To explore the endogenous changes in the progression of silicosis, metabolomics was analyzed by HPLC-QTOF-MS (Agilent, USA), and data were preprocessed using MetaboAnalyst 4.0 (https://www.metaboanalyst.ca/ ).The normalized data were then subjected to Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) in a Simca-p 14.1 workstation. Metabolites with VIP > 1 and P < 0.05 were identified as differential metabolites, and compound validation was conducted to verify the accuracy and eliminate unreliable differential metabolites. The metabolic pathways were identified by analyzing and enriching differential metabolites screened using the Pathway Analysis module in MetaboAnalyst 4.0. The MetScape plugin in Cytoscape 3.7.1 software was used for network construction of different periods and analysis of the topology of each network. Euclidean distance was used to evaluate the similarity between different periods by network topology and to determine the important metabolites in various periods [29 (link)]. A Venn diagram of metabolites in each period was displayed using the FunRich software, and changes in the same metabolite in different periods were analyzed. The diagnostic value was predicted using the ROC-based Biomarker Analysis module in MetaboAnalyst 4.0. Further details are presented in Supplemental Materials.
Hplc qtof ms
Manufactured by Agilent Technologies
Sourced in United States
The HPLC-QTOF-MS is a high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry system. It is designed to provide accurate mass measurements and high-resolution detection of a wide range of analytes.
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Lab products found in correlation
6550 ifunnel q tof, by Agilent Technologies (1 mentions)
1290 infinity uhplc, by Agilent Technologies (1 mentions)
Reverse phase c18 column, by Hill-Rom (1 mentions)
Xbridge c18 column, by Waters Corporation (1 mentions)
6210 tof mass spectrometer, by Agilent Technologies (1 mentions)
Mass hunter b4, by Agilent Technologies (1 mentions)
Agilent 1290 ultra, by Agilent Technologies (1 mentions)
4 protocols using hplc qtof ms
1
Silicosis Metabolomic Profiling and Analysis
2
Quantification of Protein-Ligand Interactions by EIC
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Extracted ion chromatography was performed with reaction mixtures containing 250 nm of MBP-STAT5b protein, 250 µm (IC20) 4-amino-furazane-3-carboxylic acid 3 , equimolar amount (250 µm ) of one heteraryl nucleophile and FA with a total volume of 100 µl. The reaction mixtures were vortexed to mix thoroughly and incubated overnight at room temperature and was analyzed using a HPLC/QTOF-MS instrument by Agilent, consisting of an Infinity 1290 UHPLC coupled to a 6550 iFunnel QTOF. After 12 h each sample was analyzed in triplicate by injecting (10 μl) into the LC/MS instrument and the ligation products were identified by their molecular weights and by comparison of the retention times of synthetic reference. Calibration curves for hit compounds 10 and 16 are given in Supplementary Figure 19 .
3
Enzymatic Characterization of BsGT1
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The catalytic activity of the BsGT1 to the three chalcone glycosides was verified by 300 μl reaction buffer containing 5 μg of purified protein, 75 μM Tris-HCl pH 8.0, 36.94 μM UDP-Glc and 0.84 mM substrates (dissolved in DMSO). The reactions were incubated at 37℃ for 3 h and extracted three times with equal volume ethyl acetate, and the organic phase was dried and added 200 μl methanol for analysis by HPLC and HPLC-QTOF-MS (Agilent Technologies, USA) at UV 254 nm with a reverse-phase C18 column (4.6 × 250 mm, 5 μm particles; Welch, China).
4
HPLC-Q-TOF/MS for Compound Identification
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For the identification of chemical compounds in Rhizoma Anemarrhenae extract and in rat plasma, an Agilent 1290 ultra-high-performance liquid chromatography-tandem quad-rupole time-of-flight mass spectrometer (HPLC-Q-TOF/MS, Agilent Technology Co., USA) was used, which consisted of a quaternary pump, an on-line degasser, a well-plate autosampler, a thermostatic column compartment and a 6210 TOF mass spectrometer. The chromatographic separations were performed at 30 °C on a Waters Xbridge C 18 column (3.5 μm, 3.0 × 100 mm, Waters, USA). The mobile phase consisted of (A) water/formic acid (100:0.1, V/V) and (B) acetonitrile/formic acid (100:0.1, V/V) with the following gradient elution: 0-3 min, 18-22 % B; 3-8 min, 22-25 % B; 8-10 min, 25-40 % B; and 10-15 min, 40-60 % B. The flow rate was set at 400 μL min -1 . The sample injection volume was 2 μL. The parameters of the ion source were as follows: acquisition mode, positive mode; capillary voltage 3500 V; drying gas (N 2 ) temperature 350 °C; drying gas flow rate, 10 L min -1 ; nebulizer gas (N 2 ) pressure 241 kPa; the fragmentor voltage 180 V and skimmer voltage 60 V. Mass spectra in the full-scan mode were recorded between m/z 100 and 1200. Agilent MassHunter B4.0 software was used for the control of equipment, data acquisition and analysis.
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