Ratiometric Detection of Ochratoxin A Using a Regenerable COF-Au-MB-Apt Signal Probe on a Thermal-Regulated Sensor Module.

Ochratoxin A (OTA) frequently contaminates grains and consequently threatens human health. Herein, we develop a regenerable signal probe and apply it to a Au-based screen-printed electrode module (SPE) for OTA determination. The signal probe, containing a structural covalent organic framework, gold nanoparticles (AuNPs), indicative methylene blue (MB), and a highly selective aptamer, is synthesized with hydrothermal and self-assembly methods. The SPE is permanently functionalized with Prussian blue (PB), AuNPs, and semicomplementary ssDNA. The signal probe, absorbed onto this SPE via hybridization, is competitively expelled by OTA, providing a ratiometric readout of ΔIMB/IPB. Probe regeneration, to erase expired COF-Au-MB-Apt after each analysis, is established with the synergy of OTA-conducted Apt-ssDNA dissociation and on-chip thermal regulation. This advantage powerfully guarantees reduplicative analyses by avoiding irreversible Apt-OTA combination and accumulation on the sensing interface. Regenerations are performed in repetitive cycles (N = 7) with 98.5% reproduction efficiency, and IMB and IPB fluctuations are calculated as 1.45 and 1.12%. This method shows log-linear OTA response in a wide range from 0.2 pg/mL to 0.6 µg/mL, and the limit of detection is 0.12 pg/mL. During natural OTA determinations, recommended readouts match well with HPLC with less than 4.82% relative error.

TPB-DMTP@S-CDs/MnO2 Fluorescence Composite on a Dual-Emission-Capture Sensor Module for Fingerprint Recognition of Organophosphorus Pesticides.

Residues of organophosphorus pesticides (OPs) raise considerable concern, while identifying OPs from unknown sources is still a challenge to onsite fluorescence techniques. Herein, a dual-emission-capture sensor module, based on a TPB-DMTP@S-CDs/MnO2 fluorescence composite, is developed for OP fingerprint recognition. TPB-DMTP@S-CDs/MnO2, synthesized by a hydrothermal method and self-assembly, is spectrographically validated as a dual-wavelength fluorescence source. OP-sensitive catalysis (acetylcholinesterase on acetylthiocholine chloride) is designed to regulate fluorescence by decomposing quenchable MnO2. A flexibly fabricated sensor module supports the optimal dual-wavelength fluorescence excitations and captures and converts fluorescence emissions into equivalent photocurrents for feasible access. The most prominent finding is that dual-fluorescence emissions alternatively respond to levels, species, and multi-pH pretreatments of OPs due to varied MnO2 sizes and distributions. Therefore, OP fingerprint recognition is conducted by refining the multidimensional information from fluorescence-triggered photocurrents and preset hydrolyzation using principal component analysis and the rule of maximum covariance. The recommended method provides a wide dynamic range (1 × 10-6 ∼ 12 µg mL-1), a good limit of detection (7.9 × 10-7 µg mL-1), 15-day stability, and good selectivity to guarantee fingerprint recognition. For laboratory and natural samples, this method credibly identifies a single kind of OPs from multiple species at trace levels (10-5 µg mL-1) and performs well in two-component and multicomponent analyses.

Advanced Electrochemical Biosensing toward Staphylococcus aureus Based on the RPA-CRISPR/Cas12a System and Conductive Nanocomposite.

Staphylococcus aureus (S. aureus) is a prevalent foodborne pathogen that poses significant challenges to food safety. Herein, a sensitive and specific electrochemical biosensor based on RPA-CRISPR/Cas12a is developed for evaluating S. aureus. In the presence of S. aureus, the extracted target DNA fragments are efficiently amplified by recombinase polymerase amplification (RPA). The designed crRNA, binding to Cas12a, effectively recognizes the target fragment cleaving hpDNA. The signal molecule of hpDNA is cleaved from the sensing interface, resulting in a reduction of current response. Under optimal experimental conditions, the developed electrochemical biosensor exhibits remarkable sensitivity in detecting S. aureus. The linear range for quantifying S. aureus in pure culture is 1.04 × 101-1.04 × 108 CFU/mL, with a detection limit as low as 3 CFU/mL. In addition, the biosensor enables the accurate and sensitive detection of S. aureus in milk within a linear range of 1.07 × 101-1.07 × 107 CFU/mL. The electrochemical biosensor enhances anti-interference capability owing to the specific amplification of RPA primers and the single-base recognition ability of crRNA. The RPA-CRISPR/Cas12a biosensor exhibits exceptional anti-interference capability, precision, and sensitivity, thereby establishing a robust foundation for real-time monitoring of microbial contamination.

Molecularly imprinted electrochemical sensor for ethyl carbamate detection in Baijiu based on "on-off" nanozyme-catalyzing process.

Ethyl carbamate (EC) is a carcinogen widely found in the fermentation process of Baijiu. Herein, we construct a molecularly imprinted polymers/MXene/cobalt (II) based zeolitic imidazolate frameworks (MIP/MXene/ZIF-67) nano-enzyme sensor for the detection of EC during Baijiu production. The ZIF-67 is synthesized in situ on the MXene nanosheets to provide a superior catalytic activity to H2O2 and amplify the electrochemical signal. The MIP is prepared by the polymerization reaction to recognize EC. Owing to the interaction between EC and EC-MIP, the interferences are effectively eliminated, greatly improving the accuracy of the expected outcome. This approach attains an ultrasensitive assay of EC ranging from 8.9 µg/L to 44.5 mg/L with detection limit of 0.405 µg/L. The accuracy of this method is confirmed by the recovery experiment with good recoveries from 95.07% to 107.41%. This method is applied in natural EC analyses, and the results are consistent with certified gas chromatograph- mass spectrometer.

Construction of Metal-Organic Framework as a Novel Platform for Ratiometric Determination of Cyanide.

Metal-organic frameworks (MOFs) are frequently utilized as sensing materials. Unfortunately, the low conductivity of MOFs hinder their further application in electrochemical determination. To overcome this limitation, a novel modification strategy for MOFs was proposed, establishing an electrochemical determination method for cyanides in Baijiu. Co and Ni were synergistically used as the metal active centers, with meso-Tetra(4-carboxyphenyl)porphine (TCPP) and Ferrocenecarboxylic acid (Fc-COOH) serving as the main ligands, synthesizing Ni/Co-MOF-TCPP-Fc through a hydrothermal method. The prepared MOF exhibited improved conductivity and stable ratio signals, enabling rapid and sensitive determination of cyanides. The screen-printed carbon electrodes (SPCE) were suitable for in situ and real-time determination of cyanide by electrochemical sensors due to their portability, low cost, and ease of mass production. A logarithmic linear response in the range of 0.196~44 ng/mL was demonstrated by this method, and the limit of detection (LOD) was 0.052 ng/mL. Compared with other methods, the sensor was constructed by a one-step synthesis method, which greatly simplifies the analysis process, and the determination time required was only 4 min. During natural cyanide determinations, recommended readouts match well with GC-MS with less than 5.9% relative error. Moreover, this electrochemical sensor presented a promising method for assessing the safety of cyanides in Baijiu.

An advanced ratiometric molecularly imprinted sensor based on metal ion reoxidation for indirect and ultrasensitive glyphosate detection in fruit.

An indirect and ultrasensitive ratiometric molecularly imprinted (MIP) sensor, based on metal ion reoxidation, is introduced for glyphosate (GLY) determination in fruit. As high-performance signal amplification substrates, carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) are conveniently modified on GCE. The artificial antibody-MIP membrane, presents typical three-dimensional structure to GLY template. Built-in reference methylene blue (MB) is directly electropolymerized on MWCNTs-Au/GCE. Particularly, Cu2+ and GLY interestingly form chelate complex, and the Cu2+ (ICu) in Cu(Ⅱ)-GLY-complex can be reoxidized, and indirectly quantizes GLY. The reference signal (IMB) presents noteworthy stability with different GLY levels, and the ratiometric readout (ICu/IMB) is recognized as a more trustworthy indicator to quantize GLY. Proposed sensor presents broad range as 1.73 âˆ¼ 400 ng/mL, and limit of detection is well found as 0.24 ng/mL (S/N = 3). Finally, as-fabricated method is verified with standard HPLC in real-fruit-sample, and the errors and recovery rates are calculated as 3.4% âˆ¼ 6.7% and 94.4% âˆ¼ 104.6%, respectively.

Competitive immunoassay using enzyme-regulated Fe3O4@COF/Fe3+ fluorescence probe for natural chloramphenicol detection.

Accurate and sensitive detection of chloramphenicol (CAP) in natural samples is essential for ensuring human health. Herein, an enzyme-regulated fluorescence sensor using Fe3O4@COF/Fe3+ probe, is developed for CAP determination. Fe3O4@COF, synthesized via hydrothermal method, exhibits dual functions as a magnetic carrier and signal probe. Bovine serum albumin conjugated-chloramphenicol, adsorbed on the surface of Fe3O4@COF, competes with CAP for antibody binding. The antibody interacts with alkaline phosphatase via the biotin-streptavidin system. Meanwhile, ascorbic acid, produced from the enzyme-catalyzed reaction dominated by alkaline phosphatase, effectively restores the fluorescence of Fe3O4@COF that is quenched by Fe3+. After experimental verification and gradual optimization, a logarithmic linear relationship between CAP concentration and fluorescence intensity is established in the range of 2 × 10-4∼10 µg mL-1, with a good limit of detection (9.2 × 10-5 µg mL-1). Proposed method exhibits excellent stability (15 days) and reusability (8 cycles), providing a sensitive and reliable method for accurate CAP detection. The readouts show good agreement with HPLC and recoveries during laboratory and natural CAP analysis.

Non-destructive assessment of equivalent umami concentrations in salmon using hyperspectral imaging technology combined with multivariate algorithms.

This study utilized equivalent umami concentrations (EUC) to characterize umami intensity in salmon with different freeze-thaw times. A rapid and non-destructive method was established to determine EUC values in salmon which is based on hyperspectral imaging (HSI) system combined with multiple characteristic variable screening methods. The established CARS-PLS model showed greater advantages in correlating the reference values of spectral data with EUC in salmon with Rc of 0.9012, Rp of 0.9009, RMSECV of 0.82, and RMSEP of 0.88. The model was employed pixel-wise to visualize the distribution of EUC with different freeze-thaw times, which demonstrated the reduction of EUC value with the increasing of freeze-thaw times. Therefore, this reseearch showed hyperspectral imaging (HSI) system combined with chemometrics possesses a substantial capability to predict and visualize the EUC of salmon, which would provide an intuitive understanding of salmon quality prediction and detection.

Collection, nucleic acid release, amplification, and visualization platform for rapid field detection of rice false smut.

Rice false smut (RFS) has brought serious food safety problems to the world. Reliable diagnostic tools are needed for the field detection of RFS. Traditional polymerase chain reaction (PCR) is inefficient due to sample transport and preparation, which cannot adapt to the needs of field detection. Herein, we successfully developed a simple, portable microfluidic test platform to rapidly detect RFS. To simplify the operation, we integrated spore purification, nucleic acid release, and amplification into one chip. A micro air pump was used to separate the spores from the impurities and complete the collection of the spores through the airflow. We rapidly lysed spores and released nucleic acids by the benzyl chloride method. The loop-mediated isothermal amplification (LAMP) products could be combined with SYBR Green I to observe the results visually. On-chip sample tests showed that the spore collection efficiency was approximately 78%. By providing on-chip detection results, the chip had 100% specificity and a detection limit of 100 copies/reaction. At the same time, the stability (CV < 5%) and quantitative ability (R2 = 0.989) of the chip were also guaranteed. Through the visual detection of large samples, the on-chip detection results were highly concordant with the classical RT-PCR detection results, and the detection timeliness was greatly enhanced. Compared with RT-PCR, the single-sample detection time was shortened by about twenty minutes. The proposed micro-diagnostic tool did not require any large end-point detection instruments and avoided the complicated operation of nucleic acid extraction. As a result, in the future, our microfluidic chip could be used for rapid and real-time monitoring and early warning of rice false smut spores in rice paddies.

Programmable-Printing Paper-Based Device with a MoS2 NP and Gmp/Eu-Cit Fluorescence Couple for Ratiometric Tetracycline Analysis in Various Natural Samples.

Paper-based fluorescence devices, with smartphone aids, bring considerable operation convenience for tetracycline (TC) sensing. Nevertheless, they must meet the challenge in real determination against complicated backgrounds. Considering that, we present a programmable-printing paper-based device and then apply it to TC determination for various natural samples. MoS2 NPs and Gmp/Eu-Cit are synthetized as composite probes. A static quenching process is found with MoS2 NP fluorescence at 430 nm, while significant magnification of Gmp/Eu-Cit emission is obtained at 617 nm, establishing a valuable ratiometric indicator. Remarkably, two-stage programmable printing maximizes the proposed sensing capability. A transitive device, containing a gradually changing amount of a certain probe, is prepared to sense TC. With a homemade smartphone application and 3D-printed measurement chamber, the corresponding signals are examined to explore optimal setups. These setups are automatically processed to prepare the final-version device, not requiring manual operations. Benefitting from this interesting feature, the proposed device gains many rewards in performances. It effectively senses TC in a wide range from 12.7 nM to 80 µM and simultaneously provides naked eye-legible signals and smartphone-based readouts with confident selectivity and stability. This device is consequently applied for various samples of soil, river water, milk, and serum and meets well with HPLC-MS and recovery tests.

Rapid detection of Atlantic salmon multi-quality based on impedance properties.

To establish a rapid, convenient, and low-cost method to assess the quality of Atlantic salmon, we analyzed the impedance between 10-1 and 105 Hz for Atlantic salmon/rainbow trout, chilled/frozen-thawed salmon, and fresh/stale salmon. We combined chemometrics with impedance properties to create a multi-quality index for Atlantic salmon. The accuracy of all three models established can reach 100% in distinguishing Atlantic salmon from rainbow trout and distinguishing chilled salmon from frozen-thawed salmon. We applied a partial least squares method to create a quantitative prediction model of bioimpedance spectroscopy and the value of total volatile basic nitrogen. The correlation coefficients of the training and test sets were 0.9447 and 0.9387. Our results showed that the combination of impedance properties and chemometrics was a simple and effective application to evaluate Atlantic salmon quality.

A colorimetric hydrogen sulfide sensor based on gellan gum-silver nanoparticles bionanocomposite for monitoring of meat spoilage in intelligent packaging.

A colorimetric hydrogen sulfide (H2S) sensor based on gellan gum capped silver nanoparticles was developed to real-time monitor meat spoilage. The colorimetric sensor strategy was attributed to the ultrastrong binding ability of Ag with H2S to form Ag2S. The sensor enables the analysis of H2S with a limit of detection (LOD) of 0.81 µM, and it exhibited excellent selectivity toward H2S against other volatile components generated from chicken breast and silver carp during spoilage. By these virtues, the sensor presented visible color changes from yellow to colorless by in situ and nondestructively sensing H2S generated from chicken breast and silver carp in a packaging system. This strategy provided a simple but useful, non-destructive, robust, cost-effective, and user-friendly platform to real time monitor meat spoilage for intelligent food packaging.

Simultaneous measurement of total acid content and soluble salt-free solids content in Chinese vinegar using near-infrared spectroscopy.

Total acid content (TAC) and soluble salt-free solids content (SSFSC) in Chinese vinegar are 2 important indicators in the assessment of its quality. This paper shows the feasibility to determine TAC and SSFSC in Chinese vinegar by near-infrared (NIR) spectroscopy. Synergy interval partial least square (Si-PLS) algorithm was performed to calibrate the regression model. The number of PLS factors and the number of intervals were optimized simultaneously by cross-validation. The performance of the model was evaluated according to root mean square error of prediction (RMSEP) and correlation coefficient (R) in the prediction set. The optimum Si-PLS model for TAC was achieved with RMSEP = 0.264 g/100 mL and R(p) = 0.9655; the optimum Si-PLS model for SSFSC was achieved with RMSEP = 1.93 g/100 mL and R(p) = 0.9302. The overall results demonstrated that NIR spectroscopy combined with Si-PLS could be utilized to determinate TAC and SSFSC in Chinese vinegar, and NIR spectroscopy has a potential to be used in vinegar industry.