All reagents and solvents were of reagent grade from commercial suppliers (Sigma Aldrich and etc.). 1H (400 MHz) NMR spectra were acquired on a Bruker-400 spectrometer. Fluorescence spectra were recorded on a Shimadzu RF-5301PC fluorometer. Absorption spectra were measured on Varian Cary 100 Bio UV-Visible spectrophotometer. A microplate reader (PerkinElmer Victor2, USA) was used for resazurin and CCK-8 assay. LC-MS data were collected on a Shimadzu LCMS-2010 system. GC-TCD studies were performed on an Agilent 7820A system. CO-RMs (CORM-2, CORM-3, CORM-A1 and CORM-401) and H2S probe (AzMC) were purchased from Sigma-Aldrich and were used without purification. Pure CO gas was purchased from Airgas company. iCORM-2 and iCORM-3 were prepared according to the literature procedures.12 (link), 54 (link) CO prodrugs (CO-103 and CO-111)27 , 55 (link) and H2S probe (DNS-Az)56 were synthesized according to the literature procedures.
2010 lcms system
Manufactured by Shimadzu
Sourced in Japan
The 2010 LCMS system is a liquid chromatography-mass spectrometry (LC-MS) instrument designed for analytical applications. It is capable of separating and detecting a wide range of compounds in complex samples. The system consists of a liquid chromatography unit and a mass spectrometer, providing both separation and high-sensitivity detection capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Luna c18 analytical column, by Phenomenex (3 mentions)
C18 guard column, by Phenomenex (3 mentions)
Lc 20ad binary solvent pumps, by Shimadzu (3 mentions)
Lcms solutions software version 3, by Shimadzu (3 mentions)
Sil 20a ht, by Shimadzu (2 mentions)
Spd m20a diode array detector, by Shimadzu (2 mentions)
Corm 2, by Merck Group (1 mentions)
Cary 100 bio uv visible spectrophotometer, by Agilent Technologies (1 mentions)
Corm a1, by Merck Group (1 mentions)
Rf 5301pc fluorometer, by Shimadzu (1 mentions)
Corm 3, by Merck Group (1 mentions)
7820a system, by Agilent Technologies (1 mentions)
Victor2, by PerkinElmer (1 mentions)
Drx 400 spectrometer, by Bruker (1 mentions)
Dionex ultimate 3000, by Thermo Fisher Scientific (1 mentions)
Hp 5890 gas chromatograph, by Hewlett-Packard (1 mentions)
Pda detector, by Shimadzu (1 mentions)
Lc ms grade acetonitrile, by Merck Group (1 mentions)
7 protocols using 2010 lcms system
1
Synthetic Protocols and Characterization
2
Comprehensive Analytical Characterization Protocol
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1H- and 13C-NMR (400 and 100 MHz, respectively) spectra were acquired on a Bruker DRX400 spectrometer (Bruker, Karlsruhe, Germany). Data processing was performed using MestReNova 6.2 (Mestrelab Research SL, San Diego, CA, USA). NMR spectra were recorded in DMSO-d6 and referenced to the residual non-deuterated solvent signal at δH 2.50 ppm. GC-FID and GC-O analyses were performed by using an HP 5890 gas chromatograph (Hewlett-Packard, San Diego, CA, USA). GC-MS (EIMS, 70 eV) analyses were carried out on a GC Agilent 7890B gas chromatograph equipped with a 5977A MSD mass selective detector (Agilent Technologies Inc., Wilmington, DE, USA). MS data were recorded between 40–400 u, and processed by Mass Hunter software. Wiley library 10ª edition with MS NIST 2011 (Ringoes, NJ, USA) was used to help in the compound identification. The analytical UHPLC-PDA-ELSD was performed on a Thermo Scientific Dionex UltiMate 3000 (Donierstr, Germany) system equipped with an autosampler, quaternary pump, PDA (Photodiode Array Detector) detector and ELSD detector (SEDEX, Alfortville Cedex, France). HPLC-ESI/MS was performed in a Shimadzu LCMS-2010 system (Shimadzu, Tokyo, Japan) equipped with a UV-VIS detector (SPD-10A) and two pumps (LC-10AD) coupled in-line with a MS-2010 mass spectrometer. The equipment also included an on-line DGU-14A degasser and a Rheodyne injection valve with a 5 μL loop.
3
LCMS Analysis of Alkaloid Purities
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The purities of dihydrosanguinarine
and sanguinarine were analyzed by an LCMS2010 system (Shimadzu, Japan)
using a TSK-gel ODS-80Tm 4.6 × 250 mm column. The samples were
analyzed under the following conditions: a column temperature of 40
°C; a flow rate of 0.5 mL/min; and a gradient schedule of 0–15
min of AcCN/H2O = 45:55, 18.5–24.5 min of AcCN/H2O = 80:20, and 28–33 min AcCN/H2O = 45:55
(containing 1% trifluoroacetic acid) in positive SIM-SCAN mode ranging
from m/z 100–500. The purities
were calculated based on % peak area for each sample.
and sanguinarine were analyzed by an LCMS2010 system (Shimadzu, Japan)
using a TSK-gel ODS-80Tm 4.6 × 250 mm column. The samples were
analyzed under the following conditions: a column temperature of 40
°C; a flow rate of 0.5 mL/min; and a gradient schedule of 0–15
min of AcCN/H2O = 45:55, 18.5–24.5 min of AcCN/H2O = 80:20, and 28–33 min AcCN/H2O = 45:55
(containing 1% trifluoroacetic acid) in positive SIM-SCAN mode ranging
from m/z 100–500. The purities
were calculated based on % peak area for each sample.
4
LCMS Compound Purity Verification
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The purity and identity
of all compounds
was verified using a Shimadzu 2010 LCMS system, consisting of a LC-20AD
binary solvent pumps, a DGU-20A degasser unit, a CTO-20A column oven,
and a SIL-20A HT auto sampler. A Shimadzu SPD-M20A diode array detector
was used for detections. A full spectra range of 190–600 nm
was obtained during analysis. Chromatographic separations were obtained
using a Phenomenex Luna C18 analytical column (5 μm, 50 ×
4.6 mm i.d.) The column was protected by a Phenomenex C18 column guard
(5 μm, 4 × 3.0 mm i.d.). All equipment was controlled and
integrated by Shimadzu LCMS solutions software version 3. Mobile phases
for LCMS analysis were HPLC grade or LCMS grade obtained from Sigma-Aldrich
and Fisher Scientific. The mobile phases consisted of a mixture LCMS
grade Acetonitrile/water (both with 0.1% formic acid for a pH of 2.7).
The initial setting for analysis was set at 5% acetonitrile (v/v),
then was linearly increased to 95% acetonitrile over 6 min. The gradient
was then held at 95% acetonitrile for 2 min and then linearly decreased
to 5% over 0.10 min and held until stop for an additional 1.90 min.
The total run time was equal to 12 min. The total flow rate was set
to 0.5 mL/min. The column oven and flow cell temperature for the diode
array detector was set at 30 °C. The auto sampler temperature
was held at 15 °C. 5uL was injected for analysis.
of all compounds
was verified using a Shimadzu 2010 LCMS system, consisting of a LC-20AD
binary solvent pumps, a DGU-20A degasser unit, a CTO-20A column oven,
and a SIL-20A HT auto sampler. A Shimadzu SPD-M20A diode array detector
was used for detections. A full spectra range of 190–600 nm
was obtained during analysis. Chromatographic separations were obtained
using a Phenomenex Luna C18 analytical column (5 μm, 50 ×
4.6 mm i.d.) The column was protected by a Phenomenex C18 column guard
(5 μm, 4 × 3.0 mm i.d.). All equipment was controlled and
integrated by Shimadzu LCMS solutions software version 3. Mobile phases
for LCMS analysis were HPLC grade or LCMS grade obtained from Sigma-Aldrich
and Fisher Scientific. The mobile phases consisted of a mixture LCMS
grade Acetonitrile/water (both with 0.1% formic acid for a pH of 2.7).
The initial setting for analysis was set at 5% acetonitrile (v/v),
then was linearly increased to 95% acetonitrile over 6 min. The gradient
was then held at 95% acetonitrile for 2 min and then linearly decreased
to 5% over 0.10 min and held until stop for an additional 1.90 min.
The total run time was equal to 12 min. The total flow rate was set
to 0.5 mL/min. The column oven and flow cell temperature for the diode
array detector was set at 30 °C. The auto sampler temperature
was held at 15 °C. 5uL was injected for analysis.
5
LC-MS Analysis of Metabolite Profiles
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LC-electrospray ionization/MS analysis was carried out using a Shimadzu 2010 LCMS system with Shimadzu autosampler, Shimadzu binary LC pumps, and Shimadzu PDA detector. The same HPLC column was used for the LC/MS analysis as for the HPLC method. The LC elution conditions were modified for the analysis to the following: Pump A: 0.1% formic acid in water; Pump B: 0.1% formic acid in acetonitrile; time 0 min (10% B), 38 min (30% B), and 60 min (30% B) at a flow rate of 0.3 mL/min. The analysis was done in negative ion mode, with nebulizing gas (nitrogen) flow at 1.5 L/min, collision energy 1.4 kV, mass range 150–600 amu, heat block temperature 200°C, and curved desolvation line heater temperature 250°C.
6
Identification and Purity of Compounds via LCMS
7
LCMS Characterization of Crude Compounds
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