A UPLC-MS/MS-Based Metabolomics analysis of the Pharmacological Mechanisms of Rabdosia Serra Against Cholestasis?超高效液相色譜-串聯(lián)質(zhì)譜/串聯(lián)質(zhì)譜聯(lián)用分析香茶菜抗膽汁淤積的藥理作用機(jī)制

Differential metabolites were identified by MS-Dial (version 4.7), general software for chromatographic deconvolution of high-resolution LC-MS spectrometry (Cui et al., 2018;?Liu et al., 2021;?Tada et al., 2019). The metabolites were intelligently identified by comparing the accurate MS and MS/MS spectra in the integrated databases, including references, and public databases: Metabolite Link (https://metlin.scripps.edu), Human Metabolome Database (https://www.hmdb.ca), Mass-bank (https:// massbank.eu/MassBank/), and Kyoto Encyclopedia of Genes and Genomes (https://www.genome.jp/kegg). Metaboanalyst (https://www.metaboanalyst.ca/) was used for metabolic pathway enrichment analysis. MS files were converted to ABF format from wiff format by ABF converter (http://www.reifycs.com/AbfConverter/). All parameters of MS-Dial are listed below. The parameters for data collection were: RT, 0-15 min; mass range, 50-1000 Da; MS1 tolerance, 0.01 Da; and MS2 tolerance, 0.05 Da. The parameters for peak detection were: minimum peak width, five scans; smoothing method, linear weighted moving average with smoothing level of two scans; mass slice width, 0.1 Da; minimum peak height, 1000 amplitude; exclusion mass list, none. The parameters for metabolite identification were: RT tolerance, 2 min; MS1 accurate mass tolerance, 0.01 Da; MS2 accurate mass tolerance, 0.05 Da; identification score cut off, 80%.

MS-DIAL是一款針對(duì)非靶向代謝組學(xué)的數(shù)據(jù)處理的第三方軟件，最大的優(yōu)勢在于能夠兼容來自不同儀器（Agilent，Bruker，LECO，Sciex， Shimadzu，Thermo和Waters）獲得的各類質(zhì)譜數(shù)據(jù)（GC-MS，GC-MS/MS，LC-MS和LC-MS/MS）。此外像netCDF（AIA）和mzML等通用的數(shù)據(jù)格式也可以被軟件識(shí)別。

Blank rat serum (1 mL) was mixed with 0.2 g of activated charcoal for 12 h to remove the endogenous matrix. The mixture was centrifuged at 14,000 rpm for 15 min, and the supernatants were incubated with activated charcoal to repeat the stripping process. The obtained supernatant was used as the bile acid-free blank serum.