State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China.
Talanta. 2013 Sep 30;114:160-6. doi: 10.1016/j.talanta.2013.04.013. Epub 2013 Apr 12.
A Ru(phen)3(2+) doped silica nanoparticle based immunochromatographic strip was developed for the rapid and quantitative detection of five common β-agonist (salbutamol (SAL), cimbuterol, terbutaline, clenbuterol, and brombuterol) residues in swine urine. The broad spectrum monoclonal antibodies generated by immunizing BALB/c mice with salbutamol conjugated cationic bovine serum albumin. The fluorescence intensities (FIs) of the strip on the test line (FIT) and control line (FIC) were determined using a strip reader. Parameters that influenced the antibody and antigen interaction on the test strip were investigated by recording FIT and FIC values, and the concept of FIT/FIC ratio was used to offset the inherent heterogeneity of the test strips and the effect of the sample matrix. Under optimal conditions, the linear range for the quantitative detection of SAL was 0.6-5.0 ng/ml with a half maximal inhibitory concentration at 1.78 ng/ml. The limit of detection for real swine urine was 0.43 ng/ml. The recovery rates of the intraassay for spiked urine at SAL concentrations of 0.8, 1.5, and 3.5 ng/mL were 88.06%±3.75%, 95.77%±5.33%, and 94.06%±7.43%, whereas those for the interassay were 84.69%±5.0%, 95.06%±9.3%, and 88.34%±7.71%, respectively. The developed quantitative method exhibited excellent agreement with a commercially available competitive enzyme-linked immunosorbent assay kit for SAL-spiked urine samples, with a correlation of coefficient of 0.95 and a slope of 0.99 (n=36). The results indicated that the developed test strip enables sensitive, reproducible, and easily implementable screening for the rapid and quantitative detection of β-agonist residues in swine urine.
基于 Ru(phen)3(2+)掺杂二氧化硅纳米颗粒的免疫层析条用于快速定量检测猪尿液中五种常见β-激动剂(沙丁胺醇(SAL)、西马特罗、特布他林、克伦特罗和溴布特罗)残留。通过用沙丁胺醇偶联阳离子牛血清白蛋白免疫 BALB/c 小鼠产生广谱单克隆抗体。使用条带读取器测定条带上测试线(FIT)和控制线(FIC)的荧光强度(FI)。通过记录 FIT 和 FIC 值研究了影响测试带上抗体和抗原相互作用的参数,并使用 FIT/FIC 比值的概念来抵消测试带的固有异质性和样品基质的影响。在最佳条件下,SAL 的定量检测线性范围为 0.6-5.0ng/ml,半抑制浓度为 1.78ng/ml。实际猪尿液的检测限为 0.43ng/ml。在 SAL 浓度为 0.8、1.5 和 3.5ng/mL 的尿液中进行的内分析回收率分别为 88.06%±3.75%、95.77%±5.33%和 94.06%±7.43%,而在间分析中的回收率分别为 84.69%±5.0%、95.06%±9.3%和 88.34%±7.71%。与市售的用于 SAL 加标尿液样品的竞争性酶联免疫吸附测定试剂盒相比,所开发的定量方法具有极好的一致性,相关系数为 0.95,斜率为 0.99(n=36)。结果表明,该测试条能够灵敏、重现且易于实施,用于快速定量检测猪尿液中的β-激动剂残留。