血清伏立康唑候选参考方法的建立和性能评价
Establishment and performance evaluation of candidate reference measurement procedures for serum voriconazole
林静 1潘清清 2沈敏 2谢阳敏 2金中淦 3潘荆珏 1刘甲娟4
作者信息
- 1. 宁波市第二医院医学实验部,浙江 宁波 315010
- 2. 美康生物科技股份有限公司参考实验室,浙江 宁波 315104
- 3. 上海市临床检验中心,上海 200126
- 4. 国科宁波生命与健康产业研究院,浙江 宁波 315010
- 折叠
摘要
目的 基于同位素稀释液相色谱-串联质谱(ID-LC-MS/MS)技术建立检测血清伏立康唑的候选参考方法.方法 以甲醇为沉淀剂,采用蛋白沉淀法(PPT)对样本进行前处理.以伏立康唑-2H3为内标,建立基于ID-LC-MS/MS的血清伏立康唑候选参考方法.评估所建立方法的线性、定量限(LOQ)、检测限(LOD)、准确度、精密度、基质效应、稳定性.结果 ID-LC-MS/MS测定血清伏立康唑的线性范围为0.2～16.0 μg·mL-1,定量限为0.08 μg·mL-1,检测限为0.016 μg·mL-1.加标回收率为97.26%～98.48%.批内变异系数(CV)<1%,批间CV<2%.相对基质效应为-1.87%～0.33%.样本分别经室温[(23±2)℃]放置4、8 h和-20℃反复冻融5次,经前处理后的样本在室温和自动进样器(8℃)分别放置4、8、12、24 h,回收率为95.63%～99.50%,检测结果稳定.相对扩展不确定度≤3.34%.结论 建立了基于ID-LC-MS/MS的血清伏立康唑候选参考方法,可用于该项目的检测标准化.
Abstract
Objective To establish a candidate reference measurement method for serum voriconazole by isotope dilution liquid chromatography-tandem mass spectrometry(ID-LC-MS/MS).Methods The samples were pretreated by protein precipitation(PPT)with methanol as precipitator.Using voriconazole-2H3 as internal standard,an ID-LC-MS/MS for the determination of voriconazole in serum was established.The linearity,limit of quantification(LOQ),limit of determination(LOD),accuracy,precision,matrix effect and stability were evaluated.Results The linear range of serum voriconazole was 0.2-16.0 μg·mL-1,the limit of quantification was 0.08 μg·mL-1,and the limit of determination was 0.016 μg·mL-1.The recoveries were 97.26%-98.48%.The within-run coefficient of variation(CV)was<1%,and the between-run CV was<2%.The relative matrix effect ranged from-1.87%to 0.33%.The samples were placed at room temperature[(23±2)℃]for 4,8 h and repeatedly frozen and thawed for 5 times at-20℃,and the samples after pre-treatment were placed at room temperature and automatic injector(8℃)for 4,8,12 and 24 h,respectively.The recoveries were 95.63%-99.50%,and the determination results were stable.The relative extended uncertainty was≤3.34%.Conclusions A candidate reference method for the determination of serum voriconazole based on ID-LC-MS/MS has been established,which can be used for standardization.
关键词
伏立康唑/同位素稀释液相色谱-串联质谱/参考方法Key words
Voriconazole/Isotope dilution liquid chromatography-tandem mass spectrometry/Reference method引用本文复制引用
基金项目
宁波市重大科技攻关任务项目(2021Z055)
宁波市重点研发计划(2023Z168)
中国合格评定国家认可中心课题(2021CNAS07)
出版年
2024
NETL
NSTL
万方数据