多黏菌素B治疗药物监测LC-MS/MS方法探索及临床初步应用  

Exploration of LC-MS/MS Method for Therapeutic Drug Monitoring and Clinical Application of Polymyxin B

作　　者：邓阳 徐兵[1,2,3] 李昕 郭思维 罗细林 李尤 DENG Yang;XU Bing;LI Xin;GUO Si-wei;Luo Xi-lin;LI You(Department of Pharmacy,The Third Hospital of Changsha,Changsha 410015,China;The Clinical Application Research Institute of Antibiotics in Changsha,Changsha 410015,China;Changsha Hospital,Hunan University of Chinese Midecine,Changsha 410015,China)

机构地区：[1]长沙市第三医院药学部,长沙410015 [2]长沙市抗菌药物临床应用研究所,长沙410015 [3]湖南中医药大学附属长沙医院,长沙410208

出　　处：《中国药学杂志》2021年第15期1249-1254,共6页Chinese Pharmaceutical Journal

基　　金：湖南省卫生健康委员会科技计划项目资助(20201375);湖南省自然科学基金青年基金项目资助(2019JJ50680);湖南省科技计划项目资助(2020SK52901)。

摘　　要：目的建立液质联用法(LC-MS/MS)测定人血浆中多黏菌素B(polymyxin B,PMB,主要活性成分PMB1和PMB2)的血药浓度,结合其全血稳定性研究制定临床采样流程,并应用到治疗药物监测(TDM)个体化给药方案设计。方法血浆样品经乙腈(含甲酸6.67%)蛋白沉淀,取上清加水稀释后进样分析;色谱柱为Shim-pack GIST C_(18)(2.1 mm×100 mm, 3μm,岛津公司);流动相为水(含0.1%甲酸)-乙腈,梯度洗脱;流速0.5 mL·min-1,柱温40℃;分别考察全血/血浆中PMB1和PMB2在不同温度、不同采血管等储存条件下的稳定性。结果 PMB1和PMB2在(0.033~18.816)和(0.034~19.872)μg·mL^(-1)线性关系良好(r=0.999 9),定量下限浓度为0.033和0.034μg·mL-1,低、中、高浓度的萃取回收率在97.2%~110.6%,基质效应在91.8%~101.1%。日内、日间RSD均小于8.2%。EDTA-K2管和肝素钠管中的PMB能在室温下稳定10 h、2~6℃冰箱保存19 h;血浆样本室温下22 h、-40℃冻存115 d或-40℃反复冻融3次均稳定。结论建立的TDM临床采样流程能确保PMB药物的稳定性及分析检测结果的准确性,可满足临床血药浓度监测需求。OBJECTIVE To establish an LC-MS/MS method for the determination of polymyxin B(PMB1 and PMB2) concentration in human plasma and build clinical sampling process based on its stability study. METHODS Protein in plasma was precipitated by acetonitrile(containing 6.67% formic acid), and supernatant was taken and diluted with water. Chromatographic separation was achieved on a Shim-pack GIST C_(18)(2.1 mm×100 mm, 3 μm)by gradient elution with 0.1% formic acid-water and acetonitrile. Flow rate was set at 0.8 mL·min-1, and column temperature at 40 ℃. The stability of whole blood samples of PMB1 and PMB2 at different temperatures and under different conditions was investigated. RESULTS The linear ranges of PMB1 and PMB2 were good at(0.033-18.816) and(0.034-19.872) μg·mL-1(r=0.999 9), and the lower limit concentration were 0.033 and 0.034 μg·mL^(-1),respectively. The extraction recoveries of low, medium and high concentration were 97.2%-110.6%, the relative standard deviation of intra-day precision and inter-day precision were less than 8.2%. With EDTA-K2 tubes as the optimal clinical blood collection, PMB1 and PMB2 in EDTA-K2 tubes and heparin sodium tubes were stable at room temperature for 10 h. Plasma samples could maintain stable at room temperature for 22 h, long-term of frozen samples after 115 days of storage and in two freeze-thaw cycle at-40 ℃, respectively. CONCLUSION The established TDM clinical sampling process fully consider the actual situation of clinical sampling for ensuring the stability of PMB and the accuracy of analysis results, which could satify the needs of clinical blood drug concentration determination.

关 键 词：多黏菌素B 全血稳定性 治疗药物监测 液质联用法 临床采样流程 

分 类 号：R965[医药卫生—药理学]