Toxins

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14 pages, 2247 KiB

Open AccessArticle

Carboxyl-Functionalized, Europium Nanoparticle-Based Fluorescent Immunochromatographic Assay for Sensitive Detection of Citrinin in Monascus Fermented Food

by Erjing Chen, Ying Xu, Biao Ma, Haifeng Cui, Chuanxin Sun and Mingzhou Zhang

Toxins 2019, 11(10), 605; https://doi.org/10.3390/toxins11100605 - 17 Oct 2019

Cited by 12 | Viewed by 4565

Abstract

A fluorescent immunochromatographic test strip (FICTS) based on the use of europium nanoparticles (EuNPs) was developed and applied to detect citrinin (CIT) in Monascus fermented food. The sensitivity of the immunoassay to detect CIT was greatly improved by the use of a specific [...] Read more.

A fluorescent immunochromatographic test strip (FICTS) based on the use of europium nanoparticles (EuNPs) was developed and applied to detect citrinin (CIT) in Monascus fermented food. The sensitivity of the immunoassay to detect CIT was greatly improved by the use of a specific monoclonal antibody to attach EuNPs to form a probe. Under optimum conditions, the visual detection limit was 2.5 ng/mL, and the detection limit of the instrument was 0.05 ng/mL. According to the results, the IC₅₀ was 0.4 ng/mL. Matrix interference from various Monascus fermented foods was investigated in food sample detection. The immunosensor also demonstrated high recoveries (86.8–113.0%) and low relative standard deviations (RSDs) (1.8–15.3%) when testing spiked Monascus fermented food. The detection results of this method showed a good correlation (R² > 0.98) with high-performance liquid chromatography (HPLC). The results showed that the FICTS method could be used as a rapid, sensitive method to detect CIT in Monascus fermented food. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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12 pages, 2356 KiB

Open AccessArticle

Development of Real-Time Immuno-PCR Based on Phage Displayed an Anti-Idiotypic Nanobody for Quantitative Determination of Citrinin in Monascus

by Wenping Huang, Zhui Tu, Zhenqiang Ning, Qinghua He and Yanping Li

Toxins 2019, 11(10), 572; https://doi.org/10.3390/toxins11100572 - 30 Sep 2019

Cited by 16 | Viewed by 4110

Abstract

Citrinin (CIT) is a mycotoxin that has been detected in agricultural products, feedstuff, and Monascus products. At present, research has been performed to develop methods for CIT detection, mainly through TLC, HPLC, biosensor, and immunoassay. The immunoassay method is popular with researchers because [...] Read more.

Citrinin (CIT) is a mycotoxin that has been detected in agricultural products, feedstuff, and Monascus products. At present, research has been performed to develop methods for CIT detection, mainly through TLC, HPLC, biosensor, and immunoassay. The immunoassay method is popular with researchers because of its speed, economy, simplicity, and ease of control. However, mycotoxins are inevitably introduced during the determination. Immunoassays require the use of toxins coupled to carrier proteins or enzymes to make competitive antigens. In this study, anti-idiotypic nanobody X27 as CIT mimetic antigen was used as non-toxic surrogate reagents in immunoassay. Therefore, the X27-based real-time immuno-PCR (rtIPCR) method had been established after optimal experiments of annealing temperature and amplification efficiency of real-time PCR, concentration of coating antibody, phage X27, and methyl alcohol. The IC₅₀ value of the established method in the present study is 9.86 ± 2.52 ng/mL, which is nearly equivalent to the traditional phage ELISA method. However, the linear range is of 0.1–1000 ng/mL, which has been broadened 10-fold compared to the phage ELISA method. Besides, the X27-based rtIPCR method has no cross-reactivity to the common mycotoxins, like aflatoxin B₁ (AFB₁), deoxynivalenol (DON), ochratoxin A (OTA), and zearalenone (ZEN). The method has also been applied to the determination of CIT in rice flour and flour samples, and the recovery was found to be in the range of 90.0–104.6% and 75.8–110.0% respectively. There was no significant difference in the results between the rtIPCR and UPLC–MS. The anti-idiotypic nanobody as a non-toxic surrogate of CIT makes rtIPCR a promising method for actual CIT analysis in Monascus products. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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9 pages, 2229 KiB

Open AccessEditor’s ChoiceCommunication

Graphene-Based Sensing Platform for On-Chip Ochratoxin A Detection

by Nikita Nekrasov, Dmitry Kireev, Aleksei Emelianov and Ivan Bobrinetskiy

Toxins 2019, 11(10), 550; https://doi.org/10.3390/toxins11100550 - 20 Sep 2019

Cited by 29 | Viewed by 6349

Abstract

In this work, we report an on-chip aptasensor for ochratoxin A (OTA) toxin detection that is based on a graphene field-effect transistor (GFET). Graphene-based devices are fabricated via large-scale technology, allowing for upscaling the sensor fabrication and lowering the device cost. The sensor [...] Read more.

In this work, we report an on-chip aptasensor for ochratoxin A (OTA) toxin detection that is based on a graphene field-effect transistor (GFET). Graphene-based devices are fabricated via large-scale technology, allowing for upscaling the sensor fabrication and lowering the device cost. The sensor assembly was performed through covalent bonding of graphene’s surface with an aptamer specifically sensitive towards OTA. The results demonstrate fast (within 5 min) response to OTA exposure with a linear range of detection between 4 ng/mL and 10 pg/mL, with a detection limit of 4 pg/mL. The regeneration time constant of the sensor was found to be rather small, only 5.6 s, meaning fast sensor regeneration for multiple usages. The high reproducibility of the sensing response was demonstrated via using several recycling procedures as well as various GFETs. The applicability of the aptasensor to real samples was demonstrated for spiked red wine samples with recovery of about 105% for a 100 pM OTA concentration; the selectivity of the sensor was also confirmed via addition of another toxin, zearalenone. The developed platform opens the way for multiplex sensing of different toxins using an on-chip array of graphene sensors. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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15 pages, 895 KiB

Open AccessEditor’s ChoiceArticle

Evaluation of High-Resolution Mass Spectrometry for the Quantitative Analysis of Mycotoxins in Complex Feed Matrices

by Tolke Jensen, Marthe de Boevre, Nils Preußke, Sarah de Saeger, Tim Birr, Joseph-Alexander Verreet and Frank D. Sönnichsen

Toxins 2019, 11(9), 531; https://doi.org/10.3390/toxins11090531 - 12 Sep 2019

Cited by 25 | Viewed by 4694

Abstract

The selective and sensitive analysis of mycotoxins in highly complex feed matrices is a great challenge. In this study, the suitability of Orbitrap^TM-based high-resolution mass spectrometry (HRMS) for routine mycotoxin analysis in complex feeds was demonstrated by the successful validation of [...] Read more.

The selective and sensitive analysis of mycotoxins in highly complex feed matrices is a great challenge. In this study, the suitability of Orbitrap^TM-based high-resolution mass spectrometry (HRMS) for routine mycotoxin analysis in complex feeds was demonstrated by the successful validation of a full MS/data-dependent MS/MS acquisition method for the quantitative determination of eight Fusarium mycotoxins in forage maize and maize silage according to the Commission Decision 2002/657/EC. The required resolving power for accurate mass assignments (<5 ppm) was determined as 35,000 full width at half maximum (FWHM) and 70,000 FWHM for forage maize and maize silage, respectively. The recovery (R_A), intra-day precision (RSD_r), and inter-day precision (RSD_R) of measurements were in the range of 94 to 108%, 2 to 16%, and 2 to 12%, whereas the decision limit (CCα) and the detection capability (CCβ) varied from 11 to 88 µg/kg and 20 to 141 µg/kg, respectively. A set of naturally contaminated forage maize and maize silage samples collected in northern Germany in 2017 was analyzed to confirm the applicability of the HRMS method to real samples. At least four Fusarium mycotoxins were quantified in each sample, highlighting the frequent co-occurrence of mycotoxins in feed. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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12 pages, 1711 KiB

Open AccessEditor’s ChoiceArticle

A Novel Magnetic Molecular Imprinted Polymer for Selective Extraction of Zearalenone from Cereal Flours before Liquid Chromatography-Tandem Mass Spectrometry Determination

by Chiara Cavaliere, Michela Antonelli, Andrea Cerrato, Giorgia La Barbera, Aldo Laganà, Michele Laus, Susy Piovesana and Anna Laura Capriotti

Toxins 2019, 11(9), 493; https://doi.org/10.3390/toxins11090493 - 27 Aug 2019

Cited by 17 | Viewed by 4134

Abstract

Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin produced by various Fusarium species and commonly occurring in corn and other cereals. Even though its acute toxicity is low, still the estrogenic activity of ZEN and metabolites is a matter of concern. In this work, [...] Read more.

Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin produced by various Fusarium species and commonly occurring in corn and other cereals. Even though its acute toxicity is low, still the estrogenic activity of ZEN and metabolites is a matter of concern. In this work, a new magnetic molecularly imprinted polymer (mMIP) for the selective extraction of ZEN from cereal flours is presented. The mMIP was synthesized previously using quercetin as dummy template, and here we wanted to test its applicability to complex food samples. Analyte determination was carried out by high-performance liquid chromatography coupled to tandem mass spectrometry. The selectivity of the mMIP and the main validation method parameters were assessed. In particular, even in samples as complex as cereals, matrix effect was negligible. Although the mMIP showed cross-selectivity towards both ZEN-related and quercetin-related compounds, nonetheless ZEN recovery was > 95% for the two lower spiking levels, and the quantification limit was 0.14 ng g⁻¹, i.e., ca. 500 times lower than the maximum limit fixed for most cereals by European law. Therefore, the material, also in comparison with a commercial sorbent, appears suitable for the application in food analysis, also to isolate ZEN at trace levels. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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11 pages, 2360 KiB

Open AccessEditor’s ChoiceArticle

Zearalenone Contamination in Corn, Corn Products, and Swine Feed in China in 2016–2018 as Assessed by Magnetic Bead Immunoassay

by Ming Li, Chuqin Yang, Yuhao Mao, Xia Hong and Daolin Du

Toxins 2019, 11(8), 451; https://doi.org/10.3390/toxins11080451 - 1 Aug 2019

Cited by 27 | Viewed by 4771

Abstract

In total, 405 samples of corn, corn products, and swine feed from China in 2016–2018 were surveyed for zearalenone (ZEN) contamination using a magnetic bead immunoassay-coupled biotin–streptavidin system (BAS-MBI). The developed BAS-MBI had a limit of detection (LOD) of 0.098 ng mL⁻¹ [...] Read more.

In total, 405 samples of corn, corn products, and swine feed from China in 2016–2018 were surveyed for zearalenone (ZEN) contamination using a magnetic bead immunoassay-coupled biotin–streptavidin system (BAS-MBI). The developed BAS-MBI had a limit of detection (LOD) of 0.098 ng mL⁻¹, with half-maximal inhibition concentration (IC₅₀) of 0.71 ng mL⁻¹ in working buffer, and an LOD of 0.98 ng g⁻¹; the detection range was from 0.98 to 51.6 ng g⁻¹ in authentic agricultural samples. The BAS-MBI has been demonstrated to be a powerful method for the rapid, sensitive, specific, and accurate determination of ZEN. The ZEN positivity rate reached the highest level of 40.6% in 133 samples in 2016; ZEN levels ranged from 1.8 to 1100.0 ng g⁻¹, with an average level of 217.9 ng g⁻¹. In 2017, the ZEN positivity rate was the lowest at 24.5% in 143 samples; ZEN levels ranged from 1.1 to 722.6 ng g⁻¹, with an average of 166.7 ng g⁻¹. In 2018, the ZEN positivity rate was 31.8% in 129 samples; ZEN levels ranged from 1.3 to 947.8 ng g⁻¹, with an average of 157.0 ng g⁻¹. About 20% of ZEN-positive samples exceeded maximum limit levels. An alternative method of ZEN detection and a valuable reference for ZEN contamination in corn and its related products in China are provided. This survey suggests the need for prevention of serious ZEN contamination, along with management for food safety and human health. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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14 pages, 689 KiB

Open AccessArticle

Biomonitoring Data for Assessing Aflatoxins and Ochratoxin A Exposure by Italian Feedstuffs Workers

by Barbara De Santis, Francesca Debegnach, Elisa Sonego, Gianmarco Mazzilli, Francesca Buiarelli, Fulvio Ferri, Paolo Giorgi Rossi, Giorgia Collini and Carlo Brera

Toxins 2019, 11(6), 351; https://doi.org/10.3390/toxins11060351 - 18 Jun 2019

Cited by 9 | Viewed by 4390

Abstract

Mycotoxins exposure by inhalation and/or dermal contact is possible in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the validity of the biomonitoring as a tool to [...] Read more.

Mycotoxins exposure by inhalation and/or dermal contact is possible in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the validity of the biomonitoring as a tool to investigate the intake of mycotoxins in a population of workers operating in an Italian feed plant. Serum samples were collected for the determination of aflatoxins B1 (AFB1), AFB1-Lysine adduct and ochratoxin A (OTA). A method based on liquid–liquid extraction coupled with high resolution mass spectrometry determination was developed and fully validated. For AFB1, a high number of non-detected samples (90%) was found and no statistical difference was observed comparing workers and control group. None of the analyzed samples showed the presence of AFB1-Lysine adduct. For OTA, the 100% of the analyzed samples was positive with a 33% of the samples showing a concentration higher than the limit of quantification (LOQ), but no statistical difference was highlighted between the average levels of exposed and control groups. In conclusion, the presence of AFB1 and OTA in serum cannot be attributable to occupational exposure. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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12 pages, 2555 KiB

Open AccessEditor’s ChoiceArticle

A Gold Growth-Based Plasmonic ELISA for the Sensitive Detection of Fumonisin B₁ in Maize

by Shengnan Zhan, Lingyan Zheng, Yaofeng Zhou, Kesheng Wu, Hong Duan, Xiaolin Huang and Yonghua Xiong

Toxins 2019, 11(6), 323; https://doi.org/10.3390/toxins11060323 - 5 Jun 2019

Cited by 18 | Viewed by 4229

Abstract

In this paper, a highly sensitive plasmonic enzyme-linked immunosorbent assay (pELISA) was developed for the naked-eye detection of fumonisin B₁ (FB₁). Glucose oxidase (GOx) was used as an alternative to horseradish peroxidase as the carrier of the competing antigen. GOx [...] Read more.

In this paper, a highly sensitive plasmonic enzyme-linked immunosorbent assay (pELISA) was developed for the naked-eye detection of fumonisin B₁ (FB₁). Glucose oxidase (GOx) was used as an alternative to horseradish peroxidase as the carrier of the competing antigen. GOx catalyzed the oxidation of glucose to produce hydrogen peroxide, which acted as a reducing agent to reduce Au³⁺ to Au on the surface of gold seeds (5 nm), This reaction led to a color change in the solution from colorless to purple, which was observable to the naked eye. Various parameters that could influence the detection performance of pELISA were investigated. The developed method exhibited a considerably high sensitivity for FB₁ qualitative naked-eye detection, with a visible cut-off limit of 1.25 ng/mL. Moreover, the proposed pELISA showed a good linear range of 0.31–10 ng/mL with a half maximal inhibitory concentration (IC₅₀) of 1.86 ng/mL, which was approximately 13-fold lower than that of a horseradish peroxidase- (HRP)-based conventional ELISA. Meanwhile, the proposed method was highly specific and accurate. In summary, the new pELISA exhibited acceptable accuracy and precision for sensitive naked-eye detection of FB₁ in maize samples and can be applied for the detection of other chemical contaminants. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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9 pages, 2366 KiB

Open AccessArticle

Rapid and Simple Detection of Ochratoxin A using Fluorescence Resonance Energy Transfer on Lateral Flow Immunoassay (FRET-LFI)

by Hyun-Kyung Oh, Hyou-Arm Joung, Minhyuk Jung, Hohjai Lee and Min-Gon Kim

Toxins 2019, 11(5), 292; https://doi.org/10.3390/toxins11050292 - 23 May 2019

Cited by 19 | Viewed by 5027

Abstract

The detection of mycotoxins is crucial because of their toxicity in plants, animals, and humans. It is very important to determine whether food products are contaminated with mycotoxins such as ochratoxin A (OTA), as mycotoxins can survive heat treatments and hydrolysis. In this [...] Read more.

The detection of mycotoxins is crucial because of their toxicity in plants, animals, and humans. It is very important to determine whether food products are contaminated with mycotoxins such as ochratoxin A (OTA), as mycotoxins can survive heat treatments and hydrolysis. In this study, we designed a fluorescence resonance energy transfer (FRET)-based system that exploits antibody-antigen binding to detect mycotoxins more rapidly and easily than other currently available methods. In addition, we were able to effectively counteract the matrix effect in the sample by using a nitrocellulose membrane that enabled fluorescence measurement in coffee samples. The developed FRET on lateral flow immunoassay (FRET-LFI) system was used to detect OTA at a limit of detection (LOD) of 0.64 ng∙mL⁻¹, and the test can be completed in only 30 min. Moreover, OTA in coffee samples was successfully detected at a LOD of 0.88 ng∙mL⁻¹_, overcoming the matrix effect, owing to the chromatographic properties of the capillary force of the membrane. We believe that the developed system can be used as a powerful tool for the sensitive diagnosis of harmful substances such as mycotoxins and pesticides for environmental and food quality control monitoring. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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12 pages, 1442 KiB

Open AccessEditor’s ChoiceArticle

Rapid Determination of Ochratoxin A in Grape and Its Commodities Based on a Label-Free Impedimetric Aptasensor Constructed by Layer-by-Layer Self-Assembly

by Mina Nan, Yang Bi, Huali Xue, Sulin Xue, Haitao Long, Lumei Pu and Guorui Fu

Toxins 2019, 11(2), 71; https://doi.org/10.3390/toxins11020071 - 28 Jan 2019

Cited by 31 | Viewed by 3811

Abstract

A simple and sensitive label-free impedimetric aptasensor for rapid determination of ochratoxin A (OTA) has been developed, which was based on the combination between thiolated aptamer and gold nanoparticles by layer-by-layer self-assembly. Because of the interaction between aptamer and OTA, the relative normalized [...] Read more.

A simple and sensitive label-free impedimetric aptasensor for rapid determination of ochratoxin A (OTA) has been developed, which was based on the combination between thiolated aptamer and gold nanoparticles by layer-by-layer self-assembly. Because of the interaction between aptamer and OTA, the relative normalized electron-transfer resistance (ΔR_ct) values obtained by electrochemical impedance spectroscopy (EIS) was proportional to the concentration of OTA and showed a good linear relationship from 0.1 to 10.0 ng/mL, with a lower detection limit (0.030 ng/mL) than one-step thiolated DNA aptasensor. The established method was successfully applied to detect and analyze OTA in table wine and grape juice, and the recovery was 90.56%–104.21% when PVP effective removed of phenolic substances. The label-free impedimetric aptasensor was used for rapid detection and quantitation of OTA in the inoculated grapes with the Aspergillus Nigri (H1), and the production of OTA (62.4 μg/kg, 20 μg/kg) far exceeded the maximum levels of 2 μg/kg after inoculation for three days. The developed method exhibited a good specificity, high sensitivity, time-efficient, and it could be applied to detect the OTA concentration in grape and its commodities. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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10 pages, 1329 KiB

Open AccessEditor’s ChoiceArticle

Aptamer-Based Fluorometric Ochratoxin A Assay Based on Photoinduced Electron Transfer

by Han Zhao, Xinying Xiang, Mingjian Chen and Changbei Ma

Toxins 2019, 11(2), 65; https://doi.org/10.3390/toxins11020065 - 24 Jan 2019

Cited by 28 | Viewed by 4156

Abstract

This study describes a novel quencher-free fluorescent method for ochratoxin A (OTA) detection based on the photoinduced electron transfer (PIET) between guanine and fluorophore. In the absence of OTA, carboxyfluorescein (FAM)-labeled aptamer can partly hybridize with the complementary strand of OTA aptamer (OTA-cAPT), [...] Read more.

This study describes a novel quencher-free fluorescent method for ochratoxin A (OTA) detection based on the photoinduced electron transfer (PIET) between guanine and fluorophore. In the absence of OTA, carboxyfluorescein (FAM)-labeled aptamer can partly hybridize with the complementary strand of OTA aptamer (OTA-cAPT), which contains four guanines at its 3′-end. As a result, the fluorescence of FAM is quenched due to PIET and stacked guanines. In the presence of OTA, FAM-labeled OTA aptamer can bind specifically to OTA, and thereby the high fluorescence intensity of the dye can be maintained. Under the optimal conditions, the method had a detection limit of 1.3 nM. In addition, the method we proposed is highly sensitive and specific for OTA. Furthermore, the method was proven to be reliable based on its successful application in the detection of OTA in red wine samples. Therefore, this promising, facile, and quencher-free method may be applied to detect other toxins by using other appropriate aptamers. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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11 pages, 3361 KiB

Open AccessEditor’s ChoiceArticle

Simultaneous Lateral Flow Immunoassay for Multi-Class Chemical Contaminants in Maize and Peanut with One-Stop Sample Preparation

by Du Wang, Jianguo Zhu, Zhaowei Zhang, Qi Zhang, Wen Zhang, Li Yu, Jun Jiang, Xiaomei Chen, Xuefang Wang and Peiwu Li

Toxins 2019, 11(1), 56; https://doi.org/10.3390/toxins11010056 - 20 Jan 2019

Cited by 50 | Viewed by 4662

Abstract

Multi-class chemical contaminants, such as pesticides and mycotoxins, are recognized as the major risk factors in agro products. It is thus necessary to develop rapid and simple sensing methods to fulfill the on-site monitoring of multi-class chemical contaminants with different physicochemical properties. Herein, [...] Read more.

Multi-class chemical contaminants, such as pesticides and mycotoxins, are recognized as the major risk factors in agro products. It is thus necessary to develop rapid and simple sensing methods to fulfill the on-site monitoring of multi-class chemical contaminants with different physicochemical properties. Herein, a lateral flow immunoassay via time-resolved fluorescence was developed for the rapid, on-site, simultaneous, and quantitative sensing aflatoxin B₁ (AFB₁), zearalenone (ZEA), and chlorothalonil (CTN) in maize and peanut. The sample preparation was optimized to a single step, combining the grinding and extraction. Under optimal conditions, the sensing method lowered the limits of detection (LOD) to 0.16, 0.52, and 1.21 µg/kg in maize and 0.18, 0.57, and 1.47 µg/kg in peanut with an analytical range of 0.48–20, 1.56–200, and 3.63–300 µg/kg for AFB₁, ZEA and CTN, respectively. The protocol could be completed within 15 min, including sample preparation and lateral flow immunoassay. The recovery range was 83.24–110.80%. An excellent correlation was observed between this approach and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) for mycotoxins and gas chromatography-tandem mass spectrometry (GC-MS/MS) for pesticide in maize and peanut. This work could be applied in on-site multi-class sensing for food safety. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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13 pages, 877 KiB

Open AccessArticle

Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay to Detect Fusarium verticillioides in Poultry Feed Samples

by Aline Myuki Omori, Elisabete Yurie Sataque Ono, Melissa Tiemi Hirozawa, Igor Massahiro de Souza Suguiura, Elisa Yoko Hirooka, Maria Helena Pelegrinelli Fungaro and Mario Augusto Ono

Toxins 2019, 11(1), 48; https://doi.org/10.3390/toxins11010048 - 17 Jan 2019

Cited by 8 | Viewed by 3729

Abstract

Fumonisins are a group of toxic secondary metabolites that are produced by Fusarium verticillioides which are associated with poultry health hazard and great economic losses. The objective of the present study was to develop an immunological method to detect F. verticillioides in poultry [...] Read more.

Fumonisins are a group of toxic secondary metabolites that are produced by Fusarium verticillioides which are associated with poultry health hazard and great economic losses. The objective of the present study was to develop an immunological method to detect F. verticillioides in poultry feed samples. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a polyclonal antibody against 67 kDa protein of the F. verticillioides 97K exoantigen was developed to detect this fungus. Antibody anti-67 kDa protein showed cross-reactivity against F. graminearum (2–7%) and F. sporotrichioides (10%), but no or low cross-reactivity against Aspergillus sp. and Penicillium sp. exoantigens. The detection limit for the 67 kDa protein of F. verticillioides was 29 ng/mL. Eighty-one poultry feed samples were analyzed for Fusarium sp. count, 67 kDa protein of F. verticillioides and fumonisin concentrations. Eighty of the 81 feed samples (98.6%) showed Fusarium sp. contamination (mean 6.2 x 10⁴ CFU/g). Mean 67 kDa protein and fumonisin concentration in the poultry feed samples was 21.0 µg/g and 1.02 µg/g, respectively. The concentration of 67 kDa protein, as determined by ic-ELISA correlated positively (p < 0.05) with fumonisin levels (r = 0.76). These results suggest that this ic-ELISA has potential to detect F. verticillioides and predict fumonisin contamination in poultry feed samples. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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12 pages, 1273 KiB

Open AccessArticle

Survey of Deoxynivalenol Contamination in Agricultural Products in the Chinese Market Using An ELISA Kit

by Ming Li, Mingna Sun, Xia Hong, Jinsheng Duan and Daolin Du

Toxins 2019, 11(1), 6; https://doi.org/10.3390/toxins11010006 - 24 Dec 2018

Cited by 15 | Viewed by 4198

Abstract

A total of 328 agricultural product samples highly suspected to be contaminated, from flour companies, feed companies, and livestock farms throughout China, were surveyed for deoxynivalenol (DON) contamination using a self-assembly enzyme-linked immunosorbent assay (ELISA) kit. An ELISA kit for DON was developed [...] Read more.

A total of 328 agricultural product samples highly suspected to be contaminated, from flour companies, feed companies, and livestock farms throughout China, were surveyed for deoxynivalenol (DON) contamination using a self-assembly enzyme-linked immunosorbent assay (ELISA) kit. An ELISA kit for DON was developed with a 4.9 ng mL⁻¹ limit of detection (LOD) in working buffer and a 200 ng g⁻¹ LOD in authentic samples. The DON contamination detection rate was 88.7%, concentrations ranged from 200.9 to 6480.6 ng g⁻¹, and the highest DON contamination was found in distillers’ dried grains with solubles with an average of 3204.5 ng g⁻¹. Wheat bran and wheat were found to be the most commonly contaminated samples, and the corn meal samples had the lowest average DON level. This ELISA kit is a powerful alternative method for the rapid, sensitive, specific, accurate, and high-throughput determination of DON and can meet the maximum requirement levels. This survey suggests that DON contamination in the Chinese market is serious, and the contamination risk deserves attention. Essential preventive measures should be implemented to ensure food safety and human health. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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10 pages, 2177 KiB

Open AccessArticle

Carbon Quantum Dots Encapsulated Molecularly Imprinted Fluorescence Quenching Particles for Sensitive Detection of Zearalenone in Corn Sample

by Manyu Shao, Ming Yao, Sarah De Saeger, Liping Yan and Suquan Song

Toxins 2018, 10(11), 438; https://doi.org/10.3390/toxins10110438 - 28 Oct 2018

Cited by 35 | Viewed by 5253

Abstract

An eco-friendly and efficient one-step approach for the synthesis of carbon quantum dots (CDs) that encapsulated molecularly imprinted fluorescence quenching particles (MIFQP) and their application for the determination of zearalenone (ZEA) in a cereal sample are described in this study. CDs with high [...] Read more.

An eco-friendly and efficient one-step approach for the synthesis of carbon quantum dots (CDs) that encapsulated molecularly imprinted fluorescence quenching particles (MIFQP) and their application for the determination of zearalenone (ZEA) in a cereal sample are described in this study. CDs with high luminescence were first synthesized, and then encapsulated in the silica-based matrix through a non-hydrolytic sol-gel process. The resulting ZEA-imprinted particles exhibited not only an excellent specific molecular recognition of ZEA, but also good photostability and obvious template binding-induced fluorescence quenching. Under the optimized conditions, the fluorescence intensity of MIFQP was inversely proportional to the concentration of ZEA. By validation, the detection range of these fluorescence quenching materials for ZEA was between 0.02 and 1.0 mg L⁻¹, and the detection limit was 0.02 mg L⁻¹ (S/N = 3). Finally, the MIFQP sensor was successfully applied for ZEA determination in corn with recoveries from 78% to 105% and the relative standard deviation (RSD %) was lower than 20%, which suggests its potential in actual applications. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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14 pages, 628 KiB

Open AccessArticle

Multi-Occurrence of Twenty Mycotoxinsin Pasta and a Risk Assessment in the Moroccan Population

by Youssef Bouafifssa, Lara Manyes, Mohamed Rahouti, Jordi Mañes, Houda Berrada, Abdellah Zinedine and Mónica Fernández-Franzón

Toxins 2018, 10(11), 432; https://doi.org/10.3390/toxins10110432 - 26 Oct 2018

Cited by 28 | Viewed by 4327

Abstract

In the present study, the multi-occurrence of twenty (20) mycotoxins in pasta samples consumed in Morocco was assessed. For this, a modified Quick, Easy, Cheap Effective, Rugged, and Safe method was validated. The mycotoxins studied were identified and quantified [...] Read more.

In the present study, the multi-occurrence of twenty (20) mycotoxins in pasta samples consumed in Morocco was assessed. For this, a modified Quick, Easy, Cheap Effective, Rugged, and Safe method was validated. The mycotoxins studied were identified and quantified by liquid chromatography–tandem mass spectrometry (LC–MS/MS) and gas chromatography–tandem mass spectrometry (GC-MS/MS). The validated method was applied to one hundred and six (n = 106) pasta samples purchased from several areas in the country. The analytical results showed that 99 out of 106 total samples (93.4%) were contaminated with at least one mycotoxin. Nine mycotoxins (Aflatoxin B1, Enniatin B, Enniatin B1, Enniatin A1, Zearalenone, Deoxynivalenol, 3-Acetyl-Deoxynivalenol, T-2, and HT-2 toxins) were present in the pasta samples. Enniatin B and Enniatin B1 were the predominant mycotoxins. The Zearalenone, Deoxynivalenol, HT-2, and T-2 toxins were present in 51.8%, 43.5%, 34.9%, and 16% of samples, respectively. Aflatoxin B1 was detected in only 2 samples. Risk exposure assessment concluded that mycotoxin levels found in pasta do not pose a significant human health risk for the Moroccan population. This is the first paper drafted on the multi-occurrence of mycotoxins in pasta from this country. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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16 pages, 1181 KiB

Open AccessArticle

Multiwalled Carbon Nanotube for One-Step Cleanup of 21 Mycotoxins in Corn and Wheat Prior to Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry Analysis

by Dongmei Jiang, Dizhe Wei, Liuqing Wang, Shuai Ma, Yuanfang Du and Meng Wang

Toxins 2018, 10(10), 409; https://doi.org/10.3390/toxins10100409 - 10 Oct 2018

Cited by 28 | Viewed by 3854

Abstract

One-step solid-phase extraction (SPE) using a multiwalled carbon nanotube (MWCNT) for simultaneous analysis of 21 mycotoxins, including nine trichothecenes, zearalenone (ZEN) and its derivatives, four aflatoxins, and two ochratoxins, in corn and wheat was developed. Several key parameters affecting the performance of the [...] Read more.

One-step solid-phase extraction (SPE) using a multiwalled carbon nanotube (MWCNT) for simultaneous analysis of 21 mycotoxins, including nine trichothecenes, zearalenone (ZEN) and its derivatives, four aflatoxins, and two ochratoxins, in corn and wheat was developed. Several key parameters affecting the performance of the one-step SPE procedure—types of MWCNT, combinations with five sorbents (octadecylsilyl (C₁₈), hydrophilic–lipophilic balance (HLB), mixed-mode cationic exchange (MCX), silica gel, and amino-propyl (NH₂)), and filling amounts of the MWCNTs—were thoroughly investigated. The combination of 20 mg carboxylic MWCNT and 200 mg C₁₈ was proven to be the most effective, allowing the quantification of all analyzed mycotoxins in corn and wheat. Under the optimized cleanup procedure prior to ultraperformance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) analysis, the method was validated by analyzing samples spiked at the limit of quantification (LOQ), two-times LOQ, and 10-times LOQ. Satisfactory linearity (r² ≥ 0.9910), high sensitivity (LOQ in different ranges of 0.5–25 μg L⁻¹), good recovery (75.6–110.3%), and acceptable precision (relative standard deviation (RSD), 0.3–10.7%) were obtained. The applicability of the method was further confirmed using raw samples of corn and wheat. In conclusion, the established method was rapid, simple and reliable for simultaneous analysis of 21 mycotoxins in corn and wheat. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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14 pages, 3442 KiB

Open AccessArticle

Development of a Sensitive Enzyme-Linked Immunosorbent Assay and Rapid Gold Nanoparticle Immunochromatographic Strip for Detecting Citrinin in Monascus Fermented Food

by Shih-Wei Wu, Yao-An Yu, Biing-Hui Liu and Feng-Yih Yu

Toxins 2018, 10(9), 354; https://doi.org/10.3390/toxins10090354 - 2 Sep 2018

Cited by 19 | Viewed by 5278

Abstract

Antibodies against citrinin (CTN) were generated from rabbits, which were injected with CTN-keyhole limpet hemocyanin (KLH). This work involved the development of a sensitive competitive direct enzyme-linked immunosorbent assay (cdELISA) and a rapid gold nanoparticle immunochromatographic strip (immunostrip) method for analyzing CTN in [...] Read more.

Antibodies against citrinin (CTN) were generated from rabbits, which were injected with CTN-keyhole limpet hemocyanin (KLH). This work involved the development of a sensitive competitive direct enzyme-linked immunosorbent assay (cdELISA) and a rapid gold nanoparticle immunochromatographic strip (immunostrip) method for analyzing CTN in Monascus-fermented food. CTN at a concentration of 5.0 ng/mL caused 50% inhibition (IC₅₀) of CTN-horseradish peroxidase (CTN-HRP) binding to the antibodies in the cdELISA. The capable on-site detection of CTN was accomplished by a rapid antibody-gold nanoparticle immunostrip with a detection limit of 20 ng/mL and that was completed within 15 min. A close inspection of 19 Monascus-fermented foods by cdELISA confirmed that 14 were contaminated with citrinin at levels from 28.6–9454 ng/g. Further analysis with the immunostrip is consistent with those results obtained using cdELISA. Both means are sensitive enough for the rapid examination of CTN in Monascus-fermented food products. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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12 pages, 2285 KiB

Open AccessArticle

Feasibility of A Novel On-Site Detection Method for Aflatoxin in Maize Flour from Markets and Selected Households in Kampala, Uganda

by Alex Paul Wacoo, Deborah Wendiro, Sarah Nanyonga, Joseph F. Hawumba, Wilbert Sybesma and Remco Kort

Toxins 2018, 10(8), 327; https://doi.org/10.3390/toxins10080327 - 11 Aug 2018

Cited by 17 | Viewed by 5390

Abstract

In sub-Saharan Africa, there is a high demand for affordable and accessible methods for on-site detection of aflatoxins for appropriate food safety management. In this study, we validated an electrochemical immunosensor device by the on-site detection of 60 maize flour samples from six [...] Read more.

In sub-Saharan Africa, there is a high demand for affordable and accessible methods for on-site detection of aflatoxins for appropriate food safety management. In this study, we validated an electrochemical immunosensor device by the on-site detection of 60 maize flour samples from six markets and 72 samples from households in Kampala. The immunosensor was successfully validated with a linear range from 0.7 ± 0.1 to 11 ± 0.3 µg/kg and limit of detection (LOD) of 0.7 µg/kg. The maize flour samples from the markets had a mean total aflatoxin concentration of 7.6 ± 2.3 µg/kg with approximately 20% of the samples higher than 10 µg/kg, which is the maximum acceptable level in East Africa. Further down the distribution chain, at the household level, approximately 45% of the total number contained total aflatoxin levels higher than the acceptable limit. The on-site detection method correlated well with the established laboratory-based HPLC and ELISA-detection methods for aflatoxin B₁ with the correlation coefficients of 0.94 and 0.98, respectively. This study shows the feasibility of a novel on-site detection method and articulates the severity of aflatoxin contamination in Uganda. Full article

(This article belongs to the Special Issue Advanced Methods for Mycotoxins Detection)

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