We re-dissolved dry protein-free lipid-free extracts in 600 μL of 20 mM deuterated phosphate buffer (pH 7.2) containing 2 × 10−5 M sodium 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS) as an internal standard. The 1H NMR spectra were obtained using an AVANCE III HD 700 MHz NMR spectrometer (Bruker BioSpin, Ettlingen, Germany) equipped with a 16.44 Tesla Ascend cryomagnet at the Center of Collective Use “Mass spectrometric investigations” SB RAS as described in [26 (link)].
Avance 3 hd 700 mhz nmr spectrometer
Manufactured by Bruker
Sourced in Germany
The AVANCE III HD 700 MHz NMR spectrometer is a high-performance nuclear magnetic resonance (NMR) spectrometer designed for advanced research applications. It operates at a magnetic field strength of 16.4 Tesla, providing high-resolution spectroscopic data. The spectrometer is equipped with a superconducting magnet and advanced electronic components for precise signal detection and processing.
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4 protocols using avance 3 hd 700 mhz nmr spectrometer
1
Protein-free Lipid-free Extract Analysis
2
HPLC Purification and NMR Analysis of Organic Compounds
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Reaction products after 2 h were extracted with ethyl acetate and then separated with reverse‐phase HPLC. HPLC was performed in Agilnet 1260 infinity LC equipped with a diode array detector and a ZORBAX SB C‐18 column at 25°C, with an aqueous solvent system (flow rate, 0.6 ml/min) containing 35% acetonitrile. Absorbance was monitored at 254 nm. The separated fractions were dissolved in acetone‐d6. NMR was performed with Bruker Avance III HD 700 MHz NMR spectrometer.
1H NMR (acetone‐d6) for Fraction 3: δ 6.23 (d, J = 2.3 Hz, H5), δ 6.78 (dd, J = 8.8, 2.5 Hz, H4), δ 6.97 (d, J = 8.8, 2.5 Hz, H1), δ 7.61 (d, J = 8.8, H3), δ 7.65 (d, J = 8.8, H2).
1H NMR (acetone‐d6) for Fraction 4: δ 6.48 (d, J = 2.4 Hz, H10), δ 6.77, δ 6.85 (m, H1, H9, H11), δ 6.93 (d, J = 8.8 Hz, H4), δ 6.97 (d, J = 2.3 Hz, H5), δ 7.06 (s, H6), δ 7.43 (s, H7), δ 7.57 (m, H2, H3, H8).
1H NMR (acetone‐d6) for Fraction 3: δ 6.23 (d, J = 2.3 Hz, H5), δ 6.78 (dd, J = 8.8, 2.5 Hz, H4), δ 6.97 (d, J = 8.8, 2.5 Hz, H1), δ 7.61 (d, J = 8.8, H3), δ 7.65 (d, J = 8.8, H2).
1H NMR (acetone‐d6) for Fraction 4: δ 6.48 (d, J = 2.4 Hz, H10), δ 6.77, δ 6.85 (m, H1, H9, H11), δ 6.93 (d, J = 8.8 Hz, H4), δ 6.97 (d, J = 2.3 Hz, H5), δ 7.06 (s, H6), δ 7.43 (s, H7), δ 7.57 (m, H2, H3, H8).
3
Multimodal Materials Characterization
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FTIR spectra were recorded with an iZ10 spectrophotometer (Nicolet) with a resolution of 6 cm−1 (KBr pellets).
Solid-state 13C CP/MAS-TOSS NMR spectrum was obtained using an AVANCE III HD 700MHz NMR spectrometer (Bruker, Karlsruhe, Germany) running at a spinning speed of 12 kHz and using a Bruker probe head (PA BBO 700S3 BB-H-D-05 Z) equipped with a 3.2 mm MAS assembly.
Element contents were investigated via a Vario EL elemental analyzer (Elementar Analysensysteme GmbH, Inc, Langenselbold, Germany) with a reduction temperature of 700 °C and a combustion temperature of 950 °C. The results were the average of the 2 times repeated tests.
Solid-state 13C CP/MAS-TOSS NMR spectrum was obtained using an AVANCE III HD 700MHz NMR spectrometer (Bruker, Karlsruhe, Germany) running at a spinning speed of 12 kHz and using a Bruker probe head (PA BBO 700S3 BB-H-D-05 Z) equipped with a 3.2 mm MAS assembly.
Element contents were investigated via a Vario EL elemental analyzer (Elementar Analysensysteme GmbH, Inc, Langenselbold, Germany) with a reduction temperature of 700 °C and a combustion temperature of 950 °C. The results were the average of the 2 times repeated tests.
4
NMR and GPC Characterization of Polysaccharide Conjugates
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All 1 H, 13 C and 1 H-13 C correlation spectra were measured using Bruker Avance III HD 700 MHz NMR spectrometer (Bruker, USA) equipped with a triple-resonance cryoprobe optimized for 13 C detection at 298 K in D2O. The 1 H-13 C multiplicity-edited heteronuclear single quantum correlation (HSQC, JH-C = 145 Hz) and 1 H-13 C heteronuclear multiple bond correlation (HMBC, n JH-C = 10 Hz) experiments were used based on the previous experience. (Münster et al., 2017) (link) Molecular weight distribution was analyzed by the gel permeation chromatography (GPC) using a Waters HPLC Breeze chromatographic system (Waters, USA) set up with a Waters 2414 refractive index detector (drift tube T = 60 °C), Tosoh TSK gel GMPWXL column (300 mm × 7.8 mm × 13 µm, column T = 30 °C). A mixture of 0.1 M NaNO3 and 0.05 M Na2HPO4 was employed as a mobile phase. Calibration was carried out using pullulan polysaccharide calibration kit SAC-10 (Agilent Technologies, USA) in a span of Mw 342-805 000 g/mol.
Zeta potential (ζ) and hydrodynamic radii of conjugate nano-assemblies were determined in demineralized water and 0.15 M NaCl solution by dynamic light scattering (DLS) method using a Zetasizer Nano ZS90 instrument (Malvern Instruments, UK). The measurements were performed at 25 °C on a DTS1070 cell using the Smoluchowski model.
Zeta potential (ζ) and hydrodynamic radii of conjugate nano-assemblies were determined in demineralized water and 0.15 M NaCl solution by dynamic light scattering (DLS) method using a Zetasizer Nano ZS90 instrument (Malvern Instruments, UK). The measurements were performed at 25 °C on a DTS1070 cell using the Smoluchowski model.
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