液相色谱-串联质谱法检测人血浆中碘帕醇

Development of LC-MS/MS method for determination of plasma iopamidol

目的建立一种稳定的检测人血浆碘帕醇的液相色谱-串联质谱法(LC-MS/MS),并对该方法进行性能评价。方法以氘代同位素作为内标,人血浆样品经直接沉淀法前处理后进样,用XSelect^(TM) HSS PFP色谱柱进行分离,用含0.1%甲酸-5mmol/L九氟戊酸-10%乙腈的水溶液等度洗脱;正离子电喷雾离子化扫描检测,多反应监测(MRM)扫描分析,建立检测血浆碘帕醇的LC-MS/MS方法。根据CLSI EP10-A和CLSI C62-A,评价该方法的线性范围、正确度、精密度、选择性、基质效应、稳定性。结果 LC-MS/MS检测血浆碘帕醇的线性范围为1～1 000μgI/mL(以含碘量计算);低、中、高浓度(3、50、800μgI/mL)血浆质控品检测结果与理论靶值的偏移为-6.1%～7.5%,重复性以不精密度表示,CV<6.6%(4.6%～6.6%),期间精密度CV<8.5%(6.6%～8.5%),方法回收率为92.5%～95.9%,基质效应为88.8%～95.2%;方法选择性良好,溶血和脂血因素均不影响检测结果。血浆样品在室温下放置最好不要超过48 h,在-80℃下反复冻融不超过2次,经前处理后的血浆样品4℃自动进样器中放置不超过72 h,碘帕醇可稳定检测。结论建立并评价了一种可靠的检测人血浆碘帕醇的LC-MS/MS方法。

Objective To develop a stable LC-MS/MS method for the determination of iopamidol in human plasma and evaluate the performance of the method. Methods Deuterium isotope was used as internal standard. Plasma samples were pretreated by direct precipitation,separated by XSelectTMHSS PFP column and equally eluted by 10% acetonitrile solution containing 0.1% fomic acid and 5 mmol/L nonafluoropentanoic acid. The detection was performed by ionic scanning with positive-ion electrospray and the results were analyzed by multiple reaction monitoring( MRM) mode. According to the CLSI EP10-A and CLSI C62-A standards,the linear range,accuracy,precision,specificity,matrix effect and stability of the method were evaluated. Results The linear range of iopamidol determined by LC-MS/MS was 1 to 1 000 μg I/mL( calculated with iodine content). The deviations between the determined results and the theoretical target values were from -6.1% to 7.5% in low,medium and high concentrations of plasma samples( 3,50 and 800 μg I/m L).The repeatability was expressed as imprecision with CV less than 6.6%( 4.6% to 6.6%). The intermediate precision was expressed with CV less than 8.5%( 6.6% to 8.5%). The recoveries were from 92.5% to 95.9%. The matrix effects of the method were from 88.8% to 95.2%. The results of the method showed excellent selectivity,since hemolysis and lipemia did not affect the results. In terms of this method,plasma samples had better kept at room temperature for no more than 48 hours and repeatedly freeze-thaw from -80 ℃ for no more than 2 times,so that iopamidol could almost still be stably detected. Iopamidol was also stable in extracted samples at 4℃ in the autosampler for 72 hours. Conclusion A reliable LC-MS/MS method for determination of iopamidol in human plasma was established and evaluated.