Development of a microfluidic device to enrich and detect zearalenone in food using quantum dot-embedded molecularly imprinted polymers.

Mycotoxins are secondary metabolites of certain moulds, prevalent in 60-80% of food crops and many processed products but challenging to eliminate. Consuming mycotoxin-contaminated food and feed can lead to various adverse effects on humans and livestock. Therefore, testing mycotoxin residue levels is critical to ensure food safety. Gold standard analytical methods rely on liquid chromatography coupled with optical detectors or mass spectrometers, which are high-cost with limited capacity. This study reported the successful development of a microfluidic "lab-on-a-chip" device to enrich and detect zearalenone in food samples based on the fluorescence quenching effect of quantum dots and selective affinity of molecularly imprinted polymers (MIPs). The dummy template and functional polymer were synthesized and characterized, and the detailed microfluidic chip design and optimization of the flow conditions in the enrichment module were discussed. The device achieved an enrichment factor of 9.6 (±0.5) in 10 min to quantify zearalenone spiked in food with high recoveries (91-105%) at 1-10 mg kg-1, covering the concerned residue levels in the regulations. Each sample-to-answer test took only 20 min, involving 3 min of manual operation and no advanced equipment. This microfluidic device was mostly reusable, with a replaceable detection module compatible with fluorescence measurement using a handheld fluorometer. To our best knowledge, the reported device was the first application of an MIP-based microfluidic sensor for detecting mycotoxin in real food samples, providing a novel, rapid, portable, and cost-effective tool for monitoring mycotoxin contamination for food safety and security.

Determination of thiram in fruit juices using a bacterial cellulose nanocrystal-based SERS substrate.

Pesticide residues in agri-foods have risk to human health and one solution is to develop simple and accurate methods for rapid detection. We developed a SERS sensor composed of gold nanoparticles (AuNPs) and bacterial cellulose nanocrystal (BCNC) to detect thiram in fruit juice. BCNC-SO3H was used as a stabilizer to support AuNPs via electrostatic repulsion, fabricating a BCNC-AuNPs SERS substrate with uniformly distributed AuNPs. This BCNC-AuNPs SERS substrate was applied to determine thiram residues in peach juice, apple juice, and grape juice with the limits of detection of 0.036 ppm, 0.044 ppm, and 0.044 ppm, respectively. The whole test took 12 min including sample preparation and analysis. The detection limits meet the maximum residue levels of thiram in fruit juices required by China, Europe and North America, indicating that this BCNC-based substrate could serve as a satisfactory SERS sensor for pesticide residue monitoring in the food supply chain.

Application of atomic force microscopy in the characterization of fruits and vegetables and associated substances toward improvement in quality, preservation, and processing: nanoscale structure and mechanics perspectives.

Fruits and vegetables are essential horticultural crops for humans. The quality of fruits and vegetables is critical in determining their nutritional value and edibility, which are decisive to their commercial value. Besides, it is also important to understand the changes in key substances involved in the preservation and processing of fruits and vegetables. Atomic force microscopy (AFM), a powerful technique for investigating biological surfaces, has been widely used to characterize the quality of fruits and vegetables and the substances involved in their preservation and processing from the perspective of nanoscale structure and mechanics. This review summarizes the applications of AFM to investigate the texture, appearance, and nutrients of fruits and vegetables based on structural imaging and force measurements. Additionally, the review highlights the application of AFM in characterizing the morphological and mechanical properties of nanomaterials involved in preserving and processing fruits and vegetables, including films and coatings for preservation, bioactive compounds for processing purposes, nanofiltration membrane for concentration, and nanoencapsulation for delivery of bioactive compounds. Furthermore, the strengths and weaknesses of AFM for characterizing the quality of fruits and vegetables and the substances involved in their preservation and processing are examined, followed by a discussion on the prospects of AFM in this field.

Simultaneous Determination of 147 Pesticide Residues in Traditional Chinese Medicines by GC-MS/MS.

Determination of pesticide residues remains a challenge in traditional Chinese medicines in which complex compounds may interfere with analysis signals. This study reports the development of a simple, effective, and high-throughput method combining gas chromatography-tandem mass spectrometry (GC-MS/MS) with either QuEChERS or solid phase extraction (SPE) to determine 147 pesticide residues in traditional Chinese medicines simultaneously. In SPE, the mixture of n-hexane and ethyl acetate (1:1, v/v) was selected to extract 147 pesticides in honeysuckle, and the extracted pesticides were determined by GC-MS/MS. The limits of detection for all pesticides were within 0.01-0.05 mg/kg. The recoveries were within 70-120% and the relative standard deviations were below 20% for over 90% pesticides. The coefficients of determination were up to 0.999 for the linearity between MS signals and different concentrations of pesticides (20-200 ng/mL). The analytical performance was confirmed in determining pesticide residues in dried tangerine peel. SPE achieved comparable recoveries for all pesticides compared to the QuEChERS method.

Determination of Dencichine in Panax notoginseng in the Forest and Field Using High Performance Liquid Chromatography.

Dencichine is a nonprotein amino acid, an effective ingredient in Panax notoginseng with hemostatic and anti-inflammatory effects. There are few studies on the effects of regions and cultivation models on the accumulation of dencichine. In the current study, the content of dencichine in P. notoginseng collected from its global cultivation and trading center Yunnan, China, (>640 samples) was determined using an optimized high-performance liquid chromatography method coupled with a diode array detector but without derivatization. The recovery rate of this method was 80-110%, the relative standard deviation was <10%, and the limits of detection and quantification were 0.003% (w/w) and 0.01% (w/w), respectively. The content of dencichine in each part of P. notoginseng was as follows: rootlets (39.59%) > main roots (29.91%) > leaves (16.21%) > stems (14.29%). For leaves, P. notoginseng in the forest (5.52 ± 2.26 mg/g) was significantly higher than that in the field (3.93 ± 1.72 mg/g) but opposite for main roots. The origins and altitudes made different contributions to the accumulation of dencichine in P. notoginseng. This study provides an effective analytical method to determine dencichines in various parts of P. notoginseng from different origins and altitudes and supports quality control and product development of P. notoginseng.

Rapid detection of three mycotoxins in animal feed materials using competitive ELISA-based origami microfluidic paper analytical device (µPAD).

We report the development of a competitive ELISA-based origami microfluidic paper-based analytical device (µPAD) for the detection of mycotoxins in animal feed material. The µPAD was patterned using the wax printing technique with the design of a testing pad in the middle and two absorption pads at the side. Anti-mycotoxin antibodies were effectively immobilized on chitosan-glutaraldehyde-modified sample reservoirs in the µPAD. The determination of zearalenone, deoxynivalenol, and T-2 toxin in corn flour was successfully achieved by performing competitive ELISA on the µPAD in 20 min. Colorimetric results were easily distinguished by the naked eye with a detection limit of 1 µg/mL for all three mycotoxins. The µPAD integrated with competitive ELISA holds potential for practical applications in the livestock industry for rapid, sensitive, and cost-effective detection of different mycotoxins in animal feed materials.

Development of an automated solid phase extraction instrument for determination of lead in high-salt foods.

In this study, an instrument for solid phase extraction of heavy metals was developed. Compared to conventional separation equipment, it has advantages of small size, less reagent consumption, time saving, easy operation, and good reproducibility. After analyzing the partition coefficient and adsorption capacity of the exchange resin, the Chelex-100 resin achieved a strong adsorption of lead at pH of 6.0. Compared to the matrix modifier method, solid phase extraction eliminated the interference of salts and the results were more accurate and reliable. The accuracy and stability of this instrument were verified by inter-laboratory testing of the quality control samples of soy sauce (CFAPA-QC1728-4), and the results from six laboratories were within the standard range with relative standard deviation (RSD) of 2.2%-2.8%. Recovery of three spiked samples was within 80.0%-100.0% and RSD was of 2.8%-9.3%, suggesting that this instrument can be used to separate and enrich lead in high-salt foods.

Rapid determination of thiram on apple using a flexible bacterial cellulose-based SERS substrate.

A flexible nanocomposite composed of bacterial cellulose (BC) and gold nanoparticles (AuNPs) was developed as a SERS substrate to determine thiram on apple surface by two collection methods namely "paste-and-peel" and "wiping". Enhancement factor of this SERS substrate for sensing thiram residues was determined to be 2.8 × 105. Compared to the benchtop Raman spectrometer, portable Raman spectroscopic device showed a lower sensitivity towards thiram residues with limit of detection at 0.98 ppm, satisfying maximum residue level of thiram on apple required by both Europe and America. A good linear correlation of SERS peak intensity at 1368 cm-1 and different concentrations of thiram (1-50 ppm) revealed a coefficient up to 0.99. This flexible BC-based SERS substrate has a great analytical performance in sensitivity, reproducibility and stability, and is suitable for rapid detection (<8 min) and quantitative analysis of pesticides on food surface.

Rapid screening and quantification of heavy metals in traditional Chinese herbal medicines using monochromatic excitation energy dispersive X-ray fluorescence spectrometry.

Traditional Chinese herbal medicines are subject to heavy metal contamination. Standard detection methods are too complicated, time-consuming, and expensive for routine analysis, so low-cost methods are in high demand for rapid on-site screening. This study reports a high-sensitivity X-ray fluorescence (HS-XRF) method to determine As, Pb, and Cd residues simultaneously in herbal medicines. It couples monochromatic excitation energy dispersive X-ray fluorescence spectrometry and the fast fundamental parameters method. Each test takes only 10-30 min and costs 1/10th to 1/5th of the standard method. The detection limits, precision and accuracy were evaluated using different approaches, and application notes in practice are also proposed. This study is the first attempt to establish and evaluate HS-XRF in analyzing multiple heavy metals in herbal medicines. This rapid screening method would promote the testing efficiency and thus improve the monitoring of heavy metal contamination in herbal medicines.

Molecularly Imprinted Polymers Coupled with Surface-Enhanced Raman Spectroscopy to Detect Chemical Hazards in Foods.

Preparation of molecularly imprinted polymers coupled with surface-enhanced Raman spectroscopy (MIP-SERS) sensor and its application in detecting chemical hazards in food matrices are described. Sample cleaning is achieved by molecularly imprinted solid-phase extraction (MISPE), and target molecules are detected by SERS. Procedures of MIP synthesis, MISPE preparation, SERS substrate preparation, spectral collection, data analysis, and food analysis application are described.

Inhibition of AI-2 Quorum Sensing and Biofilm Formation in Campylobacter jejuni by Decanoic and Lauric Acids.

Campylobacter jejuni is a major bacterial cause of human diarrheal diseases worldwide. Despite its sensitivity to environmental stresses, C. jejuni ubiquitously distributes throughout poultry production chains. Biofilm formation mediated by quorum sensing is suggested to be critical to the survival of C. jejuni in agroecosystem. C. jejuni possesses LuxS, the enzyme involved in the production of autoinducer-2 (AI-2) signaling molecules. In this study, two fatty acids, namely decanoic acid and lauric acid, were identified to be effective in inhibiting AI-2 activity of C. jejuni. Both decanoic acid and lauric acid at 100 ppm inhibited ∼90% AI-2 activity (P < 0.05) of C. jejuni without bacterial inactivation. The biofilm biomass of two C. jejuni strains was reduced by 10-50% (P < 0.05) after treatment by both fatty acids, while increased biofilm formation was observed for one C. jejuni strain. In addition, both fatty acids effectively reduced the motility of all tested C. jejuni strains. These findings can aid in developing alternative C. jejuni control strategies in agri-food and clinical settings.

Development of a Microfluidic Paper-Based Immunoassay for Rapid Detection of Allergic Protein in Foods.

Food allergy affects up to 10% of the population, causing moderate to severe systemic symptoms and even death. Detecting allergens in food is the most critical and effective measure to reduce food allergy incidents, but the most commonly used kits, lateral flow immunoassay (LFI), are still relatively high in cost for the food industry. Microfluidic paper-based analytical devices (µPADs) demonstrate the potential to address this challenge by substituting the multiple parts/materials in LFI with solely (cellulose) paper as the platform. This study reports the development of a single-piece lateral flow µPAD and demonstrates its capability of detecting allergic protein in various food samples within a 15 min including sample preparation. A confined reagent deposition on the testing zone of the patterned paper was achieved by blade cutting. Surface modification of the cellulose was optimized to enable the complete release of recognizing conjugates and elimination of background noise. The geometry of patterns was optimized to meet the liquid sample's requirement in flow rate. This LFI-format µPAD can detect as low as 1 ppm ovalbumin in standard solution, 0.01% (w/w) egg white protein in spiked cake mix, and the egg content in other commercial food products. Much simpler fabrication procedures and a lower material cost are required by the µPAD than that by conventional LFIs. With the potential to scale up, this study provides an alternative food monitoring option to many sectors throughout the agri-food chain and contributes to improving food safety and public health.

Development of paper-based microfluidic device for the determination of nitrite in meat.

This study employed the use of a microfluidic paper-based analytical device (µPAD) to determine the concentration of nitrite in pork and enhanced the limit of detection by analyzing the coffee-ring effect. The µPAD was fabricated by designing and embedding wax channels onto the cellulose-based filter paper through printing and subjecting the paper to heat treatment to allow wax penetration. Nitrite concentration was determined by monitoring the colorimetric reaction that occurred between nitrite and the added Griess reagent. The limit of detection of this device for nitrite in pork was determined to be 19.2 mg kg-1 by analyzing the inner-chamber reaction, while it could be as low as 1.1 mg kg-1 if the coffee-ring region was analyzed. The overall analysis could be completed within 15 min. This µPAD-based method has potential applications to routinely screen the nitrite concentration of meat products and ensure food safety and consumer health.

Identification of donkey meat in foods using species-specific PCR combined with lateral flow immunoassay.

Food authenticity is a global issue and has raised increasing concerns in the past decades. DNA-based methods are more favourable than the conventional protein-based techniques and have been applied to species identification and meat fraud detection. To effectively identify donkey meat for meat product authentication, a highly specific and robust method that coupled polymerase chain reaction (PCR) with lateral flow immunoassay (LFI) was developed. Donkey-specific PCR primers were designed by targeting at the mitochondrial D-loop gene and the specificity was verified in silico and in vitro against 22 species involved in meat authentication. A limit of detection of 0.0013 ng µL-1 DNA extract was achieved and as low as 0.001% w/w (raw) and 0.01% w/w (cooked) donkey meat in beef were successfully detected using the developed PCR-LFI. LFI strip-based visualization of PCR products allowed for a 10-fold higher sensitivity than conventional gel electrophoresis and significantly reduced the analysis time for the post-PCR analysis. This PCR-LFI is highly suitable for rapid identification of donkey or incorporating into multiplex screening protocol for other meat authentication in the laboratories of both regulatory agencies and commercial services.

Detecting Chemical Hazards in Foods Using Microfluidic Paper-Based Analytical Devices (µPADs): The Real-World Application.

Food safety remains one of the most important issues in most countries and the detection of food hazards plays a key role in the systematic approach to ensuring food safety. Rapid, easy-to-use and low-cost analytical tools are required to detect chemical hazards in foods. As a promising candidate, microfluidic paper-based analytical devices (µPADs) have been rarely applied to real food samples for testing chemical hazards, although numerous papers have been published in this field in the last decade. This review discusses the current status and concerns of the µPAD applications in the detection of chemical hazards in foods from the perspective of food scientists, mainly for an audience with a background in mechanical and chemical engineering who may have interests in exploring the potential of µPAD to address real-world food safety issues.

Rapid detection and quantification of 2,4-dichlorophenoxyacetic acid in milk using molecularly imprinted polymers-surface-enhanced Raman spectroscopy.

We report the development of a molecularly imprinted polymers-surface-enhanced Raman spectroscopy (MIPs-SERS) method for rapid detection and quantification of a herbicide residue 2,4-dichlorophenoxyacetic acid (2,4-D) in milk. MIPs were synthesized via bulk polymerization and utilized as solid phase extraction sorbent to selectively extract and enrich 2,4-D from milk. Silver nanoparticles were synthesized to facilitate the collection of SERS spectra of the extracts. Based on the characteristic band intensity of 2,4-D (391 cm-1), the limit of detection was 0.006 ppm and the limit of quantification was 0.008 ppm. A simple logarithmic working range (0.01-1 ppm) was established, satisfying the sensitivity requirement referring to the maximum residue level of 2,4-D in milk in both Europe and North America. The overall test of 2,4-D for each milk sample required only 20 min including sample preparation. This MIPs-SERS method has potential for practical applications in detecting 2,4-D in agri-foods.

Rapid detection of methicillin-resistant Staphylococcus aureus in pork using a nucleic acid-based lateral flow immunoassay.

Methicillin-resistant Staphylococcus aureus (MRSA) is considered as one of the leading causes of food poisonings worldwide. Due to the high prevalence and extensive challenges in clinical treatment, a rapid and accurate detection method is required to differentiate MRSA from other S. aureus isolated from foods. Since the methicillin resistance of S. aureus is due to the acquisition of the mecA gene from staphylococcal chromosome cassette, the presence of the mecA gene is interpreted as a marker for the identification of MRSA. In this study, a low-cost lateral flow immunoassay (LFI) strip was used to detect the mecA amplicons subsequent to polymerase chain reaction (PCR). The specificity of this PCR-LFI assay was tested between MRSA and methicillin-susceptive S. aureus. Both the test line and control line were shown up on the LFI strip for MRSA, whereas only the control line developed for methicillin-susceptive S. aureus. The detection limit of PCR-LFI assay was 20fg for genomic DNA (100 times more sensitive than gel electrophoresis) and 2×100CFU per 100g of pork products after enrichment at 37°C for 48h. The total detection time of using LFI was 3min, which was faster than the conventional electrophoresis (~45min). With the performance of PCR-LFI, 7 out of 42 S. aureus isolates were identified to be MRSA from imported pork products, which was consistent to the standardized minimum inhibitory concentration assay. This mecA-based PCR-LFI strip can be used for rapid and accurate detection of MRSA isolated from commercial pork products.