高效液相色谱-质谱联用法测定人血浆中羟氯喹的浓度

Determination of hydroxychloroquine in human plasma by HPLC-MS/MS

目的：建立人血浆中羟氯喹浓度测定的HPLC-MS/MS法。方法：采用Kinetex C18色谱柱（2.6μm,50 mm×2.1mm）;流动相为0.6%甲酸水溶液（pH3.2）-甲醇（80∶20）;流速0.5 ml·min-1;柱温为25℃;电喷雾电离（ESI）正离子检测,检测对象：羟氯喹m/z336.2→247.2;内标氯喹m/z320.2→247.2。20名男性健康志愿者单剂量口服硫酸羟氯喹片200 mg（100mg/片）,在设计的时间点取静脉血,用建立的方法测定血药浓度。结果：羟氯喹和内标氯喹的保留时间分别在1.3和1.5 min,羟氯喹在0.200~100μg·L-1范围内线性关系良好（r=0.993 0）,最低定量浓度为0.2μg·L-1;方法回收率为98.20%~102.75%,批内、批间RSD小于10%（n=6）。主要药动学参数：tmax（3.85±1.04）h,t1/2（32.3±14.1）h,Cmax（44.1±27.6）μg·L-1,AUC0-72 h（617±88.9）μg·h·L-1,AUC0-∞（749±108）μg·h·L-1。结论：该方法灵敏度高、结果准确、样品处理简单、分析快速,适用于羟氯喹制剂的血药浓度测定。

OBJECTIVE To establish a method to detect the concentration of hydroxychloroquine（HCQ） in human plasma by HPLC-MS/MS. METHODS Chromatograph was carried out on a Kinetex C18 column. The mobile phase was 0. 6G formic acid water （pH 3.2）- methyl alcohol（80.-20）. The flow rate was set at 0.5ml.min-1. Column temperature was 25 ~C. SRM. HCQ ~M ± HI ± m/z 336.2---~247.2~ internal standard CQEM ± HI ± m/z 320. 2---~247.2. Twenty healthy volunteers were given 200 mg of hydroxychloroquine sulfate tablets for each. Hydroxychloroquine concentration in plasma was determined by HPLC-MS/ MS method. RESULTS The retention time of HCQ and internal standard CQ were approximately 1.3 min and 1.5 rain, re- spectively. A good linearity was shown in concentration range 0. 200-100 /lg.L-1 for HCQ. The LLOQ was 0. 2 μg·L-l. The method recovery was 98. 20%-102.75% The inter- or intra- precision was less than 10% in replicate analysis （n = 6）. The pharmacokinetic parameters of hydroxychloroquine were as follows：tmx were （3.85 ± 1.04） h, tl/2 （32. 3 ± 14. 1） h·Cmax （44. 1 ± 27. 6） μg·L-1 ,AUGI72h（617± 88. 9） μg·h·L 1 ,AUG （749± 108） μg·h·L-1 o CONCLUSION The method is proved to be sensitive, robust, convenient, analysis rapid, and suitable for the clinical investigation and monitoring of the drug concentration in the plasma.