College of Life Science, Chongqing Normal University, Chongqing 401331, PR China.
Chongqing Institute of Biotechnology, Chongqing 401123, PR China.
J Pharm Biomed Anal. 2018 Jan 5;147:417-424. doi: 10.1016/j.jpba.2017.06.068. Epub 2017 Jul 20.
Monitoring and rapid evaluation of nitrofurantoin metabolite, 1-aminohydantoin (AHD), are important for food safety and human health. Herein, we established the monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and quantum dots (QDs)-fabricated fluorescence-linked immunosorbent assay (FLISA). Monoclonal antibody specific to nitrophenyl derivative of AHD was derived from hybridoma cell lines 3.2.4/5A8. For another, CdTe core QDs with emission wavelength of 605nm were also synthesized. The performances of the proposed ic-ELISA and FLISA were further examined and the corresponding results were also validated by standard LC-MS/MS analysis. The obtained results indicated that both ic-ELISA and FLISA exhibited good dynamic linear detection for NPAHD over the range from 0.1 to 3.0ngmL. Meanwhile, proposed immunosorbent assays are characterized by satisfactory recovery rates of 81.5-113.7%. The experimental data suggested these two immunoassays could be facile, cost-effective and rapid tools for the prospective quantitative method for AHD analysis in food matrix.
监测和快速评估硝基呋喃代谢物 1-氨基乙内酰脲(AHD)对于食品安全和人类健康非常重要。为此,我们建立了基于单克隆抗体的间接竞争酶联免疫吸附测定(ic-ELISA)和量子点(QDs)制备的荧光酶联免疫吸附测定(FLISA)。针对 AHD 的硝苯基衍生物的单克隆抗体是从杂交瘤细胞系 3.2.4/5A8 中衍生出来的。此外,还合成了发射波长为 605nm 的 CdTe 核 QDs。进一步考察了所提出的 ic-ELISA 和 FLISA 的性能,并通过标准 LC-MS/MS 分析验证了相应的结果。获得的结果表明,ic-ELISA 和 FLISA 均对 NPAHD 表现出良好的动态线性检测范围为 0.1 至 3.0ngmL。同时,所提出的免疫分析具有令人满意的回收率 81.5-113.7%。实验数据表明,这两种免疫分析方法可作为食品基质中 AHD 分析的有前途的定量方法的简便、经济高效和快速工具。