4.6. Hydroxyl Radical Detection Using EPR Spin Trapping Spectroscopy

Formation of HO^● by differentiating U-937 cells was measured by EPR spectrometer, MiniScope MS400 (Magnettech GmbH, Berlin, Germany). U-937 cells were treated with 50 μM PMA in absence or presence of 5 mM mannitol as a scavenger and incubated for 20 min at 37 °C and 5% CO₂. PMA differentiated cells were incubated with 50 mM POBN containing 170 mM ethanol for 20 min at 37 °C. EPR spectra were recorded under the following parameters: microwave power (10 mW), modulation amplitude (1 G), modulation frequency (100 kHz), sweep width (100 G), scan rate (1.62 G s⁻¹). Incubation steps were skipped for baseline recording.