QuEChERS-液相色谱-串联质谱法快速测定肝细胞癌血液中3种小分子代谢物  

Rapid determination of three small-molecule metabolites in the hepatocellular carcinoma blood using QuEChERS-liquid chromatography-mass spectrometry

作　　者：崔浩亮 邓晓晴 齐海娟 王洪朋 王丽 李挥 哈婧[1] CUI Hao-liang;DENG Xiao-qing;QI Hai-juan;WANG Hong-peng;WANG Li;LI Hui;HA Jing(College of Chemical and Pharmaceutical Engineering,Hebei University of Science and Technology,Shijiazhuang 050018,China;Hebei Insitute of Medical Equipment and Drugs Packaging Material Inspection,Shijiazhuang 050061,China)

机构地区：[1]河北科技大学化学与制药工程学院,石家庄050018 [2]河北省医疗器械与药品包装材料检验研究院,石家庄050061

出　　处：《药物分析杂志》2019年第5期830-838,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：河北省自然科学基金项目(H2019329002)

摘　　要：目的:通过建立QuEChERS(快速、简易、廉价、有效、稳定、安全)前处理方法,结合液相色谱-三重四极杆质谱联用技术,实现肝细胞癌血液中3种小分子代谢物的同时检测。方法:血清样品经乙腈提取,无水硫酸镁盐析及N-丙基乙二胺(PSA)吸附剂净化后,取上清液采用Phenomenex Luna Omega 3μm PS C_(18) 100?(150 mm×2.1 mm)色谱柱,以甲醇-0.1%乙酸水溶液为流动相,梯度洗脱。电喷雾离子源正离子扫描,多反应监测模式(MRM)进行定量分析。结果:十四酰胺的检测下限为0.6 ng·g^(-1),定量下限为2ng·g^(-1);油酸酰胺的检测下限为0.3 ng·g^(-1),定量下限为1 ng·g^(-1)。3种小分子代谢物质量浓度在10～100ng·mL^(-1)范围内呈线性,其中目标物线性相关系数(R^2)均大于0.991 7,基质效应(ME)在0.79～1.55之间;3个水平加样回收率在77.5%～105.1%之间,日内精密度(n=5)在6.9%～11.8%之间,日间精密度在7.4%～11.1%之间;利用该方法扩大样本量检测,3种小分子代谢物在健康组血清中的浓度(2.591、0.221、7.485 mg·kg^(-1))和肝癌组血清中的浓度(9.599、0.266、5.063 mg·kg^(-1))差异显著(P<0.01)。结论:该方法简单快捷,准确可靠,适合于大批量肝细胞癌血液样品中3种小分子代谢物的快速检测。Objective:To establish an LC-MS method for the simultaneous determination of 3 small-molecule metabolites in the hepatocellular carcinoma blood by establishing quEChERS(quick,easy,cheap,effective, rugged,safe)coupled with liquid chromatography-mass spectrometer.Methods:Serum samples were extracted with acetonitrile and anhydrous sodium sulfate were used for salting-out process.After purified by PSA,the supernatant was applied on a Phenomenex Luna Omega 3 μm PS C18 100A(150 mm×2.1 mm)column with methanol-0.1% acetic acid solution as mobile phase and subjected to gradient elution.For quantitation,electrospray ionization(ESI)source was applied and the mass spectrometer was operated in multiple reaction monitoring(MRM)modes.Results:The limits of detection(LOD)and the limits of quantification(LOQ)for n-tetradecanamide were 0.6 ng·g^-1 and 2 ng·g^-1 respectively.LOD and LOQ for oleamide were 0.3 ng·g^-1 and 1 ng·g^-1 respectively.There was a good linear relationship in the range of 10-100 ng·mL^-1 of the 3 small-molecule metabolites.The target linear correlation coefficient(R^2)was higher than 0.991 7.The matrix effect(ME)was between 0.79 and 1.55.The method recovery was between 77.5% and 105.1% in three spike levels.The intraday precision(n=5)was between 6.9% and 11.8% and daytime precision was between 7.4% and 11.1%.Using this method to expand the sample size detection, the concentration of three small-molecule metabolites in healthy group (2.591,0.221,7.485 mg·kg^-1)and cancer group(9.599,0.266,5.063 mg·kg^-1)were significantly different(P<0.01).Conclusion:This method is simple,accurate and reliable,and is suitable for the rapid detection of three small-molecule metabolites in large quantities of the hepatocellular carcinoma blood samples.

关 键 词：血液 QUECHERS 小分子代谢物 液相色谱-串联质谱法 快速检测 

分 类 号：R917[医药卫生—药物分析学]