细胞顶空气中典型醛酮化合物定量分析方法研究  

Quantitative Analysis of Typical Aldehydes and Ketones in Headspace Air of Cell

作　　者：尹志红 陈钊铭 陈星 杨子峰[3,4] 李征途[3,4] 叶枫[3,4] 张志娟 方明亮 李雪 YIN Zhi-Hong;CHEN Zhao-Ming;CHEN Xing;YANG Zi-Feng;LI Zheng-Tu;YE Feng;ZHANG Zhi-Juan;FANG Ming-Liang;LI Xue(Institute of Mass Spectrometry and Atmospheric Environment,Jinan University,Guangzhou 510632,China;Guangdong Provincial Engineering Research Center for On-line Source Apportionment System of Air Pollution,Guangzhou 510632,China;State Key Laboratory of Respiratory Disease,Guangzhou Medical University,Guangzhou 511436,China;National Clinical Research Center for Respiratory Disease,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China;School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450008,China;School of Civil and Environmental Engineering,Nanyang Technological University,Singapore 637457,Singapore)

机构地区：[1]暨南大学质谱仪器与大气环境研究所,广州510632 [2]广东省大气污染在线源解析系统工程技术研究中心,广州510632 [3]广州医科大学呼吸疾病国家重点实验室,广州511436 [4]广州医科大学附属第一医院,国家呼吸系统疾病临床医学研究中心,广州510120 [5]河南中医药大学药学院,郑州450008 [6]南洋理工大学土木与环境工程学院,新加坡637457

出　　处：《分析化学》2021年第8期1375-1383,I0005-I0008,共13页Chinese Journal of Analytical Chemistry

基　　金：“广东特支计划”本土创新创业团队项目(No.2019BT02Z546);呼吸疾病国家重点实验室开放课题项目(No.SKLRD-OP-201913);中国科学院地球科学领域核心关键技术攻关预研项目(No.282021000003)资助。

摘　　要：采用2,4-二硝基苯肼衍生柱收集挥发性醛酮类组分,转化为稳定的醛酮2,4-二硝基苯腙有机物,采用超高效液相色谱-高分辨质谱联用技术进行分析,建立了22种典型醛酮类化合物定量分析方法。2,4-二硝基苯腙有机物在高(40-1000μg/L)、中(8-400μg/L)、低(0.08-8μg/L)浓度范围内标准曲线线性良好,相关系数R^(2)>0.998,检出限范围为0.003-0.09μg/L;高、中、低浓度范围的标准溶液的日内及日间精密度(RSD)均小于10%;回收率在95.1%-109.6%之间,RSD≤5.9%。将本方法用于分析A549细胞以及甲型H1N1流感病毒(IAV)感染的A549细胞顶空气中醛酮类物质,检测到19种醛酮类物质,通过正交偏最小二乘-判别分析法分析检测结果,发现两组醛酮代谢物存在显著差别,并筛选出11种潜在的醛酮生物标记物.Aldehydes and ketones in breath can reflect the oxidative stress response of the cells in the body.A better understanding of the aldehydes and ketones emitted from the cells is the key to revealing the mechanism of breath test for the diagnosis of oxidative stress-related diseases.In this study,a quantitative method was developed for 22 typical aldehyde and ketone metabolites.Standard compounds were transformed into stable DNPH-carbonyl adducts by using 2,4-dinitrophenylhydrazine(DNPH)impregnated cartridge.These adducts were further eluted and analyzed by ultra high performance liquid chromatography coupled with high resolution mass spectrometry.The standard curves of 22 DNPH-carbonyl adducts showed linearity at high(40-1000μg/L),medium(8-400μg/L)and low(0.08-8μg/L)levels,respectively.The coefficient of correlation(R^(2))was>0.998 and the detection limits were 0.003-0.09μg/L.The method demonstrated good precision,e.g.intra-day RSD<3.92%and inter-day RSD<5.3%,as well as satisfactory recoveries of 95.1%-109.6%(RSD<5.9%).The method was further applied to the analysis of aldehydes and ketones in the headspace of A549 cells and influenza A(H1N1)infected A549 cells.The experimental data were analyzed by using orthogonal partial least squares discriminant analysis,and a total of 19 aldehydes and ketones were quantified and 11 potential biomarkers were screened for the diagnosis of IAV infection.

关 键 词：呼气疾病诊断 细胞顶空气 醛酮化合物 2 4-二硝基苯肼衍生化 超高效液相色谱-高分辨质谱联用技术 

分 类 号：O657.63[理学—分析化学] Q2-33[理学—化学]