Blue oxTMB Research Articles

Reliable ratiometric colorimetric monitoring of dopamine in practice based on the catalytic signal amplification of nano CeO2/CuO modified carboxylated chitosan

To reliably monitor dopamine (DA), a common neurotransmitter, a kind of nano CeO2/CuO modified carboxylated chitosan nanozyme (CeO2/CuO@CC) was constructed under microwave-assisted conditions. The proposed CeO2/CuO@CC exhibited superior synergistic- enhancement oxidase-like activity, with high specific surface area and superior stability. It could accelerate the oxidization of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxTMB in the air. What’s more, the presence of trace DA could exclusively change the enzyme-like activity of CeO2/CuO@CC, accompanied with visual color changes from blue to colorless. Under the optimized conditions, there expressed double ratiometric linear relationships between lg(A652/A285)/lg(A370/A285) and cDA, with a large linear range between 6.0 × 10−9 and 1.0 × 10−5 M of DA. When applied for visual detection of DA in real samples, the detection limit was low to 2.0 × 10−9 M with RSD % less than 4.14 %. The mechanisms for enhanced oxidase-mimic activity of CeO2/CuO@CC and its selective detection of DA were proposed. This work will offer an insight to design efficient nanozyme for visual detection of bio-analytes in practice.

CoMnOx Nanoflower-Based Smartphone Sensing Platform and Virtual Reality Display for Colorimetric Detection of Ziram and Cu2.

Transition metal doping is an ideal strategy to construct multifunctional and efficient nanozymes for biosensing. In this work, a metal-doped CoMnOx nanozyme was designed and synthesized by hydrothermal reaction and high-temperature calcination. Based on its oxidase activity, an "on-off-on" smartphone sensing platform was established to detect ziram and Cu2+. The obtained flower-shaped CoMnOx could exhibit oxidase-, catalase-, and laccase-like activities. The oxidase activity mechanism of CoMnOx was deeply explored. O2 molecules adsorbed on the surface of CoMnOx were activated to produce a large amount of O2·-, and then, O2·- could extract acidic hydrogen from TMB to produce blue oxTMB. Meanwhile, TMB was oxidized directly to the blue product oxTMB via the high redox ability of Co species. According to the excellent oxidase-like activity of CoMnOx, a versatile colorimetric detection platform for ziram and Cu2+ was successfully constructed. The linear detection ranges for ziram and Cu2+ were 5~280 μM and 80~360 μM, and the detection limits were 1.475 μM and 3.906 μM, respectively. In addition, a portable smartphone platform for ziram and Cu2+ sensing was established for instant analysis, showing great application promise in the detection of real samples including environmental soil and water.

Oxidized TMB mediated dual-optical sensing-based paper analytical device for high-throughput visual monitoring of 2,4-dichlorophenoxyacetic acid

High-throughput and accurate monitoring of pesticide residues is extremely crucial, but remains challenging. Herein, oxidized 3,3′,5,5′-tetramethylbenzidine (oxTMB) mediated dual-optical sensing-based paper analytical device (PAD) was unveiled for on-site visual detection of 2,4-dichlorophenoxyacetic acid (2,4-D) in real samples. Colorimetric blue oxTMB (652 nm) can be reduced to colorless TMB by ascorbic acid (AA). Meanwhile, oxTMB was demonstrated for the first time to switch oxidation and condensation reactions between o-phenylenediamine and AA to produce yellow-emission (564 nm) and blue-emission (429 nm) probes. Thus, colorimetric and ratiometric fluorescent dual-mode detection system was developed for 2,4-D detection by combining with alkaline phosphatase (ALP) initiating AA generation (ascorbic acid-2-phosphate as the substrate). Notably, the developed method can correct itself to improve the accuracy due to the dual-response signals. Using the principle, ALP-loaded PAD (array 3 × 10) was designed for efficient and high-performance detection of 2,4-D in turnip and cauliflower with good sensitivity (0.09 μg/mL for colorimetry, 0.06 μg/mL for fluorometry), satisfactory recovery (98.0 %–107.2 %), and high consistency (relative error ≤ 3.1 %). This work demonstrated a promising strategy for high-throughput visual detection of 2,4-D, which may provide a new pathway for the further development of dual-mode sensing platforms to efficiently detect other targets for food safety.

Covalent organic frameworks surface-anchored with silver nanoparticles for colorimetric detection of dopamine based on its oxidase-mimicking activity

Sensitive detection of dopamine (DA) is essential for the diagnosis of nervous system diseases. Herein, a nanozyme-based colorimetric platform for DA determination was proposed. Using an imine-type covalent organic framework (COF-TD) as a host matrix, a nanozyme (AgNPs@COF-TD) was fabricated by anchoring silver nanoparticles (AgNPs) on the surface of COF-TD via in situ reduction of silver ions. Various characterization results confirmed the successful synthesis of flower-like AgNPs@COF-TD with surface-anchored AgNPs, which could expose more active sites. The as-prepared AgNPs@COF-TD exhibited satisfactory oxidase-mimicking activity with a low Km, which could catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to blue oxidized TMB (oxTMB) with significant absorbance at 657 nm. The introduction of DA not only reduced blue oxTMB to colorless TMB but also competed with TMB for the active sites of AgNPs@COF-TD, making a significant decrease in absorbance. Therefore, a label-free sensing strategy based on the oxidase-mimicking activity of AgNPs@COF-TD was successfully established for the colorimetric detection of DA, with a detection limit down to 0.2 μM. The practical applicability of this assay was further validated by analyzing DA-spiked human urine samples. This work provided a facile approach to prepare COF-based nanozymes, and demonstrated their great potential in colorimetric detection and bioanalysis.

Ultrafine FeCuAgCeGd–based high–entropy nanozyme: Preparation, catalytic mechanism, and point–of–care detection of dopamine in human serum

Point–of–care testing (POCT) plays vital role in clinic field, especially in resource–limited areas. The nanozyme–based colorimetric platform is the potential strategy for POCT. Therefore, a high–entropy allay nanozyme (HEAzyme) was prepared via formaldehyde–assisted metal–ligand crosslinking method using multi–metal (FeCuAgCeGd) species. The obtained FeCuAgCeGd–HEAzyme showed coordination nanoparticles with ultrafine size (average size was approximately 5 nm). The peroxidase–like (POD–like) activity of FeCuAgCeGd–HEAzyme was verified through density functional theory (DFT) calculations, demonstrating the capability of FeCuAgCeGd–HEAzyme to catalyze H2O2 and generate hydroxyl radicals (•OH). These radicals can oxidize colorless 3,3′,5,5′–tetramethylbenzidine (TMB) to blue oxTMB. Dopamine (DA) scavenges •OH, resulting in an “on–off” effect for TMB oxidation. Subsequently, an FeCuAgCeGd–HEAzyme colorimetric sensor was developed for DA detection, which features a low detection limit of 0.77 µM and a linear range of 5.0–70.0 µM. The sensor demonstrated a recovery ratio in spiked serum samples ranging from 97.7 % to 104.3 % with high stability and favorable selectivity. Furthermore, an FeCuAgCeGd–HEAzyme POCT platform was designed for on–site detection of DA, achieving a reliable linear detection range (5.0–70.0 µM) and a low detection limit (5.69 µM). This platform offers a facile and efficient strategy for rapid, visual, and on–site DA detection in clinical settings with advantages of cost–effectiveness and ease of operation.

Portable hydrogel kit driven by bimetallic carbon dots nanozyme for H2O2-self-supplying dual-modal monitoring of atmospheric CH3SH

Due to the typical volatility of gaseous pollutant methyl mercaptan (CH3SH), the development of a facile, reliable, and accurate onsite environmental surveillance of highly toxic CH3SH faces many challenges, but it is critical to environmental atmosphere assessment and safeguarding public health. Here, we prepared a novel bimetallic carbon dots (Fe&Cu@CDs) nanozyme with high peroxidase-mimicking activity to design a portable hydrogel kit for onsite visual H2O2-self-supplying enzymatic cascade catalytic colorimetric and photothermal signal synergistic amplification dual-modal monitoring of CH3SH in atmospheric environment. Assisted by alcohol oxidase (AOX), CH3SH could be specifically converted into H2O2 for oxidizing chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) catalyzed by Fe&Cu@CDs to produce dark blue ox-TMB with absorption at 652 nm and photothermal characters. Consequently, a CH3SH concentration-dependent change both in naked-eye color and photothermal effect-triggered temperature were observed. By hybridizing AOX-assisted Fe&Cu@CDs + TMB with agarose, a H2O2-self-supplying colorimetric and photothermal signal synergistic amplification sensory hydrogel kit integrated with Color Picker APP-installed smartphone and 660 nm laser-equipped handheld thermal imager for CH3SH was proposed with acceptable results in atmospheric environment around wastepile (e.g., solid waste and food waste piles), which exhibited great potentials to further develop commercial onsite monitoring platforms in warning-early abnormal atmospheric CH3SH for safeguarding environmental health.

Enhanced detection of copper residues on citrus surfaces: An innovative copper fenton-catalysed colorimetric approach using creatinine and molybdenum trioxide

Accurate quantification of residual copper from inorganic copper reagents on the surface of citrus fruits is crucial for preventing diseases caused by excessive heavy metal intake. In this study, we developed a rapid colorimetric strategy for copper detection that is enhanced under near-neutral pH conditions, using molybdenum trioxide (MoO3) and creatinine for copper-Fenton catalysis. The introduction of MoO3 enriches the redox reactions of Cu2+/Cu+ within the complex formed by copper ions and creatinine, leading to enhanced copper-Fenton catalytic activity at near-neutral pH. In the presence of H2O2, this system oxidises colourless 3,3′,5,5′-tetramethylbenzidine (TMB) to blue oxTMB, thereby affording a sensitive colorimetric detection platform for Cu2+. The designed method offers high sensitivity for Cu2+ detection, with a linear range of 0.011.0 μg/mL and a detection limit as low as 0.007 μg/mL. This method was effectively used to analyse copper reagent residues on citrus surfaces, achieving recovery rates of 92.0%115.0%. With its high sensitivity, selectivity, simplicity, and low cost, this colorimetric method shows great potential as an effective approach for determining copper ion levels, particularly for monitoring copper residues in agricultural products.

A colorimetric sensing strategy based on chitosan-stabilized platinum nanoparticles for quick detection of α-glucosidase activity and inhibitor screening.

α-Glucosidase (α-Glu) is implicated in the progression and pathogenesis of type II diabetes (T2D). In this study, we developed a rapid colorimetric technique using platinum nanoparticles stabilized by chitosan (Ch-PtNPs) to detect α-Glu activity and its inhibitor. The Ch-PtNPs facilitate the conversion of 3,3',5,5'-tetramethylbenzidine (TMB) into oxidized TMB (oxTMB) in the presence of dissolved O2. The catalytic hydrolysis of 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G) by α-Glu produces ascorbic acid (AA), which reduces oxTMB to TMB, leading to the fading of the blue color. However, the presence of α-Glu inhibitors (AGIs) hinders the generation of AA, allowing Ch-PtNPs to re-oxidize colorless TMB back to blue oxTMB. This unique phenomenon enables the colorimetric detection of α-Glu activity and AGIs. The linear range for α-Glu was found to be 0.1-1.0 U mL-1 and the detection limit was 0.026 U mL-1. Additionally, the half-maximal inhibition value (IC50) for acarbose, an α-Glu inhibitor, was calculated to be 0.4769mM. Excitingly, this sensing platform successfully detected α-Glu activity in human serum samples and effectively screened AGIs. These promising findings highlight the potential application of the proposed strategy in clinical diabetes diagnosis and drug discovery.

Mechanism of ferrous ions reversibly inhibiting of oxidase-Like activity of Co3O4 nanowires and its application in horticultural fertilizer analysis

Nanozymes are becoming substitutes of some natural enzymes due to their excellent properties, however, little attention has been paid to the catalytic mechanisms and especially inhibition mechanism of nanozymes, which impeded advancement in nanozymes and their wide application including bioanalysis. Herein, non-toxic filamentous virus, M13 phage templated Co3O4 nanowires (NWs) was facilely prepared, which exhibits excellent oxidase (OXD)-like activity and catalytic efficiency. The catalytic mechanism of OXD-like is oxygen vacancies, confirmed by high resolution X-ray photoelectron spectrum of O 1 s, electron paramagnetic resonance spectra and reactive oxygen species scavenging experiments. The substrate of 3,3′,5,5′-tetramethylbenzidine (TMB) for OXD-like is oxidized into blue ox-TMB, and then turned to yellow diimine, depending on TMB concentration and incubation time. Then the mechanism of ferrous ions (Fe2+) inhibition of the oxidase-like activity of Co3O4 NWs was uncovered that Fe2+ competes with TMB for oxygen vacancies. The inhibition is reversible with the type of hybrid of competitive inhibition and non-competitive inhibition with the inhibition constants (KI and KI') of 0.134 mM and 1.012 mM, respectively. Finally, on the basis of the inhibitory effect, a fast, selective and sensitive Fe2+ colorimetric analysis was proposed with two linear parts and a limit of detection of 0.21 μM Fe2+ (S/N = 3), which is applicable to detect Fe2+ in horticultural fertilizers. This study may offer a new strategy for the in-depth study of inhibition mechanism of other nanozymes, which would extend the scope of nanozyme-based biosensing.

A dual-mode strategy for diacetyl detection based on colorimetry and surface-enhanced Raman scattering

Diacetyl is an important food additive with rich creamy flavor whose excessive intake may cause various diseases. Herein, for the first time, we designed a dual-mode method for diacetyl detection based on colorimetry and surface-enhanced Raman scattering (SERS) technology. Diacetyl exhibited excellent oxidase-like catalytic activity under light-mediated condition, facilitating the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to blue ox-TMB for colorimetric detection. Notably, diacetyl could control the oxidation process of TMB by simply adjusting light irradiation, unlike traditional enzymatic reactions that required terminators to stop the reaction. Interestingly, ox-TMB was an ideal Raman marker for SERS analysis. Then, a three-dimensional flower-like zinc oxide/silver (ZnO/Ag) for sensitive SERS detection was prepared, which exhibited outstanding SERS effect with a high enhancement factor of 1.89 × 108 due to effective charge transfer and synergistic effect between ZnO and Ag. Based on these, SERS method was constructed to indirectly detect diacetyl, which has not been reported due to low Raman activity of diacetyl. The linear ranges of colorimetry and SERS were 0.5–60 μM and 10−9-10−5 M respectively, with detection limits of 0.41 μM and 0.73 nM. Obviously, the dual-mode strategy combined the simplicity and practicality of colorimetry and the high sensitivity of SERS, offering great potential applications in practical samples.

Highly efficient persistent luminescent nanozymes-based luminescence-colorimetric dual-mode sensor for total antioxidant capacity assay

Total antioxidant capacity (TAC) is a vital parameter to assess the gross antioxidant activity in biological and food matrices. Accurate determination of TAC has become increasingly important for health monitoring and dietary guidance. Here, we report a persistent luminescent nanozyme (ZGO-Pt) based luminescence-colorimetric dual-mode sensor for selective determination of TAC. ZGO-Pt was fabricated based on ZnGa2O4:Cr nanoparticles (ZGO) with platinum nanoflower as synthetic additive. The prepared ZGO-Pt exhibited fascinating peroxidase-mimetic activity and persistent luminescence, and enabled effectively catalysis of the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to blue oxTMB. Meanwhile, the generated oxTMB gave strong absorption at 652 nm to quench the luminescence of ZGO-Pt at 700 nm via the inner filter effect. Antioxidants could compete with the oxidable TMB or reduce oxTMB to TMB, resulting in the simultaneous variations of blue color and luminescence to provide the basis for dual-mode assay for TAC. The highly efficient catalysis of ZGO-Pt (Km, 3.12 mM for H2O2, 0.592 mM for TMB) allows the dual-mode sensor to realize precise determination of TAC with a wide linear range (0.5–200 μM and 1–300 μM for colorimetric and luminescence mode, respectively) and low limit of detection (LOD, 0.05 μM and 0.78 μM for colorimetric and luminescence mode, respectively). The developed dual-mode sensor was successfully applied to the determination of TAC in vitamin tablet, beverages and fruits. This work provides an efficient persistent luminescent nanoparticle-based nanozyme for a facile TAC assay with further vision in biosensing and food technology.

Hydrogel-involved portable colorimetric sensor based on oxidase mimic Fe/Co-NC for acetylcholinesterase detection and pesticides inhibition assessment

Herein, we synthesized a novel N-doped carbon layer encapsulated Fe/Co bimetallic nanoparticles (Fe/Co-NC), which exhibited superior oxidase-like activity due to the facilitation of electron penetration and the formation of metal-nitrogen active sites. Fe/Co-NC could catalyze the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) to blue oxTMB. Acetylcholinesterase (AChE) could catalyze the hydrolysis of thioacetylcholine to produce reducing thiocholine, which prevented TMB from oxidation. Thus, a portable hydrogel colorimetric sensor was developed for on-site and visual monitoring of AChE with the detection limit of 0.36 U L−1, and successfully applied to detect AChE in human erythrocyte samples. Furthermore, this platform was used to investigate the inhibition of triazophos on AChE activity.

A three-dimensional network structure of metal-based nanozymes for the construction of colorimetric sensors for the detection of antioxidants.

Although many excellent nanozymes have been developed, designing and synthesizing highly active nanozymes is still challenging. Here, we developed a metal-based nanozyme (metal = Co, Fe, Cu, Zn) with a three-dimensional network structure. It possesses excellent peroxidase activity and catalyzes the reaction between H2O2 and TMB to produce blue oxTMB, while antioxidants have different reducing power on the oxidation product of TMB (oxTMB), which leads to different absorbance and color changes. Using these color reactions, different nanozymes were used to form a colorimetric sensor array with seven antioxidants, and seven antioxidants were sensitively identified. And the differences between the three nanozymes were compared by density functional theory calculations and enzyme kinetic curve results. In conclusion, the colorimetric sensor array based on metal-based nanozymes provides a good strategy for the identification and detection of antioxidants, which has a broad application prospect.

A novel N-CNDs/PAni modified molecular imprinted polymer for ultraselective and sensitive detection of ciprofloxacin in lentic ecosystems: a dual responsive optical sensor.

The research study describes the development of a hybrid nanocomposite called nitro-doped carbon nanodots/polyaniline/molecularly imprinted polymer (N-CNDs/PAni/MIP). This composite is specifically engineered to function as a durable and flexible dual-response sensor to detect and analyze pharmaceutical organic contaminants (POCs). Powder X-Ray diffraction (PXRD), Fourier transform infrared (FT-IR), thermogravimetric analysis (TGA), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) were employed to perform an exhaustive structural and morphological analysis of N-CNDs/PAni/MIP. N-CNDs/PAni/MIP emitted blue luminescence under ultraviolet irradiation and exhibited typical excitation-dependent emission properties. It can act as fluorescent probe for the detection of CIPRO with high selectivity and sensitivity with an IF value of 4.2. Furthermore, N-CNDs/PAni/MIP exhibited high peroxidase-like catalytic behavior. After adding CIPRO to the N-CNDs/PAni/MIP/TMB/H2O2 system, the blue color of the solution faded due to the reduction of blue ox-TMB to colorless TMB. Based on these two phenomena, with CIPRO as the target analyte, the N-CNDs/PAni/MIP dual sensor showed a minimal detection limit of 70 pM for the fluorescent signaling platform and 3.5 nM for the colorimetric probe with a linear range of 0.038-200 nM. The fluorometric and colorimetric assays based on N-CNDs/PAni/MIP for CIPRO detection were then successfully applied to lentic water as well as to tap water samples, demonstrating the sensitivity and dependability of the instrument. Furthermore, the synthesized PVA (N-CNDs/PAni/MIP) films enable the recognition of CIPRO, and these films have the potential to be integrated into portable sensing devices, providing a practical solution for rapid and on-site detection of CIPRO in various samples.

CoO/Co-graphene quantum dots as an oxidative mimetic nanozyme for the colorimetric detection of L-cysteine.

The preparation of cobalt-based nanozymes with high oxidase-like activity still needs more efforts. In this paper, we report the synthesis of a CoO/Co-tryptophan-functional graphene quantum dot hybrid (CoO/Co-Try-GQD). Firstly, cobalt ions coordinate with the indole nitrogen on Try-GQD to form a complex, followed by thermal reduction and oxidation. The resulting hybrid presents a three-dimensional network structure, and CoO/Co nanoparticles are uniformly dispersed on the graphene sheet with an average size of 10 ± 0.24 nm. This unique structure improved the oxidase-like activity of the hybrid, enabling it to catalyze the oxidation of colorless 3,3',5,5'-tetramethylbenzidine (TMB) to rapidly produce deep blue ox-TMB with a strong absorbance at 652 nm (A652). A colorimetric method was developed for the highly sensitive determination of L-cysteine (L-cys) based on the inhibition of the hybrid's oxidase-like activity and low A652 caused by the binding of L-cys with Co atoms on CoO/Co via the Co-S bond. The A652 linearly decreased with increasing L-cys concentration in the range of 0.05-2 μM, and the detection limit was 0.032 μM. Further, the established method has been successfully applied to the determination of L-cys in milk.

Peroxidase activity of a Cu-Fe bimetallic hydrogel and applications for colorimetric detection of ascorbic acid.

A Cu-Fe bimetallic hydrogel (2-QF-CuFe-G) was constructed through a simple method. The 2-QF-CuFe-G metallohydrogel possesses excellent peroxidase-like activity to catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. The catalytic mechanism was confirmed by the addition of •OH radical scavenger isopropyl alcohol (IPA), tert-butyl alcohol (TBA) and ˙OH trapping agent terephthalic acid (TA). Remarkably, the resultant blue ox-TMB system can be used to selectively and sensitively detect ascorbic acid (AA) with an LOD of 0.93 μM in the range of 4-36 μM through the colorimetric method. Moreover, the assay based on the 2-QF-CuFe-G metallohydrogel can be successfully applied to detect AA in fresh fruits.

Ni-Pt nanozyme-mediated relaxation and colorimetric sensor for dual-modality detection of norovirus

An NiPt nanozyme-mediated relaxation and colorimetric sensor is developed for dual-modality detection of norovirus (NoV). The relaxation modality is based on the “catalase-like” activity of the NiPt nanozyme, which adjusts the hydrogen peroxide (H2O2) mediated Fe (II)/Fe(III) conversion, thereby changing the relaxation signal. Poly-γ-glutamic acid (MW ≈ 1w) can enhance the relaxivity of Fe(III) (r1 = 7.11 mM−1 s−1; r2 = 8.94 mM−1 s−1). The colorimetric modality exploits the “peroxidase-like” activity of the NiPt nanozyme, which can catalyze the oxidation of colorless 3, 3′, 5, 5′-tetramethylbenzidine (TMB) to blue oxTMB in H2O2. Under optimal conditions, the relaxation modality exhibits a wide working range (1.0 × 101–1.0 × 104 fM) and a limit of detection (LOD) of 4.7 fM (equivalent to 2820 copies/μL). The spiked recoveries range from 99.593 to 106.442 %, and the relative standard deviation (RSD) is less than 5.124 %. The colorimetric modality exhibited the same working range with a lower LOD of 2.9 fM (equivalent to 1740 copies/μL) and an RSD of less than 2.611 %. Additionally, the recombinase polymerase amplification reaction enabled the detection of low NoV levels in food samples with a working range of 102–106 copies/mL and LOD of 102 copies/mL. The accuracy of the sensor in the analysis of spiked samples is consistent with the gold standard method (real-time quantitative reverse transcription-polymerase chain reaction), demonstrating the high accuracy and practical utility of the sensor.

Calcium Fluoride/Manganese Dioxide Nanocomposite with Dual Enzyme-like Activities for Uric Acid Sensing: A Comparative Study of Enzyme and Nonenzyme Methods.

The debate over enzyme methods versus nonenzyme methods in the field of nanosensing has lasted for decades despite hundreds of published studies on this topic. In this study, we first present a comparative analysis of these methods using a reaction based on the CaF2/MnO2 nanocomposite (CM Nc) with dual-enzyme activity, presenting oxidase- and peroxidase-like activities. Uric acid (UA) is a byproduct of purine metabolism in the body, and abnormal levels can cause many diseases; hence, tracking the amount of UA in human serum is crucial. The enzyme method was established using uricase and CM Nc: UA produced H2O2 when catalyzed by uricase; H2O2 was then catalyzed into reactive oxygen species (ROS) by the peroxidase activity of the CM Nc; this ROS oxidized 3,3',5,5'-tetramethylbenzidine (TMB), which was oxidized into blue oxidized TMB (oxTMB). The nonenzyme method was built on the scavenging effect of UA on the ROS, which prevented the catalytic capability of CM Nc toward TMB and induced blue oxTMB fading. The results of further tests revealed the good selectivity of the enzyme method compared to the fast response of the nonenzyme method. Additionally, both methods were effective in determining the UA concentration in human serum. The two separate methods can also independently verify each other, increasing the accuracy of the detection results in accordance with the relatively independent detection principles. This research provided theoretical backing for the practical design of multienzyme nanozyme catalysts, which can facilitate the precise detection of UA in biochemical products.

Detection of acrylamide in food based on MIL-glucose oxidase cascade colorimetric aptasensor

BackgroundMaillard reaction involves the polymerization, condensation, and other reactions between compounds containing free amino groups and reducing sugars or carbonyl compounds during heat processing. This process endows unique flavors and colors to food, while it can also produce numerous hazards. Acrylamide (AAm) is one of Maillard's hazards with neurotoxicity and carcinogenicity, these effects can trigger mutations and alternations in gene expression in human cells and accelerate organ aging. An accurate and reliable acrylamide detection method with high sensitivity and specificity for future regulatory activities is urgently needed. ResultsHerein, we constructed a colorimetric aptasensor with the hybridization of MIL-glucose oxidase (MGzyme)-cDNA and magnetic nanoparticle-aptamer (MNP-Apt) to specifically detect AAm. The incorporation of MB-Apt and AAm released MGzyme-cDNA in the supernatant, took the supernatant out, with the addition of glucose and TMB, MGzyme would oxidize glucose, the resulting •OH facilitated the oxidation of colorless TMB to blue ox-TMB. The absorbance value at 652 nm, which indicates the characteristic absorption peak of ox-TMB, exhibited a proportion to the concentration of AAm. MGzyme avoided the addition of harmful intermediate H2O2 and created an acid microenvironment for the catalytic reaction. MNP-Apt possessed the advantages of high specificity and simplified separation. Under optimal conditions, this method displayed a linear range of 0.01–100 μM with the limit of detection of 1.53 nM. With the spiked analysis data cross-verified by ELISA kit, this aptasensor was proven to specifically detect AAm at low concentrations. SignificanceThis colorimetric aptasensor was the integration of aptamer and the enzyme-cascade system, which could broaden the applicable range of enzyme-cascade system, break the limits of specific detection of substrates, eliminate the need for harmful intermediates, improve the reaction efficiency, implement the specific detection, whilst enabling the accurate detection of AAm. Given these remarkable performances, this method has shown significant potential in the field of food safety inspection.

Enzyme-free ratiometric fluorescence and colorimetric dual-signal determination of glyphosate based on copper nanoclusters (ZIF/CuNCs) combined with blue carbon dots (bCDs)

A novel ratio fluorescent and colorimetric dual-signal sensing platform for detecting glyphosate based on blue carbon dots (bCDs) combined with ZIF/CuNCs nanomaterials that encapsulate copper nanoclusters (CuNCs) in a metal-organic framework (MOF). In principle, the immobilization of Cu2+ in ZIF/CuNCs results in complexation with imidazole in ZIF, leading to fluorescence quenching of ZIF/CuNCs, while the reference fluorophore bCDs remains unaffected. In addition, the colorimetric sensing strategy was based on the efficient peroxidase-like activity of bCDs binding to Cu2+, catalyzing H2O2 to generate OH. Under this condition, TMB could be oxidized to form blue oxTMB. However, when glyphosate was involved in the system, the fluorescence of ZIF/CuNCs was restored upon due to the strong chelation between Cu2+ and glyphosate, while the peroxidase-like activity of bCDs/Cu2+ decreased and resulted in the generation of fewer oxTMB, accompanied by a lighter blue color. The sensing platform was successfully applied to the determination of glyphosate in real samples of lake water and cabbage, demonstrating reliable and sensitive performance in practical applications.