Samples were analyzed using a TSQ Quantiva Ultra triple-quadrupole mass spectrometer coupled with an Ultimate 3000 UPLC system (Thermo Fisher Scientific) equipped with a heated electrospray ionization probe. Chromatographic separation was achieved using gradient elution on a Hypercarb PGC column (2.1 × 100 mm, 1.7 μm, Thermo Fisher Scientific). Mobile phase A consisted of 5 mM ammonium bicarbonate dissolved in pure water, and mobile phase B consisted of 100% acetonitrile. A 25 min gradient with a flow rate of 250 μl/min was applied as follows: 0–1.2 min, 4% B; 1.2–19 min, 4–35% B; 19–20 min, 35–98% B; 20–22 min, 98% B; 22–25 min 4% B. The column chamber and sample tray were kept at 45°C and 10°C, respectively. Data were acquired using selected reaction monitoring in negative switch ion mode, and optimal transitions are reported as the reference. Both the precursor and fragment ion fractions were collected at a resolution of 0.7 FWHM. The source parameters were as follows: spray voltage: 3000 V; ion transfer tube temperature: 350°C; vaporizer temperature: 300°C; sheath gas flow rate: 35 arbitrary units; auxiliary gas flow rate: 12 arbitrary units; collision-induced dissociation gas pressure: 1.5 mTorr.
Tsq quantiva ultra
Manufactured by Thermo Fisher Scientific
The TSQ Quantiva Ultra is a triple-quadrupole mass spectrometer designed for sensitive and accurate quantitative analysis. It features high-performance ion optics and a robust vacuum system to deliver reliable and reproducible results.
Automatically generated - may contain errors
Lab products found in correlation
Ultimate 3000, by Thermo Fisher Scientific (3 mentions)
Fusion rp c18 column, by Phenomenex (3 mentions)
Xcalibur 3, by Thermo Fisher Scientific (3 mentions)
Hypercarb pgc column, by Thermo Fisher Scientific (2 mentions)
Ultimate 3000 uplc system, by Thermo Fisher Scientific (2 mentions)
Tracefinder 3, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
6 protocols using tsq quantiva ultra
1
Quantitative UPLC-MS/MS Analysis Protocol
2
Steady-State Labeling of Glycolytic Metabolites
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Steady-state labelling of glycolytic intermediates was accomplished by culturing KYSE450 cells (5 × 106 cells/sample) in RPMI-1640 medium (Gibco) containing 2 g/L of d -glucose (U-13C6, 99%; Sigma-Aldrich) for 6 h. All treatments were conducted in quintuplicate. Intracellular metabolites were then extracted and measured. The Dionex Ultimate 3000 UPLC system was coupled to a TSQ Quantiva Ultra triple-quadrupole mass spectrometer (Thermo Fisher, CA), equipped with a heated electrospray ionization probe. The source parameters are as follows: capillary temperature: 350 °C; heater temperature: 300 °C; sheath gas flow rate: 35; auxiliary gas flow rate: 10. Tracefinder 3.2 (Thermo, USA) was applied for metabolite identification and peak integration.
3
Drosophila Metabolite Quantification by LC-MS
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LC-MS was performed as previous reported (Han et al., 2017 (link)). The Dionex Ultimate 3000 UPLC system was coupled to a TSQ Quantiva Ultra triple-quadrupole mass spectrometer (Thermo Fisher, Waltham, MA), equipped with a heated electrospray ionization probe in negative ion mode. Extracts were separated by a Fusion-RP C18 column (2 × 100 mm, 2.5 μm, phenomenex). Data acquired in selected reaction monitoring for histamine, carcinine, and β-alanine with transitions of 112/95.2, 183/95, and 90/72, respectively. Both precursor and fragment ions were collected with resolution of 0.7 FWHM. The source parameters are as follows: spray voltage: 3000 V; ion transfer tube temperature: 350°C; vaporizer temperature: 300°C; sheath gas flow rate: 40 Arb; auxiliary gas flow rate: 20 Arb; CID gas: 2.0 mTorr. Data analysis and quantification were performed using the software Xcalibur 3.0.63 (Thermo Fisher, CA). Each sample contained 50 Drosophila heads, and the mean values from five samples were calculated.
4
Dionex UPLC-MS/MS Quantification of Biogenic Amines
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The Dionex Ultimate 3000 UPLC system was coupled to a TSQ Quantiva Ultra triple-quadrupole mass spectrometer (Thermo Fisher, CA), equipped with a heated electrospray ionization (HESI) probe in negative ion mode. Extracts were separated by a Fusion-RP C18 column (2 × 100 mm, 2.5 μm, phenomenex). Data acquired in selected reaction monitoring (SRM) for histamine, carcinine, and β-alanine with transitions of 112/95.2, 183/95, and 90/72, respectively. Both precursor and fragment ions were collected with resolution of 0.7 FWHM. The source parameters are as follows: spray voltage: 3000 V; ion transfer tube temperature: 350°C; vaporizer temperature: 300°C; sheath gas flow rate: 40 Arb; auxiliary gas flow rate: 20 Arb; CID gas: 2.0 mTorr. Data analysis and quantification were performed using the software Xcalibur 3.0.63 (Thermo Fisher, CA). Each sample contained 50 Drosophila heads, and the mean values from five samples were calculated.
5
Quantifying Neurotransmitters in Drosophila
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Liquid chromatography–mass spectrometry was performed as previously reported (48 (link)). Briefly, the Dionex Ultimate 3000 UPLC system was coupled to a TSQ Quantiva Ultra triple-quadrupole mass spectrometer (Thermo Fisher Scientific, CA) and equipped with a heated electrospray ionization probe in negative ion mode. Extracts were separated by a Fusion-RP C18 column (2 × 100 mm, 2.5 μm, Phenomenex). Data were acquired in selected reaction monitoring for histamine, carcinine, and β-alanine with transitions of 112/95.2, 183/95, and 90/72, respectively. Both precursor and fragment ions were collected with a resolution of 0.7 full width at half maximum. The source parameters (spray voltage, 3000 V; ion transfer tube temperature, 350°C; vaporizer temperature, 300°C; sheath gas flow rate, 40 Arb; auxiliary gas flow rate, 20 Arb; collision induced dissociation gas, 2.0 mtorr) are set. Data analysis and quantification were performed using the Xcalibur 3.0.63 software (Thermo Fisher Scientific, CA). Each sample contained 50 Drosophila heads. Three samples were measured for each genotype, and the mean values from three samples were calculated.
6
Targeted Metabolite Profiling by UPLC-MS/MS
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Samples were analyzed using a TSQ Quantiva Ultra triple-quadrupole mass spectrometer coupled with an Ultimate 3000 UPLC system (Thermo Fisher Scientific) equipped with a heated electrospray ionization probe. Chromatographic separation was achieved using gradient elution on a Hypercarb PGC column (2.1 × 100 mm, 1.7 μm, Thermo Fisher Scientific). Mobile phase A consisted of 5 mM ammonium bicarbonate dissolved in pure water, and mobile phase B consisted of 100% acetonitrile. A 25-minute gradient with a flow rate of 250 μl/min was applied as follows: 0-1.2 min, 4% B; 1.2-19 min, 4-35% B; 19-20 min, 35-98% B; 20-22 min, 98% B; 22-25 min 4% B. The column chamber and sample tray were kept at 45°C and 10°C, respectively. Data were acquired using selected reaction monitoring in negative switch ion mode, and optimal transitions are reported as the reference. Both the precursor and fragment ion fractions were collected at a resolution of 0.7 FWHM. The source parameters were as follows: spray voltage: 3000 V; ion transfer tube temperature: 350°C; vaporizer temperature: 300°C; sheath gas flow rate: 35 arbitrary units; auxiliary gas flow rate: 12 arbitrary units; collision induced dissociation (CID) gas pressure: 1.5 mTorr.
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