Separation of STO-609 and its metabolites was achieved using a LC-QTOFMS system (Agilent Technologies, Santa Clara, CA) equipped with a 100 mm × 2.1 mm (Agilent XDB C18) column as previously described19 (link). Briefly, the column temperature was maintained at 45 °C. The flow rate was maintained at 0.3 ml/min with a gradient ranging from 2% to 98% aqueous acetonitrile containing 0.1% formic acid over a 15 min run. QTOFMS was operated in positive mode with electrospray ionization. Ultra high pure nitrogen was applied as the drying (12 L/min) and collision gas. The drying gas temperature was set at 325°C and nebulizer pressure was maintained at 35 psi. Capillary voltages were set at 3.5 kV. During mass spectrometry, real time mass correction and accurate mass were achieved by continuously measuring standard reference ions at m/z 121.0508, 922.0098 in the positive mode. The MS/MS of STO-609 and its metabolites was performed in targeted mode with a default isolation width of 4 m/z and collision energy ramp ranging from 20 to 50 V. Mass chromatograms and mass spectra were acquired by MassHunter® Workstation data Acquisition software (Agilent, Santa Clara, CA) in centroid and profile formats from m/z 100 to 1000. The acquisition rate was set as 1.5 spectra per second. Statistical analysis was conducted using Student’s independent t-test. Data are presented as mean ± s.e.m.
Lc qtof ms system
Manufactured by Agilent Technologies
Sourced in United States
The LC-QTOF/MS system is a high-performance liquid chromatography (LC) and quadrupole time-of-flight mass spectrometry (QTOF/MS) integrated solution. The system provides accurate mass measurement and high-resolution mass analysis capabilities for a wide range of applications.
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5 protocols using lc qtof ms system
1
LC-QTOFMS Separation of STO-609 and Metabolites
2
TSWE Analysis by LC-QTOF/MS
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The TSWE was analysed with an LC-QTOF/MS system (Agilent Technologies), consisting of a 1260 LC coupled to a 6530 QTOF-MS equipped with an electrospray ionisation source. LC separation was accomplished on a Zorbax Eclipse plus C18 column (2.1× 50 mm, 1.9 μm) at 35 °C. The injection volume was 2.0 μL. The mobile phases consist of solution A 0.1% v/v formic acid in water and solution B 0.1% formic acid in acetonitrile. The MS was tuned for a low mass range (up to 1700 m/z) and run positive mode for a full scan. Data were collected and processed using “MassHunter” B.05.00 Service Pack 3.
3
Targeted Metabolomic Profiling of Maternal-Cord Samples
4
Identification of Neoflavonoids via LC-QTOF/MS
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To check the presence of the neoflavonoids, the CSO was washed with 90% MeOH at a 1:1 ratio v/v. The MeOH layer was collected and submitted to chromatographic analysis using an LC-QTOF/MS system (Agilent Technologies), consisting of a 1260 LC coupled to a 6530 QTOF-MS equipped with an electrospray ionisation source. LC separation was accomplished on a Zorbax Eclipse plus C18 column (2.1× 50 mm, 1.9 μm) at 35 °C. The injection volume was 2.0 μL. The mobile phases consist of solution A 0.1% v/v formic acid in water and solution B 0.1% formic acid in acetonitrile. The MS was tuned for a low mass range (up to 1700 m/z) and run positive mode for a full scan. Data were collected and processed using “MassHunter” B.05.00 Service Pack 3.
5
Detecting FB1 Detoxification Metabolites
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To identify the detoxification final product of FB1 by carboxylesterase and transaminases, the analysis of degradation products was performed on an LC-QTOF-MS system (Agilent Technologies, Santa Clara, CA, USA). The chromatographic column was ZORBAX Extend-C18 reversed-phase column (150 × 2.1 mm, Agilent particle size of 1.8 μM). The sample volume was 2 µL, and the mobile phase was divided into phase A and phase B (phase A: 0.1% formic acid in water, phase B: acetonitrile of chromatographic grade). The mobile phase gradient conditions were as follows: 95% A for 35 min, maintain 5% A for 10 min in the middle and then 95% A for 15 min. The flow rate was controlled at 0.3 mL/min, 40 °C. Typical parameters of the ESI source were as follows: ESI positive mode, spray voltage 3200 v, nitrogen as auxiliary gas pressure 34.48 kPa, nitrogen sheath pressure 206.89 kPa, capillary temperature, and ion source temperature 350 °C. Data Analysis software was applied for further data acquisition and processing.
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