LC-MS data were acquired using a Bruker maXis 4G ESI-Q-TOF mass spectrometer coupled with a Waters Acquity UPLC system. A gradient of MeOH and H2O (containing 0.1% formic acid) was employed with a flow rate of 0.3 mL/min on an RP C-18 column (Phenomenex Kinetex 2.6 μm, 2.1 mm × 100 mm). The gradient started from MeOH/H2O (10%/90%), followed by a linear gradient to reach MeOH/ H2O (97%/3%) in 12 min, and was held for 2 min at MeOH/H2O (97%/3%). Full scan mass spectra (m/z 150–1550) were measured in positive ESI mode [38 (link)]. The mass spectrometer was operated using the following parameters: capillary, 4.5 kV; nebulizer pressure, 4.0 bar; dry gas flow, 6.0 L/min; dry gas temperature, 200 °C; scan rate, 2 Hz. Tune mix (Agilent, Santa Clara, CA, USA, ESI-L low concentration) was introduced through a divert valve at the end of each chromatographic run. The spectra acquired on Tune mix were used for mass calibration of the data prior to PCA.
Kinetex 2.6 μm
Manufactured by Phenomenex
The Kinetex 2.6 μm is a high-performance liquid chromatography (HPLC) column manufactured by Phenomenex. It features a core-shell particle design with a 2.6 μm particle size, providing efficient separation and analysis of a variety of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Maxis 4g esi q tof mass spectrometer, by Waters Corporation (2 mentions)
Tune mix, by Agilent Technologies (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Tripletof 5600, by AB Sciex (1 mentions)
1290 uhplc system, by Agilent Technologies (1 mentions)
6530 esi qtof mass spectrometer, by Agilent Technologies (1 mentions)
Maxis 4g esi qtof mass spectrometer, by Bruker (1 mentions)
Acquity uplc system, by Bruker (1 mentions)
Prominence hplc system, by Phenomenex (1 mentions)
Gemini c18 column, by Phenomenex (1 mentions)
Luna c18 column, by Phenomenex (1 mentions)
5 protocols using kinetex 2.6 μm
1
LC-MS Metabolomics Profiling Protocol
2
Quantifying Hydroxy-plastochromanol-8 in Leaves
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Leaves were injured with tweezers and frozen in liquid nitrogen. The samples (~50 mg) were ground in ethyl acetate. After centrifugation, the supernatant was filtered and evaporated on ice under a stream of N2. The residue was recovered in methanol/hexane (17/1) and analysed by HPLC, as described elsewhere [10 (link)]. The column was a Phenomenex Kinetex 2.6 μm, 100 × 4.6 mm, 100 Å. The HPLC analysis was performed in the isocratic mode with methanol/hexane (17/1) as a solvent system and with a flow rate of 0.8 mL min−1. Hydroxy-plastochromanol-8 was detected by its fluorescence at 330 nm with an excitation at 290 nm. The standard was a kind gift from Dr. J. Kruk (Krakow, Poland).
3
Optimized LC-MS Protocol for Compound Identification
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Agilent liquid chromatography and mass spectroscopy (LC/MS) system (ABSCIEX Triple TOF, 5600) with the analyzer and electron spray ionization source was used for the LC/MS analysis. The source parameters that were optimized to provide high sensitivity are negative mode, gas temperature 600°C, drying gas flow rate 0.5 mi/min, nebulizer pressure 50 psi, capillary voltage 5500 V; separation was carried out by ultra-high-pressure LC using Shimadzu UFLCXR with a phenomenex Kinetex 2.6 μm (100 × 2.1), column mobile phase used was A (water; 5 Mm ammonium formate +0.5% formic acid) and B (acetonitrile). The gradient program was as follows: 5% B for starting condition was increased up to 95% B in 20 min and decrease 5% B in 21–30 min at the final step. Injection volume was 50 μl. The standardized collision energy was 35V with collision energy of ±15V. The compounds were identified with computer-assisted evaluation of the resulting data by searching against the spectral library.
4
Metabolomics Analysis by LC-MS
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LC-MS data were acquired using an Agilent 6530 ESI-QTOF mass spectrometer with an Agilent 1290 UHPLC system. Using a flow rate of 0.3 ml/min on an RP C18 column (Phenomenex Kinetex 2.6 μm, 2.1 mm × 100 mm), acetonitrile/H2O containing 0.1% formic acid (10%/90%) was held for 1 min, followed by a linear gradient of acetonitrile and H2O (containing 0.1% formic acid) from (10%/90%) to (100%/0%) in 11 min, and finally, held for 2 min at acetonitrile/H2O (100%/0%). Full scan mass spectra (m/z 100–1700) were measured in positive ESI mode. The mass spectrometer was operated with the following parameters: nebulizer pressure, 2.76 bar; dry gas flow, 10.0 L/min; dry gas temperature, 325°C; capillary, 3.5 kV; scan rate, 1 Hz. Two internal calibration compounds, purine and hexakis (1 hr, 1 hr, 3H-tetrafluoropropoxy) phosphazine, were used at low concentration throughout the acquisition. Targeted MS/MS data were acquired using two collision energies: 10 and 20 eV. IAA standard was purchased from Research Products International Corp (IL, USA).
5
Structural Elucidation of Compounds
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