摘要
目的建立高效液相色谱-串联质谱法快速检测血浆中6种初级胆汁酸的含量。方法血浆经甲醇沉淀蛋白后,用Thermo Accucore C18色谱柱(100 mm×4.6 mm,2.6μm)进行色谱分离,流动相为甲醇-水(0.1%甲酸+5 mmol/L乙酸铵),梯度洗脱。负离子模式下多反应监测模式检测。实测6名健康成年人和6只成年SD大鼠样本。结果6种初级胆汁酸在6 min内实现基线分离,在5~2000μg/L范围内线性良好,相关系数>0.995,检出限为0.25~0.45μg/L,定量限为0.84~1.49μg/L。日内、日间加标回收率分别为86.3%~111.5%和87.7%~104.9%;日内、日间相对标准偏差分别为5.2%~11.9%和6.6%~14.3%。健康成年人和成年SD大鼠血浆中初级胆汁酸的浓度显著不同。结论该方法简单、快速、灵敏、准确,可用于临床检验和动物试验中初级胆汁酸的快速检测。
Objective To develop a rapid and accurate high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for the simultaneous determination of 6 primary bile acids in plasma.Methods Plasma was separated on a Thermo Accucore C18column(100 mm×4.6 mm;2.6μm)after being extracted by protein precipitation using methanol.The 6 primary bile acids were separated by using the mobile phase of methanol-water(0.1%formic acid+5 mmol/L ammonium acetate)running in gradient elution.The mass spectrometer was operated on an electrospray ionization source in negative ion mode for the multiple reaction monitoring analysis.Plasma from 6 healthy adults and 6 adult SpragueDawley(SD)rats was detected.Results Baseline separation of 6 primary bile acids was achieved within 6 min.The linearity of the calibration curves of the 6 primary bile acids was excellent in the range of 5-2000μg/L with a correlation coefficient higher than 0.995.The limits of detection and limits of quantitation were in the range of 0.25-0.45μg/L and 0.84-1.49μg/L,respectively.The recoveries of intra-day and inter-day were 86.3%-111.5%and 87.7%-104.9%,with relative standard deviations of 5.2%-11.9%and 6.6%-14.3%,respectively.The concentrations of primary bile acids in plasma of healthy adults and adult SD rats were significantly different.Conclusion This method is simple,rapid,sensitive,and accurate,and can meet the needs for rapid detection of primary bile acids in clinical tests and animal experiments.
作者
牟燕
吴瑛瑕
张易天
曾颖炫
周枝凤
MU Yan;WU Ying-xia;ZHANG Yi-tian(Department of Hygiene Inspection and Quarantine Science,School of Public Health,Southern Medical University,Guangzhou,Guangdong Province 510515,China)
出处
《中国公共卫生》
CAS
CSCD
北大核心
2023年第1期102-106,共5页
Chinese Journal of Public Health
基金
国家自然科学基金(21607070)。
