05%甲酸水溶液为流动相,进行梯度洗脱;流速1mL·min^-1;柱温30℃;采用大气压化学离子源单极四极杆质谱,以选择性正离子方式检测;用于定量的选择检测离子m/z分别为338.2(达那唑)、299.2(炔诺酮,内标)。结果达那唑的线性范围为0.11～550ng·mL^-1,定量限为0.11ng·mL^-1,方法回收率均大于90%,日内、日间精密度(RSD))均小于8.0%。结论本方法专属性强,灵敏度高,线性范围宽,操作简便,适用于临床药动学研究。

Determination of danazol in human plasma by liquid chromatogram-mass spectrometry

OBJECTIVE To develop a sensitive and specific LC/MS method for the determination of Danazol in human plasma. METHODS Danazol and the internal standard norethisterone were extracted with liquid-liquid extraction. The organic layer was blowed by N₂ to dryness at 60℃.The residue was reconstituted in the mobile phase. The LC/MS method was performed on a Agilent Zorbax Eclipse XDB-C₁₈(4.6mm×150mm,5μm).The mobile phase consisted of CH₃OH and 0.05% HCOOH with gradient elution. The flow rate was at 1.0 mL·min^-1; The temperature of column was 30℃.Atmospheric pressure chemical ionization source was applied and operated in positive mode. The ion of monitor was at m/z 338.2(danazol) and m/z 299.2(norethisterone,internal standard) .RESULTS The linear calibration curve was observed in the concentration range of 0.11～550 ng·mL^-1. Thelower limit of quantification was 0.11 ng·mL^-1. The relative recoveries were more than 90%. The inter and intra-day precision (RSD) were below 8.0% .CONCLUSION The method was proved to be convenient, sensitive, rapid and suitable for pharmacokinetic study.