Peptide samples were analyzed using an 1100 series HPLC system (Agilent, USA) coupled to an ESI-Q/TOF mass spectrometer (Agilent, USA). A Vydac C18 column (300 Å, 2.1 × 150 mm, Grace Vydac, USA) was used at a flow rate of 0.2 mL min−1. HPLC was performed with buffer A (0.1% FA in H2O) and buffer B (0.1% FA in acetonitrile) using the following gradient: 3% buffer B from 0–5 min; 3–50% buffer B from 5–75 min; 50–95% buffer B from 75–80 min; 95% buffer B from 80–85 min; and 95–3% buffer B from 85–90 min. The post time was 10 min. ESI-Q/TOF was performed under the following conditions: drying gas flow rate and temperature, 12 L min−1 and 300 °C, respectively; nebulizer pressure, 45 psi; capillary voltage, 3500 V; fragmentor, 175 V; collision energy slope and offset, 3.7 and 2.5 V, respectively; MS scan range and rate, 300–1500 and 3 Hz, respectively; MS/MS scan range and rate, 100–3000 and 3 Hz, respectively; and auto MS/MS, 5 precursors with active exclusion on and 2 repeat and release after 0.5 min. The collected data were used for the identification of peptide mappings.
Esi q tof mass spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The ESI-Q-TOF mass spectrometer is an analytical instrument used for the identification and characterization of chemical compounds. It combines electrospray ionization (ESI) with a quadrupole time-of-flight (Q-TOF) mass analyzer. The ESI-Q-TOF system is designed to provide high-resolution, accurate mass measurements of a wide range of molecules, including proteins, peptides, and small molecules.
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4 protocols using esi q tof mass spectrometer
1
Peptide Analysis by HPLC-ESI-Q/TOF
2
Phytochemical Profiling of S. campanulata
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The liquid chromatography–mass spectrometry (LC-ESI-QTOF-MS) analysis of the methanolic and aqueous extracts obtained from the leaves and stem bark of S. campanulata was carried out on an Agilent 1200 HPLC system (Agilent Technologies, Santa Clara, CA, USA) coupled to an ESI-Q-TOF mass spectrometer (Agilent Technologies). The analytical procedure was reported in our earlier paper [38 (link)]. Compounds were tentatively identified based on the obtained fragmentation patterns, which were compared with spectral information available in databases and the scientific literature. Additionally, the quantitative profile was evaluated based on UV spectra detected at 254 and 320 nm.
3
Artemisia Species Profiling by LC-HRMS/MS
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The liquid chromatography–tandem high-resolution mass spectrometry (LC-HRMS/MS) analysis of the methanol and chloroform extracts obtained from the roots and aerial parts of the five Artemisia species was carried out on an Agilent 1200 HPLC system (Agilent Technologies, Santa Clara, CA, USA) equipped with auto-sampler (G1329B), degasser (G1379B), binary pump (G1312C), thermostat (G1316A) and ESI-Q-TOF mass spectrometer (G6530B). The chromatographic separations were performed as follows: Phenomenex Gemini C18 column (2 mm × 100 mm, 3 μm); mobile phase 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B); gradient 5–60% B (0–45 min), 95% B (46–55 min); flow rate 0.2 mL/min; injection volume 10 μL. The following MS parameters were used: negative ionization mode; m/z range 100–1000; gas (N2) temperature 275 °C; N2 flow 10 L/min; nebulizer 35 psi; sheath gas temperature 325 °C; sheath gas flow rate 12 L/min; capillary voltage 4000 V; nozzle voltage 1000 V; skimmer 65 V; fragmentor 140 V; collision-induced dissociation energies 10 and 30 V.
4
Analytical Characterization of β-cyclocitral
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β-cyclocitral
was purchased from commercial sources (Sigma-Aldrich, Milan, Italy).
All other reagents and solvents were of the highest quality available
or were freshly distilled. Melting points (uncorrected) were measured
with a Buchi–Tottoli apparatus, and 1H, 13C, 31P, and bidimensional NMR spectra were recorded on
a Varian 400 MHz instrument unless otherwise noted. Chemical shifts
are given in parts per million (δ) relative to the solvent,
and coupling constants are in hertz. The abbreviations used are as
follows: s = singlet, d = doublet, t = triplet, q = quartet, dd =
double doublet, m = multiplet, td = triple doublet, dt = double triplet,
and ddd = double double doublet. Mass spectrometry (MS) analyses were
performed on ESI Micromass ZMD 2000. High-resolution MS (HRMS) spectra
were recorded using an ESI-Q-TOF mass spectrometer (Agilent Technologies).
Infrared spectra were recorded on a PerkinElmer FT-IR Spectrum 100
spectrometer. Flash chromatography was carried out on a silica gel
(Merck, 230–400 mesh). High-performance liquid chromatography
analysis was performed by Beckman system gold 168 using a C18 Jupiter
column (150 × 4.6 mm, 5 μm) and water/acetonitrile/TFA
eluent.
was purchased from commercial sources (Sigma-Aldrich, Milan, Italy).
All other reagents and solvents were of the highest quality available
or were freshly distilled. Melting points (uncorrected) were measured
with a Buchi–Tottoli apparatus, and 1H, 13C, 31P, and bidimensional NMR spectra were recorded on
a Varian 400 MHz instrument unless otherwise noted. Chemical shifts
are given in parts per million (δ) relative to the solvent,
and coupling constants are in hertz. The abbreviations used are as
follows: s = singlet, d = doublet, t = triplet, q = quartet, dd =
double doublet, m = multiplet, td = triple doublet, dt = double triplet,
and ddd = double double doublet. Mass spectrometry (MS) analyses were
performed on ESI Micromass ZMD 2000. High-resolution MS (HRMS) spectra
were recorded using an ESI-Q-TOF mass spectrometer (Agilent Technologies).
Infrared spectra were recorded on a PerkinElmer FT-IR Spectrum 100
spectrometer. Flash chromatography was carried out on a silica gel
(Merck, 230–400 mesh). High-performance liquid chromatography
analysis was performed by Beckman system gold 168 using a C18 Jupiter
column (150 × 4.6 mm, 5 μm) and water/acetonitrile/TFA
eluent.
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