The LC-ESI-MS system consisted of an ultra-performance liquid chromatography (UPLC) system (Ultimate 3000 RSLC, Dionex) and an electrospray ionization (ESI) source of quadrupole time-of-flight (TOF) mass spectrometer (maXis HUR-QToF system, Bruker Daltonics). The autosampler was kept at 4 °C. Separation was performed with reversed-phase liquid chromatography (RPLC) on a BEH C18 column (2.1×100 mm, Walters). The elution started from 99% mobile phase A (0.1% formic acid in ultrapure water) and 1% mobile phase B (0.1% formic acid in acetonitrile), raised to 90% B in 1 min, held at 90% B for 2 min, and then lowered to 1% B in 0.5 min. The column was equilibrated by pumping 1% B for 3.5 min. The flow rate was set at 0.4 ml/min with per injection volume 10 μl. LC-ESI-MS chromatogram were acquired under the following conditions: capillary voltage of 4500 V in positive ion mode, dry temperature at 190 °C, dry gas flow maintained at 8 l/min, nebulizer gas at 1.4 bar, and acquisition range of m/z 100–1000. Reaction products of methylglyoxal and 2,3-diaminopyridine gave rise to signals of at least 6-fold intensity as compared with that of methylglyoxal and o-phenylenediamine in mass spectrometry detection.
Maxis hur qtof system
Manufactured by Bruker
The MaXis HUR-QToF system is a high-resolution quadrupole time-of-flight mass spectrometer designed for accurate mass measurements and high-throughput analysis. It features a high-performance quadrupole mass filter and a time-of-flight analyzer to provide precise mass determination and enhanced sensitivity.
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5 protocols using maxis hur qtof system
1
LC-ESI-MS Analysis of Methylglyoxal Derivatives
2
Untargeted Metabolite Profiling by LC-ESI-MS
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In vitro materials were ground with liquid nitrogen and mixed with 1 mL of methanol. Supernatant was collected by centrifugation (12000 rpm, 1 min). The LC-ESI-MS system consisted of an ultra-performance liquid chromatography system (Ultimate 3000 RSLC, Dionex) and an electrospray ionization source of quadrupole time-of-flight mass spectrometer (maXis HUR-QToF system, Bruker Daltonics). The autosampler was set at 4°C. Separation was performed with reversed-phase liquid chromatography on a BEH C8 column (2.1 × 100 mm, Walters). The elution started from 99% mobile phase A (0.1% formic acid in ultrapure water) and 1% mobile phase B (0.1% formic acid in ACN), held at 1% B for 1.5 min, raised to 60% B in 6 min, further raised to 90% in 0.5 min, and then lowered to 1% B in 0.5 min. The column was equilibrated by pumping 1% B for 4 min. The flow rate was set to 0.4 mL/min with an injection volume of 5 μL. LC-ESI-MS chromatogram were acquired under the following conditions: capillary voltage of 4500 V in positive ion mode, dry temperature of 190°C, dry gas flow maintained at 8 L/min, nebulizer gas at 1.4 bar, and acquisition range of m/z 100–1000.
3
Extraction and LC-ESI-MS Analysis of Plant Metabolites
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The extraction was carried out by adding 40 μL of 100% DMF to 1 mg of the dry plant powder with vortexing for 30 min. After centrifugation at 16,000 × g for 10 min, the supernatant was collected for LC-ESI-MS analysis. The LC-ESI-MS system consisted of an ultra-performance liquid chromatography (UPLC) system (Ultimate 3000 RSLC, Dionex) and an electrospray ionization (ESI) source of quadrupole time-of-flight (TOF) mass spectrometer (maxis HUR-QToF system, Bruker Daltonics). The autosampler was set at 4°C. Separation was performed with reversed-phase liquid chromatography (RPLC) on a BEH C8 column (2.1 × 100 mm, Walters). The elution started from 50% mobile phase A (0.1% formic acid in deionized water) and 50% mobile phase B (0.1% formic acid in ACN), raised to 60% B in 2 min, further increased to 75% after an additional 1 min, and then raised to 100% B in 4.5 min, held at 100% B for 2.5 min, and then lowered to 50% B during the final 1 min. The column was equilibrated by pumping 50% B for 4 min. The flow rate was set 0.4 mL/min with an injection volume of 2 μL. LC-ESI-MS chromatogram was acquired in positive ion mode under following conditions: capillary voltage of 4500 V, dry temperature at 190°C, dry gas flow maintained at 8 L/min, nebulizer gas at 1.4 bar, and acquisition range of m/z 100–1000.
4
Cysteine Reaction Kinetics Analysis
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In 1 ml of solution containing 100 μM cysteine, 100 μM PVS, PVSN, or Bay 11-7082 in 20 mM NaHCO3, pH 8.4, reaction was held at 37°C for 60 min. The resulting reaction products were subjected directly to the LC–ESI–MS analyses. The LC–ESI–MS system consisted of an ultra-performance liquid chromatography system (Ultimate 3000 RSLC, Dionex) and an electrospray ionization (ESI) source of quadrupole time-of-flight (TOF) mass spectrometer (maXis HUR-QToF system, BrukerDaltonics). The samples were kept in an autosampler at 4°C. Separation was performed with reversed-phase liquid chromatography (RPLC) on a BEH C18 column (2.1 × 100 mm, Walters). The elution started from 99% mobile phase A (0.1% formic acid in ultrapure water) and 1% mobile phase B (0.1% formic acid in ACN), held at 1% B for 0.5 min, raised to 60% B in 6 min, further raised to 90% B in 0.5 min, held at 90% B for 1.5 min, and then lowered to 1% B in 0.5 min. The column was equilibrated by pumping 1% B for 4 min. The flow rate was set 0.4 ml/min with injection volume of 2 μl. LC–ESI–MS chromatogram were acquired under following conditions: capillary voltage of 4500 V in positive ion mode, dry temperature at 190°C dry gas flow maintained at 8 l/min, nebulizer gas at 1.4 bar, and acquisition range of m/z 100–1000.
5
Cucurbitacin Quantification via LC-ESI-MS
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In vitro materials were ground with liquid nitrogen and mixed with 1 mL of methanol. Supernatant was collected by centrifugation (12000 rpm, 1 min). The LC-ESI-MS system consisted of an ultra-performance liquid chromatography system (Ultimate 3000 RSLC, Dionex) and an electrospray ionization source of quadrupole time-of-flight mass spectrometer (maXis HUR-QToF system, Bruker Daltonics). The autosampler was set at 4 °C. Separation was performed with reversed-phase liquid chromatography on a BEH C8 column (2.1 × 100 mm, Walters). The elution started from 99 % mobile phase A (0.1 % formic acid in ultrapure water) and 1 % mobile phase B (0.1 % formic acid in ACN), held at 1 % B for 1.5 min, raised to 60 % B in 6 min, further raised to 90 % in 0.5 min, and then lowered to 1 % B in 0.5 min. The column was equilibrated by pumping 1 % B for 4 min. The flow rate was set to 0.4 mL/min with an injection volume of 5 μL. LC-ESI-MS chromatogram were acquired under the following conditions: capillary voltage of 4500 V in positive ion mode, dry temperature of 190 °C, dry gas flow maintained at 8 L/min, nebulizer gas at 1.4 bar, and acquisition range of m/z 100–1000. Five samples for each condition were independently measured for cucurbitacin content levels.
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