foods Review Recent Advances in Conventional Methods and Electrochemical Aptasensors for Mycotoxin Detection Jing Yi Ong 1, Andrew Pike 2 and Ling Ling Tan 1,* 1 Address Southeast Asia Disaster Prevention Research Initiative (SEADPRI-UKM), Institute for Environment and Development (LESTARI), Universiti Kebangsaan Malaysia, UKM Bangi, Selangor Darul Ehsan 43600, Malaysia; [email protected] 2 School of Natural and Environmental Sciences, Bedson Building, Newcastle University, Newcastle upon Tyne NE1 7RU, UK; [email protected] * Correspondence: [email protected]; Tel.: +603-8921-7636; Fax: +603-8927-5629 Abstract: The presence of mycotoxins in foodstuffs and feedstuffs is a serious concern for human health. The detection of mycotoxins is therefore necessary as a preventive action to avoid the harmful contamination of foodstuffs and animal feed. In comparison with the considerable expense of treating contaminated foodstuffs, early detection is a cost-effective way to ensure food safety. The high affinity of bio-recognition molecules to mycotoxins has led to the development of affinity columns for sample pre-treatment and the development of biosensors for the quantitative analysis of mycotoxins. Aptamers are a very attractive class of biological receptors that are currently in great demand for the development of new biosensors. In this review, the improvement in the materials and methodology, and the working principles and performance of both conventional and recently developed methods are discussed. The key features and applications of the fundamental recognition elements, such as antibodies and aptamers are addressed. Recent advances in aptasensors that are based on different electrochemical (EC) transducers are reviewed in detail, especially from the Citation: Ong, J.Y.; Pike, A.; Tan, L.L. perspective of the diagnostic mechanism; in addition, a brief introduction of some commercially Recent Advances in Conventional available mycotoxin detection kits is provided. Methods and Electrochemical Aptasensors for Mycotoxin Detection. Keywords: mycotoxin; electrochemical; aptasensor; biosensor Foods 2021, 10, 1437. https://doi.org/ 10.3390/foods10071437 Academic Editor: Johanna Lindahl 1. Introduction Mycotoxins have been an agricultural concern ever since the emergence of cultivation Received: 23 May 2021 activities when humans first started to farm and store their food [1]. A mycotoxin is a Accepted: 15 June 2021 secondary metabolite, mainly produced by the Fusarium and Aspergillus fungi that are found Published: 22 June 2021 on foodstuffs and feedstuffs [2]. The first mycotoxin, aflatoxin (AF), was discovered after a severe incident where 100,000 turkeys died after consuming contaminated groundnuts in Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in 1962 [3]. Since then, many types of mycotoxins such as citrinin (CIT), fumonisin (FUM), published maps and institutional affil- ochratoxin A (OTA), patulin (PAT), zearalenone (ZEN), ergot alkaloids and trichothecenes iations. such as T-2 toxin (T-2) were characterised during the 1960s and 1970s [3]. Some of the chemical structures of common mycotoxins in foodstuffs and feedstuffs are shown in Figure1[4–6]. At present, mycotoxins are still a global concern, as they are prevalent in foodstuffs and feedstuffs that are widely consumed, such as maize, wheat, peanuts, milk and sorghum. Copyright: © 2021 by the authors. Researchers have found that the maize and wheat in China, Africa and Europe are contam- Licensee MDPI, Basel, Switzerland. This article is an open access article inated by various mycotoxins to different degrees [7,8]. The prevalence of mycotoxin types distributed under the terms and differs according to the environmental conditions. In China, contamination with aflatoxin conditions of the Creative Commons B1 (AFB1), deoxynivalenol (DON) and ZEN is prevalent in maize and wheat, whereas Attribution (CC BY) license (https:// FUM and AFB1 are prevalent in maize and sorghum grain, respectively, in Africa [7,8]. creativecommons.org/licenses/by/ The situation in Europe is estimated to worsen in the next 30 years due to climate change, 4.0/). Foods 2021, 10, 1437. https://doi.org/10.3390/foods10071437 https://www.mdpi.com/journal/foods Foods 2021, 10, x FOR PEER REVIEW 2 of 34 Foods 2021, 10, 1437 2 of 31 climate change, which leads to more favourable conditions for Aspergillus flavus to pro- whichliferate leads [9]. The to more production favourable of AFs, conditions CIT, OTA for andAspergillus PAT is flavusoften tocaused proliferate by the [9 Aspergillus]. The pro- ductionspecies, ofwhile AFs, ZEN CIT, and OTA DON and are PAT produced is often caused by the byFusarium the Aspergillus species species,[6]. Mycotoxin while ZENcon- andtamination DON are of producedfoodstuffs by and the feFusariumedstuffsspecies is a potential [6]. Mycotoxin threat to contamination public health ofand foodstuffs is espe- andcially feedstuffs concerning isa in potential China, one threat of tothe public main healthglobal anddistributors is especially of maize concerning and wheat in China, [10]. oneTherefore, of the effective main global and distributorsaffordable method of maizes for and mycotoxin wheat [10 detection]. Therefore, are indispensable effective and affordablefor maintaining methods high for levels mycotoxin of global detection food aresafety. indispensable Recently, forthemaintaining classification, high toxicity, levels ofcharacteristics global food and safety. contamination Recently, the of classification, mycotoxin in toxicity, specific characteristicsfood and feeds and were contamina- reviewed tionalong of with mycotoxin several in analytical specific food methods, and feeds including were reviewed instrumental along and with sensor several techniques analytical methods,[5,6,11]. Biosensors including instrumentalwith different and recognit sensorion techniques elements [5 ,6that,11]. focus Biosensors on nanostructured with different recognitionmaterials and elements analytical that techniques focus on nanostructured such as electrochemistry, materials and electrochemiluminescence analytical techniques such asand electrochemistry, photoelectrochemistry electrochemiluminescence are the recent trend andin mycotoxin photoelectrochemistry detection [12–14]. are the recent trendHerein, in mycotoxin the current detection trends [12 in–14 mycotoxin]. detection methods, including both conven- tionalHerein, and more the currentrecently trends developed in mycotoxin methods, detection are discussed methods, in detail. including The bothcurrent conven- pre- tionalferred andmethod more is recently based on developed an electrochemical methods, are (EC)-based discussed inaptasensor. detail. The Different current preferred types of methodmolecular is basedrecognition on an electrochemicalelements that are (EC)-based used in the aptasensor. development Different of biosensors, types of molecular such as recognitionantibodies, aptamers elements that(Apts) are and used molecularl in the developmenty imprinted of polymers biosensors, (MIPs), such are as antibodies, reviewed. aptamersFurthermore, (Apts) the and key molecularlycommercially imprinted available polymers mycotoxin (MIPs), detection are reviewed. products are Furthermore, discussed thein order key commerciallyto provide an availableoverview mycotoxinof the technologies detection that products are in current are discussed use. This in review order tois provideintended an to overview pave a clear of the path technologies for the future that advancement are in current of use. mycotoxin This review detection. is intended to pave a clear path for the future advancement of mycotoxin detection. FigureFigure 1.1. The chemical structures of of ( (aa)) aflatoxin aflatoxin B B11, ,((bb) )aflatoxin aflatoxin B B2,2 (,(c)c aflatoxin) aflatoxin G G1, 1(,(d)d aflatoxin) aflatoxin G2 G, (2e,() aflatoxine) aflatoxin M1 M, (f1), (ochratoxinf) ochratoxin A, A,(g) ( gzearalenone,) zearalenone, (h) ( hdeoxynivalenol,) deoxynivalenol, (i) (patulini) patulin and and (j) (fumonisinj) fumonisin B1. B 1. 2. Conventional and Advanced Analytical Technologies 2. Conventional and Advanced Analytical Technologies The detection of mycotoxins has been explored for nearly 60 years, ever since their The detection of mycotoxins has been explored for nearly 60 years, ever since their characterisation in the early 1960s. Although around 400 mycotoxins have now been characterisation in the early 1960s. Although around 400 mycotoxins have now been discovered, only a portion have high toxicity and are also prominent in foodstuffs and discovered, only a portion have high toxicity and are also prominent in foodstuffs and feedstuffs [2]. Screening for mycotoxins is the most cost-effective way to prevent harmful feedstuffs [2]. Screening for mycotoxins is the most cost-effective way to prevent harmful mycotoxins from entering the food chain. Therefore, many analytical techniques have been mycotoxins from entering the food chain. Therefore, many analytical techniques have developed to ensure food safety and security. been developed to ensure food safety and security. Foods 2021, 10, 1437 3 of 31 Most of the analytical techniques are based on chromatographic and immunological methods. Large instruments, such as high-performance liquid chromatography (HPLC), liquid chromatography–mass spectrometry (LC-MS) and gas chromatography–mass spec- trometry (GC-MS), were routinely used during the early developmental stages of mycotoxin detection. Their key advantages are that