MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13896/m_MTBLS13896_LC-MS_negative_hilic_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13896/m_MTBLS13896_LC-MS_positive_hilic_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13896/a_MTBLS13896_LC-MS_positive_hilic_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13896/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13896/s_MTBLS13896.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13896/a_MTBLS13896_LC-MS_negative_hilic_metabolite_profiling.txtprimary200OKftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS13896<p>Thermo Scientific Compound Discoverer 3.3.2.31 is a specialized software platform for high-resolution mass spectrometry data processing.</p>MetaboLightsPublicLiquid Chromatography MS - positive - hilicLiquid Chromatography MS - negative - hilic<p>Vanquish Series UHPLC (Ultra-High Performance Liquid Chromatography) system (Thermo Scientific).Separation of metabolites were performed on an UPLC BEH Amide column. For positive and negative ionization modes, the mobile phases consisted of 0.08% formic acid and 25 mM ammonium formate in water (Phase A) and 0.08% formic acid in 95% acetonitrile (Phase B), and 25 mM ammonium acetate in water (pH = 8) (Phase A) and 95% acetonitrile in water (Phase B), respectively. Gradient elution was applied as follows: 0 min, 0% A; 5 min, 0% A; 22 min, 60% A; 27 min, 60% A; 27.1 min, 0% A. The flow rate was set at 0.2 mL/min, and the column temperature was maintained at 36 °C.</p>Real-world neonicotinoid mixture exposure reveals synergistic cytotoxicity and divergent pathway vulnerabilities in human cell lines.Peking UniversityDuo KeaiOvaryLiverCervixKidneymass spectrometry assay<p>After removing the culture medium, the cells were lysed by adding 1 mL of ice-cold LC-MS grade extraction solvent (methanol:acetonitrile:water = 2:2:1, v/v) with repeated pipetting. The resulting suspension was subjected to ultrasonication (Covaris S220) under the following program: peak power: 150, duty factor: 20, cycles: 200, duration: 20 min, while maintaining the temperature between 5 – 9°C. To precipitate proteins, the lysate was 2012 stored at -20°C for 12 h. Subsequently, the sample was centrifuged, and the supernatant was collected, filtered through a 0.22 μm PTFE membrane, and finally lyophilized under vacuum. The dried extract can be stored at -80°C for subsequent reconstitution and UPLC-MS analysis.</p>Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS13896Huan Lin. Hainan University. linhuan@hainanu.edu.cn.Ma Li. Hainan University. malihainan1127@163.com.<p>Thermo Scientific Compound Discoverer 3.3.2.31 is a specialized software platform for high-resolution mass spectrometry data processing.</p>Groupkeaiduoduo998@126.com<p>The cells were purchased from Fenghui Biotechnology (Hunan, China). HEK293T, HeLa and HepG2 cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (P/S) mixture. Chang liver cells were also cultured in MEM with 10% FBS and 1% P/S. A2780 cells were grown in RPMI-1640 with 10% FBS, 1% P/S, and 2 mM glutamine. Cells were seeded in 6-well plates at a density of 5 × 105 cells per well and cultured for 24 h under standard conditions (37°C, 5% CO2). The medium was then replaced with fresh medium containing either NEOs/metabolites or 0.1% DMSO (control group), followed by an additional 48 h of incubation.</p>MetabolomicsHuman cervical cancer cellsMetabolomicsuntargeted analysisLiverNeonicotinoidsOrbitrap Exploris 120Homo sapiensVanquish Series UHPLC system (Thermo Scientific)CervixKidneyOvaryHuman ovarian carcinoma cell lineUPLC-MS/MSHuman embryonic kidney epithelial cell linehuman hepatocellular carcinoma cell lineHuman cervical cancer cellsMetabolomicsuntargeted analysisLiverNeonicotinoidsOrbitrap Exploris 120Homo sapiensVanquish Series UHPLC system (Thermo Scientific)CervixKidneyOvaryHuman ovarian carcinoma cell lineUPLC-MS/MSHuman embryonic kidney epithelial cell linehuman hepatocellular carcinoma cell line<p>Untargeted metabolomics analysis was performed on an Orbitrap Exploris 120 mass spectrometer (Thermo Scientific).Signals were acquired in both positive and negative ion scan modes, with a mass scan range of m/z 70-800; electrospray ionization (ESI) source, positive ion voltage 3400 V, negative ion voltage 3000 V, ion source heating temperature 320 °C, and collision energy of 20-60 V.</p>falseReal-world neonicotinoid mixture exposure reveals synergistic cytotoxicity and divergent pathway vulnerabilities in human cell lines<p>In this study, five human cell lines (A2780, Chang liver, HEK293T, HeLa, and HepG2) were used as models and exposed to mixtures of neonicotinoids (NEOs) and their metabolites. The mixture concentrations were prepared based on human biomonitoring data to reflect the actual detected levels of NEOs and their metabolites in human urine. Each cell line was treated with the corresponding human biomonitoring-matched mixture at specified concentrations, alongside a 0.1% DMSO solvent control group. After 48 hours of exposure, intracellular metabolites were extracted and subjected to untargeted metabolomic analysis using an Orbitrap Exploris 120 mass spectrometer in both positive and negative ion modes. During the acquisition process, pooled quality control samples (prepared by mixing equal volumes of all samples) were interspersed to monitor system stability.</p>2026-06-222026-02-13MTBLS13896