LC-MS analysis was performed using a Agilent Q-TOF mass spectrometer in line with a Agilent 1290 HPLC system. The 5 μl purified TMSiPhe-incorporated β-arrestin1 protein was loaded onto a reverse phase column (30 0SB-C8, 2.1 × 50 mm, 3.5 μm particle) (Agilent Technologies, SantaClara, CA). The proteins were then eluted over a gradient: 2% B for 2 min to waste, then turned LC to MS, 2–50% B in 6 min, 50–90% B in 4 min, 90% B sustained for 4 min, then decreased to 2% in 1.1 min, (where B is 100% Acetonitrile, 0.1% formic acid, A is water with 0.1% formic acid) at a flow rate of 0.2 mL/min.and the elution was introduced online into the Q-TOF mass spectrometer (Agilent Technologies, SantaClara, CA) using electrospray ionization. MS data were analyzed by MassHunter biocomfirm software.
Q tof mass spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Q-TOF mass spectrometer is an analytical instrument used for the identification and quantification of chemical compounds. It combines a quadrupole (Q) mass analyzer with a time-of-flight (TOF) mass analyzer to provide high mass accuracy and resolution. The Q-TOF mass spectrometer is capable of performing accurate mass measurements, allowing for the determination of molecular formulas and the identification of unknown compounds.
Automatically generated - may contain errors
Lab products found in correlation
Silica gel, by Qingdao Haiyang Chemical (3 mentions)
Masshunter software, by Agilent Technologies (2 mentions)
Agilent 1260 infinity capillary pump, by Agilent Technologies (2 mentions)
1290 hplc system, by Agilent Technologies (2 mentions)
Ec 400, by JEOL (2 mentions)
Ec 500, by JEOL (2 mentions)
Ec 600, by JEOL (2 mentions)
Kinetex 2.6μ xb c18 100a, by Phenomenex (1 mentions)
Agilent 6540 q tof mass spectrometer, by Agilent Technologies (1 mentions)
Masshunter qualitative analysis b 06, by Agilent Technologies (1 mentions)
Agilent 1290 infinity uplc system, by Agilent Technologies (1 mentions)
Masshunter qualitative analysis software, by Agilent Technologies (1 mentions)
As400, by Agilent Technologies (1 mentions)
Milli q system, by Merck Group (1 mentions)
Lc ms ms system, by Agilent Technologies (1 mentions)
Photodiode array detector, by Waters Corporation (1 mentions)
Av 500 spectrometer, by Bruker (1 mentions)
Lc 6ad, by Shimadzu (1 mentions)
Pack ods a column, by YMC (1 mentions)
Lc 1290, by Agilent Technologies (1 mentions)
33 protocols using q tof mass spectrometer
1
LC-MS Analysis of TMSiPhe-Incorporated β-Arrestin1
2
Comprehensive Chemical Analysis of SAHA Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine that the isolated compounds were generated by SAHA, the EtOAc extracts were accurately analyzed by ultra-high-performance liquid chromatography coupled with Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, United States). Chromatographic analysis was performed by using a C18 reverse-phase analytical column: Phenomenex Kinetex 2.6 μ XB-C18 100A (Torrance, CA, United States). UHPLC-Q-TOF parameters were as follows: the column oven temperature was set at 40°C and the injection volume was 5 μL, with a flow rate of 0.4 mL/min. The mobile phase consisted of 0.4% acetic acid in water and 100% acetonitrile; the gradient of the latter increased linearly from 10 to 60% (v/v) at 10 min, to 100% at 15 min, to 60% at 18 min, and to 10% at 20 min. Samples were analyzed in the fast polarity switching mode at a fragmentation voltage of 175 V, over the range of m/z 100–1700. A drying gas flow rate of 11 L/min under a temperature of 350°C and a capillary voltage of 3.5 kV were used.
3
Optimized QTOF Mass Spectrometry Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MS data were recorded using an Agilent 6540 QTOF mass spectrometer (Agilent Technologies) equipped with a QTOF mass spectrometer and a jet stream electrospray ion source. Data acquisition were controlled by Mass Hunter Qualitative Analysis B.06 and Quantitative Analysis B.04 (Agilent Technologies). The optimized operating parameters in the negative ion modes were as follows: nebulizing gas (N2) flow rate, 8.0 L/min; nebulizing gas temperature, 300°C; jet stream gas flow, 8 L/min; sheath gas temperature, 350°C; nebulizer pressure, 45 psi; capillary voltage, 3000 V; skimmer, 65 V; Octopole RFV, 600 V; and collision energy, 15 eV. Mass spectra were recorded across the range m/z 100–1300 with accurate mass measurement of all mass peaks. Deprotonated molecular ions of the 10 analytes were selected as their respective extraction ions in quantitative analysis using the extracting ion mode.
4
Metabolite Identification Using LC-HRMS-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The chemical structures of key metabolites were further confirmed using tandem mass spectrometry (LC-HRMS-MS) methods with chromatographic conditions identical to those used for their discovery. LC-HRMS-MS analyses were performed on an Agilent QTOF mass spectrometer for patient samples and/or, reference blood samples with collision energy conditions optimized to obtain the highest quality product ion spectra. The resulting product ion spectra were then compared to MS-MS spectra available in public spectral databases such as METLIN [26] (link), MassBank [27] (link) and Stemina's own SteminaMetDB database.
5
UPLC-QTOF Analysis of RSV Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A previously validated method
(linearity, precision, accuracy, limits of detection, and quantification)
was used to analyze RSV and derived metabolites.30 (link) Briefly, the analyses were performed on an Agilent 1290
Infinity UPLC system coupled to a 6550 accurate-mass quadrupole-time-of-flight
(QTOF) mass spectrometer (Agilent Technologies, Waldbronn, Germany)
using an electrospray interface (Jet Stream Technology), using chrysin
as an internal control of the ionization signal. Spectra were acquired
in the m/z range of 100 to 1100
in a negative polarity mode and at an acquisition rate of 1.5 spectra/s.
Data were processed using Mass Hunter Qualitative Analysis software
(version B.06.00, Agilent), which lists and rates possible molecular
formulas consistent with the accurate mass measurement and the actual
isotopic pattern. A target screening strategy was applied to qualitatively
analyze possible metabolites that could be present after RSV consumption.
In addition, targeted MS/MS analysis provided additional information
to achieve a reliable compound identification. MS/MS product ion spectra
were collected at m/z 50–800
range using a retention time window of 1 min, collision energy of
20 V, and an acquisition rate of 4 spectra/s.
(linearity, precision, accuracy, limits of detection, and quantification)
was used to analyze RSV and derived metabolites.30 (link) Briefly, the analyses were performed on an Agilent 1290
Infinity UPLC system coupled to a 6550 accurate-mass quadrupole-time-of-flight
(QTOF) mass spectrometer (Agilent Technologies, Waldbronn, Germany)
using an electrospray interface (Jet Stream Technology), using chrysin
as an internal control of the ionization signal. Spectra were acquired
in the m/z range of 100 to 1100
in a negative polarity mode and at an acquisition rate of 1.5 spectra/s.
Data were processed using Mass Hunter Qualitative Analysis software
(version B.06.00, Agilent), which lists and rates possible molecular
formulas consistent with the accurate mass measurement and the actual
isotopic pattern. A target screening strategy was applied to qualitatively
analyze possible metabolites that could be present after RSV consumption.
In addition, targeted MS/MS analysis provided additional information
to achieve a reliable compound identification. MS/MS product ion spectra
were collected at m/z 50–800
range using a retention time window of 1 min, collision energy of
20 V, and an acquisition rate of 4 spectra/s.
6
NMR Spectroscopy and Mass Spectrometry Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All reagents were purchased from
Sigma-Aldrich, Merck, Himedia, and used directly without further purification.
Column chromatography was performed using 60–120 mesh silica
gels. Reactions were monitored by thin-layer chromatography (TLC)
on silica gel 60 F254 (0.25 mm). 1H NMR and 13C NMR spectra were recorded at 400 and 100 MHz, respectively,
with a Varian AS400 spectrometer and 600 and 151 MHz with Brucker
spectrometers, using tetramethylsilane (TMS) as an internal standard
with CDCl3, dimethyl sulfoxide (DMSO)-d6. The coupling constants (J values)
and chemical shifts (δppm) were reported in hertz
(Hz) and parts per million (ppm), respectively, downfield from tetramethylsilane
using residual chloroform (d = 7.24 for 1H NMR, d = 77.23 for 13C NMR) as an internal
standard. Multiplicities are reported as follows: s (singlet), d (doublet),
t (triplet), m (multiplet), and br (broadened). High-resolution mass
spectroscopy (HRMS) images were recorded on an Agilent Q-TOF mass
spectrometer with Z-spray source using built-in software
for analysis of the recorded data. 1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine (PE) was purchased from Avanti Polar
Lipids (Alabaster, AL). Ultrapure water (Milli-Q system, Millipore,
Billerica, MA) was used for the preparation of buffers.
Sigma-Aldrich, Merck, Himedia, and used directly without further purification.
Column chromatography was performed using 60–120 mesh silica
gels. Reactions were monitored by thin-layer chromatography (TLC)
on silica gel 60 F254 (0.25 mm). 1H NMR and 13C NMR spectra were recorded at 400 and 100 MHz, respectively,
with a Varian AS400 spectrometer and 600 and 151 MHz with Brucker
spectrometers, using tetramethylsilane (TMS) as an internal standard
with CDCl3, dimethyl sulfoxide (DMSO)-d6. The coupling constants (J values)
and chemical shifts (δppm) were reported in hertz
(Hz) and parts per million (ppm), respectively, downfield from tetramethylsilane
using residual chloroform (d = 7.24 for 1H NMR, d = 77.23 for 13C NMR) as an internal
standard. Multiplicities are reported as follows: s (singlet), d (doublet),
t (triplet), m (multiplet), and br (broadened). High-resolution mass
spectroscopy (HRMS) images were recorded on an Agilent Q-TOF mass
spectrometer with Z-spray source using built-in software
for analysis of the recorded data. 1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine (PE) was purchased from Avanti Polar
Lipids (Alabaster, AL). Ultrapure water (Milli-Q system, Millipore,
Billerica, MA) was used for the preparation of buffers.
7
LC-MS/MS Analysis of Peptide Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The peptide sample was analyzed with an LC-MS/MS system (Agilent Technologies, Santa Clara, CA, USA) consisting of a nanopump (G2226A, Agilent) with a four-channel micro vacuum degasser (G1379B, Agilent), a microfluidic chip cube (G4240-64000, Agilent) interfaced to a Q-TOF mass spectrometer (6530, Agilent), a capillary pump (G1376A, Agilent) with degasser (G1379B, Agilent), and an auto-sampler with thermostat (G1377A, Agilent). All modules were controlled by Mass Hunter software (version B.06.00, Agilent). A microfluidic reversed-phase HPLC chip (Zorbax 300SB-C18, 5-μm particle size, 0.75 × 150 mm) was used for peptide separation. A mixture of 96.9% water, 3% acetonitrile, and 0.1% formic acid (v/v) was used as the sample loading solution and solvent. Buffer B was 99.9% ACN, 0.1% formic acid (v/v). Samples were loaded on a trap-column at a flow rate of 3 µL/min for 5 min and eluted through a separation column at a flow rate of 300 nL/min. The gradient was from 15 to 85% of buffer B within 30 min.
8
Chickpea Peptide Fractionation and Identification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chickpea peptide was fractionated by Agilent 1260 Infinity Capillary Pump and coupled to Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). Theoretical pI and molecular weight of the identified proteins was estimated using ExPASY pI/Mw tool. The obtained raw data was cross-referenced to the plant species sub-directory within the Swiss-Prot database (UniProt, EBI, Cambridgeshire, UK) and analyzed in Mascot search against NCBI database.
9
Analytical Characterization of Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ultraviolet spectral were obtained on a Waters 2695 high performance liquid chromatography (HPLC) instrument with a photo diode array detector (Waters, Massachusetts, USA). Preparative HPLC was carried out on a Shimadzu LC-6AD instrument (Shimadzu, Kyoto, Japan) with a UV–Vis detector (SPD-20A), using a YMC-Pack-ODS-A column (250 × 20 mm, 5 μm) (YMC, Kyoto, Japan). The carbohydrates were measured by ion chromatography using an amperometric detector (Metrohm, Herisau, Switzerland) with a Hamilton RCX-30 column (250 × 4.6 mm, 7 μm) (Hamilton, Nevada, USA). Nuclear magnetic resonance (NMR) spectra were obtained using Bruker AV-500 spectrometers (500 MHz for 1H NMR and 125 MHz for 13C NMR) (Bruker, Zurich, Switzerland). Chemical shifts (δ) are given in ppm, with TMS as an internal standard, and coupling constants (J) are in Hz. High-resolution electrospray ionisation-mass spectrometry (HRESIMS) spectra were obtained using an Agilent 6540 ultra-performance liquid chromatography (UPLC) with high-resolution quadrupole time-of-flight (Q-TOF) mass spectrometer (Agilent, California, USA), using an RRHD Eclipse Plus C18 column (150 × 2.1 mm, 1.8 μm) (YMC, Kyoto, Japan)36 (link).
10
Molecular Networking for Untargeted Metabolomics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An LC-MS/MS data file (.d) created from the Agilent QTOF mass spectrometer were converted to .mgf file formats with an Agilent Qualitative 10.0 and uploaded to the GNPS Web platform (http://gnps.ucsd.edu ., accessed on 20 December 2021) for the classical molecular networking generation. MS-Cluster (0.1 Da tolerance) and a 0.02 Da tolerance for fragment ions were applied to create the consensus parent mass spectra. A network was generated where there were more than six matched fragment ions and the edges were filtered to have a minimal cosine score of 0.7. A maximum size of a molecular family was also set to 100. The output molecular networking was visualized and analyzed using Cytoscape 3.9.0. The GNPS data can be found at https://gnps.ucsd.edu/ProteoSAFe/status.jsp?task=e9f7b2ac30bd4ddabcc847f44b089a4b , accessed on 20 December 2021.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!