A highly selective dual-signal response ratiometric fluorescence sensing strategy for malachite green in fish based on carbon dots/copper nanoclusters nanocomposite.

Malachite green (MG), a widely used antiparasitic agent, poses health risks to human due to its genotoxic and carcinogenic properties. Herein, a stable dual-emission fluoroprobe of carbon dots/copper nanoclusters is prepared for highly selective detection of MG based on the inner filter effect. This probe exhibits characteristic emission bands at 435 and 625 nm when excited at 376 nm. After adding MG, the both emission signals were significantly quenched, and the ratio of fluorescence intensity (F435/F625) was linearly related to the concentration of MG in the range of 0.05-40 µmol L-1 with a limit of detection of 18.2 nmol L-1. Meanwhile, the two signals exhibit linear relationships with the concentration of MG, respectively, and the corresponding detection results were consistent. The fluoroprobe was successfully used for the detection of MG in fish samples with the recoveries ranging from 96.0% to 103.8% and a relative standard deviation of <3.3%.

Highly-Selective fluorescent Fe3O4@PPy aptasensor.

Label-free nucleic acid fluorescent probes are gaining popularity due to their low cost and ease of application. However, the primary challenges associated with label-free fluorescent probes stem from their tendency to interact with other biomolecules, such as RNA, proteins, and enzymes, which results in low specificity. In this work, we have developed a simple detection platform that utilizes Fe3O4@PPy in combination with a label-free nucleic acid probe, 1,1,2,2-tetrakis[4-(2-bromo-ethoxy)phenyl]ethene (TTAPE) or Malachite Green (MG), for highly selective detection of metal ions, acetamiprid, and thrombin. Fe3O4@PPy not only adsorbs aptamers through electrostatic interactions, π-π bonding, and hydrogen bonding, but also quenches the fluorescence of the TTAPE/MG. Upon the addition of target compounds, the aptasensor separates from Fe3O4@PPy through magnetic separation. Moreover, by changing different aptamers, the aptasensor was applied to detect metal ions, acetamiprid, and thrombin, with the turned-on photoluminescence (PL) emission intensity recorded and showing linearity to the concentrations of targets. The robustness of method was demonstrated by applying it to real samples, which included vegetables (for detecting acetamiprid with LODs of 0.02 and 0.04 ng/L), serum samples (for detecting thrombin with LODs of 5.5 and 4.3 nM), and water samples (for detecting Pb2+ with an LOD of 0.17 nM). Therefore, due to its impressive selectivity and sensitivity, the Fe3O4@PPy aptasensor could be utilized as a universal detection platform for various clinical and environmental applications.

[A fluorescence method based on malachite green/aptamer for detection of ochratoxin A in traditional Chinese medicines].

A label-free fluorescence method based on malachite green/aptamer was developed for the detection of ochratoxin A(OTA) in traditional Chinese medicines. Malachite green itself exhibits weak fluorescence. Upon interaction with the aptamer specific to OTA, the G-quadruplex structure of the aptamer provides a protective microenvironment for malachite green, which significantly enhances its fluorescence signal. After OTA is added, preferential binding occurs between the aptamer and OTA, and malachite green will be released from the aptamer, which weakens the fluorescence signal. According to this principle, this paper established a fluorescence method with the aptamer of OTA as the recognition element and malachite green as the fluorescent probe for the detection of OTA in traditional Chinese medicines. The key experimental factors such as the concentrations of metal ions, aptamer, and malachite green were optimized to improve the performance of the method. OTA was detected under the optimal experimental conditions, and the results showed that with the increase in OTA concentration, the fluorescence signal gradually weakened. Within the range of 20-1 000 nmol·L~(-1), the OTA concentration was linearly correlated with the fluorescence signal ratio ΔF/F(ΔF=F_0-F, where F_0 is the fluorescence signal of aptamer/malachite green, and F is the fluorescence signal of OTA/aptamer/malachite green), with R~2 of 0.995. The limit of detection of the established method was 7.1 nmol·L~(-1). Furthermore, three substances structurally similar to OTA and two mycotoxins that may coexist with OTA were selected for experiments, which aimed to examine the cross-reactivity and specificity of the established method. The cross-reactivity experiments demonstrated that the interferers did not significantly affect the fluorescence signal of the detection system. The specificity experiments revealed that when mycotoxins were mixed with OTA, the fluorescence signal generated by the mixture closely resembled that of OTA itself. The results indicated that even in the presence of interferents, the established method remained unaffected and demonstrated excellent specificity. Additionally, this method exhibited remarkable reproducibility and stability. In the case of simple centrifugation and dilution of traditional Chinese medicine samples(Puerariae Lobatae Radix, Sophorae Flavescentis Radix, and Periplocae Cortex), the OTA detection method was applicable, with recovery rates ranging from 91.5% to 121.3%. Notably, this approach does not need complex pretreatment of traditional Chinese medicines while offering simple operation, low detection costs, and short detection time. Furthermore, by incorporating aptamers into the quality evaluation of traditional Chinese medicines, this method expands the application scope of aptamers.

Dual-mode aptasensor based on P-CeO2NR@Mxene and exonuclease I-assisted target recycling for malachite green detection.

Malachite green (MG) has been illicitly employed in aquaculture as a parasiticide, however, its teratogenic and carcinogenic effects pose a significant human health threat. Herein, a dual-mode colorimetric and electrochemical aptasensor was fabricated for MG detection, capitalizing on the robust catalytic and peroxidase-like activity of P-CeO2NR@Mxene and good capture efficiency of a tetrahedral DNA nanostructure (TDN) designed with multiple aptamers (m-TDN). P-CeO2NR@Mxene-modified complementary DNA (cDNA) served as both colorimetric and electrochemical probe. m-TDN was attached to AuE to capture MG and P-CeO2NR@Mxene/cDNA. The superior aptamer and MG binding to cDNA regulated signals and enabled precise MG quantification. The further introduced Exo I enabled aptamer hydrolysis, releasing MG for further binding rounds, allowing target recycling amplification. Under the optimal conditions, the aptasensor reached an impressively low detection limit 95.4 pM in colorimetric mode and 83.6 fM in electrochemical mode. We believe this dual-mode approach holds promise for veterinary drug residue detection.

Smartphone-based dual inverse signal MOFs fluorescence sensing for intelligent on-site visual detection of malachite green.

The development of intelligent, sensitive, and visual methods for the rapid detection of veterinary drug residues is essential to ensure food quality and safety. Here, a smartphone-based dual inverse signal MOFs fluorescence sensing system was proposed for intelligent in-site visual detection of malachite green (MG). A UiO-66-NH2@RhB-dual-emission fluorescent probe was successfully synthesized in one step using a simple one-pot method. The inner filter effect (IFE) quenches the red fluorescence, while hydrogen bonding interaction enhances the blue fluorescence, enabling highly sensitive, accurate, and visual detection of MG dual inverse signals through fluorescence analysis. The probe showed great linearity over a wide range of 0.1-100 µmol/L, with a limit of detection (LOD) of 20 nmol/L. By integrating smartphone photography and RGB (red, green, and blue) analysis, accurate quantitative analysis of MG in water and actual fish samples can be achieved within 5 min. This developed platform holds great promise for the on-site detection of MG in practical applications, with the advantages of simplicity, cost-effectiveness, and rapidity. Consequently, it may open up a new pathway for on-site evaluation of food safety and environmental health.

A highly sensitive surface-enhanced Raman scattering sensor with MIL-100(Fe)/Au composites for detection of malachite green in fish pond water.

Concerns about food safety have been arisen due to the improper use of chemicals in aquaculture. Malachite green (MG) has attracted attention because of its illegal usage and its potential negative impacts on the environment and public health. Surface-enhanced Raman scattering (SERS) platforms coupled with different SERS substrates have been employed for rapid analysis of MG residues in food. However, the most commonly used SERS substrates were non-reusable and showed limited detection sensitivity. In this study, a novel SERS substrate with a good recyclability and a high sensitivity was prepared by electrostatically assembling together a metal-organic framework material called materials of institute lavoisie-100(Fe) (MIL-100(Fe)) and Au NPs. The lowest detectable concentration of MG was 10-13 M based on the optimal substrate. The SERS sensor was applied for the detection of the trace MG in fish pond water, which was accomplished with the correlation coefficients R2 = 0.991-0.996 in a concentration range of 10-6-10-13 M. Moreover, MIL-100(Fe)/Au was recycled at least five times, realizing a "detection to degradation", showing great potential for food contamination monitoring due to its distinguished performance.

Enhanced artificial intelligence for electrochemical sensors in monitoring and removing of azo dyes and food colorant substances.

It is necessary to determine whether synthetic dyes are present in food since their excessive use has detrimental effects on human health. For the simultaneous assessment of tartrazine and Patent Blue V, a novel electrochemical sensing platform was developed. As a result, two artificial azo colorants (Tartrazine and Patent Blue V) with toxic azo groups (-NN-) and other carcinogenic aromatic ring structures were examined. With a low limit of detection of 0.06 µM, a broad linear concentration range 0.09µM to 950µM, and a respectable recovery, scanning electron microscopy (SEM) was able to reveal the excellent sensing performance of the suggested electrode for patent blue V. The electrochemical performance of an electrode can be characterized using cyclic and differential pulse voltammetry, and electrochemical impedance spectroscopy. Moreover, the classification model was created by applying binary classification assessment using enhanced artificial intelligence comprises of support vector machine (SVM) and Genetic Algorithm (GA), respectively, a support vector machine and a genetic algorithm, which was then validated using the 50 dyes test set. The best binary logistic regression model has an accuracy of 83.2% and 81.1%, respectively, while the best SVM model has an accuracy of 90.3% for the training group of samples and 81.1% for the test group (RMSE = 0.644, R2 = 0.873, C = 205.41, and = 5.992). According to the findings, Cu-BTC MOF (copper (II)-benzene-1,3,5-tricarboxylate) has a crystal structure and is tightly packed with hierarchically porous nanomaterials, with each particle's edge measuring between 20 and 37 nm. The suggested electrochemical sensor's analytical performance is suitable for foods like jellies, condiments, soft drinks and candies.

Polyphenol characterisation and diverse bioactivities of native Australian lilly pilly (Syzygium paniculatum) extract.

Magenta lilly pilly (Syzygium paniculatum) is an Australian native tree that produces berry fruits that are rich in phytochemicals reportedly beneficial to human health. Here we explored the biological activities of polyphenol-enriched extracts from the magenta lilly pilly fruit, benchmarking it against traditional sources including purple sweet potato and blackberry. We show that the extracts exert potent antioxidant and neuroprotective properties as well as antimicrobial activity against Staphylococcus aureus. The phenolic composition of lilly pilly was investigated using liquid chromatography coupled to mass spectrometry (HPLC-DAD-MS), revealing anthocyanins to be the primary component in high abundance compared to traditional anthocyanin-containing plants. Three anthocyanins from lilly pilly, along with their glycosylation patterns and stability, were characterised. Altogether, our results demonstrate the potential to exploit magenta lilly pilly fruits as a high-yielding source of phenolics with beneficial biological properties of potential interest for multiple downstream applications.

A Rapid Tricolour Immunochromatographic Assay for Simultaneous Detection of Tricaine and Malachite Green.

In this research, we designed a rapid tricolour immunochromatographic test strip with double test lines (TS-DTL) and two-colour AuNP probes, which realised the simultaneous detection of tricaine mesylate (TMS) and malachite green (MG). Through a distinct tricolour system (red T1 line, blue T2 line and purple C line), a visual identification of TMS (0.2 µg/mL) and MG (0.5 µg/mL) was quickly achieved on site, which improved the accuracy of naked eye observations. The LODs of TMS in aquaculture water, fish and shrimp were 11.0, 29.6 and 61.4 ng/mL, respectively. MG LODs were 47.0 ng/mL (aquaculture water), 82.8 ng/mL (fish) and 152.4 ng/mL (shrimp). The LOD of MG was close to the similar TS methods. However, visual detection of TMS could meet the requirements of the residue limit (1 µg/mL) of TMS in the USA, and the quantitative detection of TMS was over 16 times lower than the USA standard. The developed platform was rapid (~20 min, HPLC~3 h) and accurate, which was verified using a traditional HPLC method. The recovery rates ranged from 82.2% to 108.6% in three types of real samples, indicating a potential application in on-site fast screening or multiple detection for TMS and MG residues in aquatic products.

Branched poly(ethylenimine) carbon dots-MnO2 nanosheets based fluorescent sensory system for sensing of malachite green in fish samples.

Malachite green (MG) is an organic dye compound that is frequently used as a fungicide and antiseptic in aquaculture. However, human or animal exposure to MG causes carcinogenic, teratogenic and mutagenic effects. Herein, a novel fluorescent assay was designed for the detection of MG using manganese dioxide nanosheets (MnO2 NS) as an energy acceptor to quench the fluorescence of branched poly(ethylenimine) carbon dots (BPEI-CDs) via Förster resonance energy transfer. When butyrylcholinesterase is introduced to form thiocholine in the presence of S-butyrylthiocholine iodide, MnO2 NS can be recovered by thiocholine to Mn2+, resulting in restoration of the fluorescence of BPEI-CDs. Exploiting these changes in fluorescence intensity in the above system, a fluorescence probe was successfully developed for the quantitative detection of MG. Besides, this assay was applied to fish samples, verifying the high potential for practical application of the proposed sensor for the monitoring of MG in aquatic products.

Tolerance and decolorization potential of duckweed (Lemna gibba) to C.I. Basic Green 4.

With growing human culture and industrialization, many pollutants are being introduced into aquatic ecosystems. In recent years, dyes have become a major water pollutant used in the manufacture of paints and other production purposes. In this research, the potential of duckweed (Lemna gibba) plant was investigated spectrophotometrically as an obvious bioagent for the biological decolorization of the organic dye C.I. Basic Green 4 (Malachite Green, BG4). Photosynthetic efficiency analysis showed that the photosynthetic apparatus of L. gibba is very tolerant to BG4. Significant induction of reactive oxygen species (ROS) scavenging enzymes was observed after 24h of biodecolorization process in L. gibba treated with 15 and 30 mg/l BG4. The experimental results showed that L. gibba has a strong ability to extract BG4 from contaminated water and the best results were obtained at 25-30°C and pH 8.0. We conclude that duckweed L. gibba can be used as a potent decolorization organism for BG4.

Development and applicability of a multi-residue method for dyes, including new residue markers, to detect drug misuse in aquaculture.

A qualitative and quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the sensitive and exhaustive analysis of residues from triarylmethane dyes, triarylmethane-derivative dyes, phenothiazines, phenoxazines and xanthenes in aquaculture samples. For a wider and more robust detection of dye misuse on farms, other residue markers were also included the leuco forms of brilliant green, crystal violet and malachite green; one direct metabolite of Victoria pure blue BO and methylene blue and three bile acids, which are endogenous markers of the effects of dye contamination in fish. We optimised the extraction method by comparing several extraction solvents and sample solvents reported in the literature to have the best extraction efficiency. The residues were determined using a positive electrospray ionisation source. We assessed the parameters of this LC-MS/MS method by evaluating the matrix effects, identification and quantitative parameters according to the criteria stipulated in the European Commission Decision No. 2002/657/EC. A study on the applicability of the method was conducted on various aquaculture species and on a positive catfish.

Quantitative SERS sensor based on self-assembled Au@Ag heterogeneous nanocuboids monolayer with high enhancement factor for practical quantitative detection.

Accurate and rapid quantitative detection of pesticide and pollutant levels in the actual sample can aid in protecting food security, environmental security, and human health. A high Raman enhancement factor and good repeatability of the surface-enhanced Raman spectroscopy (SERS) substrates are favorable to quantitative analysis. Herein, a quantitative SERS sensor based on constructed self-assembled plasmonic Au@Ag heterogeneous nanocuboids (Au@Ag NCs) monolayer was developed. The sensor was used to quantitatively detect the trace pesticides extracted from pear surfaces and pollutants in fishpond water. Densely packed Au@Ag NCs fabricated into large-scale monolayer films were chemically functionalized using 4-methyl-thiobenzoic acid (4-MBA) at the organic/aqueous interface, in which plentiful nanogaps contribute to increase hotspots. Their sharp corners and edges make the sensor have high SERS performance through providing abundant "hot spots." The obtained optically SERS-based sensor with uniform liquid-state interfacial nanoparticle arrays appeared to have nice SERS performance and uniformity using crystal violet (CV) as a probe molecule. In particular, the proposed SERS sensor was applied for quantitative detection of thiabendazole (TBZ) extracted from pear surfaces and malachite green (MG) in fishpond water down to levels of 0.0105 nM and 0.87 nM for SERS assay respectively. As a result, our proposed SERS quantitative detection strategy is quite preferred to on-site analysis and supervision of contaminant in food samples.

Film wrap nanoparticle system with the graphene nano-spacer for SERS detection.

Film wrap nanoparticle system (FWPS) is proposed and fabricated to perform SERS effect, where the Ag nanoparticle was completely wrapped by Au film and the double-layered graphene was selected as the sub-nano spacer. In this system, the designed nanostructure can be fully rather than partly used to generate hotspots and absorb probe molecules, compared to the nanoparticle to nanoparticle system (PTPS) or nanoparticle to film system (PTFS). The optimal fabricating condition and performance of this system were studied by the COMSOL Multiphysics. The simulation results show that the strongly large-scale localized electromagnetic field appears in the whole space between the Ag nanoparticle and Au film. The experimental results show that the FWPS presents excellent sensitivity (crystal violet (CV): 10-11 M), uniformity, stability and high enhancement factor (EF: 2.23×108). Malachite green (MG; 10-10 M) on the surface of fish and DNA strands with different base sequence (A, T, C) were successfully detected. These advanced results indicate that FWPS is highly promising to be applied for the detection of environmental pollution and biomolecules.

Designed Eu(III)-functionalized nanoscale MOF probe based on fluorescence resonance energy transfer for the reversible sensing of trace Malachite green.

An ingenious nanoscale fluorescent sensor derived from Eu3+-postfunctionalized MIL-53 (Al) (Eu3+@MIL-53 [Al]) was fabricated though a simple and effective approach. Malachite green (MG) effectively turned off the luminescence of Eu3+@MIL-53 (Al) via fluorescence resonance energy transfer (FRET), thus enabling MG sensing. The developed probe exhibited instantaneous reusability after being cleaned with deionized water. The fluorescence intensity, quenching efficiency, and crystal structure of the recoverable sensor after five recycling processes were unchanged compared with those of the original sample. Moreover, the potential mechanism of MG detection was revealed in detail. This work represents the first attempt to determine MG in aquaculture water and products by using metal-organic frameworks (MOFs). The Eu3+@MIL-53 (Al) probe proved to be a remarkable fluorescence probe for MG with high selectivity, sensitivity, and excellent regeneration capability. It provides a promising functional platform for the recognition of illegal MG addition to aquaculture water and products.

Generation of functional single-chain fragment variable from hybridoma and development of chemiluminescence enzyme immunoassay for determination of total malachite green in tilapia fish.

To determine malachite green (MG) and its major metabolite, leucomalachite green (LMG) residual levels in tilapia fish, chemiluminescent enzyme immunoassay (CLEIA) was developed based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein. At first, VH and VL gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against LMG, and then thoroughly by database-assisted sequence analysis. Finally, the productive VH and VL were assembled to an intact scFv sequence and engineered to produce scFv-AP fusion protein. The fusion protein was further identified as a bifunctional reagent for immunoassay, then a sensitive one-step CLEIA against LMG was developed with a half-maximal inhibitory concentration (IC50) and limit of detection (LOD) of 1.3 and 0.04 ng/mL, respectively. The validation results of this novel competitive CLEIA was in line with those obtained by classical HPLC method for determination of total MG in spiked and field incurred samples.

Fluorescence detection of malachite green in fish tissue using red emissive Se,N,Cl-doped carbon dots.

Facile detection of malachite green (MG), a toxic dye, in aquaculture is urgently demanded for environment and food safety. Herein, we design a novel fluorescent probe, namely red emissive Se,N,Cl-doped carbon dots (CDs), to accurately determinate MG. CDs are prepared by hydrothermal treatment of selenourea and o-phenylenediamine in HCl solution. This material exhibits excitation-independent dual emissions at 625 and 679 nm, with a high quantum yield of 23.6%. A selective and sensitive fluorescent sensor toward MG is established based on inner filter effect, because both the excitation and emission light of CDs can be strongly absorbed by MG. The fluorescence quenching of CDs is linear to the MG concentration over the range of 0.07-2.50 µM with a low detection limit of 21 nM. Trace-level analysis of MG in fish tissue is successfully explored, demonstrating the great potential of the proposed sensor for MG monitoring in aquatic products.

Detection of Malachite Green in Water Using Edge Excited Label Free Fluorescent Probe NCQDs.

The fluorescent properties of nitrogen doped carbon quantum dots (NCQDs) prepared through microwave assisted green method has been used as label free fluorescent probe for selective and sensitive detection of malachite green (MG) in water. The optical responses revealed that the NCQDs are highly stable and have good fluorescent quantum yield. The NCQDs were used to detect the Malchite Green in Mili Q water. Reduction in the fluorescence response was monitored in the range 17.12-128.43 µM of MG dissolved in Mili Q water. Linear response was observed in the range, 10-80 µM. The calculated value of limit of detection is 5.16 µM and the sensitivity is (0.03536 ± 0.00001) µM-1. The future application of this work is that it can be employed to detect MG in the tap water and other natural sources of water.

Analysis of trace malachite green, crystal violet, and their metabolites in zebrafish by surface-coated probe nanoelectrospray ionization mass spectrometry.

Malachite green (MG) and crystal violet (CV) are the typical triphenylmethane dyes, which are recalcitrant molecules exerting mutagenic and carcinogenic effects on living organisms. Characterization of the residues of MG, CV, and their metabolites in biological organisms is of importance, especially for in vivo and in situ characterization. In this study, a method for determination of trace MG, CV, and their leuco metabolites in zebrafish by surface-coated probe nanoelectrospray ionization mass spectrometry (SCP-nanoESI-MS) was developed. A microscale solid-phase microextraction (SPME) probe was developed and used for extraction and enrichment of trace MG, CV, and their leuco metabolites in zebrafish after exposure. After that, the loaded SPME probe was directly employed for nanoESI-MS analysis under ambient and open-air conditions. Under the optimum conditions, the method demonstrated good linearity, with correlation coefficient values (r2) no less than 0.9925. The limits of detection and quantification were 0.014-0.023 ng mL-1 and 0.046-0.077 ng mL-1, respectively. By using the proposed method, the bioaccumulation of MG and CV in zebrafish was investigated, and the distribution of MG, CV, and their leuco metabolites in different organs of zebrafish was studied. MG, CV, and their leuco metabolites were all found in zebrafish tissues including brain, muscle, heart, and kidney after exposure, with highest concentration in intestine followed in ovary.

Synthesis and SERS application of gold and iron oxide functionalized bacterial cellulose nanocrystals (Au@Fe3O4@BCNCs).

Bacterial cellulose nanocrystals (BCNCs) are biocompatible cellulose nanomaterials that can host guest nanoparticles to form hybrid nanocomposites with a wide range of applications. Herein, we report the synthesis of a hybrid nanocomposite that consists of plasmonic gold nanoparticles (AuNPs) and superparamagnetic iron oxide (Fe3O4) nanoparticles supported on BCNCs. As a proof of concept, the hybrid nanocomposites were employed to isolate and detect malachite green isothiocyanate (MGITC) via magnetic separation and surface-enhanced Raman scattering (SERS). Different initial gold precursor (Au3+) concentrations altered the size and morphology of the AuNPs formed on the nanocomposites. The use of 5 and 10 mM Au3+ led to a heterogenous mix of spherical and nanoplate AuNPs with increased SERS enhancements, as compared to the more uniform AuNPs formed using 1 mM Au3+. Rapid and sensitive detection of MGITC at concentrations as low as 10-10 M was achieved. The SERS intensity of the normalized Raman peak at 1175 cm-1 exhibited a log-linear relationship for MGITC concentrations between 2 × 10-10 and 2 × 10-5 M for Au@Fe3O4@BCNCs. These results suggest the potential of these hybrid nanocomposites for application in a broad range of analyte detection strategies.