Inova 300 mhz spectrometer

Manufactured by Agilent Technologies

The INOVA 300 MHz spectrometer is a nuclear magnetic resonance (NMR) instrument designed for analytical applications. It operates at a frequency of 300 MHz and is capable of performing various NMR experiments to analyze the structure and properties of chemical compounds.

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4 protocols using inova 300 mhz spectrometer

Protein NMR Sample Preparation

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The protein was concentrated
to ∼2 mM using 10 kDa molecular
mass cutoff spin filters (Amicon), made to 7% D₂O for locking,
and loaded into a 5 mm Shigemi low-volume NMR tube. 1D ¹³C NMR spectra were all recorded with ¹H decoupling on
a Varian Inova 300 MHz spectrometer unless otherwise noted. The T₁ relaxation time was measured for the chromophore ¹³C_ζ resonance using standard inversion recovery
techniques. The best duty cycle in the 1D spectra was achieved empirically
using a 50° pulse and a delay time of 4.0 s. All experiments
were performed with the temperature control set to 25 °C. The
typical samples for NMR contained 40 mM buffer (HEPES for high-pH
samples and MES for lower-pH samples) and 40 mM NaCl. Phosphate and
citrate buffers and higher salt concentrations were also used for
a limited set of spectra and found to give identical results for a
given pH.

Oltrogge L.M., Wang Q, & Boxer S.G. (2014). Ground-State Proton Transfer Kinetics in Green Fluorescent Protein. Biochemistry, 53(37), 5947-5957.

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Optimized Synthesis and Characterization of Compounds

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All reagents and solvents for synthesis were purchased from commercial sources and used without purification. All glassware was flame-dried prior to use. Thin layer chromatography (TLC) was carried out on aluminium plates coated with 60 F254 silica gel. TLC plates were visualized under UV light (254 nm or 365 nm) or stained with 5% phosphomolybdic acid. Normal phase column chromatography was carried out using a Yamazen Smart Flash AKROS system. Analytical reverse-phase high pressure liquid chromatography (RP-HPLC) was carried out on Shimadzu LC20 HPLC system with an analytical C18 column. Semi-preparative HPLC was carried out on the same system with a semi-preparative C18 column. The mobile phases for were H₂O with 0.1% formic acid (A) and acetonitrile with 0.1% formic acid (B) if not mentioned otherwise. NMR spectra were recorded on a Bruker Avance Neo 400 MHz or Varian INOVA 300 MHz spectrometer in specified deuterated solvents. High resolution electrospray ionization mass spectrometry (HRMS-ESI) was carried out on a Thermo Scientific Q Exactive Focus system. The purities of compounds were confirmed by NMR and analytical HPLC-UV as ≥ 95%.

Ma X.R., Xu L., Xu S., Klein B.J., Wang H., Das S., Li K., Yang K.S., Sohail S., Chapman A., Kutateladze T.G., Shi X., Liu W.R, & Wen H. (2021). Discovery of Selective Small Molecule Inhibitors for the ENL YEATS Domain. Journal of medicinal chemistry, 64(15), 10997-11013.

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Physicochemical Characterization of Nanoparticles

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Zeta potential of nanoparticles suspension in water (1 mg mL⁻¹) was measured in a folded capillary zeta cell using a Zetasizer Nano-ZS apparatus (Malvern Instruments). All measurements were done in triplicates with 10 scans each. TEM analysis was performed using a FEI Tecnai G2 on samples dispersed in ethanol and deposited on carbon-coated copper grids. Average size and size distribution of nanoparticles were analyzed by Fiji software41 (link) using TEM images, analyzing a minimum of 50 nanoparticles per sample. FT-IR was performed in a Bruker Vertex 70 spectrophotometer. Samples in ethanol dispersion were deposited onto NaCl FT-IR cards and dried at 70 °C. Spectra were obtained using a resolution of 4 cm⁻¹, 64 scans and a spectral window from 4000 to 400 cm⁻¹. Dissolution ¹H NMR was used to dissolve the MSNs and obtain a clear spectrum of the surface groups, based on a previously reported method.42 (link) Approximately 10–15 mg of MSNs were dissolved in 662 µL of D₂O and 38 µL of NaOD. The mixture was incubated overnight at 37 °C under stirring. The spectra were obtained in a Varian Inova 300 MHz Spectrometer using 128 scans per sample.

Fulaz S., Devlin H., Vitale S., Quinn L., O’Gara J.P, & Casey E. (2020). Tailoring Nanoparticle-Biofilm Interactions to Increase the Efficacy of Antimicrobial Agents Against Staphylococcus aureus. International Journal of Nanomedicine, 15, 4779-4791.

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Photophysical Characterization of Schiff Base Probe

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The solvents were received from commercial sources and puri ed by standard methods. For all spectroscopic studies, CDCl 3 was used. 2-hydroxy-1-nepthaldehyde, p-toluidine, MgCl 2 , AlCl 3 , CdCl 2 , NiCl 2 , CoCl 2 , KCl, AgCl, NaCl, ZnCl 2 , CrCl 3 , HgCl 2 , CaCl 2 , Mn(acetate) 2 , Pb(acetate) 2 , and Cu(acetate) 2 were purchased from Sigma Aldrich. The photophysical properties of the probe were studied using a spectro urometer (RF-5301 PC, Shimadzu, Japan) with having Xenon lamp of 150-watt and a 1.0 cm quartz cell. A Bio-Rad FTS 3000 MX Fourier Transmission Infrared spectrometer was used to record the FT-IR spectra of SB and its Cu-complex in the range of 400-4000 cm - 1 with KBr disks. Varian, INOVA 300 MHz spectrometer was used to record the 1 H-NMR. The solution of the samples was prepared in CDCl 3 in 5 nm OD NMR tubes.

Khan S., Muhammad M., Kamran A.W., Al-Saidi H.M., Alharthi S.S, & Algethami J.S. (2023). An ultrasensitive colorimetric and fluorescent "turn-on" chemosensor based on Schiff base for the detection of Cu²⁺ in the aqueous medium. Environmental monitoring and assessment, 195(6).

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