【摘要】 目的:建立一种同时、快速检测人血浆中甲磺酸伊马替尼(imatinib mesylate, IM)及其代谢产物浓度的二维液相色谱法,并应用于临床。方法:血浆样品经过蛋白沉淀、离心后直接进样,通过使用二维液相色谱仪进行检测。一维萃取柱:型号为Aston SX1(3.5 mm×25 mm, 5μm),流动相为V(甲醇)∶V(乙腈)∶V(磷酸铵水溶液)=25∶20∶55,流速=0.8 mL/min;中间柱:型号为Aston SCB(4.6 mm×10 mm, 3.5μm);二维分析柱:型号为Aston SCB(4.6 mm×125 mm, 5μm),流动相为V(乙腈)∶V(磷酸铵盐溶液)=38∶62,pH=6.9,流速=1.2 mL/min;柱温:40℃,检测波长:264 nm,进样量:500μL。结果:色谱图显示IM及其代谢产物与杂质分离良好;IM与N-去甲基伊马替尼(N-Desmethyl imatinib, NDI)血药浓度线性范围分别是13.44～1 400.00 ng/mL(R~2=0.9999)与13.17～1 372.00 ng/mL(R~2=0.9999),线性关系均良好,定量下限变异系数分别为5.79%和2.77%,灵敏度良好;二者均可在12.80 min内完全出峰,方法回收率均高于90%,日内、日间精密度均小于6%,稳定性试验相对标准偏差均小于5%。所建立方法对10位口服IM的胃肠间质瘤患者进行了IM及NDI血药浓度检测,结果IM血药浓度结果为300.34～1 392.07 ng/mL不等;NDI血药浓度在122.44～446.34 ng/mL之间。结论:该方法简便、快捷、灵敏度高、重现性好,结果准确且成本低廉,且适用于临床IM血药浓度检测。更多还原

【Abstract】 Objective: To establish a system of two-dimensional liquid chromatography with ultraviolet detection for simultaneously and rapidly detecting the concentration of imatinib mesylate(IM) and its metabolites in human plasma, and apply it in clinical practice. Methods: Plasma samples were injected directly onto the system after protein precipitation and centrifugation, and were detected by two-dimensional liquid chromatography. The first-dimension column was in version Aston SX1(3.5 mm×25 mm, 5 μm), with the mobile phase of a solution of methanol, acetonitrile and ammonium phosphate aqueous solution [V(methanol)∶V(acetonitrile)∶V(ammonium phosphate aqueous solution)=25∶20∶55], and the flow rate of(0.8 mL/min); the middle column was in version Aston SCB(4.6 mm×10 mm, 3.5 μm); the second-dimension column was in version Aston SCB(4.6 mm×125 mm, 5 μm), with the mobile phase of a solution of acetonitrile and ammonium phosphate salt solution [V(acetonitrile)∶V(ammonium phosphate salt solution)=38∶62, PH=6.9], and the flow rate of 1.2 mL/min; the column temperature was 40 ℃; the detection wavelength was 264 nm; and the injection volume was 500 μL. Results: The chromatogram showed that IM and its metabolites were well separated from impurities. The linear ranges of plasma IM and N-Desmethyl imatinib(NDI) concentrations were 13.44～1,400.00 ng/mL(R~2=0.9999) and 13.17～1,372.00 ng/mL(R~2=0.9999), respectively, indicating a good linear relationship; the variation coefficients of the lower limit of quantification were 5.79% and 2.77%, respectively, suggesting a good sensitivity; both IM and NDI could completely peak within 12.80 min, with the recovery rates of over 90%, with intra-day and inter-day precision of less than 6%, with stability of less than 6%(relative standard deviation). The established method was performed on 10 gastrointestinal stromal tumor patients who received IM orally. The concentration of plasma IM ranged from 300.34 to 1,392.07 ng/mL, and that of plasma NDI was between 122.44 and 446.34 ng/mL. Conclusion: The method is simple, fast, highly sensitive, reproducible and accurate with low cost, which is suitable for clinical detection of plasma IM concentration.更多还原