3D nanocake-like Au-MXene/Au pallet structure-based label-free electrochemical aptasensor for paraquat determination.

3D nanocake-like Au-MXene and Au pallet (Au-MXene/AuP) nanocomposite-modified screen-printed carbon electrodes (SPCEs) were utilized to construct an ultrasensitive label-free electrochemical aptasensor through a self-assembly procedure for trace paraquat (PQ) residue detection. Benefiting from the excellent electrochemical (EC) performances (e.g., high conductivity and large surface area) of Au-MXene nanocomposites and AuP substrate, the developed Apt/Au-MXene/AuP/SPCE-based EC aptasensor displayed excellent specificity and anti-interference ability, good repeatability, and stability. A linear relationship between the log value of the change in current intensity [lg (ΔI)] and the log value of the concentration of PQ [lg (CPQ)] was obtained in the range 0.05-1000 ng/mL. The limit of detection was 0.028 ng/mL, and the sensitivity was 255.5 µA/(µM·cm2). Practical applications in malt and mint samples confirmed the accuracy of the EC aptasensor in complex matrices for PQ detection, providing a universal analytical tool for other trace pesticides in different food samples by simply replacing the corresponding aptamers.

Oral Intake of Inosine 5'-Monophosphate in Mice Promotes the Absorption of Exogenous Fatty Acids and Their Conversion into Triglycerides though Enhancing the Phosphorylation of Adenosine 5'-Monophosphate-Activated Protein Kinase in the Liver, Leading to Lipohyperplasia.

Inosine 5'-monophoaphate (IMP) is a food additive that promotes serious lipohyperplasia in the liver of C57/KsJ-db/db (db/db) mice. Thus, IMP taken orally by healthy mice might also damage their health. To date, how IMP affects health after being taken by healthy animals is still unclear. Therefore, we investigated the health of C57BL/6J mice affected by IMP intake. Our data revealed that C57BL/6J mice administered 255 µM IMP daily via oral gavage for 4 months caused hyperlipidemia and an increase in body fat rate. The expressions of acetyl-CoA carboxylase 1 (ACC1) and phosphorylated acetyl-CoA carboxylase 2 (ACC2) in hepatocytes increased though the administration of IMP, promoting the phosphorylation of adenosine 5'-monophosphate-activated protein kinase (AMPK). The conversion of acetyl-CoA into triglycerides (TGs) was promoted by ACC1. These TGs were transported from the hepatocytes to avoid the development of non-alcoholic fatty liver disease (NAFLD), causing a deficiency of acetyl-CoA in the liver, and then, the increased phosphorylated ACC2 promoted the cytoplasm fatty acids entering the mitochondria and conversion into acetyl-CoA through the fatty acid ß-oxidation pathway, causing a deficiency in fatty acids. Therefore, the liver showed enhanced absorption of exogenous fatty acids, which were converted into TGs, causing lipohyperplasia. In conclusion, an excessive IMP intake promotes metabolic dysfunction in adipose tissue.

Multi-mycotoxin detection and human exposure risk assessment in medicinal foods.

Mycotoxin contamination in medicinal foods has attracted increasing global attention. In this study, a simple and sensitive ultrasonication assisted one-step extraction based ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was developed for simultaneous detection of multi-mycotoxins in five kinds of medicinal foods rich in starch. Under optimal conditions, the developed technique displayed excellent analytical performances. Limits of detection and quantitation for the six mycotoxins were 0.04-0.25 ng/mL and 0.10-0.67 ng/mL, respectively. Average recoveries at three fortified levels ranged from 75.33 % to 118.0 %. Real-world application in 103 batches of medicinal foods displayed that 58 samples were positive with one or more mycotoxins at an occurrence rate of 56.31 % (58/103). Coix seed gave the highest positive rate of 96.15 %, followed by Lily (90 %), Chinese yam (50 %), Lotus seed (34.04 %) and Malt (30 %). Zearalenone had the highest positive rate of 28.16 % with contents in 5 Coix seeds exceeding the maximum residue limit (MRL), followed by aflatoxin B1 of 27.18 % (28/103) with contents in 7 Coix seed and 10 Lotus seeds over its MRL, and ochratoxin A (OTA) of 11.65 % with contents in 1 Lotus seed and 5 Lily samples greater than its MRL. Exposure risk assessment indicated that Coix seed and Lotus seeds that were susceptible to aflatoxins posed great threats to human health. Long-term consumption of Lily that was easily contaminated with OTA were also harmful. This work provides a robust platform for multi-mycotoxin monitoring in medicinal foods to protect the consumers from potential health risks.

Ultrasensitive electrochemical aptasensor with Nafion-stabilized f-MWCNTs as signal enhancers for OTA detection.

In this study, an ultrasensitive electrochemical (EC) aptasensor with Nafion-stabilized functionalized multi-walled carbon nanotubes (f-MWCNTs) as signal enhancers was established for ochratoxin A (OTA) determination. Herein, f-MWCNTs were prepared through functionalization with nitric acid. The incorporation of Nafion promoted a good dispersion of f-MWCNTs and prevented their leaching on the electrode, making a robust stability of the aptasensor. The Nafion-f-MWCNTs composites were used as the sensing substrates to largely enhance the electroactive surface area and the conductivity of the electrode, realizing a significant signal amplification. Carboxyl groups on the surface of f-MWCNTs readily exposed from Nafion membrane to couple with streptavidin, facilitating the immobilization of biotinylated aptamers to achieve selective recognition towards OTA. When OTA existed, aptamers preferentially combined with it, causing a noticeable decline in the current response. Under optimum conditions, a good linear relationship between the current changes and the logarithm of OTA concentration was observed from 0.005 ng/mL to 10 ng/mL, with a limit of detection low to 1 pg/mL for OTA. The specific, sensitive, and reproducible aptasensor succeeded in application in malt samples, confirming a great promise for more contaminants and providing a universal platform in complex matrices by simply replacing the corresponding aptamers.

Development of a Nafion-MWCNTs and in-situ generated Au nanopopcorns dual-amplification electrochemical aptasensor for ultrasensitive detection of OTA.

Trace detection of ochratoxin A (OTA) in foods is essential to mitigate risks to human health. Herein, a label-free electrochemical (EC) aptasensor based on dual-signal amplification of Nafion dispersed multi-walled carbon nanotubes (Nafion-MWCNTs) and Au nanopopcorns was developed for ultrasensitive detection of OTA. Nafion solution prevented the leaching of MWCNTs, and the Nafion-MWCNTs modified screen-printed carbon electrode (SPCE) acted as the sensing substrate which facilitated the uniform distribution of the electrodeposited Au nanopopcorns. The in-situ generated Au nanopopcorns could not only load a large amount of aptamers for specific identification of OTA, but also promote the electron transfer of the sensing platform. The incorporation of Nafion-MWCNTs and Au nanopopcorns realized dual-amplification of the aptasensor due to the enhanced conductivity and the increased electroactive surface area of the electrode. The modified electrodes were characterized through scanning electron microscopy, X-ray photoelectron spectroscopy and EC evaluation. Under optimal conditions, the electrochemical impedance spectroscopy (EIS) was measured for the determination of OTA. The as-fabricated Au nanopopcorns/Nafion-MWCNTs impedimetric aptasensor displayed excellent sensitivity with a detection limit as low as 1 pg/mL and a wide linear range of 1 pg/mL-10 ng/mL for OTA. Practical application of the aptasensor in the spiked malt samples achieved satisfactory recoveries of 89.82-95.65 %, which was also successfully verified to detect OTA in eleven batches of actual malt samples collected from the local market. The creative aptasensor is simple, cost-effective, sensitive, and accurate, showing great promise for on-site monitoring of other trace contaminants in foods by simply replacing the aptamers.

Single-Molecule Real-Time Sequencing to Explore the Mycobiome Diversity in Malt.

This study determined the composition of fungal communities and characterized the enriched fungal species in raw and roasted malts via the third-generation PacBio-based full-length single-molecule real-time (SMRT) sequencing of the full-length amplicon of the internal transcribed spacer (ITS) region. In total, one kingdom, six phyla, 23 classes, 56 orders, 120 families, 188 genera, 333 species, and 780 operational taxonomic units (OTUs) were detected with satisfactory sequencing depth and sample size. Wickerhamomyces (56%), Cyberlindnera (15%), Dipodascus (12%), and Candida (6.1%) were characterized as the dominant genera in the raw malts, and Aspergillus (35%), Dipodascus (21%), Wickerhamomyces (11%), and Candida (3.5%) in the roasted malts. Aspergillus proliferans, Aspergillus penicillioides, and Wickerhamomyces anomalus represented the crucial biomarkers causing intergroup differences. Correlation analysis regarding environmental factors indicated that the water activity (aw) of the samples affected the composition of the fungal communities in the malts. In practice, special attention should be paid to the mycotoxin-producing fungi, as well as other fungal genera that are inversely correlated with their growth, to ensure the safe use of malt and its end products. IMPORTANCE Fungal contamination and secondary metabolite accumulation in agricultural products represent a global food safety challenge. Although high-throughput sequencing (HTS) is beneficial for explaining fungal communities, it presents disadvantages, such as short reads, species-level resolution, and uncertain identification. This work represents the first attempt to characterize the fungal community diversity, with a particular focus on mycotoxin-producing fungi, in malt via the third-generation PacBio-based full-length SMRT sequencing of the ITS region, aiming to explore and compare the differences between the fungal communities of raw and roasted malts. The research is beneficial for developing effective biological control and conservation measures, including improving the roasting conditions, monitoring the environmental humidity and aw, and effectively eliminating and degrading fungi in the industry chain according to the diverse fungal communities determined, for the safe use of malts and their end products, such as beers. In addition, the third-generation SMRT sequencing technology allows highly efficient analysis of fungal community diversity in complex matrices, yielding fast, high-resolution long reads at the species level. It can be extended to different research fields, updating modern molecular methodology and bioinformatics databases.

Development of a graphene oxide nanosheet and double-stranded DNA structure based fluorescent "signal off" aptasensor for ochratoxin A detection in malt.

A "signal off" fluorescent aptasensor based on graphene oxide (GO) nanosheet and double-stranded DNA structure was fabricated for OTA detection. In the absence of OTA, the aptamer and its complementary DNA (cDNA) formed double-stranded conjugates that could coexist with GO, presenting fluorescence responses. Then, the presented OTA was captured by the aptamers, causing the release of cDNA-FAM probes. The free probes were adsorbed by GO, leading to an OTA concentration-dependent fluorescence quenching through fluorescence resonance energy transfer. Under optimum conditions, the fluorescent aptasensor exhibited outstanding sensitivity with a LOD of 11 pg/mL and a wide dynamic range of 0.04-30 ng/mL. The selectivity of the aptasensor was confirmed against other four mycotoxins, and the feasibility and reliability were verified by determining OTA in the spiked malt samples with satisfactory recovery of 95.50%-112.05%. This aptasensing platform could be adapted to measure other mycotoxins by replacing the aptamer sequences for food safety.

Transfer behavior of pesticides from honeysuckle into tea infusions: Establishment of an empirical model for transfer rate prediction.

Affected by some external conditions and internal factors, pesticides can be transferred from tea into its infusion, causing subsequent damage to humans as tea infusion is generally consumed. This study aimed to explore the inherent regularity in transfer behavior of 23 pesticides belonging to different classes from honeysuckle to its tea infusion, and to understand the effects of external brewing conditions and internal physicochemical parameters of the pesticides on their transfer rates. Results indicated that the transfer rates (Rt) of pesticides from honeysuckle into tea solutions increased with prolonged brewing time, or adding a cover on a container, but decreased with increasing the times of infusion. In addition, the transfer potential of these pesticides greatly depended on their physicochemical properties but not their type. The pesticides with high water solubility and low water partition coefficient (LogKow, e.g., omethoate) were more easily transferred into tea infusions than those with low water solubility and high LogKow (e.g., chlorpyrifos). Compared the tea brewing in a covered container, the empirical models obtained in an uncovered cup predicted the transfer behavior and drinking risk of pesticides potentially introduced into honeysuckle and its tea infusion. The linear equation was as follow: Rt = 10.756 LogWS + 7.517, R = 0.8771. In practice, honeysuckle should be brewed in an uncovered cup within a short brewing time, and the first tea infusion should be abandoned to reduce the transfer percentage of pesticides. This study provided beneficial references for pesticide application in honeysuckle plantation to establish realistic maximum residue limits of multi-pesticides in honeysuckle tea and related products.

Recent advances in synthesis and modification of carbon dots for optical sensing of pesticides.

Serious threat from pesticide residues to the ecosystem and human health has become a global concern. Developing reliable methods for monitoring pesticides is a world-wide research hotspot. Carbon dots (CDs) with excellent photostability, low toxicity, and good biocompatibility have been regarded as the potential substitutes in fabricating various optical sensors for pesticide detection. Based on the relevant high-quality publications, this paper first summarizes the current state-of-the-art of the synthetic and modification approaches of CDs. Then, a comprehensive overview is given on the recent advances of CDs-based optical sensors for pesticides over the past five years, with a particular focus on photoluminescent, electrochemiluminescent and colorimetric sensors regarding the sensing mechanisms and design principles by integrating with various recognition elements including antibodies, aptamers, enzymes, molecularly imprinted polymers, and some nanoparticles. Novel functions and extended applications of CDs as signal indicators, catalyst, co-reactants, and electrode surface modifiers, in constructing optical sensors are specially highlighted. Beyond an assessment of the performances of the real-world application of these proposed optical sensors, the existing inadequacies and current challenges, as well as future perspectives for pesticide monitoring are discussed in detail. It is hoped to provide powerful insights for the development of novel CDs-based sensing strategies with their wide application in different fields for pesticide supervision.

A CdSe@CdS quantum dots based electrochemiluminescence aptasensor for sensitive detection of ochratoxin A.

In this work, a CdSe@CdS quantum dots (QDs) based label-free electrochemiluminescence (ECL) aptasensor was developed for the specific and sensitive detection of ochratoxin A (OTA). Chitosan (CHI) could immobilize abundant QDs on the surface of an Au electrode as the luminescent nanomaterials. Glutaraldehyde was used as the crosslinking agent for coupling a large number of OTA aptamers. Thanks to the excellent stability, good biocompatibility, and strong ECL intensity of CdSe@CdS QDs, as well as the quick reactions of the generated SO4•- in the electrolyte, strong ECL signals were measured. Because of the specific recognition of aptamer toward OTA, the reduced ECL signals caused by OTA in the samples were recorded for quantify the content of OTA. After optimizing a series of crucial conditions, the ECL aptasensor displayed superior sensitivity for OTA with a detection limit of 0.89 ng/mL and a wide linear concentration range of 1-100 ng/mL. The practicability and viability were verified through the rapid and facile analysis of OTA in real Lily and Rhubarb samples with recovery rates (n = 3) of 98.1-105.6% and 97.3-101.5%, respectively. The newly-developed QDs-based ECL aptasensor provided a new universal analytical tool for more mycotoxins in safety assessment of foods and feeds, environmental monitoring, and clinical diagnostics.

Aptasensors for mycotoxins in foods: Recent advances and future trends.

Mycotoxin contamination in foods has posed serious threat to public health and raised worldwide concern. The development of simple, rapid, facile, and cost-effective methods for mycotoxin detection is of urgent need. Aptamer-based sensors, abbreviated as aptasensors, with excellent recognition capacity to a wide variety of mycotoxins have attracted ever-increasing interest of researchers because of their simple fabrication, rapid response, high sensitivity, low cost, and easy adaptability for in situ measurement. The past few decades have witnessed the rapid advances of aptasensors for mycotoxin detection in foods. Therefore, this review first summarizes the reported aptamer sequences specific for mycotoxins. Then, the recent 5-year advancements in various newly developed aptasensors, which, according to the signal output mode, are divided into electrochemical, optical and photoelectrochemical categories, for mycotoxin detection are comprehensively discussed. A special attention is taken on their strengths and limitations in real-world application. Finally, the current challenges and future perspectives for developing novel highly reliable aptasensors for mycotoxin detection are highlighted, which is expected to provide powerful references for their thorough research and extended applications. Owing to their unique advantages, aptasensors display a fascinating prospect in food field for safety inspection and risk assessment.

Fungal communities in Nelumbinis semen characterized by high-throughput sequencing.

For a long period, Nelumbinis semen has been widely used as a medicinal and edible product. However, it is susceptible to contamination with toxigenic fungi and aflatoxins during the growth, collection, transportation, and storage processes, causing serious health threats to humans and huge economic losses. Effectively monitoring the fungal communities is of great importance to avoid aflatoxins contamination in Nelumbinis semen. High-throughput sequencing (HTS) is a new technology to evaluate fungal communities so as to overcome the limitations of the traditional cultural ways. In this study, the ITS2 based Illumina-MiSeq platform was developed to evaluate the fungal communities in normal and moldy Nelumbinis semen by using the HTS technology. Two different primer pairs were introduced to compare their performance in amplifying the target gene. The primer pair that produced more reads was selected to analyze the results. In all the nine tested Nelumbinis semen samples, 2 phyla, 5 classes, 6 orders, 8 families, 9 genera and 4 species were detected. A total of 9 genera were spotted, of which, Aspergillus (0.04%-72.93%) and Rhizopus (0.002%-48.12%) were the most dominant. ANOISM analysis showed no significant differences in the normal and moldy groups. The use of HTS technology can detect the fungal communities in complex Nelumbinis semen samples, providing an early warning for toxigenic fungi and aflatoxins contamination to warrant their quality and safety.

Development of a quantum dot nanobead-based fluorescent strip immunosensor for on-site detection of aflatoxin B1 in lotus seeds.

Owing to the serious threat of aflatoxin B1 (AFB1) to public health, development of a reliable method for accurate determination of it is extremely necessary and urgent. In this study, a simple, rapid and highly-sensitive quantum dot nanobeads (QBs) based lateral flow fluorescent strip immunosensor was developed for on-site detection of AFB1 in edible and medicinal lotus seeds. Carboxylated QBs were used as the fluorescent markers to prepare the fluorescent probe through coupling QBs with anti-AFB1 antibodies. Bovine serum albumin (BSA)-AFB1 antigens and goat anti-mouse IgG antibodies were coated on the nitrocellulose (NC) membrane to prepare the test (T) and control (C) lines, respectively. Qualitative analysis of AFB1 was realized by naked eye, and the quantitative determination was achieved with a portable strip reader. Results showed that the newly-developed test strip sensor could achieve rapid detection of AFB1 within 15 min, allowing a limit of detection (LOD) of 1 ng/mL (2 µg/kg) and a linear range of 1-19 ng/mL (2-38 µg/kg). Recovery rates from the fortified lotus seeds with low, medium and high spiking concentrations (2.5, 5 and 10 µg/kg) ranged from 94.0% to 116.0% with relative standard deviations less than 10%. All the results were confirmed by a standard LC-MS/MS method. The QBs-based fluorescent strip immunosensor with high sensitivity, easy operation, and low cost provided a preferred solution for rapid, on-site screening and highly-sensitive quantitation of AFB1 in a large number of lotus seed samples.

Development of a multi-channel magnetic bead micro-probe assay for high-throughput detection of zearalenone in edible and medicinal Coix seed.

A multi-channel magnetic bead micro-probes assay (MBPA) based on indirect competitive principle was developed for high-throughput detection of zearalenone (ZEA) in edible and medicinal Coix seed. This strategy introduced magnetic beads as the carriers, the specific primary antibodies as the capture probes for targets and the secondary antibodies functionalized goat anti-mouse immunoglobulin G labeled fluorescein isothiocyanate as the fluorescence signal probes. Through the competitive reaction of ZEA in Coix seed samples and that covalently coupled on the surface of MBs with their specific antibodies, as well as fast magnetic separation and sensitive fluorescence detection, the developed MBPA strategy allowed low limit of detection (2.03 ng/mL) with broad dynamic range (2.03-440.67 ng/mL), as well as excellent accuracy with the average recovery rate of 96.39% and relative standard deviation (RSD) of 5.48% for ZEA. 36 samples could realize simultaneous analysis in one operation within less than 20 min only needing 50 µL of solution and 30 s of sampling, avoiding large consumption of time and organic solvents. Multiple centrifugation and cleanup steps were omitted because of magnetic separation, avoiding the loss of targets. Diverse capture and fluorescent probes can be randomly bound onto the surface of MBs, making the MBPA strategy a promising tool for on-site high-throughput monitoring of various trace hazard factors in food safety, and environmental monitoring.

Recent progress in immunosensors for pesticides.

Immunosensors for rapid detection of pesticide residues has attracted considerable interest in the past few years for healthcare and environmental monitoring. And the publications have grown exponentially over the past decades, making it a trending hot-spot. Therefore, this review first examines the current situation regarding pesticide residue in various foods, feeds, traditional Chinese medicines and environmental samples. Then, the primary focus is on the recent development of the proposed immunosensors for pesticide detection in the past five years, with particular emphasis on the fluorescence, colorimetric, chemiluminescence, electrochemiluminescence, surface plasmon resonance, surface-enhanced Raman spectroscopy, electrochemical and piezoelectric sensors. Beyond a highlight of the real-world application and advantages of these emerging immunosensors for pesticide inspection, this paper also discusses their potential limits and current challenges, as well as future perspectives. This review will provide powerful insights to researchers for the future development of immunosensors, as well as their broader application in different fields, such as analytical chemistry, food safety control, clinical diagnostics, and environmental monitoring.

Exploration of the safe water content and activity control points for medicinal and edible lotus seeds from mildew.

Affected by the inner properties and the external environmental conditions, medicinal and edible lotus seeds are susceptible to mildew with fungal infection under suitable temperature and humidity conditions, leading to the production and contamination of various mycotoxins, along with threats to its quality and safety. In this study, the changes of water content (Cw) and water activity (Aw) of lotus seeds stored at 25 °C and different relative humidity conditions, as well as the correlation between them and mildew of this edible and medicinal material were studied, aiming to explore the safe Cw and Aw control points for screening out the suitable storage conditions from mildew. Blank (without fungal conidia) and experimental (artificially added with Aspergillus flavus conidia) groups of lotus seeds were stored at 25 °C and relative humidity of 40%, 50%, 60% and 70% for about 30 days, respectively. The mildew was observed and the changes of Cw, Aw, together with the production of aflatoxins were measured. Results showed that no mildew was found and aflatoxins were not detected in lotus seeds when they were stored for 30 days at 25 °C and relative humidity of 40%, 50% and 60% with Cw < 12% and Aw < 0.6. While, when the relative humidity was up to 70%, the Cw and Aw values rose quickly, and the Cw exceeded the officially-permitted level (14%). Although no mildew was observed, AFB1 was still detected, increasing the potential risk of lotus seeds regarding aflatoxins. For warranting the quality with economic and safe storage, lotus seeds are suggested to be stored at 25 °C and relative humidity lower than 60% with 12% and 0.6 as the safe Cw and Aw control points, respectively, to prevent medicinal and edible products from mildew and the contamination of aflatoxins.

Development of a ZnCdS@ZnS quantum dots-based label-free electrochemiluminescence immunosensor for sensitive determination of aflatoxin B1 in lotus seed.

In this study, we designed a ZnCdS@ZnS quantum dots (QDs)-based label-free electrochemiluminescence (ECL) immunosensor for sensitive determination of aflatoxin B1 (AFB1). A Nafion solution assembled abundant QDs on the surface of a Au electrode as ECL signal probes, with specially coupled anti-AFB1 antibodies as the capturing element. As the reduction reaction between S2O82- in the electrolyte and QDs on the electrode led to ECL emission, the decreased ECL signals resulting from target AFB1 in the samples were recorded for quantification. We evaluated electrochemical impedance spectroscopy and ECL measurements along each step in the construction of the proposed immunosensor. After systematic optimization of crucial parameters, the ECL immunosensor exhibited a good sensitivity, with a low detection limit of 0.01 ng/mL for AFB1 in a wide concentration range of 0.05-100 ng/mL. Testing with lotus seed samples confirmed the satisfactory selectivity, stability, and reproducibility of the developed ECL immunosensor for rapid, efficient, and sensitive detection of AFB1 at trace levels in complex matrices. This study provides a powerful and universal analytical platform for a variety of analytes that can be used in broad applications for real-time analysis, such as food and traditional Chinese medicine safety testing, environmental pollution monitoring, and disease diagnostics. Graphical abstract Development of a ZnCdS@ZnS quantum dots based label-free electrochemiluminescence immunosensor for sensitive detection of aflatoxin B1 in lotus seed.

A self-assembled electrochemical immunosensor for ultra-sensitive detection of ochratoxin A in medicinal and edible malt.

Trace residue of mycotoxins in complex medicinal and edible food matrices has brought huge challenges for the development of ultrasensitive analytical methods. Here, a green electrochemical immunosensor for the ultrasensitive detection of ochratoxin A (OTA) was fabricated by self-assembling a compact 2-mercaptoacetic (TGA) monolayer on the surface of the working Au electrode to form the Au/TGA/bovine serum aibumin (BSA)-OTA/anti-OTA monoclonal antibody composite probes for selective and ultra-sensitive detection of OTA based on indirect competitive principle and differential pulse voltammetry analysis. The electrochemical impedance spectroscopy and cyclic voltammetry methods were introduced to characterize the assemble situation of the TGA-modified Au electrode and optimize some critical parameters for the green electrochemical immunoseonsor. Under the optimal conditions, the developed immunosensor exhibited much lower limit of detection (0.08 ng/mL) in the range of 0.1-1.0 ng/mL for OTA compared with other direct or disposable electrochemical immunosensors. Real application in the spiked malt samples verified high accuracy with no matrix interferences of the proposed immunoseonsor. This is a meaningful study on a self-assembled electrochemical immunoseonsor for ultra-sensitive and rapid detection of OTA in malt samples, which suggested a general simple-to-use sensing platform and prospect as an economical and green tool for ultra-sensitive detection of much more trace-level of toxic small molecules in other complex matrices to ensure their quality and safety.

Development of a high-performance liquid chromatography with tandem mass spectrometry method for identifying common adulterant content in saffron (Crocus sativus L.).

OBJECTIVES: This study aimed to develop an efficient and reliable method for estimating common adulterants in saffron by detecting their characteristic components to warrant its efficacy and regular use as a highly valuable medicinal herb. METHODS: A selective and sensitive high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) method was developed to estimate the common adulterants in saffron from corn stigma, chrysanthemum and safflower through the simultaneous determination of specific constituents including allantoin, chlorogenic acid (ChA) and hydroxysafflor yellow A (HSYA). Peak identification of each target compound was confirmed from product ions obtained using multiple reaction monitoring triggered enhanced product ions mass chromatogram. Method validation in terms of linearity, sensitivity, reproducibility, accuracy and stability was systematically performed according to official guidelines. KEY FINDINGS: Satisfactory separation of the three components was achieved on a C18 column (4.6 × 250 mm, 5 µm) with methanol-acetonitrile-ammonium acetate (3.0 mm) as the mobile phase at gradient elution. The identification of these specific constituents was accomplished using the multiple reaction monitoring mode in combination with enhanced product ion supplementary confirmation. The established method was validated in terms of linearity, sensitivity, reproducibility, accuracy and recovery, which were found satisfactory for sensitive detection of the three target compounds. CONCLUSIONS: By detecting the specific constituents allantoin, ChA and HSYA in one run, the adulterants of corn stigma, chrysanthemum and safflower can be effectively identified and estimated in saffron. This is the first report on developing a simple, sensitive and operational method for the identification and estimation of common adulterants of saffron, that was forwarded for broaden application.

Simultaneous determination of 19 mycotoxins in lotus seed using a multimycotoxin UFLC-MS/MS method.

OBJECTIVES: In relevance to the internal components and improper environmental conditions, lotus seeds are susceptible to fungal contamination and mycotoxins residue, leading to harmful impacts on the quality and safety, as well as their pharmaceutical efficacy and clinical use. It is necessary and urgent to assess various mycotoxins residue in lotus seeds. This study aimed to develop a sensitive method for accurate assessment of multimycotoxin residues in lotus seeds. METHODS: A simple and reliable modified ultrasonication-assisted extraction, QuEChERS purification based ultrafast liquid chromatography tandem mass spectrometry (UFLC-MS/MS) method was successfully developed for ultrasensitive determination of 19 multiclass mycotoxins in starch-rich lotus seeds. Four extraction modes and three clean-up sorbents for improving the recoveries of mycotoxins were optimized. Limits of detection (LODs) and quantification, linearity, precision, accuracy, and matrix effect were studied for method validation. For simultaneous qualitation and quantification, the 19 chemically diversified mycotoxins were well separated on a CAPCELL CORE C18 column (100 mm × 2.1 mm, 2.7 µm) and detected in positive/negative electrospray ionization mode within 7 min. KEY FINDINGS: The validated method exhibited satisfactory linearity (r > 0.995), ultragood selectivity (LODs of 0.1-15.0 µg/kg), excellent precision (RSDs <13.0%) and convincing accuracy (recoveries between 79.4% and 131.6% with RSDs <14.4%). Matrix effect, between 54.5% and 113.6%, appeared especially for aflatoxins B1 and B2 , deoxynivalenol and T-2 toxins. Matrix-matched curve-based quantification showed that 26 (57.8%) out of 45 lotus seed samples were contaminated with one or more mycotoxins, and ochratoxin A, aflatoxin B2 , aflatoxin B1 and citrinin were the most prevalent mycotoxins. CONCLUSIONS: This study reports for the first time the incidence of a wide range of 19 mycotoxins in lotus seeds and the proposed method will get broad application for more trace components in other complex matrices.