1(College of Food Science and Technology,Nanchang University,Nanchang 330031,China); 2(State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047,China); 3(Jiangxi-OAI Joint Research Institute,Nanchang University,Nanchang 330047,China)
Abstract: This study aims to prepare a novel core-shell heterostructure magnetic gold nanoparticles (MGNPs) with high magnetic and optical strength,and then it was used as a bifunctional probe to establish a new immunochromatographic method (ICA) for highly-sensitive detection of clenbuterol (Clen) in swine urine.A facile emulsion self-assembly strategy was used to synthesize MGNPs,and the EDC method was used to prepare a bifunctional signal probe by coupling anti-Clen monoclonal antibody onto the surface of MGNPs.Moreover,the competition inhibition curve for the quantitative detection of Clen was established,and the dynamic detection range and detection sensitivity of the developed MGNP-ICA were then determined.The accuracy and precision of the MGNPs-ICA were also been evaluated by a standard recovery experiment,and the reliability of the MGNPs-ICA was estimated by a commercial ELISA kit.Results showed that the as-prepared MGNPs have high saturation magnetization (62.2% of the initial iron oxide nanoparticles) and strong absorbance (8 times higher than that of traditional 30 nm colloidal gold nanoparticles).Taking advantage of their excellent magnetic properties and optical sensing capacity,the detection range of MGNPs-ICA for Clen was 0.078-40 ng/mL.Furthermore,the half-maxinal inhibitory concentration was 1.225 ng/mL with the limit of detection 0.041 ng/mL.And this assay could be finished in 15 min.The recoveries of intra- and inter-assay of MGNPs-ICA range from 93.66% to 114.80% with the coefficient of variation (CV) of below 13.45%.In addition,this method had a good correlation with the detection results obtained from commercial ELISA kits.
LOZANO M S,CHAVEZ J F,LOPEZ F A,et al.Horse meat sold as beef and consequent clenbuterol residues in the unregulated Mexican marketplace[J].Food Control,2020,110:107 028.
[3]
TANG Yiwei,GAO Jingwei,LIU Xiuying,et al.Ultrasensitive detection of clenbuterol by a covalent imprinted polymer as a biomimetic antibody[J].Food Chemistry,2017,228:62-69.
[4]
DUFAYET L,GORGIARD C,VAYSSETTE F,et al.Death of an apprentice bodybuilder following 2,4-dinitrophenol and clenbuterol intake[J].International Journal of Legal Medicine,2020,134:1 003-1 006.
XU J,LI Y,GUO J,et al.Fluorescent detection of clenbuterol using fluorophore functionalized gold nanoparticles based on fluorescence resonance energy transfer[J].Food Control,2014,46:67-74.
[7]
朱自平.高效液相色谱检测猪肉盐酸克伦特罗的方法优化[J].现代食品,2017(16):95-99.
[8]
LIA L A,TOMLINSON J A,SATZGER R D,et al.Detection of clenbuterol in bovine retinal tissue by high-performance liquid chromatography with electrochemical detection[J].Journal of Chromatography A,1997,762(1-2):275-280.
[9]
DUVIVIER W F,VAN BEEK T A V,MEIJER T,et al.Ultratrace LC-MS/MS analysis of segmented calf hair for retrospective assessment of time of clenbuterol administration in agriforensics[J].Journal of Agricultural and Food Chemistry,2015,63(2):493-499.
MA Luyao,NILGHAZ A,CHOI J R,et al.Rapid detection of clenbuterol in milk using microfluidic paper-based ELISA[J].Food Chemistry,2018,246:437-441.
[12]
PENG Dapeng,ZHANG Lingyan,SITU C,et al.Development of monoclonal antibodies and indirect competitive enzyme-linked immunosorbent assay kits for the detection of clenbuterol and salbutamol in the tissues and products of food-producing animals[J].Food Analytical Methods,2017,10(11):3 623-3 633.
[13]
HUANG Zhen,XIONG Zhijuan,CHEN Yuan,et al.Sensitive and matrix-tolerant lateral flow immunoassay based on fluorescent magnetic nanobeads for the detection of clenbuterol in swine urine[J].Journal of Agricultural and Food Chemistry,2019,67(10):3 028-3 036.
[14]
LI Yu,ZHOU Yaofeng,CHEN Xirui,et al.Comparison of three sample addition methods in competitive and sandwich colloidal gold immunochromatographic assay[J].Analytica Chimica Acta,2020,1 094:90-98.
YU Q,LIU J,ZHAO G,et al.A silica nanoparticle based 2-color immunochromatographic assay for simultaneous determination of clenbuterol and ractopamine[J].Mikrochimica Acta,2019,186(7):1-9.
[17]
HUANG Z,PENG J,HAN J,et al.A novel method based on fluorescent magnetic nanobeads for rapid detection of Escherichia coli O157:H7[J].Food Chemistry,2019,276:333-341.
[18]
GUO L,SHAO Y,DUAN H,et al.Magnetic quantum dot nanobead-based fluorescent immunochromatographic assay for the highly sensitive detection of aflatoxin B1 in dark soy sauce[J].Analytical Chemistry,2019,91(7):4 727-4 734.
CHOWDHURY A D,AGNIHOTRI N,DOONG R,et al.Label-free and nondestructive separation technique for isolation of targeted DNA from DNA-protein mixture using magnetic Au-Fe3O4 nanoprobes[J].Analytical Chemistry,2017,89(22):12 244-12 251.
HUI W,ZHANG S,ZHANG C,et al.A novel lateral flow assay based on GoldMag nanoparticles and its clinical applications for genotyping of MTHFR C677T polymorphisms[J].Nanoscale,2016,8(6):3 579-3 587.
[23]
YOU S M,LUO K,JUNG J Y,et al.Gold nanoparticle-coated starch magnetic beads for the separation,concentration and SERS-based detection of E.coli O157:H7[J].ACS Applied Materials &Interfaces,2020,12(16):18 292-18 300.
[24]
MAZUEL F,ESPINOSA A,RADTKE G,et al.Magneto-thermal metrics can mirror the long-term intracellular fate of magneto-plasmonic nanohybrids and reveal the remarkable shielding effect of gold[J].Advanced Functional Materials,2017,27(9):1 605 997.
[25]
HU Y,MENG L,NIU L,et al.Facile Facile synthesis of superparamagnetic Fe3O4@polyphosphazene@Au shells for magnetic resonance imaging and photothermal therapy[J].ACS Applied Materials &Interfaces,2013,5(11):4 586-4 591.
[26]
CHEN X,ZHU J,CHEN Z,et al.A novel bienzyme glucose biosensor based on three-layer Au-Fe3O4@SiO2 magnetic nanocomposite[J].Sensors and Actuators B.Chemical,2011,159(1):220-228.
[27]
AND H H,OSTERLOH F E.A simple large-scale synthesis of nearly monodisperse gold and silver nanoparticles with adjustable sizes and with exchangeable surfactants[J].Chemistry of Materials,2004,16(13):2 509-2 511.
[28]
FU R,JIN X,LIANG J,et al.Preparation of nearly monodispersed Fe3O4/SiO2 composite particles from aggregates of Fe3O4 nanoparticles[J].Journal of Materials Chemistry,2011,21(39):15 352-15 356.
[29]
HAO L,LENG Y,ZENG L,et al.Core-shell-heterostructured magnetic-plasmonic nanoassemblies with highly retained magnetic-plasmonic activities for ultrasensitive bioanalysis in complex matrix[J].Advanced Science,2020,7(2):1 092 433.
2020, Vol. 46 
