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

作者：Gussoni Maristella^*; Cremonini Mauro A; Vezzoli Alessandra; Greco Fulvia; Zetta Lucia

来源：Analytical Biochemistry, 2010, 400(1): 33-45.

DOI：10.1016/j.ab.2010.01.022

• H-1 NMR
• Myoglobin
• Tissue oxygenation
• In vivo
• Arenicola marina

A nuclear magnetic resonance (NMR) method was implemented to assess in vivo oxygenation levels by a quantitative determination of the H-1 NMR myoglobin (Mb) resonances. The proximal His-FS N delta H at 70-90 ppm and Val-E11 gamma CH3 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.

• 出版日期2010-5-1