基于亚胺培南稳定性考察的治疗药物监测临床采样流程建立

Establishing the clinical sampling process of therapeutic drug monitoring based on stability of imipenem

目的建立人血中测定亚胺培南浓度的HPLC法,完善其稳定性研究并将其应用到临床治疗药物监测。方法以50 mmol·L^(-1)磷酸二氢钾含0.05%甲酸-乙腈(30∶70,V∶V)为流动相,流速0.8 mL·min^(-1),用Venusil HILIC(4.6mm×250 mm,5.0μm)色谱柱进行色谱分离,检测波长为299 nm,分别考察亚胺培南全血在不同温度、不同采血管等条件下的稳定性。结果亚胺培南在0.38~71.30μg·mL^(-1)线性关系良好(r=0.999 9),定量下限为0.38μg·mL^(-1),低、中、高质控浓度的样品萃取回收率在74.4%~88.9%,日内、日间RSD均小于8.2%;以肝素钠管为最佳的临床采血管,全血中无稳定剂时亚胺培南能稳定3 h;室温下6 h、冻存42 d、反复冻融2次均稳定。结论本研究建立的治疗药物监测临床采样流程充分考虑到了临床采样的实际情况并能确保亚胺培南药物的稳定性及分析监测结果的准确性。

Objective To determine the concentration of imipenem in human plasma by high performance liquid chromatography（ HPLC）method and investigate the stability of imipenem in plasma,then applied to therapeutic drug monitoring. Methods HPLC was performed on a HILIC column（ 4. 6 mm × 250 mm,5. 0 μm）,phosphate buffer 50 mmol · L^-1 containing 0. 05% formic acid and acetonitrile（ 30 ∶ 70,V∶ V） as the mobile phase. The column temperature was set at 40 ℃,detection wavelength at 299 nm,and flow rate at 0. 8 mL· min^-1. The stability of imipenem in whole blood at the different temperatures and blood-collected vessel was investigated. Results The linear calibration curves were obtained in the concentration range of 0. 38-71. 30μg·mL^-1（ r = 0. 999 9）. The lower limit of quantification was 0. 38μg·mL^-1. The recovery of the sample were 74. 4%-88. 9%. The intra-day and inter-day RSD were less than 8. 2%. Meanwhile,the results showed heparin sodium tube might be the best blood-collected vessel. Imipenem in whole blood could maintain stable for 3 h without stabilizer added. In addition,imipenem in the plasma could maintain stable at room temperature for 6 h,in two freeze-thaw cycles and long-term of frozen samples after 42 d of storage at -40 ℃,respectively. Conclusion This study had established the clinical sampling process of therapeuticdrug monitoring based on actual condition to ensure stability and detection accuracy of imipenem.