Development of an immunochromatographic assay for rapid and quantitative detection of clenbuterol in swine urine
Development of an immunochromatographic assay for rapid and quantitative detection of clenbuterol in swine urine
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DOI：
10.1016/j.foodcont.2013.06.021
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发表时间：
2013-12-01
期刊：
影响因子：
6
通讯作者：
Xiong, Yonghua
中科院分区：
文献类型：
--
作者：
Li, Chaohui;Luo, Wei;Xiong, Yonghua
开发了侧流试纸条测试，用于在 10 分钟的总测定时间内定量检测猪尿液中的克仑特罗 (CLE)。这种新的 CLE 定量系统的开发理念是利用测试线 (AT) 与对照线 (A(C)) 的光密度 (OD) 之比来抵消侧流测试条 (LFTSAT/AC) 固有的不均匀性和样品基质的影响。在尿样中优化了免疫反应时间、分析物浓度、反应温度和反应 pH 值以及离子强度等各种参数。基于免疫反应的测定动力学表明，根据 CLE 浓度，在孵育 5-10 分钟内实现恒定信号。使用优化的参数，在 10 分钟的总测定时间内，使用 LFTSAT/AC 定量检测 CLE 的线性范围在 100 pg/mL 至 2500 pg/mL 之间。 CLE LFTSAT/AC 显示半数最大抑制浓度 (IC50) 为 460+/-60 pg/mL (n = 5) 和可靠的相关系数 (R-2 = 0.996)。 CLE 测试条与马布特罗、异丙肾上腺素、沙丁胺醇、特布他林、溴布特罗、莱克多巴胺和班布特罗表现出不显着的交叉反应 (Cr)。猪尿液的检测限 (LOD) 为 220 pg/mL，加标样品的回收率在 500 pg/mL 时分别为 102.35% +/- 7.66%，在 1000 pg/mL 时为 95.07% +/- 7.14%，在 1500 pg/mL 时为 100.65% +/- 10.21%。 500-1500 pg/mL CLE 浓度下的测定内和测定间精度显示出变异系数 (CV)
A lateral flow strip test was developed for quantitative detection of clenbuterol (CLE) in swine urine in a 10 min total assay time. This new quantitative system for CLE was developed based on the concept of using the ratio of the optical densities (ODs) of the test line (AT) to that of the control line (A(C)) to offset the inherent heterogeneity of lateral flow test strips (LFTSAT/AC) and the effect of sample matrix. Various parameters such as immunoreaction time, the concentration of analyte, reaction temperature and pH of the reactions, as well as the ionic strength were optimized in urine samples. The assay kinetics based on the immunological reactions indicated achievement of constant signal within 5-10 min of incubation depending upon the CLE concentration. Using the optimized parameters, the linear range for the quantitative detection of CLE using the LFTSAT/AC was between 100 pg/mL and 2500 pg/mL over a 10 min total assay time. The CLE LFTSAT/AC showed a half maximal inhibitory concentration (IC50) of 460+/-60 pg/mL (n = 5) and a reliable coefficient of correlation (R-2 = 0.996). The CLE test strip exhibited insignificant cross-reaction (Cr) to mabuterol, isoproterenol, salbutamol, terbutaline, brombuterol, ractopamine and bambuterol. The limit of detection (LOD) for swine urine was 220 pg/mL, and recoveries for spiked sample were 102.35% +/- 7.66% at 500 pg/mL, 95.07% +/- 7.14% at 1000 pg/mL and 100.65% +/- 10.21% at 1500 pg/mL, respectively. The intra-and inter-assay precision at 500-1500 pg/mL CLE concentrations showed coefficients of variation (CVs) at