An integrative UHPLC-MS/MS untargeted metabonomics combined with quantitative analysis of the therapeutic mechanism of Si-Ni-San
An integrative UHPLC-MS/MS untargeted metabonomics combined with quantitative analysis of the therapeutic mechanism of Si-Ni-San
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DOI：
10.1016/j.ab.2018.10.023
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发表时间：
2019-02-15
影响因子：
2.9
通讯作者：
Xiong, Zhili
中科院分区：
文献类型：
--
作者：
Wen, Jing;Yang, Lina;Xiong, Zhili
为进一步了解四逆散的抗肝损伤作用及其机制，本研究建立并验证了一种结合大鼠血清中5种潜在生物标志物定量分析的非靶向血清代谢组学方法。代谢产物在BEH C-18柱(100 mm x 2.1 mm，1.7微米)上使用ACQUITY UHPLC-MS系统(Waters Corp.，Milford，MA，USA)进行分离和鉴定。主成分分析(PCA)用于识别潜在的生物标志物。在非靶向性代谢组学研究中，发现了与氨基酸代谢、脂肪代谢、胆汁酸生物合成和氧化-抗氧化平衡相关的主要潜在生物标志物，包括苯丙氨酸、色氨酸、甘鹅脱氧胆酸(GCDCA)和磷脂酰胆碱(LPC)。此外，这些目标生物标志物进一步在正电离模式下的多反应监测(MRM)中进行分离和定量。该方法对每个分析物都是线性的，相关系数在0.99以上。所有质控水平的日内和日间精密度(RSD)均小于13.1%，准确度(Re)在-9.5%~10.3%之间。将该方法成功地应用于正常对照组、模型组、阳性对照组(水飞蓟素组)和四逆散组大鼠血清样品的研究。
A UHPLC-MS/MS untargeted serum metabonomic method combined with quantitative analysis of five potential biomarkers in rat serum was developed and validated, to further understand the anti-liver injury effect of Si-Ni-San and its mechanism on liver injury rats in this study. The metabolites were separated and identified on BEH C-18 column (100 mm x 2.1 mm, 1.7 mu m) using the ACQUITY UHPLC-MS system (Waters Corp., Milford, MA, USA). Principal component analysis (PCA) was used to identify potential biomarkers. Primary potential biomarkers including phenylalanine, tryptophan, Glycochenodeoxycholic acid (GCDCA) and hysophosphatidylcholine (LPC), which were related to amino acid metabolism, lipid metabolism, bile acid biosynthesis and oxidation-antioxidation balance, were found in the untargeted metabonomic research. Moreover, these targeted biomarkers were further separated and quantified in multiple-reaction monitoring (MRM) with positive ionization mode. The proposed method was linear for each analyte with correlation coefficients over 0.99. The intra- and inter-day precision values (relative standard deviation, RSD) were less than 13.1% and accuracy (relative error, RE) was from -9.5% to 10.3% at all quality control (QC) levels. The validated method was successfully applied to study the serum samples of control group, model group, positive control group (silymarin group) and Si-Ni-San group in rats.