Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.
J Sci Food Agric. 2014 Jan 30;94(2):301-7. doi: 10.1002/jsfa.6258. Epub 2013 Jul 11.
A rapid one-step chemiluminescent competitive indirect enzyme-linked immunosorbent assay (CL-ciELISA) for florfenicol (FF) and its major metabolite florfenicol amine (FFA) residues in animal meat products has been developed.
The 50% binding inhibition (IC₅₀) values of the method were 0.195 µg kg⁻¹ for FFA and 0.24 µg kg⁻¹ for FF under optimum conditions. The cross-reactive rates for FF and FFA were 100.0% and 81.2%, respectively. FF and FFA were easily extracted from animal meat product with an FF/FFA extraction buffer, obtaining recoveries of 81.8-92.0% (FF) and 77.2-100% (FFA). The whole one-step CL-ciELISA test can be accomplished within 40 min in theory. The detection limits (LODs) of the assay were 0.98 µg kg⁻¹ for FF and 0.80 µg kg⁻¹ for FFA in animal meat samples. Finally, field animal meat samples were analyzed with the CL-ciELISA method, and the results correlated well with those obtained using traditional ELISA and a previously reported liquid chromatographic-tandem mass spectrometric method.
The combined results confirmed the utility of this faster one-step CL-ciELISA for simultaneous trace analysis of FF and FFA. To date, this is the most rapid developed ELISA and CL-ELISA method for detection of FF and FFA.
建立了一种快速一步化学发光竞争间接酶联免疫吸附测定法（CL-ciELISA），用于检测动物肉产品中的氟苯尼考（FF）及其主要代谢物氟苯尼考胺（FFA）残留。
在最佳条件下，该方法的 50%结合抑制（IC₅₀）值分别为 0.195µg kg⁻¹ 用于 FFA 和 0.24µg kg⁻¹ 用于 FF。FF 和 FFA 的交叉反应率分别为 100.0%和 81.2%。FF 和 FFA 可使用 FF/FFA 提取缓冲液从动物肉产品中轻松提取，回收率为 81.8-92.0%（FF）和 77.2-100%（FFA）。从理论上讲，整个一步 CL-ciELISA 测试可以在 40 分钟内完成。该测定法的检测限（LOD）为 0.98µg kg⁻¹ 用于 FF 和 0.80µg kg⁻¹ 用于 FFA 在动物肉样品中。最后，使用 CL-ciELISA 方法分析了现场动物肉样品，结果与传统 ELISA 和先前报道的液相色谱-串联质谱法的结果吻合良好。
综合结果证实了这种更快的一步 CL-ciELISA 用于同时痕量分析 FF 和 FFA 的实用性。到目前为止，这是检测 FF 和 FFA 开发的最快的 ELISA 和 CL-ELISA 方法。
