超高效液相色谱串联质谱法检测人肺组织、支气管黏膜和支气管分泌液中奈诺沙星浓度及临床应用  

A UPLC-MS/MS method for rapid determination of nemonoxacin concentration in human lung tissues,bronchial mucosa and bronchial secretions and its clinical application

作　　者：赵锦锦 徐晓勇 朱勇俊 陈志明 范亚新 胡佳丽 毋海兰 王雨 李熠 郭蓓宁 张菁 ZHAO Jinjin;XU Xiaoyong;ZHU Yongjun;CHEN Zhiming;FAN Yaxin;HU Jiali;WU Hailan;WANG Yu;LI Yi;Guo Beining;ZHANG Jing(Institute of Antibiotics,Key Laboratory of Clinical Pharmacology of Antibiotics,Ministry of Health/National Clinical Research Center for Aging and Medicine,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院抗生素研究所,国家卫健委抗生素临床药理重点实验室,国家老年疾病临床医学研究中心(华山),上海200040 [2]复旦大学附属华山医院胸外科,上海200040

出　　处：《中国感染与化疗杂志》2020年第2期146-152,共7页Chinese Journal of Infection and Chemotherapy

基　　金：国家科技部重大新药创制科技重大专项(2017ZX09304005)。

摘　　要：目的建立一种快速、准确地检测人体肺组织、支气管黏膜和支气管分泌液中奈诺沙星浓度的超高效液相色谱串联质谱(UPLC-MS/MS)方法,并将该方法应用于临床样品检测。方法色谱柱为ACE UltraCore 2.5 super C18(4.6 mm×50 mm,2.5μm),流动相为0.3%甲酸水溶液-甲醇,采用电喷雾离子源、正离子多反应监测进行分析扫描。内标为苹果酸奈诺沙星-D3。样品经乙腈沉淀蛋白后离心,取上清液稀释后进样。方法学验证中用0.9%NaCl溶液作为替代基质代替支气管黏膜和支气管分泌液。利用经方法学验证过的检测方法进行组织穿透性研究样品的检测,通过肺组织、支气管黏膜和支气管分泌液与血浆浓度的比值评价奈诺沙星的组织体液穿透性。结果该方法测定奈诺沙星在组织体液中的线性范围均为0.0200~10.0 mg/L,检测下限均为0.0200 mg/L。肺组织样品及替代基质样品的各质控浓度测定结果的批内和批间精密度均≤6.0%,准确度偏差均≤11.5%。肺组织内标归一化基质效应因子为96.5%~99.3%,变异系数均≤1.5%。0.9%NaCl溶液替代支气管黏膜及支气管分泌液的精密度和准确度偏差分别≤2.8%和≤10.3%。肺组织和替代基质中奈诺沙星的总体回收率分别为95.0%和102.8%,变异系数分别为1.0%和1.5%。肺组织穿透性研究结果显示5例需行肺叶切除术患者口服奈诺沙星500 mg,每日1次,连续服药(4±1)d后,体内药物分布达到稳态,肺组织、支气管黏膜和支气管分泌液中浓度与血浆浓度比值分别为3.72~6.56、4.03~9.51、1.18~1.88。结论该研究建立了一种准确、方便检测人肺组织、支气管黏膜和支气管分泌液中奈诺沙星浓度的UPLC-MS/MS法,其方法学验证结果均符合生物样品分析的要求。组织穿透性结果显示奈诺沙星在肺组织、支气管黏膜和支气管分泌液中穿透性良好。Objective To establish and validate a rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometric(UPLC-MS/MS)method for quantification of nemonoxacin in human lung tissues,bronchial mucosa and bronchial secretions,and use this method to analyze clinical samples.Methods Chromatographic separation was performed on an ACE UltraCore 2.5 super C18(4.6 mm×50 mm,2.5μm)column.The mobile phase consisted of 0.3%of formic acid and acetonitrile.Quantitative analysis was conducted in multiple reaction monitoring mode.Nemonoxacin-D3 was used as internal standard.Lung tissues,bronchial mucosa and bronchial secretions were subjected to acetonitrile protein precipitation.The supernatant was diluted for injection.Normal saline(0.9%NaCl)was used as surrogate matrix for bronchial mucosa and bronchial secretion in method validation.The validated method was used to determine the penetration into tissues,which was based on nemonoxacin concentration ratio between lung tissue,bronchial mucosa and bronchial secretions and plasma.Results The calibration curve for nemonoxacin in tissues were linear within the range of 0.0200-10.0 mg/L.The lower limit of quantification was 0.0200 mg/L for all samples.The inter-and intra-batch precision was≤6.0%in lung tissue and surrogate matrix samples.The inter-and intra-batch accuracy deviation was≤11.5%in lung tissue and surrogate matrix samples.The internal standard normalized matrix effect factor was 96.5%-99.3%for in lung tissues,coefficient of variation(CV)≤1.5%.The precision and accuracy deviation of normal saline as surrogate bronchial mucosa and bronchial secretions was≤2.8%and≤10.3%,respectively.The overall recovery of nemonoxacin was 95.0%for lung tissues and 102.8%for surrogate matrix,CV was 1.0%and 1.5%,respectively.Lung tissue penetration study showed that steady state was reached after oral administration of nemonoxacin 500 mg once daily for(4±1)consecutive days in 5 patients undergoing lobectomy.The concentration ratio between lung tissue,bronchial muc

关 键 词：奈诺沙星 超高效液相色谱串联质谱 肺组织 支气管黏膜 支气管分泌液 穿透性 

分 类 号：R969.1[医药卫生—药理学] R978[医药卫生—药学]