Model 6540

Manufactured by Agilent Technologies

Sourced in United States

The Agilent Model 6540 is an ultra-high-performance liquid chromatography quadrupole time-of-flight (UHPLC-QTOF) mass spectrometer. It is designed for accurate mass measurement and high-resolution accurate mass (HRAM) analysis of a wide range of compounds.

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Lab products found in correlation

Agilent 1260 lc system, by Agilent Technologies (2 mentions) Calibrant solution a, by Agilent Technologies (1 mentions) Zorbax sb c18 column, by Agilent Technologies (1 mentions) 1290 uplc system, by Agilent Technologies (1 mentions) Agilent c18 reverse phase column, by Agilent Technologies (1 mentions) G1312b binary pump, by Agilent Technologies (1 mentions) G1322a degasser, by Agilent Technologies (1 mentions)

3 protocols using model 6540

LC-QTOF Analysis of MsPDE Products

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The products of MsPDE were identified by LC/Q-TOF analysis, which was performed on Agilent 1260 LC system (Agilent Technologies, USA) coupled to an ultra high definition quadrupole time-of-flight mass spectrometer Model 6540 (Agilent Technologies, USA) equipped with a dual source electrospray ionization ion source.
The reaction products were separated on Agilent C18 reverse-phase column, with the binary mobile phase composed of 2% methanol and 98% water (containing 0.2% ammonium acetate and 0.1% acetic acid) being set at a constant flow rate of 300 μL/min and column temperature of 30 °C.
Q-TOF parameters were as follows: ionization mode, positive mode; capillary voltage: 4,000 V; drying gas: 9 L/min; nebulizer pressure: 40 psig; gas temperature: 350 °C; skimmer voltage: 65 V; octopole RF Peak voltage: 750 V, fragmentor voltage: 150 V. LC/MS accurate mass spectra were recorded across a range of 100-1000 m/z at the MS scan rate 1.5 spectra/s. Accurate mass measurements of each peak from the total ion chromatograms were obtained by an automated calibrant delivery system using a low flow of a calibrating solution (calibrant solution A, Agilent Technologies), which contains the internal reference masses purine (C₅H₄N₄) at m/z 121.0509 and HP-921 [hexakis-(1H, 1H, 3H-tetrafluoro-pentoxy) phosphazene] (C₁₈H₁₈O₆N₃P₃F₂₄) at m/z 922.0098.

Tang Q., Luo Y., Zheng C., Yin K., Ali M.K., Li X, & He J. (2015). Functional Analysis of a c-di-AMP-specific Phosphodiesterase MsPDE from Mycobacterium smegmatis. International Journal of Biological Sciences, 11(7), 813-824.

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UPLC-qTOF analysis of yeast extracts

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The yeast extracts were dissolved in methanol and the solution was filtered through 0.22 μm membrane prior to injection. Chromatography was performed on an Agilent 1290 UPLC system using a ZORBAX SB-C₁₈ column (1.8 μm, 2.1 × 150 mm, Agilent). The mobile phase consisted of 0.1% formic acid aqueous solution (v/v, solvent A) and acetonitrile/0.1% formic acid (v/v, solvent B). The flow was 0.3 mL/min, and the injection volume was 1 μL. A linear gradient with the following proportion of phase B (t_min, B%) was used: (0, 30), (17, 100), (22, 100). The UPLC was coupled with an electrospray ionization (ESI), a hybrid quadrupole time-of-flight (qTOF) mass spectrometer (model 6540, Agilent). The mass acquisition was performed on positive ionization and full scan (50–800 Da) modes. Spray parameters were as follows: gas temp. 350°C, gas flow 11 L/min, nebulizer 32 psi, sheath gas temp. 350°C, sheath gas flow 8.5 L/min, Vcap 4,000, nozzle voltage 500 V, fragment voltage 135 V.

Zhou Y., Ma Y., Zeng J., Duan L., Xue X., Wang H., Lin T., Liu Z., Zeng K., Zhong Y., Zhang S., Hu Q., Liu M., Zhang H., Reed J., Moses T., Liu X., Huang P., Qing Z., Liu X., Tu P., Kuang H., Zhang Z., Osbourn A., Ro D.K., Shang Y, & Huang S. (2016). Convergence and divergence of cucurbitacin biosynthesis and regulation in Cucurbitaceae. Nature plants, 2, 16183.

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Quantitative LC-MS/MS Analysis of Metabolites

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Liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (LC/Q-TOF) analysis was performed on an Agilent 1260 LC system [consisting of a G1322A degasser, a G1312B binary pump, a G1367E thermostated autosampler, G1316A thermostated column compartment, and a G4212B diode array detector (DAD)] (Agilent Technologies, Santa Clara, CA, USA) coupled to an ultra high definition quadrupole time-of-flight mass spectrometer Model 6540 (Agilent Technologies, Santa Clara, CA, USA) equipped with a dual source electrospray ionization ion source. The sample treatment, LC separation and Q-TOF parameters were conducted according to previously described methods (Tang et al., 2015 (link)).
The analytes were separated on an Agilent C18 reverse-phase column (100 × 1.8 mm, particle size of 3.5 μm). The binary mobile phase composed of 2% methanol and 98% water (containing 0.2% ammonium acetate and 0.1% acetic acid) was set at a constant flow rate of 300 μL/min and the column temperature was kept at 30°C, and the sample volume was 2 μL. Q-TOF parameters were as previously reported (Tang et al., 2015 (link)).

Zheng C., Ma Y., Wang X., Xie Y., Ali M.K, & He J. (2015). Functional analysis of the sporulation-specific diadenylate cyclase CdaS in Bacillus thuringiensis. Frontiers in Microbiology, 6, 908.

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