A nuclear magnetic resonance (NMR) method was implemented to assess in vivo oxygenation levels by a quantitative determination of the 1H NMR myoglobin (Mb) resonances. The proximal His-F8 NδH at 70–90 ppm and Val-E11 γCH3 resonance at –2.8 ppm, reflecting deoxygenated (deoxy-Mb) and oxygenated (met-Mb) states, were alternately recorded. The method was developed in vitro choosing a couple of NMR sequences that could each maximize the signal-to-noise ratio (SNR) while avoiding baseline rolling and suppressing the water signal. Two quantitative calibration methods were implemented for deoxy- and met-Mb samples (0.1–1 mM), respectively. The respective limit of detection (LOD) and limit of quantification (LOQ) were 0.015 and 0.05 mM for met-Mb and 0.013 and 0.042 mM for deoxy-Mb. Sequences and calibration curves were employed in vivo in Arenicola marina to obtain, for the first time, an accurate measurement of oxy- and deoxy-Mb actual concentrations. In Arenicola, the peaks at approximately 87 and –2.7 ppm, reflecting the deoxy- and oxy-Mb states, respectively, were alternately recorded during increasing hypoxia. The deoxy-Mb concentrations were obtained from the calibration curve. The oxy-Mb concentrations were calculated from the calibration of met-Mb because it was proved that oxy- and met-Mb gave the same NMR molar response. From oxy- and deoxy-Mb concentrations, the intracellular oxygen partial pressure (PiO2) trend was determined.

M. Gussoni, M. A. Cremonini, A. Vezzoli, F. Greco, L. Zetta (2010). A quantitative method to assess muscle tissue oxygenation in vivo by monitoring 1H nuclear magnetic resonance myoglobin resonances. ANALYTICAL BIOCHEMISTRY, 400, 33-45 [10.1016/j.ab.2010.01.022].

A quantitative method to assess muscle tissue oxygenation in vivo by monitoring 1H nuclear magnetic resonance myoglobin resonances

CREMONINI, MAURO ANDREA;
2010

Abstract

A nuclear magnetic resonance (NMR) method was implemented to assess in vivo oxygenation levels by a quantitative determination of the 1H NMR myoglobin (Mb) resonances. The proximal His-F8 NδH at 70–90 ppm and Val-E11 γCH3 resonance at –2.8 ppm, reflecting deoxygenated (deoxy-Mb) and oxygenated (met-Mb) states, were alternately recorded. The method was developed in vitro choosing a couple of NMR sequences that could each maximize the signal-to-noise ratio (SNR) while avoiding baseline rolling and suppressing the water signal. Two quantitative calibration methods were implemented for deoxy- and met-Mb samples (0.1–1 mM), respectively. The respective limit of detection (LOD) and limit of quantification (LOQ) were 0.015 and 0.05 mM for met-Mb and 0.013 and 0.042 mM for deoxy-Mb. Sequences and calibration curves were employed in vivo in Arenicola marina to obtain, for the first time, an accurate measurement of oxy- and deoxy-Mb actual concentrations. In Arenicola, the peaks at approximately 87 and –2.7 ppm, reflecting the deoxy- and oxy-Mb states, respectively, were alternately recorded during increasing hypoxia. The deoxy-Mb concentrations were obtained from the calibration curve. The oxy-Mb concentrations were calculated from the calibration of met-Mb because it was proved that oxy- and met-Mb gave the same NMR molar response. From oxy- and deoxy-Mb concentrations, the intracellular oxygen partial pressure (PiO2) trend was determined.
2010
M. Gussoni, M. A. Cremonini, A. Vezzoli, F. Greco, L. Zetta (2010). A quantitative method to assess muscle tissue oxygenation in vivo by monitoring 1H nuclear magnetic resonance myoglobin resonances. ANALYTICAL BIOCHEMISTRY, 400, 33-45 [10.1016/j.ab.2010.01.022].
M. Gussoni; M. A. Cremonini; A. Vezzoli; F. Greco; L. Zetta
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/88450
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