Determination of acysole in whole blood by high-performance liquid chromatography

A method is developed for the determination of a carbon monoxide antidote acysole in whole blood using hydrophilic interaction high-performance liquid chromatography with UV detection. Two methods were used for sample preparation: protein precipitation with acetonitrile and solid-phase extraction (SPE) with an acysole recovery of 95.8 and 89.8%, respectively. Chromatographic determination was performed in the isocratic mode on a Nucleodur HILIC column and with an acetonitrile: water 95: 5 mobile phase at 225 nm. The calibration graphs are linear in the concentration range 0.056–111.1 μg/mL. The limits of detection and determination of acysole were 0.050 and 0.120 μg/mL after protein precipitation with acetonitrile and 0.071 and 0.169 μg/mL with SPE, respectively.