Electrochemical Immunosensor for the Detection of Aflatoxin B₁ in Palm Kernel Cake and Feed Samples.
Electrochemical Immunosensor for the Detection of Aflatoxin B₁ in Palm Kernel Cake and Feed Samples.
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DOI：
10.3390/s17122776
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发表时间：
2017-11-30
期刊：
影响因子：
--
通讯作者：
Sukor R
中科院分区：
文献类型：
--
作者：
Azri FA;Selamat J;Sukor R
棕榈仁饼（PKC）是棕榈仁提取油后的固体残渣，可作为仅补充矿物质和维生素的单一饲料或与其他饲料如玉米粒或大豆混合用于育肥动物。饲料样品中出现的霉菌毒素（黄曲霉毒素、赭曲霉毒素、玉米赤霉烯酮和伏马菌素）会影响动物的健康，也会通过食用鸡蛋、牛奶和肉类对人类造成二次污染。其中，黄曲霉毒素B1（AFB 1）是毒性最强的，并已证实对人类和动物都是致癌物质。高效液相色谱法（HPLC）和液相色谱-质谱法（LC-MS/MS）等方法在真菌毒素的测定中很常见。然而，这些方法通常需要样品的预处理，大量的净化和熟练的操作人员。因此，在本工作中，电化学免疫传感器的快速检测黄曲霉毒素B1的间接竞争酶联免疫吸附试验（ELISA）的基础上，建立了一种方法。多壁碳纳米管（MWCNT）和壳聚糖（CS）被用作电极修饰剂的信号增强。N-乙基-N′-（3-二甲氨基丙基）-碳二亚胺（EDC）和N-羟基琥珀酰亚胺（NHS）活化纳米复合材料表面的羧基，使AFB 1-BSA抗原共价结合。AFB_1-BSA与游离AFB_1之间存在间接竞争反应，竞争一定量的抗AFB_1抗体的结合位点。在存在过氧化氢（H2 O2）和3，3 ′，5，5 ′-四甲基联苯胺（TMB）介体的情况下，观察到基于辣根过氧化物酶（HRP）的催化信号，这是二抗附着到免疫测定系统的结果。结果，通过使用差分脉冲伏安法（DPV）分析测量TMB（Ox）的还原峰。基于结果，电化学表面积从0.396 cm 2增加到1.298 cm 2，由于与MWCNT/CS的电极改性。在最佳条件下，该电化学免疫传感器的工作范围为0.0001 ~ 10 ng/mL，检测限为0.1 pg/mL。该方法用于饲料样品（PKC、玉米粒、大豆）的检测，回收率良好。该方法可用于真实的样品中黄曲霉毒素B1的筛选。
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 B1 (AFB1) 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 AFB1 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 AFB1-BSA antigen by covalent bonding. An indirect competitive reaction occurred between AFB1-BSA and free AFB1 for the binding site of a fixed amount of anti-AFB1 antibody. A catalytic signal based on horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2) and 3,3′,5,5′-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 cm2 to 1.298 cm2 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 AFB1 in real samples.
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影响因子：
6.1
作者：
Lee, NA;Wang, S;Kennedy, IR
通讯作者：
Kennedy, IR
DOI：
10.1016/j.ijfoodmicro.2007.07.019
发表时间：
2007-10-20
影响因子：
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作者：
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DOI：
10.1080/1536383x.2010.494778
发表时间：
2011-01-01
影响因子：
2.3
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通讯作者：
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影响因子：
1.5
作者：
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通讯作者：
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DOI：
10.1080/19440049.2014.978396
发表时间：
2014-12-02
影响因子：
2.9
作者：
Yibadatihan, S.;Jinap, S.;Mahyudin, N. A.
通讯作者：
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