The analyses were performed using a Thermo Scientific LCQ FLEET system consisting of an LCQ FLEET ion trap mass spectrometer, a Surveyor MS Pump/Autosampler/PDA Detector (Thermo Fisher Scientific, Waltham, MA, USA) through an ESI source. The separation was obtained by using a Gemini® C18 110 A analytical column (150 × 2.00 mm i.d., 5 μm) and the pre-column (Phenomenex, Torrance, CA, USA). The mobile phase consisted of aqueous formic acid at 0.1% and acetonitrile (solvent B) at 0.3 mL/min (the injection volume was 10 µL). A linear solvent gradient was used as follows: from 10% B to 95% B in 25 min with a final plateau of 3 min at 100% B. The ion trap operated in data-dependent, full scan (60–2000 m/z), and MSn mode to obtain fragment ions m/z with a collision energy of 35% and an isolation width of 3 m/z. The negative and positive parameters of the ion mode ESI source have been optimized to an ionization voltage of 5.0 kV, a capillary temperature of 320 °C, a capillary voltage of 32 V, a sheath gas flow rate of 25 arbitrary units (AU), and an auxiliary gas flow rate of 10 AU. Data was acquired using Thermo Excalibur 2.2 software (Thermo Fisher Scientific, MA, USA).
Lcq fleet system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The LCQ FLEET system is a liquid chromatography-mass spectrometry (LC-MS) instrument designed for qualitative and quantitative analysis. It combines a high-performance liquid chromatography (HPLC) system with a quadrupole ion trap mass spectrometer to provide comprehensive analytical capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Pre column, by Phenomenex (2 mentions)
Lcq fleet ion trap mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Surveyor ms pump autosampler pda detector, by Thermo Fisher Scientific (2 mentions)
Gemini c18 110a analytical column, by Phenomenex (2 mentions)
Proswift rp 4h column, by Thermo Fisher Scientific (2 mentions)
Ultimate 3000, by Thermo Fisher Scientific (2 mentions)
Thermo excalibur 2, by Thermo Fisher Scientific (1 mentions)
6 protocols using lcq fleet system
1
Comprehensive LC-MS/MS Analysis of Metabolites
2
Protein Detection via HPLC-MS
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Protein samples (0.1 mg/ml, 1 µl) were injected with an UltiMate3000® HPLC system (UHPLC+ focused, Dionex) into a ProSwift™ RP-4H column (1× 50 mm, Thermo) at a flow rate of 0.7 ml/min. The proteins eluted with a linear gradient of 5–100% acetonitrile (0.1% formic acid) in 6 min. The desalted samples were ionized and analyzed by a LCQ fleet™ system (Thermo) combining electro spray ionization with an ion trap mass analyzer.
3
GC-MS Analysis of Volatile and Non-Volatile Compounds
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The GC–MS analysis of the volatile fractions
and the derivatives
of the non-volatile compounds were achieved using an ISQ 7000 Single
Quadrupole GC–MS apparatus equipped with a TraceGOLD TG-5MS
GC (5% phenyl and 95% dimethyl polysiloxane) capillary column (30
m × 0.25 mm i.d.; 0.25 μm film). The experimental parameters
of the GC–MS instrument are similar to our previous published
work.35 (link)The mass spectrometric data
were obtained using an ESI-MS mass spectrometer with an LCQ FLEET
system from Thermo Scientific Company (USA) with a mass scanning range
of 0–2000. Nitrogen was used as nebulizing gas, and helium
(grade 5.8) was used as carrier gas. The sample was dissolved in HPLC-grade
acetonitrile and introduced by direct infusion with a syringe pump
employing a 250 mL syringe at a constant flow rate of 3 mL min–1.
and the derivatives
of the non-volatile compounds were achieved using an ISQ 7000 Single
Quadrupole GC–MS apparatus equipped with a TraceGOLD TG-5MS
GC (5% phenyl and 95% dimethyl polysiloxane) capillary column (30
m × 0.25 mm i.d.; 0.25 μm film). The experimental parameters
of the GC–MS instrument are similar to our previous published
work.35 (link)The mass spectrometric data
were obtained using an ESI-MS mass spectrometer with an LCQ FLEET
system from Thermo Scientific Company (USA) with a mass scanning range
of 0–2000. Nitrogen was used as nebulizing gas, and helium
(grade 5.8) was used as carrier gas. The sample was dissolved in HPLC-grade
acetonitrile and introduced by direct infusion with a syringe pump
employing a 250 mL syringe at a constant flow rate of 3 mL min–1.
4
Quantitative Mass Spectrometry Analysis of GB1
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GB1 samples were analysed using an LCQ‐Fleet system (Thermo Fisher Scientific) equipped with a 3D ion trap and ESI device. The MS instrument was connected to an UltiMate 3000 high‐performance liquid chromatography (HPLC) system (Dionex, Thermo Fisher Scientific) with a 10 × 2.1 mm HyperSep Retain PEP column (MZ Analysetechnik, Mainz, Germany). The labelling efficiency achieved for GB1 in heavy isotope‐containing media was calculated using Equation 2. where EL is the labelling efficiency, mL is the mass of the protein obtained from cells grown in labelled cultivation medium [Da], mR is the mass of the protein obtained from a reference medium [Da], Δmi is the mass difference between the common and heavy isotope [Da] and ni is the number of atoms of the element in the protein, that is 88 for N and 362 for H (only non‐exchangeable protons (Hoopes et al.,
2015 ) are considered). For 15N labelling, Δmi was 1 Da and the two pairs of media were H (reference)/N (labelled) and D (reference)/DN (labelled), for 2H labelling, Δmi was also 1 Da and the two pairs of media were H (reference)/D (labelled) and N (reference)/DN (labelled).
2015 ) are considered). For 15N labelling, Δmi was 1 Da and the two pairs of media were H (reference)/N (labelled) and D (reference)/DN (labelled), for 2H labelling, Δmi was also 1 Da and the two pairs of media were H (reference)/D (labelled) and N (reference)/DN (labelled).
5
Analytical Workflow for Compound Characterization
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The analyses were performed using a Thermo Scientific LCQ FLEET system consisting of LCQ FLEET ion trap mass spectrometer, Surveyor MS Pump/Autosampler/PDA Detector (Thermo Fisher Scientific) through an ESI source. A Gemini C18 110A analytical column (150 × 2.00 mm i. d., 5 µm) with pre-column (Phenomenex) was used for the separation. The mobile phase consisted of formic acid 0.1 % in water (solvent A) and methanol (solvent B) at 0.3 mL/ min (injected volume 10 µL). A linear solvent gradient used was as follows: from 15 % B to 100 % B in 30 min, with a final plateau of 3 min at 100 % B. The ion trap operated in data-dependent, full scan (60-2000 m/z), and MS n mode to obtain fragment ions m/z with a collision energy of 35 % and an isolation width of 3 m/z. The positive parameters of the ion mode ESI source have been op-timized to an ionization voltage of 5.0 kV, a capillary temperature of 300 °C, a capillary voltage of 33 V, a sheath gas flow rate of 30 arbitrary units (AU), and an auxiliary gas flow rate of 10 AU.
6
UHPLC-based Protein Separation and Analysis
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Protein samples (0.1 mg/ml, 1 µl) were injected with an UltiMate3000® HPLC system (UHPLC + focused, Dionex) into a ProSwift™ RP-4H column (1x 50 mm, Thermo) at a flow rate of 0.7 ml/min. The proteins eluted with a linear gradient of 5-100 % acetonitrile (0.1 % formic acid) in 6 min. The desalted samples were ionized and analyzed by a LCQ fleet ™ system (Thermo) combining electro spray ionization (ESI) with an ion trap mass analyzer.
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