Automated on-line microdialysis sampling coupled with high-performance liquid chromatography for simultaneous determination of malondialdehyde and ofloxacin in whole blood

We have developed a system that couples an on-line microdialysis (MD) system with flow injection high-performance liquid chromatography (HPLC)-fluorescence detection for simultaneous measurement of the concentrations of malondialdehyde (MDA) and ofloxacin (OFL) in whole blood samples. The sample matrix was first cleaned with an MD system using an MD probe. A continuously flowing dialysate stream was derivatized on-line and auto-injected into a separation column. MDA and OFL were separated through a reverse-phase C18 column (250 mm × 4.6 mm) at a flow rate of 0.8 mL min⁻¹ and then detected using a fluorescence detector (excitation: 532 nm; emission: 553 nm); the system's components were connected on-line using a valve control. Validation experiments demonstrated good linearity, precision, accuracy, and recovery. The precisions for the determinations of MDA and OFL, measured in terms of relative standard deviations, were 6.5% and 4.6%, respectively, for intra-day assays and 7.5% and 8.7%, respectively, for inter-day assays. The average recoveries of MDA and OFL spiked in plasma were each close to 100%. The use of this on-line MD-HPLC system permitted continuous monitoring of MDA and OFL in OFL-treated whole blood subjected to UV-A irradiation. Based on our results, the UV-A irradiation markedly increased the level of MDA in the OFL-treated whole blood.