Detailed procedures have been previously described63 (link). In brief, permethylated glycans were hydrolyzed in TFA before reduction by sodium borodeuteride and acetylation with acetic anhydride. The resulting partially permethylated alditol acetates were dissolved in hexane and analyzed by a Thermo Scientific TRACE 1310 Gas Chromatograph equipped with a Thermo Scientific Q Exactive Orbitrap mass spectrometry system. 2–3 µL of each sample was injected into an Agilent fused-silica capillary column of cross-linked DB-5MS (30 m × 0.25 mm × 0.25 µm). The GC conditions were as follows: inlet and transfer line temperatures, 290 °C; oven temperature program, 90 °C for 1 min, 8 °C/min to 290 °C for 5 min, 10 °C/min to 300 °C for 5 min; inlet helium carrier gas flow rate, 1 mL/min; split ratio, 10. The electron impact (EI)-MS conditions were as follows: ion source temperature, 300 °C; full scan m/z range, 30–750 Da; resolution, 60,000; AGC target, 1e6; maximum IT, 200 ms. Data were acquired and analyzed with Thermo TraceFinder 4.1 software package.
Q exactive orbitrap mass spectrometry system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Q Exactive Orbitrap mass spectrometry system is a high-resolution, accurate-mass (HRAM) instrument designed for a variety of analytical applications. It combines quadrupole mass filtering with the Orbitrap mass analyzer to provide high-performance mass spectrometry capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Ultimate 3000, by Thermo Fisher Scientific (3 mentions)
Acquity uplc hss t3, by Waters Corporation (2 mentions)
Xcalibur 3, by Thermo Fisher Scientific (2 mentions)
Fused silica capillary column of cross linked db 5ms, by Agilent Technologies (1 mentions)
Trace 1310 gas chromatograph, by Thermo Fisher Scientific (1 mentions)
Acquity uplc hss t3 column, by Waters Corporation (1 mentions)
4 protocols using q exactive orbitrap mass spectrometry system
1
Structural Analysis of Permethylated Glycans
2
Lipidomics Analysis by LC-MS/MS
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A Dionex Ultimate 3000 liquid chromatography system (Sunnyvale, CA, USA) (SN: 7254012) coupled to a Thermo Fisher Q Exactive Orbitrap mass spectrometry system (Waltham, MA, USA) (SN: SN02386L) were used for lipidomics analysis. The LC conditions were as follows: column, Waters Acquity UPLC HSS T3 (1.8 μm, 2.1 × 100 mm; Milford, MA, USA); mobile phase A, acetonitrile-water (60:40, v/v) containing 0.1% formic acid and 10 mM ammonium formate; mobile phase B, isopropanol-acetonitrile (90:10, v/v) containing 0.1% formic acid and 10 mM ammonium format; The gradient conditions were set as follows: 0.0–4.0 min, 30 to 60% B; 4.0–9.0 min, 60 to 100% B; 9.0–15.0 min, 100% B; 15.0–18.0 min, 100% B to 30% B. The injection volume was 5 μL, and the column temperature was 50 °C, as well as the flow rate was 0.3 mL/min.
The MS spectrometric parameters were as follows: spray voltage, 3.5 kV; sheath gas flow rate, 50 psi; auxiliary gas flow rate, 13 arb; capillary temperature, 320 °C; auxiliary gas heater temperature, 420 °C; scan modes, full MS (resolution 70,000) and ddMS2 (resolution 17,500 with stepped collision energy (10, 20, and 40 eV); and scan range, m/z 100–1200. All data were acquired using the Thermo Scientific Xcalibur 3.1 software (Waltham, MA, USA).
The MS spectrometric parameters were as follows: spray voltage, 3.5 kV; sheath gas flow rate, 50 psi; auxiliary gas flow rate, 13 arb; capillary temperature, 320 °C; auxiliary gas heater temperature, 420 °C; scan modes, full MS (resolution 70,000) and ddMS2 (resolution 17,500 with stepped collision energy (10, 20, and 40 eV); and scan range, m/z 100–1200. All data were acquired using the Thermo Scientific Xcalibur 3.1 software (Waltham, MA, USA).
3
Serum Metabolite Profiling by UPLC-MS/MS
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Aliquots of 5 µL of serum were subjected to a Dionex Ultimate 3000 liquid chromatography system (Sunnyvale, CA, USA) (SN: 7254012) equipped with a Waters Acquity UPLC HSS T3 column (1.8 μm, 2.1 × 100 mm; Milford, MA, USA) for the separation analysis. The mobile phase was represented by a gradient of eluent A (water: acetonitrile = 4:6, v/v, containing 0.1% formic acid and 10 mM ammonium formate) and eluent B (isopropanol: acetonitrile = 9:1, v/v, containing 0.1% formic acid and 10 mM ammonium formate) with flow rate of 0.3 mL/min. The gradient conditions were set as follows: 0.0–4.0 min, 30% to 60% B; 4.0–9.0 min, 60% to 100% B; 9.0–15.0 min,100% B; 15.0–18.0 min,100% B to 30% B. The column temperature was 50°C.
The mass spectrometry analysis was performed on a Thermo Fisher Q Exactive Orbitrap mass spectrometry system (Waltham, MA, USA) (SN: SN02386L, operating in both ESI+ and ESI- mode. The mass range was set between m/z 100–1200. The key detected parameters were as follows: spray voltage, 3.5 kV; sheath gas flow rate, 50 psi; auxiliary gas flow rate, 13 arb; capillary temperature, 320°C; auxiliary gas heater temperature, 420°C; scan modes, full MS (resolution of 70,000) combined ddMS2 (resolution of 17,500) scan with stepped collision energy (10, 20, and 40 eV). All data were acquired using the Thermo Scientific Xcalibur 3.1 software (Waltham, MA, USA).
The mass spectrometry analysis was performed on a Thermo Fisher Q Exactive Orbitrap mass spectrometry system (Waltham, MA, USA) (SN: SN02386L, operating in both ESI+ and ESI- mode. The mass range was set between m/z 100–1200. The key detected parameters were as follows: spray voltage, 3.5 kV; sheath gas flow rate, 50 psi; auxiliary gas flow rate, 13 arb; capillary temperature, 320°C; auxiliary gas heater temperature, 420°C; scan modes, full MS (resolution of 70,000) combined ddMS2 (resolution of 17,500) scan with stepped collision energy (10, 20, and 40 eV). All data were acquired using the Thermo Scientific Xcalibur 3.1 software (Waltham, MA, USA).
4
Lipidomic Analysis by LC-MS/MS
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A Dionex Ultimate 3000 liquid chromatography system (Sunnyvale, CA, USA) (SN: 7254012) coupled to a Thermo Fisher Q Exactive Orbitrap mass spectrometry system (Waltham, MA, USA) (SN: SN02386L) were used for lipidomics analysis. The LC conditions were as follows: column, Waters Acquity UPLC HSS T3 (1.8 μm, 2.1 × 100 mm; Milford, MA, USA); mobile phase A, acetonitrile-water (60:40, v/v) containing 0.1% formic acid and 10 mM ammonium formate; mobile phase B, isopropanol-acetonitrile (90:10, v/v) containing 0.1% formic acid and 10 mM ammonium format; The gradient conditions were set as follows: 0.0-4.0 min, 30% to 60% B; 4.0-9.0 min, 60% to 100% B; 9.0-15.0 min, 100% B; 15.0-18.0 min, 100% B to 30% B. The injection volume was 5 μL, and the column temperature was 50°C, as well as the ow rate was 0.3 mL/min.
The MS spectrometric parameters were as follows: spray voltage, 3.5 kV; sheath gas ow rate, 50 psi; auxiliary gas ow rate, 13 arb; capillary temperature, 320°C; auxiliary gas heater temperature, 420°C; scan modes, full MS (resolution 70,000) and ddMS2 (resolution 17,500 with stepped collision energy (10, 20, and 40 eV); and scan range, m/z 100-1200. All data were acquired using the Thermo Scienti c Xcalibur 3.1 software (Waltham, MA, USA).
The MS spectrometric parameters were as follows: spray voltage, 3.5 kV; sheath gas ow rate, 50 psi; auxiliary gas ow rate, 13 arb; capillary temperature, 320°C; auxiliary gas heater temperature, 420°C; scan modes, full MS (resolution 70,000) and ddMS2 (resolution 17,500 with stepped collision energy (10, 20, and 40 eV); and scan range, m/z 100-1200. All data were acquired using the Thermo Scienti c Xcalibur 3.1 software (Waltham, MA, USA).
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