Determination of Amantadine Residue in Honey by Solid‐phase Extraction and High‐performance Liquid Chromatography with Pre‐column Derivatization and Fluorometric Detection |
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Authors: | Jinzhen Zhang Jing Zhao Jinhui Zhou Xiaofeng Xue Yi Li Liming Wu Fang Chen |
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Institution: | Bee Products Quality Supervision and Testing Center, Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China |
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Abstract: | 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 (λex=470 nm, λem=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%. |
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Keywords: | amantadine honey solid‐phase extraction high‐performance liquid chromatographic (HPLC) fluorometric detection |
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