All metabolites were analyzed by liquid chromatography–mass spectrometry (LC–MS; Agilent Technologies 1200 Series Rapid Resolution HPLC system and Agilent Technologies 6210 time-of-flight mass spectrometer (TOF–MS) (Agilent Technologies, Santa Clara, CA, USA) on a SeQuant® ZIC®-pHILIC column (150 mm length, 2.1-mm internal diameter, and 5-µm particle size). Targeted proteomics was performed using an Agilent 1290 liquid chromatography system coupled to an Agilent 6460 QQQ mass spectrometer (Agilent Technologies, Santa Clara, CA) via selected reaction monitoring (SRM) at the same points of the samples of metabolites. The sample preparation and detailed method of metabolite analysis and targeted proteomics are presented in Additional file 1 : Supplementary Methods.
6210 time of flight mass spectrometer tof ms
Manufactured by Agilent Technologies
Sourced in United States
The 6210 time-of-flight mass spectrometer (TOF–MS) is an analytical instrument designed to measure the mass-to-charge ratio of ions. It utilizes time-of-flight technology to separate and detect ions based on their flight time through a vacuum chamber. The 6210 TOF–MS provides accurate mass measurements and high-resolution mass spectra for the identification and quantification of a wide range of molecular compounds.
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2 protocols using 6210 time of flight mass spectrometer tof ms
1
Metabolite and Targeted Proteomics Analysis
2
Comprehensive Analytical Characterization of Chemical Compounds
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1H NMR spectra were recorded at 400 MHz (Bruker Avance III Nanobay) with CDCl3, [D6]DMSO, or [D4]methanol as solvent. 13C NMR spectra were recorded at 100.6 MHz (Bruker Avance III Nanobay) with CDCl3, [D6]DMSO, or [D4]methanol as solvent. Low‐resolution mass spectra were recorded by injection with an Agilent 1100 Series LC/MSD type SL mass spectrometer with electrospray ionization (ESI, 70 eV) and by using a mass‐selective detector (quadrupole). Upon analyzing crude reaction mixtures, the mass spectrometer was preceded by a HPLC reverse‐phase column with a diode array UV/Vis detector. High‐resolution mass spectra were obtained with an Agilent Technologies 6210 time‐of‐flight mass spectrometer (TOFMS) equipped with ESI/APCI‐multimode source. IR spectra were measured with a Fourier‐transform infrared spectrophotometer (The IRaffinity‐1S). Melting points of crystalline compounds were measured with a Kofler Bench, type WME Heizbank of Wagner & Munz. Microwave reactions were performed with a CEM Discover microwave at fixed temperature. The purity of all tested compounds was assessed by 1H NMR spectroscopy and/or HPLC analysis, confirming a purity of ≥95 %.
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1H NMR spectra were recorded at 400 MHz (Bruker Avance III Nanobay) with CDCl3, [D6]DMSO, or [D4]methanol as solvent. 13C NMR spectra were recorded at 100.6 MHz (Bruker Avance III Nanobay) with CDCl3, [D6]DMSO, or [D4]methanol as solvent. Low‐resolution mass spectra were recorded by injection with an Agilent 1100 Series LC/MSD type SL mass spectrometer with electrospray ionization (ESI, 70 eV) and by using a mass‐selective detector (quadrupole). Upon analyzing crude reaction mixtures, the mass spectrometer was preceded by a HPLC reverse‐phase column with a diode array UV/Vis detector. High‐resolution mass spectra were obtained with an Agilent Technologies 6210 time‐of‐flight mass spectrometer (TOFMS) equipped with ESI/APCI‐multimode source. IR spectra were measured with a Fourier‐transform infrared spectrophotometer (The IRaffinity‐1S). Melting points of crystalline compounds were measured with a Kofler Bench, type WME Heizbank of Wagner & Munz. Microwave reactions were performed with a CEM Discover microwave at fixed temperature. The purity of all tested compounds was assessed by 1H NMR spectroscopy and/or HPLC analysis, confirming a purity of ≥95 %.
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