For the measurement of free PUFA (μg/ml), 100 μl of the blood sample from each piglet was precipitated using 100 μl of internal standard (deuterated ARA, DHA, and EPA obtained from Sigma-Aldrich) solution in IPA (POCh). After that, it was acidified by addition of 100 μl of 2% HCOOH. Samples were subsequently extracted using 600 μl of hexane (Avantor Baker) and the organic layer was evaporated under nitrogen. Samples were solubilized in 100 μl of 5% NH4OH in 65% MeOH and 10 μl was injected into LC/MS. Samples were then analysed using Waters Xevo TQ-S triple quadrupole MS coupled with Waters Acquity UPLC chromatograph. The column used was an ACQUITY UPLC BEH C18 Column, 1.7 μm, 2.1 mm × 50 mm thermostated in 60°C. Mobile phase A was 0.1% NH4OH in MQ water; mobile phase B was pure LC/MS grade ACN (Avantor Baker). A linear gradient was used, starting from 40% to 95% B over 1.7 min. MS was operating in negative MRM mode. Parameters were as follows: capillary voltage 3.2, desolvation temperature 500°C, desolvation gas (nitrogen) flow 900 l /h, and cone gas flow 150 l /h. Calibration and control samples were prepared by spiking PUFA standards into 45 mg/ml fatty acid-free BSA (Sigma-Aldrich, St. Louis, MO, USA) solution in 0.9% NaCl.
Xevo tq s triple quadrupole ms
Manufactured by Waters Corporation
Sourced in United States
The Xevo TQ-S is a triple quadrupole mass spectrometer (MS) designed for quantitative and qualitative analysis. It features high sensitivity, resolution, and mass accuracy for a wide range of applications. The instrument utilizes multiple mass analyzers to perform tandem mass spectrometry (MS/MS) for compound identification and quantification.
Automatically generated - may contain errors
Lab products found in correlation
Acquity uplc chromatograph, by Waters Corporation (1 mentions)
Fatty acid free bsa, by Merck Group (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Acquity uplc h class, by Waters Corporation (1 mentions)
Lc ms grade, by Carl Roth (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
Formic acid, by Merck Group (1 mentions)
5 protocols using xevo tq s triple quadrupole ms
1
Measurement of Free Plasma PUFAs
2
Targeted Serum Amino Acid Profiling
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The targeted metabolomics analyses of AAs in the serum samples were performed by combining 4 μl serum and 991 μl of solution (1.7 mM ammonium formate in 85% acetonitrile containing 0.1% formic acid) and 5 μl of IS (phenylalanine-d5, 100 ng/ml). This mixture was vortexed for 1 min. After the centrifugation (12,000 rpm for 5 min at 4°C), 5 μl of the supernatant was directly analyzed by UPLC/MS–MS. The LC–MS/MS system was made up of a Waters ACQUITY UPLC system coupled to a Waters (Milford, MA, USA) Xevo TQ-S triple quadrupole MS with flow through-needle sample management, cooling autosampler, temperature-controlled column oven, degasser, and binary pump. The detailed chromatographic separations and MS conditions can be seen in method part in supplementary materials.
3
Tryptophan and Kynurenine Metabolites Analysis
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Detection of tryptophan, kynurenine, quinolinic acid (QUIN), and KYNA was performed using a Waters Xevo TQ-S triple quadrupole MS in mice (Grk3+/+n = 8, Grk3−/−n = 7). For details see Supplementary Information.
4
Quantitative LC-MS/MS Analysis of RNA and DNA Adducts
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Analyses were performed on an ACQUITY UPLC H-class coupled to a Xevo TQ-S triple quadrupole MS (Waters). An ACQUITY UPLC BEH C18 column (50 × 2.1 mm I.D., 1.7 μm, 130 A, Waters) was used for the chromatographic separation of analytes. For the analysis of RNA, adducts were separated with a flow of 0.8 ml/min. Mobile phase A was Millipore water and B was acetonitrile (LC-MS grade, Carl Roth), both supplemented with 0.01% formic acid (Sigma-Aldrich). The initial conditions were 100% A. B was increased to 5% until a runtime of '2.5 min' and held constant until '4 min'. Then, B was further increased to 15% until '5 min', to 30% until '5.5 min', and then held constant until '6 min'. DNA adducts were separated with a flow of 0.35 ml/min. In an initial gradient, solvent B was increased to 15% until a runtime of '5 min', further increased to 30% until '6 min', and then held constant until '7 min'.
The source settings in positive ionization mode were set to: Capillary: 0.7 kV, Cone: 11 V, Source Offset: 50 V, Source Temperature: 150 °C, Desolvation Temperature: 500 °C, Cone Gas Flow: 150 l/h, Desolvation Gas Flow: 1000 l/h, Collision Gas Flow: 0.15 ml/min, and Nebuliser Gas Flow: 7 bar. The analyzer was set to high mass resolution.
The analytes were measured with cone voltage of 6 V, collision energy of 10 V, and an auto dwell time ( 37
The source settings in positive ionization mode were set to: Capillary: 0.7 kV, Cone: 11 V, Source Offset: 50 V, Source Temperature: 150 °C, Desolvation Temperature: 500 °C, Cone Gas Flow: 150 l/h, Desolvation Gas Flow: 1000 l/h, Collision Gas Flow: 0.15 ml/min, and Nebuliser Gas Flow: 7 bar. The analyzer was set to high mass resolution.
The analytes were measured with cone voltage of 6 V, collision energy of 10 V, and an auto dwell time ( 37
5
Targeted MS Analysis of Analytes
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A Waters Xevo TQ-S triple-quadrupole MS was operated in negative electrospray ionization (ESI) mode with a capillary voltage of 2.5 kV and ion-source temperature of 150 °C. The desolvation gas was nitrogen, and the flow was set to 800 l/h at a temperature of 300 °C. The collision energy for each MRM transition was optimized for each compound both manually and using the "Intellistart"function in MassLynx 4.1. The MS was run in dynamic MRM mode, and the retention time (RT) window for each compound was set to ±2 min of the expected RT. Downstream data processing was performed in MassLynx V4.1.
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