Electrochemical Immunosensor for the Detection of Aflatoxin B1 in Palm Kernel Cake and Feed Samples

被引:25
作者
Azri, Farah Asilah [1 ]
Selamat, Jinap [1 ,2 ]
Sukor, Rashidah [1 ,2 ]
机构
[1] Univ Putra Malaysia, Inst Trop Agr & Food Secur, Food Safety & Food Integr FOSFI, Serdang 43400, Selangor, Malaysia
[2] Univ Putra Malaysia, Fac Food Sci & Technol, Dept Food Sci, Serdang 43400, Selangor, Malaysia
关键词
PKC; feed; electrochemical immunosensor; AFB(1); ELISA; nanocomposite; CARBON NANOTUBES; MYCOTOXINS; FOOD;
D O I
10.3390/s17122776
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Palm kernel cake (PKC) is the solid residue following oil extraction of palm kernels and useful to fatten animals either as a single feed with only minerals and vitamins supplementation, or mixed with other feedstuffs such as corn kernels or soy beans. The occurrence of mycotoxins (aflatoxins, ochratoxins, zearalenone, and fumonisins) in feed samples affects the animal's health and also serves as a secondary contamination to humans via consumption of eggs, milk and meats. Of these, aflatoxin B-1 (AFB(1)) is the most toxically potent and a confirmed carcinogen to both humans and animals. Methods such as High Performance Liquid Chromatography (HPLC) and Liquid Chromatography-Mass Spectrometry (LC-MS/MS) are common in the determination of mycotoxins. However, these methods usually require sample pre-treatment, extensive cleanup and skilled operator. Therefore, in the present work, a rapid method of electrochemical immunosensor for the detection of AFB(1) was developed based on an indirect competitive enzyme-linked immunosorbent assay (ELISA). Multi-walled carbon nanotubes (MWCNT) and chitosan (CS) were used as the electrode modifier for signal enhancement. N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) activated the carboxyl groups at the surface of nanocomposite for the attachment of AFB(1)-BSA antigen by covalent bonding. An indirect competitive reaction occurred between AFB(1)-BSA and free AFB(1) for the binding site of a fixed amount of anti-AFB(1) antibody. A catalytic signal based on horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2) and 3,3 0,5,5 0 -tetramethylbenzidine (TMB) mediator was observed as a result of attachment of the secondary antibody to the immunoassay system. As a result, the reduction peak of TMB(Ox) was measured by using differential pulse voltammetry (DPV) analysis. Based on the results, the electrochemical surface area was increased from 0.396 cm(2) to 1.298 cm(2) due to the electrode modification with MWCNT/CS. At the optimal conditions, the working range of the electrochemical immunosensor was from 0.0001 to 10 ng/mL with limit of detection of 0.1 pg/mL. Good recoveries were obtained for the detection of spiked feed samples (PKC, corn kernels, soy beans). The developed method could be used for the screening of AFB(1) in real samples.
引用
收藏
页数:11
相关论文
共 27 条
[1]  
Adesehinwa A.O.K., 2007, BULG J AGRIC SCI, V13, P593
[2]  
Agriculture and Horticulture Development Board (AHDB), 2015, FEED MAIZ SIL
[3]  
[Anonymous], 2005, PALM OIL DEV
[4]  
Asgari M., 2013, ISRN Electrochem, V2013, P1, DOI [10.1155/2013/564784, DOI 10.1155/2013/564784]
[5]  
Azri FA., 2017, Asian J Chem, V29, P31, DOI [DOI 10.14233/AJCHEM.2017.20104, 10.14233/ajchem.2017.20104]
[6]   Biosensors based on carbon nanotubes [J].
Balasubramanian, Kannan ;
Burghard, Marko .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2006, 385 (03) :452-468
[7]   Carry-Over of Aflatoxin B1 to Aflatoxin M1 in High Yielding Israeli Cows in Mid- and Late-Lactation [J].
Britzi, Malka ;
Friedman, Shmulik ;
Miron, Joshua ;
Solomon, Ran ;
Cuneah, Olga ;
Shimshoni, Jakob A. ;
Soback, Stefan ;
Ashkenazi, Rina ;
Armer, Sima ;
Shlosberg, Alan .
TOXINS, 2013, 5 (01) :173-183
[8]  
Farah AA, 2016, INT FOOD RES J, V23, P1849
[9]  
Idrus Z, CHEAPER POULTRY FEED
[10]   Surface Modification and Characterization of Multi-Walled Carbon Nanotube [J].
Kanbur, Yasin ;
Kucukyavuz, Zuhal .
FULLERENES NANOTUBES AND CARBON NANOSTRUCTURES, 2011, 19 (06) :497-504