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

Rapid Determination of Meropenem Concentration in Human Plasma by Column-Switching-Ultra High Performance Liquid Chromatography

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 column-switching-uhra high performance liquid chromatography method for determi- ning meropenem concentration in human plasma. METHODS Meropenem plasma samples were diluted by internal standard aqueous solution（ doxofylline）, filtrated by microporous membrane, and then extracted by on-line column-switching method. The samples were chromatographed on Waters Acquity UPLC ？ BEH-C18 column using gradient elution method （ mobile phase A and extracting mobile phase C ： methanol-0. 05 mol . L^- 1 K2 HPO4 （ pH 7.0） 5 ： 95 ; mobile phase B ： methanol）. The maximum absorption wavelength of mer openem was 299 nm. The column temperature were 35 ℃. The meropenem concentration was calculated by internal standard method and external standard method, respectively. The results of the two methods were compared by paired t-test using SPSS statistical soft- ware. RESULTS The liner range of the calibration curve for meropenem was 0. 67 - 86. 00 mg . L^- 1 （ r = 0. 999 9） . The method recovery was （93.59 ± 1.15 ）% - （99. 21 ± 4. 06）% and extraction recovery was above 85.0%, relative standard deviation was below 5.0% o The SPSS statistic results indicated there was no significant difference between the concentrations calculated by the two methods and existed good correlation. The transformed equation is Plntemal = 1. 039pExtemal - 0. 468 （ r = 0. 997 0）. CONCLUSION On-line col- umn-switching method is used and it is proved to be rapid, sensitive and suitable for therapeutic drug monitoring and pharmacokinetic investigation of meropenem in human plasma.