1H-, 13C-, and 2D-NMR spectra were carried out in CDCl3 using a Bruker AM-400 MHz spectrometer (Karlsruhe, Baden-Wuerttemberg, Germany) with TMS as internal standard (IS); δ in ppm and J in Hz. ESI-MS was obtained with a TSQ Quantum Access MAX mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). Column chromatography was performed using silica gel (200–300 mesh, Qingdao Marine Chemical Ltd., Qingdao, China), RP-C18 silica gel (150–200 mesh, Merck, Darmstadt, Hesse, Germany) and Sephadex LH-20 (20–100 μm, Sigma, St Louis, MO, USA). FBS (fetal bovine serum) and DMEM (Dulbecco’s Modified Eagle Medium) were purchased from Gibco (Life Technologies, Grand Island, NY, USA). Doxorubicin was provided by Sigma-Aldrich (St Louis, MO, USA). All other chemicals and solvents were of analytical grade.
Am 400 mhz spectrometer
Manufactured by Bruker
Sourced in Germany
The AM 400 MHz spectrometer is a nuclear magnetic resonance (NMR) instrument designed for laboratory use. It operates at a frequency of 400 MHz and is capable of performing various NMR experiments to analyze the structure and properties of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Doxorubicin, by Merck Group (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Rp c18 silica gel, by Merck Group (1 mentions)
Tsq quantum access max mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Fbs fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Sephadex lh 20, by Merck Group (1 mentions)
Silica gel, by Qingdao Marine Chemical (1 mentions)
Agilent lc msd tof mass spectrometer, by Agilent Technologies (1 mentions)
Rotary evaporator, by Büchi (1 mentions)
Model 510 pump, by Waters Corporation (1 mentions)
Pl aquagel oh 40 column, by Agilent Technologies (1 mentions)
Optilab rex, by Wyatt Technology (1 mentions)
Dawn heleos 2 18, by Wyatt Technology (1 mentions)
Ultimate 3000, by Thermo Fisher Scientific (1 mentions)
Q exactive hf, by Thermo Fisher Scientific (1 mentions)
6 protocols using am 400 mhz spectrometer
1
NMR Characterization of Organic Compounds
2
Synthesis of Methoxy-Terminated PEG
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In a nitrogen atmosphere, TA (2.29 g, 10.00 mmol), mPEG (8.25 g, 15.00 mmol), Py-PTSA (0.027 g, 0.06 mmol) and toluene (100 ml) were refluxed together at 115 °C for 8 h. Afterwards, the mixture was concentrated, dissolved in CDM (100 ml), and extracted with saturated NaCl solution in turn. The CDM layer was orderly dehydrated over anhydrous MgSO4, concentrated and stirred in NaOH solution (20 ml, 1.00 mol/l) for 5 h. Finally, the OEmPEG as a white powder (5.22 g, 78.3%) was yielded by extraction with CDM twice, drying over anhydrous MgSO4 and vacuum desiccation. Its structure was determined via 1H NMR spectrum in CDCl3 using a Bruker AM-400 MHz spectrometer (Bruker Biospin).
3
Synthesis and Characterization of Organic Compounds
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Tetrahydrofuran was dried by refluxing over sodium with diphenyl ketone as an indicator. Melting points were obtained on a Yanaco MP-500 melting point apparatus (Yanaco Ltd., Osaka, Japan) and are uncorrected. 1H- and 13C-NMR spectra were recorded on a Bruker AM 400 MHz spectrometer (Bruker Company, Billerica, MA, USA) in CDCl3 with TMS as an internal standard and the chemical shifts (δ) are reported in parts per million (ppm). The IR spectra (KBr pellets) were taken on a Nicolet FTIR 920 spectrometer (Thermo Nicolet Corporation, Madison, WI, USA). HRMS measurements were carried out on an Agilent LC/MSD TOF mass spectrometer (Agilent, Santa Clara, CA, USA). TLC separations were performed on silica gel GF254 plates (Qingdao Ocean Chemical Industry, Qingdao, China), and the plates were visualized with UV light. Column chromatography was performed on silica gel zcx II (200–300 mesh) (Qingdao Ocean Chemical Industry, Qingdao, China) with petroleum-ether (PE) and ethyl-acetate (EA) (Beijing Chemical Reagent Company, Beijing, China) as the eluent.
4
Physicochemical Characterization of Novel Compounds
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The melting points are determined on Thomas Hoover apparatus and are uncorrected. The IR (KBr) spectra are recorded on Shimadzu 8300 Fourier transform infrared spectrometer. 1H NMR is recorded on Bruker AM 400 MHz spectrometer and 13C NMR is recorded on Bruker AM 100 MHz spectrometer using DMSO as solvent and TMS as an internal standard (Chemical shift in ppm). Mass spectral (MS) analysis is carried out on Agilent 6520 ESIQTOF MS with Ionization source ESIQTOF and acetonitrile as solvent (110 Volts). Elemental analyses are obtained on a Vario-EL instrument. Thin layer chromatography is conducted on 0.25 mm silica gel plates (60F254, Merck). Visualization is made with ultraviolet light. All extracted solvents are dried over Na2SO4 and evaporated with a BUCHI rotary evaporator. Reagents are obtained commercially and used as received.
5
Characterization of Lentinan-Based Polysaccharides
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The ultraviolet–visible (UV–Vis) spectra and Fourier transform infrared (FTIR) spectra of LPs and MLPs were measured by the methods of Han et al. [4 (link)]; periodate oxidation analyses were performed with the method described by Hu et al. [15 (link)]; and atomic force microscope (AFM) images were observed according to our previous method [17 (link)].
The Mw distributions of LPs and MLPs were detected using a HPSEC system mainly composed of a model 510 pump (Waters, Milford, MA, USA), a PL aquagel-OH 40 column (8 μm, 7.5 × 300 mm, Agilent), a multi-angle laser light scattering (MALLS) detector (DAWN HELEOS-II 18, Wyatt Technology Co., Santa Barbara, CA, USA), and a differential refractive index (RI) detector (Optilab rEX, Wyatt Technology Co., Santa Barbara, CA, USA). Ammonium acetate solution (0.2 mol/L) was used as the mobile phase at a flow rate of 0.7 mL/min. Five milligrams of samples were dissolved in 2 mL ammonium acetate solution, followed by filtration through a 0.22 μm filter for injection. The injection volume was 20 μL and the column temperature was 30 °C.
After having been kept in a vacuum drier for a week, samples (30–40 mg) were put into a 5 mm NMR tube and dissolved in 1.0 mL dimethyl sulfoxide (DMSO). A Bruker AM 400 MHz spectrometer (Bruker, Rheinstetten, Germany) was used to record 1H NMR spectrum with the operating frequency of 400.13 MHz at 30 °C.
The Mw distributions of LPs and MLPs were detected using a HPSEC system mainly composed of a model 510 pump (Waters, Milford, MA, USA), a PL aquagel-OH 40 column (8 μm, 7.5 × 300 mm, Agilent), a multi-angle laser light scattering (MALLS) detector (DAWN HELEOS-II 18, Wyatt Technology Co., Santa Barbara, CA, USA), and a differential refractive index (RI) detector (Optilab rEX, Wyatt Technology Co., Santa Barbara, CA, USA). Ammonium acetate solution (0.2 mol/L) was used as the mobile phase at a flow rate of 0.7 mL/min. Five milligrams of samples were dissolved in 2 mL ammonium acetate solution, followed by filtration through a 0.22 μm filter for injection. The injection volume was 20 μL and the column temperature was 30 °C.
After having been kept in a vacuum drier for a week, samples (30–40 mg) were put into a 5 mm NMR tube and dissolved in 1.0 mL dimethyl sulfoxide (DMSO). A Bruker AM 400 MHz spectrometer (Bruker, Rheinstetten, Germany) was used to record 1H NMR spectrum with the operating frequency of 400.13 MHz at 30 °C.
6
NMR and MS Analysis of Genistein-Copper Complexes
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1H-NMR spectra were obtained on a Bruker AM 400 MHz spectrometer (Karlsruhe, Germany). MS spectra were obtained on a Thermo Scientific™ Q Exactive™ HF (Waltham, MA, USA) in the positive ion mode. The samples were prepared by mixing solutions of genistein and copper(II), and then the mixed solutions were pushed through the nylon membrane with 220 nm sieve pores. The samples were analyzed by direct infusion ESI by means of a syringe pump (Thermo UltiMate 3000, Waltham, MA, USA) at a flow rate of 5 μL/min. Capillary temperature is 320 °C and spray voltage is 3.50 kV.
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