A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071, Moscow, Russia.
Anal Methods. 2024 Feb 1;16(5):721-730. doi: 10.1039/d3ay01714a.
Fluoroquinolone antibiotics are used to cure and protect bees and apiaries from infections. Consequently, they may contaminate honey and other products of beekeeping. In this study, a highly sensitive immunoenzyme assay (EIA) was for the first time developed for the determination of a fluoroquinolone flumequine (FLU) in honey. The EIA was carried out in an indirect competitive format with colorimetric detection. The analysis was characterized by a low limit of detection of 30 pg mL. The polyclonal antibodies used showed no cross-reactivity with 24 other (fluoro)quinolones; the assay was highly specific only toward FLU. Different coating FLU-protein conjugates were tested to achieve the most sensitive competitive immunodetection. A highly simplified and rapid (3-5 min) sample preparation was proposed based on the 100-300 times dilution of honey by a buffer. The developed EIA has been tested to detect FLU in honey of different origins, namely acacia, flower, buckwheat, chestnut, and linden honey. It has been demonstrated that 76.2-115.9% of FLU could be determined by the assay. The versatility, simplicity, and rapidity of the EIA enable us to propose this method as an effective tool to control the contamination of honey.
氟喹诺酮类抗生素被用于治疗和保护蜜蜂和蜂群免受感染。因此，它们可能会污染蜂蜜和其他养蜂产品。在这项研究中，首次开发了一种用于测定蜂蜜中氟喹诺酮类药物氟甲喹（FLU）的高灵敏度免疫酶分析（EIA）。EIA 采用间接竞争格式和比色检测进行。分析的特点是检测限低至 30 pg/mL。所用的多克隆抗体与 24 种其他（氟）喹诺酮类药物没有交叉反应；该测定法对 FLU 具有高度特异性。测试了不同的包被 FLU-蛋白缀合物，以实现最敏感的竞争免疫检测。根据缓冲液将蜂蜜稀释 100-300 倍，提出了一种高度简化和快速（3-5 分钟）的样品制备方法。已用该 EIA 测试了来自不同来源的蜂蜜中的 FLU，即刺槐、花、荞麦、栗和椴树蜂蜜。结果表明，该测定法可测定 76.2-115.9%的 FLU。EIA 的多功能性、简单性和快速性使我们能够提出该方法作为控制蜂蜜污染的有效工具。
