Identification of the protein in the gel spots was performed using an LCMS-IT-TOF instrument (Shimadzu, Kyoto, Japan). The pieces of interest were excised from the gel, and in-gel digestion of proteins was performed. The gel pieces were washed with 30% acetonitrile in 50 mM NH4HCO3 followed by 60% acetonitrile in 50 mM NH4HCO3, and the samples were dried using a SpeedVac concentrator (Thermo Fisher Scientific, Waltham, MA, USA). For in-gel digestion of proteins, the samples were incubated with a trypsin solution at 37 °C overnight, and the extracted trypsinized peptides were eluted stepwise with 30%, 100%, and 0% of acetonitrile in 50 mM NH4HCO3. The samples were dried and then dissolved in 50 mM NH4HCO3. Tryptic peptides were separated by reversed-phase liquid chromatography/MS using a Prominence nanoLC instrument (Shimadzu, Kyoto, Japan) for analytical separation on a PicoFritTMBetaBasic C18 column (100 mm × 75 μm; New Objective, Woburn, MA, USA). MS was performed on an LCMS-IT-TOF instrument with argon gas for ion cooling and collision-induced dissociation experiments. MS/MS data were obtained in a data-dependent manner. The Mascot search engine (Matrix Science, Boston, MA, USA) was used for protein database searching. Proteins with a statistically significant MASCOT/Mowse score (>28), indicating identity or extensive homology (P < 0.05), were considered for identification.
Lcms it tof instrument
Manufactured by Shimadzu
Sourced in Japan
The LCMS-IT-TOF is a liquid chromatography-mass spectrometry instrument that combines ion trap and time-of-flight technologies. It provides accurate mass measurement and high-resolution separation capabilities for compound identification and quantification.
Automatically generated - may contain errors
Lab products found in correlation
Drx advance 600 spectrometer, by Bruker (3 mentions)
Speedvac concentrator, by Thermo Fisher Scientific (1 mentions)
Mascot search engine, by Matrix Science (1 mentions)
Nicolet is5 ftir spectrometer, by Thermo Fisher Scientific (1 mentions)
Agilent 400 mr spectrometer, by Agilent Technologies (1 mentions)
Kieselgel 60 f254, by Merck Group (1 mentions)
Inova, by Agilent Technologies (1 mentions)
Gemini 2000, by Agilent Technologies (1 mentions)
Silica gel 60, by Kanto Chemical (1 mentions)
Ecs 400, by JEOL (1 mentions)
J 1500 spectrometer, by Jasco (1 mentions)
P 2200 digital polarimeter, by Jasco (1 mentions)
Ft ir 4600 typea spectrometer, by Jasco (1 mentions)
Jnm eca600, by JEOL (1 mentions)
Cto 20a column, by Shimadzu (1 mentions)
Hybridit tof mass spectrometer, by Shimadzu (1 mentions)
Dgu 20a3, by Shimadzu (1 mentions)
Cbm 20a, by Shimadzu (1 mentions)
Lc 20ad pump, by Shimadzu (1 mentions)
Avance nmr spectrometer, by Bruker (1 mentions)
16 protocols using lcms it tof instrument
1
Protein Identification via LC-MS/MS
2
Spectroscopic Characterization of Compounds
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IR spectra in the solid state were recorded on a Nicolet iS5 FTIR spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) using an internal reflectance attachment with diamond optical element−attenuated total reflection (ATR) with a 45° angle of incidence. The resolution was 4 cm−1; the number of scans was 32. The NMR spectra were recorded in the standard 5 mm sample tubes using an Agilent 400-MR spectrometer (Agilent Technologies, Santa Clara, CA, USA) with operating frequencies of 400.1 MHz (1H), and 100.6 MHz (13C). Registration of HRMS ESI mass spectra was carried out using an LCMS-IT-TOF instrument (Shimadzu, Japan).
3
Spectroscopic Characterization of Organic Compounds
4
LC-MS Analysis of Compound Derivatives
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LC-MS data of JC005, JC007, JC010, JC011, JC017 and JC040 were collected on a Shimadzu LCMS-IT-TOF instrument equipped with SPD-M20A PDA detector, LCMS-IT-TOF MS detector and LC-20AD binary gradient pump using a Shimpack VP-ODS (2.0 mm I.D.×150 mm) column (File S1 ). An isocratic elution with 20% H2O and 80% CH3CN at a flow rate of 0.2 mL/min over 3 min was used.
5
Comprehensive Analytical Characterization of Compounds
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Specific rotations were measured using a P-2200 digital polarimeter (l = 5 cm; JASCO, Tokyo, Japan). IR spectra were measured using a JASCO FT/IR-4600 typeA spectrometer; ECD spectroscopy was performed using a JASCO J-1500 spectrometer. ESI and high-resolution ESI mass spectra were recorded on a Shimadzu LCMS-IT-TOF instrument. FAB and HRFABMS data were recorded using a JASCO SX-102A mass spectrometer. 1H NMR spectra were recorded on JEOL ECS400 (400 MHz) and JNM-ECA 600 (600 MHz) spectrometers (JEOL). 13C NMR spectroscopy was performed on a JEOL JNM-ECA 600 (150 MHz) spectrometer. 2D-NMR experiments were carried out on a JEOL JNM-ECA 600 (600 MHz) spectrometer (JEOL).
Normal-phase silica-gel column chromatography was performed using silica gel 60 (63–210 μm; Kanto Chemical Co., Tokyo, Japan). Reverse-phase silica-gel column chromatography was performed using C18-OPN gel (140 μm; Nacalai Tesque, Kyoto, Japan). HPLC was performed using an SPD-M10Avp UV-vis detector (Shimadzu, Kyoto, Japan).
Normal-phase silica-gel column chromatography was performed using silica gel 60 (63–210 μm; Kanto Chemical Co., Tokyo, Japan). Reverse-phase silica-gel column chromatography was performed using C18-OPN gel (140 μm; Nacalai Tesque, Kyoto, Japan). HPLC was performed using an SPD-M10Avp UV-vis detector (Shimadzu, Kyoto, Japan).
6
Characterization of Organic Compounds
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Mass spectra were recorded on a Shimadzu LCMS-IT-TOF instrument (Kyoto, Japan) equipped with an ESI source in positive/negative ion mode at a resolution of 10,000 full width at half-maximum. 1H and 13C NMR spectra were recorded at 26°C with a Bruker DRX Advance-600 spectrometer (Bruker Biospin AG, Fallanden, Switzerland) at 600 MHz for 1H and 150 MHz for 13C. Chemical shifts were given in ppm downfield from internal TMS of D2O. Chemical shifts and coupling constants were calculated from a first-order analysis of the spectra. Assignments were fully supported by homo- and hetero- nuclear correlated 2D techniques, including COSY (correlation spectroscopy), HSQC (hetero-nuclear single quantum coherence) and HMBC (hetero-nuclear multiple band correlation) experiments following standard Bruker pulse programs.
7
Comprehensive Mass and NMR Spectroscopy
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Mass spectra were recorded on a Shimadzu LCMS-IT-TOF instrument (Kyoto, Japan) equipped with an ESI source in positive/negative ion mode at a resolution of 10,000 full width at half-maximum. 1H and 13C NMR spectra were recorded at 25°C with a Bruker DRX Avance 600 MHz spectrometer (Switzerland) at 600 MHz for 1H and 150 MHz for 13C. Chemical shifts in parts per million (ppm) were reported relative to the internal standard 2,2-dimethyl-2-silapentane-5-sulfonate. Chemical shifts and coupling constants were obtained from first- and second-order spectra analysis. Standard homo- and hetero-nuclear correlated 2D techniques were used to support the assignments, including correlation spectroscopy (COSY), heteronuclear single quantum coherence (HSQC), and heteronuclear multiple band correlation (HMBC) experiments.
8
HPLC-ESI-IT-TOF-MSn Analysis
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A Shimadzu LCMS-IT-TOF instrument was used to perform HPLC-ESI-IT-TOF-MSn analysis, which consists of a CBM-20A system controller, a DGU-20A3 degasser, two LC-20AD pumps, an SIL-20AC autosampler, a CTO-20A column oven, an SPD-M20A photodiode array (PDA) detector, an ESI ion source, and a hybrid IT-TOF mass spectrometer (Shimadzu, Kyoto, Japan).
9
Chromatographic and Spectroscopic Analysis of Organic Compounds
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For reaction progress testing, liquid chromatography (HPLC) was performed using a KNAUER apparatus equipped with a UV-Vis detector with application of the standard procedure [70 (link),71 (link),72 (link)]. LiChrospher RP-18 10 μm column (4 × 250 mm) was applied and methanol–water MeOH:H2O (70:30 v/v) was used as eluent at a flow rate of 1.5 cm3 min−1. Melting points were determined with the Boetius PHMK 05 apparatus and were not corrected. IR spectra (KBr pellets) were registered on a Thermo Nicolet 6700 FT-IR apparatus. 1H NMR (500 MHz) and 13C NMR (125 MHz) spectra were recorded with a Bruker AVANCE NMR spectrometer using CDCl3 as a solvent. TMS was used as an internal standard. UV-Vis spectra were determined for the 200–500 nm range through usage of spectrometer UV-5100 Biosens. HR-MS spectra were performed on a Shimadzu LCMS-IT-TOF instrument with ES ionization (heat bock and CDL temperatures of 200 °C, nebulizing gas flow of 1.5 mL/min) connected to a Shimadzu Prominence two LC-20AD pump chromatograph equipped with Phenomenex Kinetex 2.6 µm C18 100A column (acetonitrile–water mixtures, ACN:H2O, 65:35 v/v, were used as the eluent).
10
HPLC-ESI-IT-TOF Analysis of Compounds
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A Waters Alliance HPLC 2695 series instrument (Waters, Manchester, UK) was used to perform the high performance liquid chromatography (HPLC) analysis. Mobile phase: A (acetonitrile); B (H2O:CH3COOH, 100:0.2, v/v). Column: Agilent C18 (250 mm × 4.6 mm, 5 μm), maintained at 30 °C with flow rate of 1.0 mL·min−1. The detection wavelength was set at 254 nm for acquiring chromatograms. The injection volume was 20 µL. Gradient elution procedure: 0 min (20 % A) → 8 min (29% A) → 25 min (29% A) → 55 min (30% A) → 65 min (30% A) → 70 min (31% A) → 90 min (65% A) → 110 min (90% A) → 135 min (90% A).
The LCMS-IT-TOF instrument (Shimadzu, Kyoto, Japan) was equipped with an ESI source used in negative ionization mode. The interface and MS parameters were as follows: nebulizer pressure, 100 kPa; dry gas, N2 (1.5 L/min); drying gas temperature, 200 °C; spray capillary voltage, 4000 V; scan range, m/z 100–1000. Mobile phase: A (acetonitrile); B (H2O:CH3COOH, 100:0.2, v/v). Column: Agilent C18 (250 mm × 4.6 mm, 5 μm), maintained at 30 °C with flow rate of 1.0 mL·min−1. The injection volume was 20 µL. Gradient elution procedure: 0 min (20 % A) → 8 min (29% A) → 25 min (29% A) → 55 min (30% A) → 65 min (30% A) → 70 min (31% A) → 90 min (65% A) → 110 min (90% A) → 135 min (90% A).
The LCMS-IT-TOF instrument (Shimadzu, Kyoto, Japan) was equipped with an ESI source used in negative ionization mode. The interface and MS parameters were as follows: nebulizer pressure, 100 kPa; dry gas, N2 (1.5 L/min); drying gas temperature, 200 °C; spray capillary voltage, 4000 V; scan range, m/z 100–1000. Mobile phase: A (acetonitrile); B (H2O:CH3COOH, 100:0.2, v/v). Column: Agilent C18 (250 mm × 4.6 mm, 5 μm), maintained at 30 °C with flow rate of 1.0 mL·min−1. The injection volume was 20 µL. Gradient elution procedure: 0 min (20 % A) → 8 min (29% A) → 25 min (29% A) → 55 min (30% A) → 65 min (30% A) → 70 min (31% A) → 90 min (65% A) → 110 min (90% A) → 135 min (90% A).
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