title: A novel immunochromatographic assay based on a time-resolved chemiluminescence strategy for the multiplexed detection of ractopamine and clenbuterol.
abstract: A novel multiplexed immunochromatographic assay (ICA) based on a time-resolved chemiluminescence (CL) strategy was developed for quantitative detection of β-agonists, by utilizing ractopamine (RAC) and clenbuterol (CLE) as the models. Different from conventional multiplexed ICA methods which usually require two or more test lines, this strategy was developed for detection of two β-agonists by using only one test line on the nitrocellulose membrane. In this study, horseradish peroxidase and alkaline phosphatase were used as the signal probes to label RAC antibody and CLE antibody, respectively. The two CL reactions with flash type and glow type kinetics characteristics were triggered simultaneously by injecting the coreactants, then the signals for RAC and CLE detections were recorded at 3 s and 300 s after coreactants injection, respectively. Owing to the utilization of CL detection, this protocol showed ideal sensitivity for quantitation. Under the optimal conditions, the detection limits for RAC and CLE were 0.17 ng mL(-1) and 0.067 ng mL(-1) (S/N = 3), respectively. The whole assay process can be accomplished within 20 min without complicated sample pretreatment. The proposed method was successfully applied for the detection of RAC and CLE in spiked swine urine. It opens up a new pathway for designing a low cost, time-efficiency and multiplexed strategy for rapid screening and field assay.
authors.affiliation: Key Laboratory of Luminescence and Real-Time Analytical Chemistry, Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Wang
authors.affiliation: Key Laboratory of Luminescence and Real-Time Analytical Chemistry, Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China.
authors.forename: Xiaoxiao
authors.initials: X
authors.lastname: Su
authors.affiliation: Key Laboratory of Luminescence and Real-Time Analytical Chemistry, Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Ouyang
authors.affiliation: Key Laboratory of Luminescence and Real-Time Analytical Chemistry, Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory of Luminescence and Real-Time Analytical Chemistry, Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China. Electronic address: fuzf@swu.edu.cn.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Fu
mesh_terms: D000318:Adrenergic beta-Agonists / Q000032:analysis*; D000469:Alkaline Phosphatase / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D002976:Clenbuterol / Q000032:analysis*; D006735:Horseradish Peroxidase / Q000737:chemistry; D007700:Kinetics; D008163:Luminescent Measurements; D010627:Phenethylamines / Q000032:analysis*
chemical_list: D000318:Adrenergic beta-Agonists; D010627:Phenethylamines; C060897:ractopamine; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase; D002976:Clenbuterol
keywords: Chemiluminescence; Clenbuterol; Immunochromatographic assay strip; Multiplexed immunoassay; Ractopamine