Multi-NMR spectra were recorded on either a Bruker 300 MHz, a Bruker 400 MHz, or a Varian Unity Inova 500 MHz spectrometer, and resonances are given in parts per million (p.p.m.) relative residual solvent. 1H NMR spectra were obtained as DMSO-d6 or CDCl3 solutions as indicated (reported in p.p.m.), using chloroform as the reference standard (7.25 p.p.m.) or DMSO-d6 (2.50 p.p.m.), unless otherwise stated. Peak multiplicities are reported using the following abbreviations: s=singlet, d=doublet, t=triplet, m=multiplet, br=broadened, dd=doublet of doublets, dt=doublet of triplets. J coupling constants, when given, are reported in hertz (Hz). HRMS spectra were recorded on an Agilent 6,220 ESI TOF mass spectrometer using flow injection analysis. Mass spectrum data were also obtained by liquid chromatography mass spectrometry (LCMS) on an Agilent instrument using atmospheric pressure chemical ionization (APCI) or electrospray ionization (ESI). Selected infrared spectra were recorded from neat compounds on a Bruker ALPHA FT-IR.
6220 esi tof mass spectrometer
Manufactured by Agilent Technologies
The 6220 ESI-TOF mass spectrometer is a laboratory instrument designed for high-resolution, accurate mass measurements. It utilizes electrospray ionization (ESI) and time-of-flight (TOF) mass analysis to provide precise molecular mass data for a wide range of analytes. This product is intended for use in analytical laboratories for applications such as small molecule analysis, proteomics, and metabolomics.
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Lab products found in correlation
1260 hplc, by Agilent Technologies (2 mentions)
Plrp s column, by Agilent Technologies (2 mentions)
Alpha ftir, by Bruker (1 mentions)
Unity inova 500 mhz spectrometer, by Agilent Technologies (1 mentions)
Nicolet is5 spectrometer, by Thermo Fisher Scientific (1 mentions)
Ultrashield plus 400 mhz spectrometer, by Bruker (1 mentions)
Agilent 1260 infinity hplc, by Agilent Technologies (1 mentions)
Eclipse xdb c8, by Agilent Technologies (1 mentions)
Masshunter acquisition software, by Agilent Technologies (1 mentions)
6 protocols using 6220 esi tof mass spectrometer
1
Nuclear Magnetic Resonance Spectroscopy Protocol
2
Characterization of Chemical Compounds
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Commercially available reagents and solvents (analytical grade) were used without further purification unless otherwise stated. The anhydrous solvents were obtained from an MBraun MB-SPS 800 Dry Solvent System. 1H, 13C, and two-dimensional NMR spectra were recorded on a Bruker Ultrashield Plus 400 MHz spectrometer at 25°C. High-resolution mass spectra were acquired on an Agilent 6220 ESI-TOF mass spectrometer. Optical rotation (OR) was performed with a Schmidt & Haensch UniPol L 1000 at 589 nm and a concentration (c) expressed in g/100 mL. Infrared (IR) spectra were acquired on Nicolet iS5 spectrometer (Thermo Fisher).
3
Protein Mass Spectrometry Analysis
4
HPLC Analysis of Polyphenolic Compounds
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An Agilent 1260 Infinity HPLC (Agilent Technologies, Santa Clara, CA, USA) was used for the purity check of the cyanidin glycosides, (−)epicatechin, catechin, and thiolysis products by an Eclipse XDB‐C8 (4.6 × 150 mm, 5 μm) column (Agilent Technologies, Santa Clara, CA, USA) according to the method of Bräunlich et al. (2013 (link)) with a diode array detector (DAD). Anthocyanin standard stock solutions were prepared in methanol containing 0.1% formic acid and were used to identify the compounds. Wavelengths of (−)epicatechin at 280 nm and anthocyanin glycosides at 520 nm were determined. The mass spectra of thiolytic products were acquired in positive mode using electrospray ionization on an Agilent 6220 ESI‐TOF mass spectrometer. The mDP was calculated by the following formula (Ci et al., 2018 (link)):
5
Protein Mass Spectrometry Analysis
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Purified protein samples were diluted to 10 μM in dialysis buffer lacking reducing agent or glycerol prior to analysis on an Agilent 6220 ESI-TOF mass spectrometer equipped with an Agilent 1260 HPLC. Separation and desalting was performed on an Agilent PLRP-S Column (1,000A, 4.6 x 50 mm, 5 μm). Mobile the phase A was 0.1% formic acid in water and mobile phase B was acetonitrile with 0.1% formic acid. A constant flow rate of 0.250 mL/min was used. Ten μL of the protein solution was injected and washed on the column for the first 3 min at 5% B, diverting non-retained materials to waste. The protein was then eluted using a linear gradient from 5% B to 100% B over 7 min. The mobile phase composition was maintained at 100% B for 5 min and then returned to 5% B over 1 min. The column was then re-equilibrated at 5%B for the next 4 min. Data was analyzed using Agilent MassHunter Qualitative Analysis software (B.06.00, Build 6.0.633.0 with Bioconfirm). The charge state distribution for the protein produced by electrospray ionization was deconvoluted to neutral charge state using Bioconfirm’s implementation of MaxEnt algorithm, giving a measurement of average molecular weight. The average molecular weight of the proteins were predicted using ExPASy Compute pI/Mw tool (http://web.expasy.org/compute_pi/ ).
6
HPLC-MS Analysis of GS-MQ Compound
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Synthetic GS-MQ or APR-246 treated de-proteinized JH-EsoAd1 cell lysates were resolved on an ODS-Hypersil C18 HPLC column (2.1 × 100 mm, 5 μm, Hewlett-Packard) in 10 mM NH4HCO3 pH 8, using a 0–80% acetonitrile gradient over 10 min. Flow rate was 0.1 ml min−1. Fractions of 25 μl were collected and acidified with 2% formic acid for mass spectrometric analysis. All analyses were done on an Agilent 6220 ESI-TOF mass spectrometer. Mass spectrometry data was acquired and reference mass corrected via a dual-spray electrospray ionization (ESI) source. Mass spectra were created by averaging the scans across each peak and background subtracted against the first 10 s of the total ion current. Acquisition was performed using the Agilent Mass Hunter Acquisition software version B.02.01 (B2116.30) and analysed using Mass Hunter version B.03.01. GS-MQ was detected at a 445.17 m/z peak.
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