A UHPLC system equipped with a Vanquish binary pump H system (Thermo Fisher Scientific, Waltham, MA, USA) coupled with auto-sampler and column compartment. Chromatographic separation was performed on a Phenomenex KINETEX® C18 column (100 mm × 2.1 mm, 1.7 μm particle size; Torrance, CA, USA) and the injection volume was 5 μL. The column temperature was set to 40 °C, and the flow rate was 0.3 mL/min. The mobile phase consisted of 0.1% v/v formic acid in water (A) and 0.1% v/v formic acid in acetonitrile (B). The gradient parameters were set as follows: Five percent solvent B was maintained initially for 1 min, followed by a linear increase to 100% solvent B over 9 min and then sustained at 100% solvent B for 1 min, with a gradual decrease to 5% solvent B over 3 min. The total run time was 14 min. The MS data were collected in the range of 100–1500 m/z (under negative- and positive-ion mode) using an Orbitrap Velos ProTM system, which is combined with an ion trap mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) coupled with a HESI-II probe. The probe heater and capillary temperatures were set to 300 °C and 350 °C, respectively. The capillary voltage was set to 2.5 kV in negative mode (positive mode, 3.7 Kv).
Orbitrap velos protm system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Orbitrap Velos Pro system is a high-performance mass spectrometer designed for advanced proteomics and metabolomics applications. It features an Orbitrap mass analyzer that provides high mass resolution, mass accuracy, and sensitivity. The system is capable of fast scanning and data acquisition, enabling comprehensive analysis of complex biological samples.
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Lab products found in correlation
5 protocols using orbitrap velos protm system
1
UHPLC-MS/MS Analysis of Metabolites
2
Metabolite Profiling of Culture Supernatants
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Analytical samples for metabolite profiling were prepared by diluting culture supernatant broth extracts (0.2 ml) with HPLC-grade water (0.2 ml). The UHPLC-LTQ-Orbitrap-MS/MS system used was equipped with a Vanquish binary pump H system (Thermo Fisher Scientific, USA) coupled with an autosampler and a column compartment. Chromatographic separation was performed on a Phenomenex KINETEX C18 column (100 mm × 2.1 mm, 1.7μm; Torrance, USA) with an injection volume of 5 μl. The column temperature was set to 40°C, and the flow rate was 0.3 ml/min. The mobile phases consisted of 0.1% v/v formic acid in water (A) and 0.1% v/v formic acid in acetonitrile (B). The gradient parameters were set as described by Kwon et al. [19 (link)]. The MS data were collected in the range of 100–1500 m/z (under negative- and positive-ion mode) using an Orbitrap Velos ProTM system, which is combined with an Ion-Trap Mass Spectrometer (Thermo Fisher Scientific) coupled with a HESI-II probe. The probe heater and capillary temperatures were set to 300°C and 350°C, respectively. The capillary voltage was set to 2.5 kV in negative mode (positive mode, 3.7 kV).
3
Metabolite Profiling of Culture Supernatants
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Analytical samples for metabolite profiling were prepared by diluting culture supernatant broth extracts (0.2 ml) with HPLC-grade water (0.2 ml). The UHPLC-LTQ-Orbitrap-MS/MS system used was equipped with a Vanquish binary pump H system (Thermo Fisher Scientific, USA) coupled with an autosampler and a column compartment. Chromatographic separation was performed on a Phenomenex KINETEX C18 column (100 mm × 2.1 mm, 1.7μm; Torrance, USA) with an injection volume of 5 μl. The column temperature was set to 40°C, and the flow rate was 0.3 ml/min. The mobile phases consisted of 0.1% v/v formic acid in water (A) and 0.1% v/v formic acid in acetonitrile (B). The gradient parameters were set as described by Kwon et al. [19 (link)]. The MS data were collected in the range of 100–1500 m/z (under negative- and positive-ion mode) using an Orbitrap Velos ProTM system, which is combined with an Ion-Trap Mass Spectrometer (Thermo Fisher Scientific) coupled with a HESI-II probe. The probe heater and capillary temperatures were set to 300°C and 350°C, respectively. The capillary voltage was set to 2.5 kV in negative mode (positive mode, 3.7 kV).
4
UHPLC-MS Analysis of Metabolites
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Ultrahigh performance liquid chromatography (UHPLC) was performed on a Vanquish binary pump H system (Thermo Fisher Scientific, Waltham, MA, USA) coupled with an auto-sampler and column compartment. Chromatographic separation was carried out on Phenomenex KINETEX® C18 column (100 mm × 2.1 mm, 1.7 μm particle size, Torrance, CA, USA) and the injection volume was 5 μL. The column temperature was set to 40 °C and the flow rate was 0.3 mL/min. The mobile phase consisted of 0.1% formic acid in water (Solvent A) and 0.1% formic acid in acetonitrile (Solvent B). The gradient parameters were set as follows: 5% solvent B was maintained initially for 1 min followed by a linear increase to 100% solvent B over 9 min and then sustained at 100% solvent B for 1 min with a gradual decrease to 5% solvent B over 3 min. The total run time was 14 min. The MS data were collected in the range of 100–1000 m/z (under a negative-ion and positive-ion mode) using an Orbitrap Velos ProTM system, which is combined with an ion trap mass spectrometer (Thermo Fisher Scientific) coupled with a Heated Electrospray Ionization (HESI-II) probe. The probe heater and capillary temperatures were set to 300 °C and 350 °C, respectively. The capillary voltage was set to 3.7 kV in a positive mode (negative mode, 2.5 kV). Leucine encephalin was utilized as reference lock mass (m/z 554.2615).
5
Comprehensive Metabolite Profiling of AME
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The dried AME were re-dissolved in 80% MeOH for ultra-high-performance liquid chromatography-linear trap quadrupole-orbitrap-tandem mass spectrometry (UHPLC-LTQ-Orbitrap- MS/MS) analysis to determine the components of AME.
UHPLC system consisting of a Vanquish binary pump H system (Thermo Fisher Scientific Co., Ltd., Waltham, MA, USA), autosampler, column compartment, and a detector. Sample separation was performed on a Phenomenex KINETEX® C18 column (100 mm × 2.1 mm, 1.7 μm) and the column temperature was set to 40 °C, and the flow rate was 0.3 mL/min. The MS data were collected in the range of 100–1500 m/z (under both negative- and positiveion modes) using an Orbitrap Velos ProTM system, which was combined with an ion-trap mass spectrometer (Thermo Fisher Scientific) coupled with a HESI-II probe.
UHPLC system consisting of a Vanquish binary pump H system (Thermo Fisher Scientific Co., Ltd., Waltham, MA, USA), autosampler, column compartment, and a detector. Sample separation was performed on a Phenomenex KINETEX® C18 column (100 mm × 2.1 mm, 1.7 μm) and the column temperature was set to 40 °C, and the flow rate was 0.3 mL/min. The MS data were collected in the range of 100–1500 m/z (under both negative- and positiveion modes) using an Orbitrap Velos ProTM system, which was combined with an ion-trap mass spectrometer (Thermo Fisher Scientific) coupled with a HESI-II probe.
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