Abstract:

To develop a rapid, effective, and accurate method for the simultaneous determination of chloramphenicol, thiamphenicol, and florfenicol in human blood by pass-through cleanup solid-phase extraction combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), novel nano-titania-coated modified magnetic graphene oxide (TiO₂-Mag-GO) is used as the PRiME pass-through cleanup solid-phase extraction sorbent for the cleanup of blood phospholipids. The chromatographic separation was performed on an Eclipse Plus C18 column (100 mm×2.1 mm, 1.8 μm) using 0.08% (v/v) aqueous ammonia solution and acetonitrile having aqueous ammonia (0.08%, v/v) as mobile phases. The tandem mass spectrometer was operated using electrospray ion source (ESI) in the multiple reaction monitoring (MRM) mode. The results showed that the linearities were in the range of 0.1-10.0 μg/L with the determination coefficients (r²) greater than 0.9990 for chloramphenicol, thiamphenicol, and florfenicol. The limits of quantification (LOQs) (S/N>10) in the blood samples were between 0.056 and 0.082 μg/L and the recoveries ranged from 90.0% to 105% at three spiked levels. Precision values expressed as relative standard deviations (RSDs) were in the range of 1.2%-6.6%. The developed method can be used for routine analyses to determine chloramphenicol, thiamphenicol, and florfenicol in clinical studies.

Key words: chloramphenicol, florfenicol, human blood, liquid chromatography-tandem mass spectrometry (LC-MS/MS), nano-titania-coated modified magnetic graphene oxide (TiO₂-Mag-GO), pass-through cleanup, thiamphenicol