Simultaneous determination of ten macrolides drugs in feeds by high performance liquid chromatography with evaporation light scattering detection

Abstract

A sensitive and reproducible method based on high performance liquid chromatography with evaporation light scattering detection (ELSD) was developed for the simultaneous determination of 10 macrolides drugs such as azithromycin, tulathromycin, spiramycin, tilmicosin, tylosin, erythromycin, clarithromycin, roxithromycin, midecamycin and josamycin in feeds. Feed samples were extracted with a sodium borate buffer solution (pH 10.0) − ethyl acetate. The dry extracts were dissolved in a phosphate buffer solution (pH 8.0), and then applied to an Oasis HLB solid-phase extraction cartridge for cleanup. The residues were reconstituted in 0.5 mL of the mobile phase. By optimizing the main operational parameters of ELSD and chromatographic conditions, all target compounds were well separated on an Ecosil C₈-SH column (250 mm × 4.6 mm, 5 μm) using a gradient elution program. The calibration curves showed good linearity (r > 0.9985) in the range of 1–200 μg mL⁻¹ for ten analytes. The average recoveries of all analytes from five kinds of feeds spiked at three levels were between 60.2% and 112%, with intra-day and inter-day relative standard deviations below 11% and 15%, respectively. The limits of detection ranged from 0.4 to 0.8 mg kg⁻¹ for ten macrolides.