Journal
ANALYTICA CHIMICA ACTA
Volume 1040, Issue -, Pages 143-149Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2018.08.014
Keywords
AuNCs-MLFIA sensor; Arg/ATT/AuNCs; Clenbuterol; Ractopamine; Multiplex detection
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Funding
- National Natural Science Foundation of China [31672600]
- Sanming Project of Medicine in Shenzhen [SZSM201611068]
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A multiplex lateral flow immunoassay sensor based on highly luminescent green-emitting Au nanoclusters (AuNCs-MLFIA sensor) was successfully established for the simultaneous and quantitative determination of clenbuterol (Clen) and ractopamine (RAC) in swine urine. The antigens of Clen and RAC were dispersed on a nitrocellulose membrane as two test lines, and the Au nanoclusters were synthesized from 6-aza-2-thiothymine and L-arginine to obtain highly green luminescence and ultra-small nanoparticles (Arg/ATT/AuNCs). Free carboxyl groups on Arg/ATT/AuNCs enabled conjugation with biomolecules to afford an indicator for the biosensor. The AuNCs-MLFIA sensor is based on the indirect competition assay and could successfully detect samples within 18 min without sample pretreatment, qualitative results can be obtained by visual inspection under a UV lamp. The limits of detection of Clen and RAC by the naked eye were both 0.25 mu g L-1. In addition, the AuNCs-MLFIA sensor allowed quantitative detection combined with a portable fluorescence reader. The half-maximal inhibitory concentrations of Clen and RAC were 0.06 and 0.32 mu g L-1, respectively, with detection limits of 0.003 and 0.023 mu g L-1. Thirty blind-spiked swine urine samples were analyzed by the AuNCs-MLFIA sensor and liquid chromatography-tandem mass spectrometry, and the results of the two methods showed a significant correlation. The newly developed AuNCs-MLFIA sensor overcomes several limitations of conventional LFIA sensors, including their low sensitivity, limitation to quantify analytes, and single-analyte detection. (C) 2018 Elsevier B.V. All rights reserved.
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