• Vol 10, No 4 (2019)
  • Chemical Engineering

CdS Nanoparticle-based Biosensor Development for Aflatoxin Determination

Moh Hayat, Endang Saepudin, Yasuaki Einaga, Tribidasari A Ivandini

Corresponding email: ivandini.tri@sci.ui.ac.id


Cite this article as:
Hayat, M., Saepudin, E., Einaga, Y., Ivandini, T.A., 2019. CdS Nanoparticle-based Biosensor Development for Aflatoxin Determination. International Journal of Technology. Volume 10(4), pp. 787-797
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Moh Hayat Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Indonesia, Kampus UI Depok 16424, Indonesia
Endang Saepudin Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Indonesia, Kampus UI Depok 16424, Indonesia
Yasuaki Einaga Department of Chemistry, Keio University, Japan
Tribidasari A Ivandini Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Indonesia, Kampus UI Depok 16424, Indonesia
Email to Corresponding Author

Abstract
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An anodic stripping voltammetry detection method with lateral flow immunochromatographic assay for determining aflatoxin M1 was developed. Cadmium sulfide nanoparticles (CdSNPs) were employed as a label for aflatoxin antibody. Conjugation between the CdSNPs and aflatoxin antibody was performed as the first step. The CdSNP-antibody conjugate was then immobilized on a test strip on a conjugate pad. When the conjugate interacted with the sample containing aflatoxin, it was captured by the aflatoxin in the test zone. The CdSNPs retained in the test zone were then measured by anodic stripping voltammetry using a boron-doped diamond electrode. A linear concentration range of 0–70 ppb aflatoxin, with a sensitivity of 20 µA/mM and a detection limit of 30 ppb, was achieved by this method.

Aflatoxin; Anodic stripping voltammetry; Boron-doped diamond; CdS nanoparticles

Introduction

Aflatoxin is a secondary metabolite, mainly produced by Aspergillus flavus and Aspergillus parasiticus, which contaminate agricultural products, namely nuts, cereals and animal feeds (Bennett & Klich, 2003). Six important types of aflatoxin have been identified and are denoted as B1, B2, G1, G2, M1 and M2 (Wacoo et al., 2014). Among these types, the International Agency for Research on Cancer classified aflatoxin B1 (AFB1) as a group 1 carcinogenic substance due to its ability to disrupt the induction of certain enzymes and inhibit RNA synthesis (IARC, 2002). Meanwhile, aflatoxin M1 (AFM1) is a hydrolyzed metabolite of AFB1, produced by the mammary gland; hence, it is mainly found in milk and dairy products, such as cheese and yoghurt (Martins & Martins, 2000). Due to its stability during pasteurization and other heat treatment (Wang et al., 2009), as well as its hepatotoxicity and carcinogenic effects (Badea et al., 2004), the aflatoxin content must be controlled. The regulated upper limits for AFB1 and total aflatoxins (B1, B2, G1 and G2) are 30 and 50 µg/kg respectively, while that for AFM1 in milk is 1.0 µg/kg. (IARC, 2002).

In recent years, many analytical techniques for quantitative aflatoxin determination have been studied, including thin layer chromatography (TLC), gas chromatography (GC) and high-performance liquid chromatography (HPLC) (Roseanu et al., 2010). These techniques are sufficiently sensitive for aflatoxin; however, GC and HPLC operation is time-consuming and requires a skilled analyst, complex sample pre-treatment and expensive instrumentation, which affect aflatoxin detection in food products. Beside TLC, another fast detection method is immunochemistry. This can be used to detect aflatoxin in solid food (Betina, 1985; Shepard, 2009) as well as dairy products (Anfossi et al., 2008; Li et al., 2009). Immunochemistry has advantages with respect to its selectivity and sensitivity. One immunochemistry development is lateral flow immunoassay. Furthermore, enzyme-linked immunosorbent assay, which depends on a specific interaction between an antigen toxin target and its antibody, is also widely used to detect aflatoxin (Zhang et al., 2016). However, this method has several drawbacks, such as the requirement of an incubation period, together with time-consuming washing and mixing (Badea et al., 2004). Accordingly, the development of a new strategy for aflatoxin determination is urgently required.

To achieve a more suitable aflatoxin detection technique, several fast methods based on biosensors or immunosensor applications have recently been developed, including flow-injection immunoassay (Badea et al., 2004); a microcomb electrode modified with a self-assembly horseradish peroxidase (HRP) and anti-AFB1 (Liu et al., 2006); and electrochemical methods (Piermarini et al., 2007; Parker & Tothill, 2009; Linting et al., 2012; Chauhan et al., 2015; Zhang et al., 2016; Gouda et al., 2016; Kong et al., 2018; Peng et al., 2018). Additionally, Paniel et al. (2010) developed an electrochemical immunosensor for AFM1 in food products based on a competitive immunoassay with HRP as the label. A fast and simple immunochromatography test was also developed to detect AFB1 in less than 10 min. The developed AFB1 sensor test also shows no cross-reaction with ochratoxin A. (Moon et al., 2012).

Electrochemical measurements were have been performed by anodic stripping voltammetry (ASV) using a boron-doped diamond (BDD) as the working electrode. ASV is generally considered to be the most suitable method for trace metal detection, since this offers certain advantages for metal analysis, including excellent selectivity and sensitivity, a low detection limit, simple operation and a relatively low-cost process (Ivandini et al., 2012). BDD is superior to other conventional solid electrodes due to its wide potential window, low capacitance, chemical inertness, mechanical durability and exceptional biocompatibility (Fujishima et al., 2005, Ivandini & Einaga, 2017). The use of the ASV technique for immunochromatographic test strips using BDD electrodes has been reported (Wicaksono et al., 2014; Ivandini et al., 2015; Hayat et al., 2016).

The above methods mainly yield only qualitative results. The electrochemical method offers accurate quantitative results but requires coupling with another relatively complex method. In this research, a method for determining aflatoxin was developed which combines the speed and reproducibility of the immunochromatography technique with the excellent selectivity and sensitivity of the electrochemical method. As a label, cadmium sulfide nanoparticles (CdSNPs) were used.  These are suitable as labels in immunochromatographic sensors, since they are inexpensive and exhibit low potential oxidation, good electrical conductivity (Zheng et al., 2015), and high sensitivity (Du et al., 2012). Although no research has been conducted on the use of CdS as an aflatoxin label, CdSNPs have been investigated as labels for several biosensors, such as a cyanide ion sensor (Salariya et al., 2017), DNA (Svitkova et al., 2017), and glucose (Huang et al., 2005).  Nanoparticles can be prepared by several methods, including the microwave technique (Usman et al., 2018), the hydrothermal method (Fadli et al., 2017), the alkaline hydrolysis method (Khalil et al., 2018), and microemulsions (Mirgorod & Efimova, 2007).  The advantages of the synthesis of nanoparticles in microemulsions are that the process occurs in mild conditions, and that the size and shape of the nanoparticles can be controlled by the micelle structure and dynamics (Mirgorod & Efimova, 2007).

Conclusion

ASV of Cd2+ in 0.1 M HClO4 showed positive results when applied to CdSNP-based aflatoxin biosensors. A deposition potential of ?2.5 V with a deposition time of 700 s and a scan rate of 100 mV/s generated an oxidation peak at ?0.6 V. A linear correlation was observed between the decrease in the current peak and the increase in aflatoxin concentration in the range of 0–70 ppb, indicating that the method is promising for aflatoxin determination. The test strip was measured by anodic stripping voltammetry using a boron-doped diamond electrode. A linear concentration range of 0–70 ppb aflatoxin, with a sensitivity of 20 µA/mM and a detection limit of 30 ppb, was achieved by this method. 

Acknowledgement

This work was funded by Hibah Tugas Akhir Mahasiswa Doktor Universitas Indonesia, under Contract No. 1334/UN2.R3.1/HKP05.00/2018.

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