Blood samples were collected by a physician in the fasting state before and after the 12-week intervention. Serum samples were used to blindly analyze serum metabolites at the Gustave Roussy Cancer Campus facility (Villejuif, France) using mass spectrometers coupled to multiple different liquid or gas phase chromatography methods. Bile acids metabolomics were obtained using a UHPLC/MS—RRLC 1260 system (Agilent Technologies, Waldbronn, Germany) coupled to a Triple Quadrupole 6410 (Agilent Technologies). Short chain fatty acids, oxylipin, and lipids metabolomics were assessed by a UHPLC/QUAD+—RRLC 1260 system (Agilent Technologies, Waldbronn, Germany) coupled to a 6500+ QTRAP (Sciex, Darmstadt, Germany). Polyamines metabolomics were quantified using a UHPLC/QQQ—RRLC 1260 system (Agilent Technologies, Waldbronn, Germany) coupled to a Triple Quadrupole 6410 (Agilent Technologies). Concerning the level of identification, for the targeted metabolomics it is identified (level 1, validated by standards injections on the Orbitrap, and multiple reaction monitoring MRM developments for the LCQQQ and GCQQQ), and for the metabolomic profiling it is putative. Intra batch correction was performed based on quality control pool and processed on R software. The details of each method were previously described [26 (link),27 (link)].
Triple quadrupole 6410
Manufactured by Agilent Technologies
Sourced in Germany, United States
The Triple Quadrupole 6410 is a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) system designed for a wide range of analytical applications. It features two quadrupole mass analyzers for enhanced selectivity and sensitivity in quantitative and qualitative analysis.
Automatically generated - may contain errors
Lab products found in correlation
Rrlc 1260 system, by Agilent Technologies (9 mentions)
Mass hunter quantitative software, by Agilent Technologies (5 mentions)
Zorbax eclipse plus c18 column, by Agilent Technologies (2 mentions)
Qtrap 6500, by AB Sciex (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
1290 infinity uplc system, by Agilent Technologies (1 mentions)
7100 capillary electrophoresis system, by Agilent Technologies (1 mentions)
Polyimide, by Molex (1 mentions)
1260 infinity isocratic pump, by Agilent Technologies (1 mentions)
Kromasil, by Merck Group (1 mentions)
1290 infinity lc, by Agilent Technologies (1 mentions)
Masshunter workstation quantitative analysis software, by Agilent Technologies (1 mentions)
Xdb c18, by Agilent Technologies (1 mentions)
Heptafluorobutyric acid hfba, by Merck Group (1 mentions)
Kinetex c18 column, by Phenomenex (1 mentions)
19 protocols using triple quadrupole 6410
1
Comprehensive Metabolomic Analysis of Serum Samples
2
Quantification of Saponin Aglycones by UPLC-MS
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Saponins aglycones were analysed on a 1290 Infinity UPLC system connected to tandem mass spectrometer 6410 Triple Quadrupole (Agilent Technologies, Santa Clara, CA, USA). The atmospheric pressure chemical ionization source was set to negative ionization mode and mass analyser was optimized for single ion monitoring. Data acquisition and processing were performed with MassHunter (Agilent Technologies). Separation was performed on an Acquity UPLC BEH C18 column (100 mm × 2.1 mm I.D., 1.7 µm, Waters) with linear elution gradient as described in [17 (link)]. The total amount of saponin aglycones was measured using internal calibration with oleanolic acid and the determined concentration was adjusted to dry plant mass. The limit of quantification for the oleanolic acid was estimated form regression parameters and was 0.25 mg/g.
3
Quantification of RBC Metabolites
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An Agilent 6410 triple quadrupole fitted to an electrospray ionizer (ESI) with an Agilent 1200 rapid resolution liquid chromatographer (RRLC) was utilized. The most optimized chromatographic and mass parameters were selected for RBC and its related metabolites. Fragment ions for RBC and their metabolites were formed inside the second hexapole (collision cell) by collision induced dissociation technique using high purity nitrogen. The chosen chromatographic conditions for resolution of RBC metabolites are shown in Table 1 .
4
CE-MS and CE-UV Analysis of Compounds
5
Phytochemical Profiling of P. grandiflorum
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PD from P. grandiflorum extract was analyzed on an LC–MS system (Agilent Technologies 6410 Triple quadrupole) equipped with a heated ESI interface. The sample solution was prepared by dissolving P. grandiflorum extract in MeOH/DW (1:3, v/v) solution at 1000 ppm. A C18 reversed‐phase column (Kromasil, 3.0 μm, 3.0 mm × 150 mm; Merck) was used for separation. The column temperature was 26°C. The mobile phase was a binary gradient with 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The gradient elution was 0 min, 10% B; 1–6 min, 15% B; 7–20 min, 15% B; 21–30 min, 25% B; 31–35 min, 100% B; and the flow rate was 0.4 mL/min. The MS detector was operated in negative ion mode with the following settings: capillary voltage of 4 kV, gas temperature of 350°C, nebulizer of 40 psi, and fragmentor of 135 V.
6
Quantification of Glycerolipid-bound OPDA
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In tissue quantification of glycerolipid-bound 12-Oxo-phytodienoic acid (OPDA) was performed as previously described (Nilsson et al., 2012 (link)). For the quantification of free OPDA and JA released by tissue, 6 leaf discs were placed in a glass tube with 2 ml dH2O and kept on light agitation on an orbital shaker. Discs were removed at indicated times and samples were acidified with 50 µl 1.6 M HCl follow by phase separation through the addition of 2 ml ethyl acetate. The organic phase was collected and the original sample was re-extracted with 2 ml of ethyl acetate before drying under a stream of nitrogen gas. Samples were dissolved in 50 µl methanol, run on an Agilent 1290 Infinity LC coupled to an Agilent 6410 triple quadrupole mass selective detector and the amounts of JA and free OPDA were quantified as described (Pan, Welti & Wang, 2010 (link)), except that hexadeuterated JA (OlChemim Ltd, Olomouc, Czech Republic) was used as internal standard.
7
Quantitative Mass Spectrometry Analysis
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The mass spectrometry detection system consisted of an Agilent Technologies 6410, triple quadrupole with ESI in positive ion mode. The mass spectrometry was operated in multiple-reaction monitoring (MRM) mode. The desolvation gas (nitrogen) and flow is operated at 350°C and 12 L/min, respectively. The nebulizer pressure was 60 psi and the capillary voltage was 5 kV. The MS collision gas was highly pure N2 (>99.9995).
The dwell time, fragmentor voltage and collision energy were optimized for all the compounds (Table 1). After separation by HPLC, the peak area corresponding to the mass-to-charge ratio (m/z) transition for the quantifier ion was measured as relative to the m/z transition for its IS. The m/z transition for the qualifier ion was also monitored to add specificity to the results. Additionally, for endogenous phospholipids monitoring, m/z 184.1>184.1 transition was used (Table 1). The integration peak area of the MRM transitions of each analyte was calculated using MassHunter Workstation Quantitative Analysis software (Agilent Technologies, Madrid, Spain).
The dwell time, fragmentor voltage and collision energy were optimized for all the compounds (Table 1). After separation by HPLC, the peak area corresponding to the mass-to-charge ratio (m/z) transition for the quantifier ion was measured as relative to the m/z transition for its IS. The m/z transition for the qualifier ion was also monitored to add specificity to the results. Additionally, for endogenous phospholipids monitoring, m/z 184.1>184.1 transition was used (Table 1). The integration peak area of the MRM transitions of each analyte was calculated using MassHunter Workstation Quantitative Analysis software (Agilent Technologies, Madrid, Spain).
8
Seasonal Monitoring of PPCPs in Wastewater
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The determination of PPCPs took place during the fall season (Sep to Nov 2015). 8-h composites samples were collected twice per week (n=10) by grabbing a volume of about 100 mL 3 times per day (at 9, 13 and 17h), in order to buffer diurnal and weekly variations. Samples were immediately taken to the laboratory and a solid phase extraction (SPE) was carried out within 24h. Analysis of the target compounds was performed by an Agilent 6410 Triple Quadrupole LC/MS-MS at the University of Catania (Catania, Italy) according to the procedure reported by Sgroi et al. (2016) (link) and here described in Supplementary material (Text S2, Table S1, Table S2, Table S3).
9
RRLC-MS/MS Quantitative Analysis
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Targeted analysis was performed on a RRLC 1260 system (Agilent Technologies) coupled to a Triple Quadrupole 6410 (Agilent Technologies) equipped with an electrospray source operating in negative mode. Gas temperature was set to 350° C with a gas flow of 12 L/min. Capillary voltage was set to 4.0 kV [26 (link)]. Peak detection and integration of analytes were performed using the Agilent Mass Hunter quantitative software (B.07.01), exported as tables and processed within R software.
10
RRLC-MS/MS for Targeted Metabolite Analysis
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Targeted analysis was performed on a RRLC 1260 system (Agilent) coupled to a Triple Quadrupole 6410 (Agilent) equipped with an electrospray source operating in positive mode. The gas temperature was set at 350°C with a gas flow of 12l/min. The capillary voltage was set at 3.5kV. 10μL of sample were injected on a Column Zorbax Eclipse plus C18 (100mm x 2.1mm, particle size 1.8μm) from Agilent technologies, protected by a guard column XDB-C18 (5mm × 2.1mm, particle size 1.8μm) and heated at 40°C. The gradient mobile phase consisted of water with 2mM of DBAA (A) and acetonitrile (B). The flow rate was set to 0.2 ml/min, and gradient as follow: initial condition is 90% phase A and 10% phase B, maintained during 4 min. Molecules are then eluted using a gradient from 10% to 95% phase B over 3 min. The column was washed using 95% mobile phase B for 3 minutes and equilibrated using 10% mobile phase B for 3 min. The autosampler was kept at 4°C. The collision gas was nitrogen. The scan mode used was the MRM for biological samples. Peak detection and integration of the analytes were performed using the Agilent Mass Hunter quantitative software (B.07.01).
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