高效液相色谱法同时测定伊马替尼及主要代谢物去甲基伊马替尼的血药浓度

Simultaneous determination of concentrations of imatinib and N-desmethyl imatinib in human plasma by HPLC

目的:建立同时测定人血浆中伊马替尼及其主要代谢物去甲基伊马替尼的高效液相色谱法。方法:用1 mol·L^(-1)NaOH碱化血桨样品后经乙酸乙酯-正己烷(75∶25)萃取,选择达沙替尼为内标,浓缩、复溶后采用HPLC分析。色谱柱为ZORBAX SB-C18柱反相柱(250 mm×4.6 mm,5μm),流动相为乙腈-磷酸二氢钾缓冲液(25 mmol·L^(-1)KH_2PO_3,H_2PO_3调p H值至3)=27∶73(V/V),检测波长为267 nm,流速为1 mL·min^(-1),柱温为35℃,进样量为20μL。结果:伊马替尼和去甲基伊马替尼血药浓度线性范围分别为0.10~5.00和0.01~0.5μg·mL^(-1),二者线性均良好(r分别为0.998和0.997),最低定量下限分别为0.10和0.01μg·mL^(-1)。伊马替尼和去甲基伊马替尼低、中、高浓度的提取回收率分别为86.02%,87.31%,88.21%和81.64%,84.78%,88.74%。二者批内和批间RSD均小于15%。结论:该方法简便、快速、灵敏度高,能同时满足检测伊马替尼及其代谢物血药浓度,可用于临床常规血药浓度监测、药动学研究及体外代谢研究。

Objective： To establish a method for determining the concentrations of imatinib and its metabolites N-desmethyl imatinib in plasma by HPLC. Methods： Blood samples were pretreated by 1 mol·L^-1 NaOH using ethyl acetate n-hexane(75∶ 25） with dasatinib as an internal standard,then concentrated in vacuum,and the reconstituted solution was detected by HPLC. The analytical column was ZORBAX SB-C18(250 mm ×4. 6 mm,5 μm）,mobile phase was acetonitrile-potassium dihydrogen phosphate buffer(0. 025 mol·L^-1 KH2PO3,pH value was adjusted to about 3. 0 by H2PO3）(27∶ 73,V/V） at a flow rate of 1. 0 mL·min^-1. The detection wavelength was set at 267 nm,the temperature was kept at 35 ℃,and the injection volume was 20 μL. Results：The calibration curves had good linearity in the rang of 0. 10 5. 00 μg·mL^-1 for imatinib and 0. 010 0. 05μg·mL^-1 for N-desmethyl imatinib(r = 0. 998 and 0. 997,respectively）. The limits of quantification of imatinib and N-desmethyl imatinib were 0. 1 μg·mL^-1 and 0. 01 μg·mL^-1,respectively. The extraction recoveries of low,medium and high concentration control samples were 86. 02%,87. 31%,88. 21% for imatinib and 81. 64%,84. 78%,88. 74% for N-desmethyl imatinib,respectively. The intra-day and inter-day RSD values were less than 15%. Conclusion： The method is simple,rapid and sensitive for drug monitoring and pharmacokinetic study on imatinib and N-desmethyl imatinib.