Determination of Amantadine Residue in Honey by Solid-phase Extraction and High-performance Liquid Chromatography with Pre-column Derivatization and Fluorometric Detection

摘要

Amantadine (AMA) is an anti-viral drug used in apiculture to protect honeybee against the sacbrood virus (Morator aetatulae). This study described a reliable high-performance liquid chromatographic (HPLC) method for analyzing AMA in honey using a solid-phase extraction (SPE) cartridge (Plexa PCX) for purification, 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) as a pre-column derivatization agent, and fluorometric detection (λ_ex=470 nm, λ_em=530 nm). The chromatographic separation was performed on an XDB C18 column (150×4.6 mm i.d.) using 0.1% trifluoroacetic acid/acetonitrile (35:65,V/V) as the mobile phase at a flow rate of 1.0 mL·min?1 with a run time of 20 min. Under these optimal conditions, a linear relationship was observed in the range of 0.025–1.0 μg· mL?1 with a good correlation coefficient (0.998) and low limit of detection (0.0080 μg·g?1), the recoveries were all above 90%, and the intra-day and inter-day precision (RSD) ranged from 3.4%–5.1%.