1 μM wild-type or C668S mutant dehydrogenase domain (untreated or after incubation with VAS2870 or VAS3947) dissolved in a mixture of acetonitrile/water/formic acid 0.1% was analysed by ESI-MS using a LCQFleet Thermo Scientific Ion Trap.
Ion trap
Manufactured by Thermo Fisher Scientific
The Ion Trap is a type of mass spectrometer used for the analysis and identification of chemical compounds. It operates by trapping and analyzing ions within an electric field, allowing for the precise measurement of their mass-to-charge ratios. This information can then be used to determine the molecular structure and composition of the analyzed sample.
Automatically generated - may contain errors
3 protocols using ion trap
1
MS Analysis of Dehydrogenase Domain
2
General Synthetic Methodology for Organic Compounds
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General: All solvents and reagents were obtained from commercial suppliers and were used directly without further purification. The reaction progress was monitored on a TLC plate (Merck, silica gel 60 F254, Darmstadt, Germany). 1H, 13C, and 19F NMR spectra were recorded on a Bruker Advance 400 MHz spectrometer using deuterated chloroform or dimethyl sulfoxide (DMSO). Chemical shifts (δ) are reported in parts per million (ppm) up-field from tetramethylsilane (TMS) as an internal standard, and s, d, t, and m are presented as singlet, doublet, triplet, and multiplet, respectively. Coupling constants (J) are reported in hertz (Hz). Liquid chromatography-mass spectra (LC-MS) were recorded on an Agilent (single quad) or Thermo Scientific ion trap. Abbreviations: AA: ascorbic acid; ACN: acetonitrile; DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene; DCM: dichloromethane; DMAP: dimethylamino pyridine; DMF: dimethylformamide; EtOAc: ethyl acetate; MeOH: methanol; PE: petroleum ether; TCEP: tris[2-carboxyethyl]phosphine hydrochloride; TEA: triethyl amine; THF: tetrahydrofuran; TLC: thin-layer chromatography.
3
NMR and LC/MS Compound Characterization
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NMR measurements
for characterization of the compounds were performed
using either a Bruker UltraShield 400 MHz or a Varian Mercury 400
MHz spectrometer using 400 MHz for 1H and 100 MHz for 13C measurements. Proton and carbon chemical shifts were documented
in parts per million (ppm) downfield from trimethylsilane (TMS), using
the resonance frequency of the deuterated solvent as the internal
standard. Abbreviations reported for the multiplets are s: singlet;
t: triplet; and m: multiplet. Liquid chromatography/mass spectrometry
(LC/MS) measurements were performed on a system using Shimadzu LC-10AD
VP liquid chromatography pumps equipped with an Alltima C18 3 μm
(50 mm × 2.1 mm) reversed-phase column, a diode array detector
(Thermo Finnigan Surveyor PDA Plus detector), and an Ion-Trap (Thermo
Scientific LCQ Fleet). This system used a water–acetonitrile
mobile phase enriched with 0.1% v/v formic acid.
for characterization of the compounds were performed
using either a Bruker UltraShield 400 MHz or a Varian Mercury 400
MHz spectrometer using 400 MHz for 1H and 100 MHz for 13C measurements. Proton and carbon chemical shifts were documented
in parts per million (ppm) downfield from trimethylsilane (TMS), using
the resonance frequency of the deuterated solvent as the internal
standard. Abbreviations reported for the multiplets are s: singlet;
t: triplet; and m: multiplet. Liquid chromatography/mass spectrometry
(LC/MS) measurements were performed on a system using Shimadzu LC-10AD
VP liquid chromatography pumps equipped with an Alltima C18 3 μm
(50 mm × 2.1 mm) reversed-phase column, a diode array detector
(Thermo Finnigan Surveyor PDA Plus detector), and an Ion-Trap (Thermo
Scientific LCQ Fleet). This system used a water–acetonitrile
mobile phase enriched with 0.1% v/v formic acid.
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