基于超高效液相色谱-质谱的药物性肝损伤患者血清代谢组学研究

摘要

利用超高效液相色谱与四级杆-轨道阱高分辨质谱仪联用技术( UPLC-MS/MS)结合多变量统计分析方法,开展健康者与肿瘤药物、中药、他汀类降脂药、抗菌药引发肝损伤患者的血清代谢组学研究。血清样本采用冰乙腈沉淀蛋白,HSS T3-C18色谱柱,水(含0.01％甲酸)与乙腈为流动相梯度洗脱,液相色谱串联质谱分析测定。基于健康者与药物性肝损伤初诊病例代谢轮廓的正交偏最小二乘判别分析( OPLS-DA)以及偏最小二乘法判别分析( PLS-DA),寻找114个与药物性肝损伤早期诊断密切相关的氨基酸类生物标志物,其中有38个代谢物在两组间具有显著差异(p<0.001)。对各组分离贡献大的化合物进行串联质谱分析,经 Human Metabolome Database( HMDB)等数据库检索,进行质谱信息匹配,鉴定出与药物性肝损伤相关的可能生物标志物苯丙氨酸和二甲基鸟苷。%Ultra high performance liquid chromatography coupled with a Q Exactive hybrid quadrupole-orbitrap mass spectrometry ( UPLC-MS/MS) and multivariate statistical analysis was applied to metabonomic study of drug-induced liver injury. This investigation focused on the liver injury in clinic which was induced by antitumor drug, traditional Chinese medicine, statins and antibiotics. After precipitated by cold acetonitrile, the serum samples were separated on a HSS T3-C18 column using water (containning 0. 01% formic acid) and acetonitrile as mobile phase by gradient elution. Based on metabonomic profiles analysis of the orthogonal projections to latent structures discriminant analysis ( OPLS-DA ) and partial least squares-discriminant analysis ( PLS-DA) , 114 endogenous metabolites had been found to have significant differences between drug-induced liver injury patients and healthy volunteers, 38 endogenous metabolites have significant difference (p<0. 001). According to the results of the OPLS-DA, MS/MS data and human metabolome database ( HMDB) , phenylalanine and dimethylguanosine had been identified as potential biomarkers related to drug-induced liver injury for early discovery and diagnosis.