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目的建立超高效液相色谱柱切换技术快速测定人血浆中的美罗培南浓度的方法。方法美罗培南血浆样品稀释、过滤,在线柱切换提取,分析色谱柱为Acquity UPLC BEH-C18柱。流动相A和提取流动相C:甲醇-0.05 mol·L-1K2HPO4(pH7.0)5∶95;流动相B:甲醇。柱温:35℃。梯度洗脱。美罗培南最大吸收波长299 nm。分别用内标法和外标法对测定结果进行计算,用SPSS统计软件对计算结果进行配对t检验,考察2种计算结果是否存在差异。结果人血浆美罗培南线性范围为0.67~86.00 mg·L-1(r=0.999 9),方法学回收率为(93.59±1.15)%~(99.21±4.06)%,提取回收率大于85%,日内日间精密度RSD小于5.0%。内、外标法配对t检验两者无显著差异,转换方程为ρInternal=1.039ρExternal-0.468(r=0.997 0)。结论采用在线柱切换技术快速、灵敏、高回收率、重现性好,适用于人血浆中美罗培南浓度监测和药动学研究。
Objective To establish a rapid method for determination of Meropenem in human plasma by ultra performance liquid chromatography (HPLC) column switching technique. Methods Meropenem plasma samples were diluted, filtered and switched on-line. The analytical column was Acquity UPLC BEH-C18 column. Mobile phase A and mobile phase C: methanol -0.05 mol·L-1K2HPO4 (pH 7.0) 5:95; mobile phase B: methanol. Column temperature: 35 ° C. Gradient elution. Meropenem maximum absorption wavelength of 299 nm. The results were calculated by internal standard method and external standard method, respectively, and SPSS statistical software was used to paired t-test results to see if there was any difference between the two methods. Results The linear range of meropenem in human plasma was 0.67-86.00 mg · L-1 (r = 0.999 9). The recovery rate was (93.59 ± 1.15)% ~ (99.21 ± 4.06)%, and the recovery rate was more than 85% The daytime precision RSD is less than 5.0%. There was no significant difference between the internal standard and the external standard paired t test, and the conversion equation was ρInternal = 1.039ρExternal -0.468 (r = 0.997 0). Conclusion The on-line column-switching technique is rapid, sensitive, high recovery and reproducible. It is suitable for monitoring the concentration of meropenem and pharmacokinetics in human plasma.