He Tong, Cui Peng Lei, Zhang Shuai, Fan Yu Hang, Jin Qiu Shi, Wang Jian Ping
College of Veterinary Medicine, Hebei Agricultural University, Baoding, Hebei 071000, China.
College of Science, Hebei Agricultural University, Baoding, Hebei 071000, China.
Food Chem X. 2023 Sep 6;19:100867. doi: 10.1016/j.fochx.2023.100867. eCollection 2023 Oct 30.
With the increasing focus on food security, a screening method with high-throughput, ultra-sensitivity, and user-friendly operation is urgently needed for monitoring of sulfonamides residues in animal-derived foods. In this study, the sulfonamides' receptor dihydropteroate synthase of was subjected to saturate mutation, and a mutant with higher affinities for sulfonamides was obtained. The mutant was then used as recognition material to establish a fluorescence polarization assay for determination of 35 sulfonamides in pork. Due to the use of an enhanced fluorescent tracer containing two fluorophore molecules, the sensitivities for the 35 sulfonamides were improved for 2.8-8.6 folds (LODs 0.03-1.16 ng/mL) in comparison with using conventional fluorescent tracer. The present method outperformed all previous fluorescence polarization (immuno)assays for sulfonamides due to its broader spectrum, higher sensitivity, and shorter assay time. Furthermore, this is the first study reporting an enhanced fluorescence polarization assay for determination of small molecule substance.
随着对食品安全的关注度不断提高，迫切需要一种高通量、超灵敏且操作简便的筛选方法来监测动物源性食品中的磺胺类药物残留。在本研究中，对磺胺类药物的受体二氢蝶酸合酶进行饱和突变，获得了对磺胺类药物具有更高亲和力的突变体。然后将该突变体用作识别材料，建立了一种荧光偏振分析法，用于测定猪肉中的35种磺胺类药物。由于使用了含有两个荧光团分子的增强型荧光示踪剂，与使用传统荧光示踪剂相比，对这35种磺胺类药物的灵敏度提高了2.8至8.6倍（检测限为0.03至1.16 ng/mL）。由于其更宽的检测范围、更高的灵敏度和更短的分析时间，本方法优于以往所有用于磺胺类药物的荧光偏振（免疫）分析方法。此外，这是第一项报道用于测定小分子物质的增强型荧光偏振分析方法的研究。
