pH-Controllable Redox Responsive Amphiphilic Viologens for Switchable Emulsions.

A pH and redox dual responsive amphiphilic viologen is synthesized, which can be reversibly transformed among the zwitterionic (SVa), monovalent anionic (SV+), and divalent anionic (SVH2+) forms upon pH variation, exhibiting pH-controllable redox responsive properties. Switchable Pickering emulsions with different droplet size and viscosity are prepared by the mixture of hydrophilic silica nanoparticles and the viologens (SV+ or SVH2+) at acidic conditions, while such combination yielded an oil-in-dispersion emulsion at neutral pH value. Not only can rapid reversible demulsification/stabilization of the Pickering emulsions be achieved by redox reactions, but the rate of redox-demulsification can also be controlled by pH trigger. The dual-responsive amphiphilic viologens have potential applications in developing intelligent colloid materials and molecular logic systems.

Highly Efficient Fabrication of Fluorescent "Turn-On" Lateral Flow Strips for Highly Sensitive Detection of Small Molecules Based on Self-Assembly of AuAg Nanoclusters.

Currently, fluorescent "turn-on" lateral flow assay (FONLFA) has shown enhanced "naked eye" detection sensitivity for small molecules, while it is urgent to adopt biocompatible fluorescent nanomaterials and needs new strategies to simplify the preparation process. In this study, a highly effective method was proposed to produce FONLFA strips for the detection of small molecules. The gold-silver nanoclusters (AuAgNCs) were immobilized onto the nitrocellulose membrane of the strips by the self-assembly of poly(sodium 4-styrenesulfonate), antigen, and AuAgNCs. The immobilization process entails a straightforward mixing of the three components, taking merely 1 min, thereby bypassing the necessity for chemical modification of fluorescent nanomaterials. The strategy offers a significantly simplified process, which substantially enhances the efficiency of the strip fabrication. Utilizing this method, a FONLFA was developed for carbendazim with a visual limit of detection (vLOD) reduced by 40-fold compared with the conventional colorimetric lateral flow assay (LFA). Furthermore, the approach demonstrates versatility by enabling the immobilization of AuAgNCs and streptavidin, which facilitates the development of aptamer-based FONLFAs. The designed aptamer-based FONLFA for kanamycin exhibited a 50-fold reduction in the vLOD compared with conventional colorimetric LFAs. Therefore, FONLFA holds promising potential for widespread applications in the analysis of small molecules.

Sensitive colorimetric and fluorescence dual-mode detection of thiophanate-methyl based on spherical Fe3O4/GONRs composite nanozyme.

Pesticide detection based on nanozyme is largely limited in terms of the variety of pesticides. Herein, a spherical and well-dispersed Fe3O4/graphene oxide nanoribbons (Fe3O4/GONRs) composite nanozyme was applied to firstly develop an enzyme-free and sensitive colorimetric and fluorescence dual-mode detection of thiophanate-methyl (TM). The synthesized Fe3O4/GONRs possess excellent dual enzyme-like activities (peroxidase and catalase) and can catalyze H2O2 to oxidize 3,3',5,5'-tetramethylbenzidine (TMB) into oxidized TMB (oxTMB). We found that Fe3O4/GONRs can adsorb TM through the synergistic effect of multiple forces, thereby inhibiting the catalytic activities of nanozyme. This inhibition can modulate the transformation of TMB to oxTMB, producing dual responses of absorbance decrease (oxTMB) and fluorescence enhancement (TMB). The limits of detection (LODs) of TM were 28.1 ng/mL (colorimetric) and 8.81 ng/mL (fluorescence), respectively. Moreover, the developed method with the recoveries of 94.8-100.8% also exhibited a good potential application in the detection of pesticides residues in water and food samples.

Deep learning-assisted flavonoid-based fluorescent sensor array for the nondestructive detection of meat freshness.

Gas sensors containing indicators have been widely used in meat freshness testing. However, concerns about the toxicity of indicators have prevented their commercialization. Here, we prepared three fluorescent sensors by complexing each flavonoid (fisetin, puerarin, daidzein) with a flexible film, forming a fluorescent sensor array. The fluorescent sensor array was used as a freshness indication label for packaged meat. Then, the images of the indication labels on the packaged meat under different freshness levels were collected by smartphones. A deep convolutional neural network (DCNN) model was built using the collected indicator label images and freshness labels as the dataset. Finally, the model was used to detect the freshness of meat samples, and the overall accuracy of the prediction model was as high as 97.1%. Unlike the TVB-N measurement, this method provides a nondestructive, real-time measurement of meat freshness.

Colorimetric and Reverse Fluorescence Dual-Signal Readout Immunochromatographic Assay for the Sensitive Determination of Sibutramine.

Later flow immunochromatographic assay has been widely used in clinical, environmental, and other diagnostic applications owing to its high sensitivity and throughput. However, most immunoassays operate in the "turn-off" mode for detecting targets of low molecular weight. The signal intensity decreases as the analyte concentration increases, which poses a challenge for achieving ultrasensitive detection at low concentrations and is counterintuitive to new users. In this work, a fluorometric immunochromatographic assay (FICA) is developed to simultaneously read "turn-on" fluorescent and "turn-off" colorimetric signals, where ZnCdSe/ZnS quantum dots act as fluorescence donors and gold nanoparticles (AuNPs) act as quenchers. The fluorescent signal (excitation/emission wavelengths of 365/525 nm) is positively correlated with analytes' concentration. Taking sibutramine (SBT) as the analysis target, the visual limit of detection for SBT reached 3.9 ng/mL, and the limit of Quantitation was 5.0 ng/mg in spiked samples. The developed FICA achieves a high sensitivity in SBT detection, which is much lower than that of the colloidal gold-based immunochromatographic assay. This dual-function detection mode has great potential to be used as a rapid on-site semiquantitative method, providing an alternative mode for the determination of low levels of target analytes.

Rapid and ultra-sensitive lateral flow assay for pathogens based on multivalent aptamer and magnetic nanozyme.

Ultra-sensitive LFA methods for pathogen detection commonly depended on tedious and time-consuming nucleic acid amplification. Here, a high affinity multivalent aptamer (multi-Apt) for S. aureus was obtained through exquisite engineering design. The scaffold and conformation of the multi-Apt were found to be key factors in the detection signal of aptsensors. After optimization, the binding affinity of the multi-Apt to S. aureus was improved by more than 8-fold from 135.9 nM to 16.77 nM. By the joint use of the multi-Apt and a multifunctional nanozyme Fe3O4@MOF@PtPd, a fast and ultra-sensitive LFA for S. aureus was developed (termed MA-MN LFA). In this method, a Fe3O4@MOF@PtPd nanozyme was modified with vancomycin and could efficiently capture and separate S. aureus. Moreover, the multi-Apt worked together with the nanozyme to bind with S. aureus to form a ternary complex at the same time, which simply the fabrication of LFA strip. The developed MA-MN LFA could detect S. aureus as low as 2 CFU/mL within 30 min and a wide linear range of 10-1 × 108 CFU/mL was obtained. The detection is easily operated, fast (can be completed within 30 min) and versatile for Gram-positive pathogens, thus has great potential as a powerful tool in pathogen detection.

A Polyamidoamine-Based Electrochemical Aptasensor for Sensitive Detection of Ochratoxin A.

Sensitive detection of ochratoxin A (OTA) is significant and essential because OTA may pose risks to human and animal health. Here, we developed an electrochemical aptasensor for OTA analysis using polyamidoamine (PAMAM) dendrimers as a signal amplifier. As a carrier, PAMAM has numerous primary amino groups that can be coupled with thiolated complementary strand DNA (cDNA), allowing it to recognize aptamers bound to the surface of horseradish peroxidase (HRP)-modified gold nanoparticles (AuNPs), thereby improving the sensitivity of the aptasensor. When monitoring the positive samples, OTA was captured by the aptamer fixed on the HRP-conjugated AuNP surface by specific recognition, after which the formed OTA-aptamer conjugates were detached from the electrode surface, ultimately decreasing the electrochemical signal monitored by differential pulse voltammetry. The novel aptasensor achieved a broad linear detection range from 5 to 105 ng L-1 with a low detection limit of 0.31 ng L-1. The proposed aptasensor was successfully applied for OTA analysis in red wine, with recovery rates ranging from 94.15 to 106%. Furthermore, the aptasensor also exhibited good specificity and storage stability. Therefore, the devised aptasensor represents a sensitive, practical and reliable tool for monitoring OTA in agricultural products, which can also be adapted to other mycotoxins.

A colorimetric sensor array based on nanoceria crosslinked and heteroatom-doped graphene oxide nanoribbons for the detection and discrimination of multiple pesticides.

Nanozymes have demonstrated high potential in constructing colorimetric sensor array for pesticides. However, rarely array for pesticides constructed without bio-enzyme were reported. Herein, nanoceria crosslinked graphene oxide nanoribbons (Ce-GONRs) and heteroatom-doped graphene oxide nanoribbons (Ce-BGONRs and Ce-NGONRs) were prepared, demonstrating excellent peroxidase-like activities. A colorimetric sensor array was developed based on directly inhibiting the peroxidase-like activities of the above three nanozymes, which realized the discrimination and quantitative analysis of six pesticides. In the presence of pesticides including carbaryl (Car), fluroxypyr-mepthyl (Flu), thiophanate-methyl (Thio), thiram (Thir), diafenthiuron (Dia) and fomesafen (Fom), the peroxidase-like activities of three nanozymes were inhibited to different degrees, resulting in different fingerprint responses. The six pesticides in the concentration range of 0.1-50 µg/mL and two pesticides mixtures at varied ratios could be detected and discriminated, and minimum detection limit for pesticides was 0.022 µg/mL. In addition, this sensor array has been successfully applied for pesticides discrimination in lake water and apple samples. This work provided a new strategy of constructing simple and sensitive colorimetric sensor array for pesticides based on directly inhibiting the catalytic activities of nanozymes.

Fluorescence "turn-on" sensing for five PDE5 inhibitors in functional food based on bimetallic nanoclusters.

Some phosphodiesterase type-5 (PDE5) inhibitors are active ingredients of prescription drugs that are widely used in the treatment of erectile dysfunction (ED). Recently, a large number of substances with this activity have been developed. Illegal addition of PDE5 inhibitors to foods could lead to cardiovascular diseases and even death, which poses a serious threat to food safety, therefore an on-site rapid screening method is urgently needed. Herein, a host functionalized bimetallic nanoclusters, CD/Au Ag NCs, were synthesized through self-assembly of 6-Aza-2-thiothymine gold nanoclusters (ATT-Au NCs), Arginine silver nanoclusters (Arg-Ag NCs) and carboxymethyl ß-cyclodextrin (ß-CMCD). The introduction of Rhodamine 6G (R6G) could quench the fluorescence of CD/Au Ag NCs based on the inner filter effect (IFE) and fluorescence resonance energy transfer effect (FRET). Importantly, it was discovered that several PDE5 inhibitors exhibited a higher binding affinity to ß-CMCD and could displace R6G binding with CD cavity, which disrupted the fluorescence quenching effects and resulted in the fluorescence recovery of CD/Au Ag NCs. This fluorescence turn-on signal could be utilized for the detection of PDE5 inhibitors. At present, emerging PDE5 inhibitor analogues pose a great challenge to food safety due to their unknown efficacy and safety. The proposed method holds the advantages of high sensitivity, simple probe synthesis, easy operation, and simultaneous detection of multiple PDE5 inhibitors. Meanwhile, the successful application in functional food sample demonstrated its high application potential in multiple PDE5 inhibitors screening.

Highly resistant and sensitive colorimetric immunochromatographic assay for sibutramine (SBT) illegally adulterated into diet food based on PDA/AuNP labelling.

A gold nanoparticle (AuNP) based immunochromatographic assay strip is a valuable tool for monitoring chemicals in foods. However, the sensitive ICA strip for SBT is rarely reported due to the fact that monoclonal antibodies (mAbs) against SBT with high affinity are commercially unavailable. Herein, a monoclonal antibody against SBT was prepared through a designed hapten with a carboxyl end-capped space arm. The obtained mAb showed high affinity for SBT and N-desmethylsibutramine, a metabolite of SBT. Furthermore, a series of core-shell NPs, polydopamine (PDA) coated AuNPs (PDA/AuNPs) with controlled shell thickness and packing density were synthesized. The obtained PDA/AuNP-mAb conjugate demonstrated high tolerance to salt and good stability in a wide pH range, which is beneficial for improving the matrix interference common in ICA. As a result, PDA/AuNP-based ICA could quantify SBT in the range of 3.39-69.60 ng mL-1 with a limit of detection (LOD) of 0.98 ng mL-1. This novel ICA improved the sensitivity of the traditional AuNP-based ICA by nearly 12 times. Method validation was conducted with spiked samples of slimming food and human serum and compared with HPLC-MS/MS. Consistent results indicated that high sensitivity, accuracy, and reliability of the PDA/AuNP-based ICA approach were achieved. To the best of our knowledge, this study reported the most sensitive immunoassay for SBT thus far.

A Versatile Strategy of Designing Gold Nanoparticle-cDNA Nanoprobes in Developing Aptamer-Based Lateral Flow Assay for Small-Molecule Detection.

Aptamer-based lateral flow assay (Apt-LFA) has shown promising applications for small-molecule detection. However, the design of the AuNP (gold nanoparticle)-cDNA (complementary DNA) nanoprobe is still a big challenge due to the moderate affinity of the aptamer to small molecules. Herein, we report a versatile strategy to design a AuNPs@polyA-cDNA (poly A, a repeat sequence with 15 A bases) nanoprobe for small-molecule Apt-LFA. The AuNPs@polyA-cDNA nanoprobe contains a polyA anchor blocker, complementary DNA segment to DNA on the control line (cDNAc), partial complementary DNA segment with aptamer (cDNAa), and auxiliary hybridization DNA segment (auxDNA). Using adenosine 5'-triphosphate (ATP) as a model target, we optimized the length of auxDNA and cDNAa and achieved a sensitive detection of ATP. In addition, kanamycin was used as a model target to verify the universality of the concept. Therefore, this strategy can be easily extended to other small molecules; therefore, high application potential in Apt-LFAs can be envisaged.

Ratiometric fluorescent detection of carbendazim in foods based on metallic nanoclusters self-assembled nanocomplex.

Indicator replacement assay combining with fluorescence resonance energy transfer (FRET) effect has attractive performance in sensing small molecules, however, there wasn't application in pesticide molecule sensing reported so far. In this work, we prepared a nanocomplex (NCP), AuAgNCs-CD, through self-assembly of gold nanoclusters (AuNCs), silver nanoclusters (AgNCs) and carboxymethyl-ß-cyclodextrin (CM-ß-CD) by one-step method. The emission of AuNCs was significantly enhance. It was found that FRET between the AuAgNCs-CD and rhodamine B (RhB) existed after AuAgNCs-CD combined with RhB. And carbendazim (CBZ) could induce anti-FRET effect through competing with RhB and binding to AuAgNCs-CD. Thus, this phenomenon was utilized to develop a ratiometric fluorescent detection of CBA. This method was applied in food sample detection and reliable results were obtained. Due to high sensitivity, rapidness and good selectivity, this ratiometric fluorescent method was expected to hold high application potential in monitoring CBZ in foods.

Highly Sensitive Fluorescence Detection of Three Organophosphorus Pesticides Based on Highly Bright DNA-Templated Silver Nanoclusters.

It is still challenging to achieve simultaneous and sensitive detection of multiple organophosphorus pesticides (OPs). Herein, we optimized the ssDNA templates for the synthesis of silver nanoclusters (Ag NCs). For the first time, we found that the fluorescence intensity of T base-extended DNA-templated Ag NCs was over three times higher than the original C-riched DNA-templated Ag NCs. Moreover, a "turn-off" fluorescence sensor based on the brightest DNA-Ag NCs was constructed for the sensitive detection of dimethoate, ethion and phorate. Under strong alkaline conditions, the P-S bonds in three pesticides were broken, and the corresponding hydrolysates were obtained. The sulfhydryl groups in the hydrolyzed products formed Ag-S bonds with the silver atoms on the surface of Ag NCs, which resulted in the aggregation of Ag NCs, following the fluorescence quenching. The fluorescence sensor showed that the linear ranges were 0.1-4 ng/mL for dimethoate with a limit of detection (LOD) of 0.05 ng/mL, 0.3-2 µg/mL for ethion with a LOD of 30 ng/mL, and 0.03-0.25 µg/mL for phorate with a LOD of 3 ng/mL. Moreover, the developed method was successfully applied to the detection of dimethoate, ethion and phorate in lake water samples, indicating a potential application in OP detection.

An ultra-sensitive photothermal lateral flow immunoassay for 17ß-estradiol in food samples.

Hormone residues in food and drinking water endanger human health, therefore, on-site analysis techniques of superior performance are important for monitoring this risk. In this study, an ultra-sensitive photothermal lateral flow immunoassay (LFIA) for quantification of 17ß-estradiol (E2) has been developed. Anti-E2 antibody modified black phosphorus-Au (BP-Au) nanocomposite was developed as a photothermal contrast signal probe and the temperature at test-zone was recorded with an infrared camera. Under the irradiation of 808 nm laser at test-zone, it gave temperatures negatively related to the concentrations of E2 in samples. Under optimal detecting conditions, the developed photothermal LFIA exhibited a limit of detection of 50 pg mL-1, over 100-fold more sensitive than visual LFIA, and a linear range of 3 orders of magnitude. This method has been successfully applied to water, milk, and milk powder samples.

Label-Free Fluorescent Turn-On Glyphosate Sensing Based on DNA-Templated Silver Nanoclusters.

In this work, a label-free fluorescent detection method for glyphosate, based on DNA-templated silver nanoclusters (DNA-Ag NCs) and a Cu2+-ion-modulated strategy, was developed. In the presence of Cu2+, the fluorescence of the DNA-Ag NCs was quenched. Glyphosate can restore the fluorescence of DNA-Ag NCs. By analyzing the storage stability of the obtained DNA-Ag NCs using different DNA templates, specific DNA-Ag NCs were selected for the construction of the glyphosate sensor. The ultrasensitive detection of glyphosate was achieved by optimizing the buffer pH and Cu2+ concentration. The sensing of glyphosate demonstrated a linear response in the range of 1.0-50 ng/mL. The limit of detection (LOD) was 0.2 ng/mL. The proposed method was successfully applied in the detection of glyphosate in a real sample, indicating its high application potential for glyphosate detection.

Accelerated CRISPR/Cas12a-based small molecule detection using bivalent aptamer.

CRISPR/Cas holds great promise for biosensing applications, however, restricted to nucleic acid targets. Here, we broaden the sensing target of CRISPR/Cas to small molecules via integrating a bivalent aptamer as a recognition component. Using adenosine 5'-triphosphate (ATP) as a model molecule, we found that a bivalent aptamer we selected could shorten the binding time between the aptamer and ATP from 30 min to 3 min, thus dramatically accelerating the detection of ATP. The accelerated bivalent aptamer binding to ATP was mainly ascribed to the extended conformation of the aptamer, which was stabilized through linking with a 5 T bases connector on specific loops of the monovalent aptamer. To facilitate on-site detection, we integrated lateral flow assay (LFA) with the CRISPR/Cas sensing strategy (termed BA-CASLFA) to serve as a visual readout of the presence of ATP. In addition, in the CASLFA platform, due to the unique characteristics of LFA, the thermal step of Cas12a inactivation can be omitted. The BA-CASLFA could output a colorimetric "TURN ON" signal for ATP within 26 min, which could be easily discriminated by the naked eye and sensitively quantified by the portable reader. Furthermore, we showed the versatility of BA-CASLFA for detecting kanamycin using a kanamycin bivalent aptamer obtained through the same design as the ATP bivalent aptamer. Therefore, this strategy is amenable to serve as a general sensing strategy for small molecular targets. The above work opened a new way in developing CRISPR-based on-site sensors for clinic diagnosis, food safety, and environmental analysis.

Review in isothermal amplification technology in food microbiological detection.

Food-borne diseases caused by microbial contamination have always been a matter of great concern to human beings. Hence, the research on these problems has never stopped. With the development of microorganism amplification technology, more and more detection methods have come into our vision. However, traditional detection technologies presents more or less drawbacks, such as complicated operation, low accuracy, low sensitivity, long-time detection, and so on. Therefore, more convenient, accurate, and sensitive measurement for the microorganism are needed. Isothermal amplification technology is one of the alternative approach containing the above mentioned advantages. This work mainly summarizes the principles of loop-mediated isothermal amplification (LAMP) and rolling circle amplification (RCA) which belong to isothermal amplification. Meanwhile, the application of LAMP and RCA in food microorganism detection is introduced.

Highly selective and sensitive colorimetric detection for glyphosate based on ß-CD@DNA-CuNCs enzyme mimics.

A colorimetric assay based on an enzyme-inhibition strategy is promising for the on-site detection of pesticide residues. However, very few works of pesticide detection were reported based on the inhibition toward nanozymes although nanozymes have demonstrated many advantages in sensing various targets. Herein, a facile colorimetric detection for Glyp was developed based on ß-CD@DNA-CuNCs enzyme mimics. The ß-CD@DNA-CuNCs with high peroxidase-like activity was synthesized using random DNA double strands as template and ß-CD as surface ligand. ß-CD@DNA-CuNCs could catalyze the H2O2-3,3',5,5'-tetramethylbenzidine (TMB) system. The oxidation product OxTMB with a blue color and presented a large absorption signal at 652 nm. However, Glyp could destroy the synergic effect between redox doublet (Cu2+/Cu+) on the ß-CD@DNA-CuNCs surface, resulting in the inhibition of the peroxidase-like activity. Colorimetric detection for Glyp could be established by detecting the changes of absorption signal at 652 nm. The linear range was 0.02-2 µg/mL and the detection limit was 0.85 ng/mL (3δ/s). The method was applied in measuring Glyp spiked in lake water and various food samples. This method had rapidness, high sensitivity, and selectivity advantages, indicating the high application potential in monitoring Glyp residue in food.

Non-thiolated nucleic acid functionalized gold nanoparticle-based aptamer lateral flow assay for rapid detection of kanamycin.

A novel Apt-LFA has been established for kanamycin based on non-thiolated nucleic acid-modified colloidal gold nanoprobe (AuNPs@polyA-DNA). The improvement in nucleic acid hybridization speed and efficiency was verified by modifying AuNPs with polyA-DNA strands instead of thiolated oligonucleotides (SH-DNA) strands. Moreover, the AuNPs@polyA-DNA was explored to apply in an Apt-LFA. The experimental factors including the concentration of the aptamer, the concentration of SA-DNAT conjugate, the incubation time, and temperature were carefully investigated. In addition, the kanamycin aptamer was modified by extending several bases at its end to modulate the hybridization complementary strand, which was found to significantly improve the performance of Apt-LFA. Under optimal experimental conditions, the Apt-LFA can detect kanamycin in honey with a LOD of 250 ng mL-1 by the naked eyes. A linear range of 50-1250 ng mL-1 was obtained with a LOD of 15 ng mL-1 in honey by a portable reader. The Apt-LFA was successfully applied to the detection of kanamycin in honey with recoveries of 95.1-105.2%.

A simplified fluorescent lateral flow assay for melamine based on aggregation induced emission of gold nanoclusters.

A novel lateral flow assay (LFA) was developed based on the aggregation induced emission (AIE) of gold nanoclusters (AuNCs) for the detection of melamine. We compared the interaction between melamine and AuNCs prepared with 6-Aza-2-thiothymine (ATT) and its four structural analogs as ligands and found that the ATT-AuNCs showed the best AIE induced by melamine. Then the ATT-AuNCs were directly immobilized on the test line of LFA strips as reporters. The sensing platform exhibited a linear response to melamine from 1 to 100 µM (R2 = 0.997) with a detection limit of 217 nM. Moreover, this LFA strip could discriminate 1.0 µM melamine clearly by the naked eyes within 3 min. The recoveries of 91.6-101.5% were attained in milk and baby formula samples. This strip inherited the superior characteristics of low background interference of aggregation-induced fluorescence recognition and LFA technology, thus effectively avoiding the use of antibodies, aptamers, and other biological elements.