MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14349/m_MTBLS14349_LC-MS_negative_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14349/m_MTBLS14349_LC-MS_positive_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14349/a_MTBLS14349_LC-MS_negative_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14349/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14349/s_MTBLS14349.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14349/a_MTBLS14349_LC-MS_positive_reverse-phase.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14349<p>The identification of metabolites is based on the Psngm Database (PerSonalbio Next-Generation Metabolomics Database). This database includes self-built standards, the mzCloud database (https://www.mzcloud.org/), LIPID MAPS (https://www.lipidmaps.org/), HMDB (https://hmdb.ca/), MoNA (https://mona.fiehnlab.ucdavis.edu/), NIST_2020_MSMS, and an AI-predicted MSMS spectrum database. The main parameters for searching the database are as follows: MS1 identification tolerance 0.01, MS2 identification tolerance 0.05, smoothing level 3, Minimum peak height 10000, Minimum peak width 5, Mass slice width 0.05, and Identification score cut-off 70.</p>MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phase<p>The ACQUITY UPLC HSS T3 chromatographic column (100A, 1.8 µm, 2.1 mm × 100 mm) was used, with a flow rate of 0.4 mL/min.</p><p>Temperature: 40℃, automatic sampler: 8℃, injection volume: 2 μL.</p><p>Positive and negative mode mobile phases: Mobile phase A is 0.1% formic acid water, and mobile phase B is acetonitrile (containing 0.1% formic acid).</p>Multi-omics profiling unravels novel oncogenic mechanisms in colorectal cancer initiation.jiang feiXiangya Hospital, Central South Universityfecesmass spectrometry assay<p>1. Weigh 20 mg of the sample into a 2 mL centrifuge tube.</p><p>2. Add 300 μL of pre-cooled methanol (containing 5 ppm 2-chloropphenylalanine), add 2 steel balls, and vortex for 30 seconds;</p><p>3. Place it in a high-throughput tissue grinder at 55 Hz and grind for 60 seconds. Repeat this step once;</p><p>4. Put it into the ultrasonic cleaner and ultrasonicate for 10 minutes;</p><p>5. Freeze in a -20℃ refrigerator for 30 minutes;</p><p>6. Centrifuge at 12,000 rpm and 4 ° C for 10 minutes. Take the supernatant and pass it through a 0.22 μm filter membrane. Add the filtrate to the test bottle.</p><p>7. Take 10-20 μL of each sample filtrate and mix them into one QC sample. QC samples are used for the evaluation instrument device stability and data reliability.</p>Mus musculus, NCBITaxon, http://purl.obolibrary.org/obo/NCBITaxon_10090https://www.ebi.ac.uk/metabolights/MTBLS14349jiang fei. Xiangya Hospital, Central South University. Xiangya Hospital, Central South University, Kaifu, Changsha, Hunan, China. feijiangfj@csu.edu.cn.<p>Import the downloaded .raw format data into the software MS-DIAL (version 4.9.221218). Based on this software, perform operations such as peak extraction, alignment, filtering, and qualitative identification of metabolites. Filter out the peaks that are not detected in more than 50% of the QC samples. For the undetected peaks, fill in the missing values using the Gap filling algorithm of the software and perform normalization.</p>Treatmentfeijiangfj@csu.edu.cn<p>The origin of samples is feces of mouse. Stored in -80 degree.</p>MetabolomicsProteoWizard msconvert//purl.obolibrary.org/obo/NCBITaxon_10090Metabolomicscolorectal canceruntargeted analysisThermo Scientific Orbitrap Exploris 120Thermo Scientific Vanquish Flex UHPLC Systemuntargeted metabolite profilingdata-independent acquisitionThermo Fisher Scientific (China)XCMSexperimental blankfeces//purl.obolibrary.org/obo/NCBITaxon_10090ProteoWizard msconvertMetabolomicscolorectal canceruntargeted analysisThermo Scientific Vanquish Flex UHPLC SystemThermo Scientific Orbitrap Exploris 120untargeted metabolite profilingdata-independent acquisitionThermo Fisher Scientific (China)XCMSexperimental blankfeces<p>The Thermo Orbitrap Exploris 120 mass spectrometer was used for calibration and calibration respectively under the control of the software Xcalibur (version 4.7, Thermo).</p><p>Data acquisition of DDA mass spectrometry in negative ion mode. Using HESI source, spray voltage 3.5 kV/-3.0 kV, sheath gas 40 arb, auxiliary gas 10 arb, capillary temperature 320 ° C, auxiliary air temperature 300 ° C, first-level resolution 60,000, scanning range 70-1000 m/z, AGC Target Standard, Max IT 100 ms, screen the top 4 ions in response for secondary fragmentation, dynamic exclusion time 4 s, secondary resolution15,000, HCD collision energy 30%, AGC Target Standard, Max IT Auto.</p><p>All formal samples and QC samples are loaded onto the machine in accordance with the above-mentioned chromatography and mass spectrometry methods. For formal injection, 2 to 4 needles of QC samples are advanced for system balance, during the sample injection process, a QC sample is injected every 5 to 10 samples. They are used for subsequent procedures data evaluation and quality control.</p>falseUntargeted metabolome of mouse fecesUntargeted metabolome of mouse feces. Feces were divided into two groups, control and experimental.2026-04-222026-04-22MTBLS14349