用LC-MS/MS法测定感染小鼠血浆中去甲万古霉素及万古霉素的浓度及药代动力学研究  

Determination the concentration of norvancomycin and vancomycin in infected mice plasma by LC-MS/MS for the study of their pharmacokinetic properties

作　　者：冀希炜[1] 梁家彬 李耘[1] JI Xi-wei;LIANG Jia-bin;LI Yun(Institute of Clinical Pharmacology,Peking University First Hospital,Beijing 100191,China)

机构地区：[1]北京大学第一医院临床药理研究所,北京100191

出　　处：《中国临床药理学杂志》2023年第23期3455-3459,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的建立一种快捷、简便、灵敏的液质联用方法测定小鼠血浆中万古霉素及去甲万古霉素的浓度并考察2种药物在感染小鼠体内的药代动力学特征。方法万古霉素及去甲万古霉素互为内标,血浆样品经过乙腈沉淀蛋白,离心后取上清液进行分析。色谱柱:Kinetex Biphenyl 100A(2.1 mm×50.0 mm,5μm);流动相:甲醇-0.2%甲酸水溶液;梯度洗脱;流速:0.4 mL·min^(-1);进样量:10μL。电喷雾正离子检测方式,多反应监测。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应、残留效应、稀释效应和稳定性。感染小鼠分别给予14、140及320 mg的万古霉素或去甲万古霉素并在不同时间点采集血样用于考察2种药物的药代动力学特征。结果去甲万古霉素及万古霉素均在50.0~3000.0 ng·mL^(-1)内,专属性及线性关系良好,定量下限均为50.0 ng·mL^(-1),批内及批间精密度分别为≤4.33%、≤18.26%以及≤5.03%、≤15.19%,回收率分别为80.61%~109.38%和83.94%~97.34%,批内及批间准确度分别为-10.52%~10.70%、-8.07%~4.19%和-6.18%~4.27%、-4.65%~4.98%,基质效应分别为71.46%~84.20%和85.64%~93.11%,残留效应、稀释效应和稳定性等方法学考察的结果均符合要求。结论本文建立了一种灵敏、快速、稳定的液质联用方法,可用于考察去甲万古霉素及万古霉素在小鼠体内的药代动力学特征。Objective To establish a rapid,sensitive method for the determination of vancomycin and norvancomycin in mice plasma and to investigate the pharmacokinetic(PK)characters of them in infected mice.Methods Propranolol was taken as the internal standard,the plasma samples were extracted by acetonitrile to precipitate protein.After centrifugation,the supernatant was analyzed by liquid chromatographymass spectroscopy(LC-MS/MS).Chromatographic column:Kinetex Biphenyl 100A(2.1 mm×50.0 mm,5μm);mobile phase composed of methanol-water containing 0.2%formic acid in gradient elution mode;flow-rate of 0.4 mL·min^(-1);injection volume:10μL.A triple quadrupole tandem mass spectrometer with electrospray ionization source was used as detector and operated by multiple reaction monitoring(MRM)in the positive ion mode.The specificity,standard curve,lower limit of quantification,precision and recovery,matrix effect,residual effect,dilution effect,and stability of this method were examined.The infected mice received three single oral doses(14,140 and 320 mg·kg^(-1))of vancomycin and norvancomycin and plasma samples were collected at different time points.Results The calibration curves of vancomycin and norvancomycin were linear between 50.0 and 3000.0ng·mL^(-1);the quantitation limit was 50 ng·mL^(-1);the intra-and inter-precision were≤4.33%,≤18.26%and≤5.03%,≤15.19%,respectively;the extraction recovery were 80.61%to 109.38%and 83.94%to 97.34%,respectively;the intra-and inter-accuracy were-10.52%to 10.70%,-8.07%to 4.19%and-6.18%to4.27%,-4.65%to 4.98%,respectively;and the matrix effects were 71.46%to 84.20%,and 85.64%to93.11%,respectively.The results of methodological study of residual effect,dilution effect and stability conformed to the requirements.Conclusion A reliable LC-MS/MS method with high sensitivity,simple sample preparation and short elution time was developed for characterizing the PK profiles of vancomycin and norvancomycin in mice plasma.

关 键 词：万古霉素 去甲万古霉素 液质联用 血药浓度 药代动力学 

分 类 号：R978.1[医药卫生—药品]