Electron paramagnetic resonance (EPR) spectra of the oxidovanadium(IV) complexes were measured using a Bruker ELEXYS E 500 operating at the X-band frequency (9.7 GHz). The solid compounds 1 and 2 dissolved in water and a few drops of DMSO, were added to the samples to ensure good glass formation at liquid nitrogen temperature [101 (link)]. A microwave frequency of 6.231 GHz, power of 10 mW and modulation amplitude of 8 G was used. Anisotropic spectra were recorded on frozen solutions at 77 K using quartz Dewar and glass capillary tubes at room temperature. An analysis of the EPR spectra was carried out using the WinEPR SimFonia software package, version 1.26b [102 ].
Elexys e 500
Manufactured by Bruker
Sourced in Germany
The Elexys E-500 is a high-performance electron paramagnetic resonance (EPR) spectrometer designed and manufactured by Bruker. It is capable of performing continuous wave (CW) and pulsed EPR measurements at X-band frequencies.
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5 protocols using elexys e 500
1
Electron Paramagnetic Resonance of Oxidovanadium(IV) Complexes
2
Characterization of Metal Complexes
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Nuclear magnet resonance measurements were recorded with a JEOL (400 YH magnet) Resonance 400 MHz. Infrared spectrum was collected with a Bruker IFS 66 v/S FT-IR spectrometer, in which the DLaTGS detector was employed with 2 cm -1 resolution. Sample solution was prepared inside a gastight IR cell with CaF 2 or KBr as windows. For UV-visible spectrum, the Carry 5000 spectrometer was used and the sample solution was placed in a gas-tight UV-visible cuvette. The mass spectrum was collected in a negative mode with a Thermo Finnigan LCQ Deca XP Max LC/MS spectrometer through the direct injection mode. EPR spectra were recorded at X-band at 9.28 GHz, modulation frequency 100 kHz and modulation amplitude 5 Gauss on a Bruker ELEXYS E500 using an ER049X SuperX microwave bridge in a Bruker SHQ0601 cavity equipped with an Oxford Instruments continuous flow cryostat and an ITC 503 temperature controller (Oxford Instruments). A 1 mM CuSO 4 and 10 eq. of EDTA dissolved in water was employed as a standard sample for spin counting.
3
EPR Spectroscopy of PNSE Samples
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EPR
measurements were performed following an experimental procedure recently
set up.34 (link),35 (link) Samples were measured using an X-band (9
GHz) Bruker Elexys E-500 spectrometer equipped with a superhigh sensitivity
probe head. PNSE was introduced in a flame-sealed glass capillary
coaxially inserted in a standard 4 mm quartz sample tube. Measurements
were carried out at room temperature, with the following settings:
sweep width, 140 G; resolution, 1024 points; modulation frequency,
100 kHz; modulation amplitude, 1.0 G, and receiver gain, 60 dB. The
amplitude of the field modulation was preventively checked to be low
enough to avoid detectable signal overmodulation. A microwave power
of ∼0.6 mW was used to avoid microwave saturation of the resonance
absorption curve. To improve the signal-to-noise ratio, 16 scans were
accumulated. As concerning power saturation experiments, the microwave
intensity was gradually increased from 0.004 to 127 mW. The g value and the spin density were determined using Mn2+-doped MgO as an internal standard.35 (link) Spin density values must be considered as order of magnitude estimates,
since sample hydration was not controlled.
measurements were performed following an experimental procedure recently
set up.34 (link),35 (link) Samples were measured using an X-band (9
GHz) Bruker Elexys E-500 spectrometer equipped with a superhigh sensitivity
probe head. PNSE was introduced in a flame-sealed glass capillary
coaxially inserted in a standard 4 mm quartz sample tube. Measurements
were carried out at room temperature, with the following settings:
sweep width, 140 G; resolution, 1024 points; modulation frequency,
100 kHz; modulation amplitude, 1.0 G, and receiver gain, 60 dB. The
amplitude of the field modulation was preventively checked to be low
enough to avoid detectable signal overmodulation. A microwave power
of ∼0.6 mW was used to avoid microwave saturation of the resonance
absorption curve. To improve the signal-to-noise ratio, 16 scans were
accumulated. As concerning power saturation experiments, the microwave
intensity was gradually increased from 0.004 to 127 mW. The g value and the spin density were determined using Mn2+-doped MgO as an internal standard.35 (link) Spin density values must be considered as order of magnitude estimates,
since sample hydration was not controlled.
4
EPR Spectroscopy of Lipid Vesicles
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EPR spectra were recorded with a 9 GHz Bruker Elexys E-500 spectrometer (Bruker, Rheinstetten, Germany). The samples (20 μL) were inserted in glass capillaries with an internal diameter of 1.2 mm, which were flame-sealed. Then, capillaries containing MLV suspensions were placed in a standard 4 mm quartz sample tube containing light silicone oil for thermal stability and analysed with no further treatment. All the measurements were performed at 25 °C. Spectra were recorded using the following instrumental settings: sweep width, 100 G; resolution, 1024 points; time constant, 20.48 ms; modulation frequency, 100 kHz; modulation amplitude, 1.0 G; incident power, 6.37 mW. Several scans, at least eight, were accumulated to improve the signal-to-noise ratio.
5
Lipid Bilayer Dynamics: EPR Analysis
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The EPR spectra of nPC-SL in POPC/POPS MLV suspensions were recorded in the absence or in the presence of LL-III at a L/P mole ratio of 10. The total lipid concentration in each sample was 10 mM. Samples were prepared in 10 mM sodium cacodylate buffer in the absence or in the presence of 1 mM CaCl 2 . EPR experiments were performed using a 9 GHz Bruker Elexys E-500 spectrometer (Bruker, Rheinstetten, Germany). Capillaries containing the samples (B25 mL) were placed in a standard 4 mm quartz sample tube. The instrumental settings were as follows: sweep width, 100 G; resolution, 1024 points; modulation frequency, 100 kHz; modulation amplitude, 1.0 G; time constant, 20.5 ms; and incident power, 5.0 mW. Several scans, typically 8, were accumulated to improve the signal-tonoise ratio. A quantitative analysis of nPC-SL spectra was performed by determining the acyl chain order parameter, S, and the nitrogen isotropic hyperfine coupling constant, a 0 N , as described in the literature. 22 The differential order parameters, DS, and differential nitrogen isotropic hyperfine coupling constants, Da 0 N , were calculated subtracting the values determined for POPC/POPS MLVs alone from the values determined for each sample. The experiments were performed in triplicates.
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