Search

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 Fe 3 O 4 @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. Fe 3 O 4 @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 Fe 3 O 4 @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 Pb 2+ with an LOD of 0.17 nM). Therefore, due to its impressive selectivity and sensitivity, the Fe 3 O 4 @PPy aptasensor could be utilized as a universal detection platform for various clinical and environmental applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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 (F 435 /F 625 ) 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%., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

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-CeO 2 NR@Mxene and good capture efficiency of a tetrahedral DNA nanostructure (TDN) designed with multiple aptamers (m-TDN). P-CeO 2 NR@Mxene-modified complementary DNA (cDNA) served as both colorimetric and electrochemical probe. m-TDN was attached to AuE to capture MG and P-CeO 2 NR@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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Developing a multifunctional material that can detect and remove carcinogens in water environments, simultaneously monitor their toxic metabolites in living organisms is significant for environmental remediation and human health. However, most research only focused on detection or adsorption carcinogens due to the difficulty of integrating multiple functions into one material, let alone monitoring their toxic metabolites. Here, a multifunctional Tb/Eu@TATB-HOF (1) is first developed to monitor two carcinogens, malachite green (MG) and its metabolites leucomalachite green (LMG), and simultaneously remove MG from the contaminated water. 1, as the dual-emission fluorescence sensor, can achieve ultrasensitive and highly visualized sensing for MG and LMG with different response modes. Even in actual samples, 1 still exhibits satisfactory sensing performances. As the adsorbent, 1 displays good recyclability and high adsorption capacity for MG. The sensing and adsorption mechanisms are explored through experiments and theoretical calculations. This work not only provides a novel insight for environmental remediation and human health through detection and removal of carcinogens, simultaneously monitoring their toxic metabolites, but first reveals the enormous potential of HOFs as multifunctional materials simultaneously for fluorescence sensing and adsorption., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In this study, a hydrothermal process was utilized to grow mixed-valence CuFe 2 O 4 /Cu 0 nanosheets on carbon fiber paper, forming a three-dimensional hierarchical electrode (CuFe 2 O 4 /Cu 0 @CFP). The ordered array structure, coupled with the porous bowl-like structure, enhances the exposure of more electrode active sites and facilitates analyte penetration, thus enhancing the electrode sensing performance. As a binder-free sensor, the CuFe 2 O 4 /Cu 0 @CFP sensor exhibited remarkable sensitivity in detecting Malachite Green (MG), Sunset Yellow (SY) and Tartrazine (TA) over wide concentration ranges: 0.1-300 μM for MG (R 2  = 0.994), 0.005-200 μM for SY (R 2  = 0.996), and 0.005-300 μM for TA (R 2  = 0.995) with low detection limits of 0.033 μM for MG, 0.0016 μM for SY, and 0.0016 μM for TA (S/N = 3), respectively. Additionally, the 3D CuFe 2 O 4 /Cu 0 @CFP sensor detected MG, SY, and TA in a mixed solution with satisfactory results. It also performs well in beverage, fruit juice powder, and jelly samples, with results matching those from HPLC., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Additive abuse in fishery, such as tricaine methanesulfonate (MS222), ciprofloxacin (CPFX), and malachite green (MG), threatens public human health and interferes with the ecological equilibrium of water resources. However, the majority of the present detection methods suffer from high costs, complex operations, and poor portability. Therefore, real-time and rapid detection of the above additive by mobile devices is becoming increasingly important. Here we report the fabrication and performance of an entirely electrochemical system with USB-stick size for simultaneous detection of MS222, CPFX, and MG, as well as pH and permittivity. The limits of detections are 0.17, 0.67, and 0.28 µg/mL, while the resolution ratios are 10 %, 10 %, and 5 % for MS222, MG, and CPFX, respectively. For both pH and permittivity, they have linear regressions measured by brightness and capacitance of the sample respectively, at the range of 1.5-9 (pH) and 10-20 (permittivity). The interference experiments, using target analytes (40 μg/mL) and 15 interfering analytes (80 μg/mL), demonstrated the anti-interference performance of the sensor patches. The field studies on carps, catfishes, and chubs indicated that the developed integrated portable system could be used for real sample analysis with high performance., Competing Interests: Declaration of Competing Interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service, and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled, “Fully integrated sensor array for additives, permittivity, and pH monitoring for fishery”., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Rapid and quantitative detection of malachite green (MG) in aquaculture products is very important for safety assurance in food supply. Here, we develop a point-of-care testing (POCT) platform that combines a flexible and transparent surface-enhanced Raman scattering (SERS) substrate with deep learning network for achieving rapid and quantitative detection of MG in fish. The flexible and transparent SERS substrate was prepared by depositing silver (Ag) film on the polydimethylsiloxane (PDMS) film using laser molecular beam epitaxy (LMBE) technique. The wrinkled Ag NPs@PDMS film exhibits high SERS activity, excellent reproducibility and good mechanical stability. Additionally, the fast in situ detection of MG residues onfishscales was achieved by using the wrinkled Ag NPs/PDMS film and a portable Raman spectrometer, with a minimum detectable concentration of 10 -6 M. Subsequently, a one-dimensional convolutional neural network (1D CNN) model was constructed for rapid quantification of MG concentration. The results demonstrated that the 1D CNN quantitative analysis model possessed superior predictive performance, with a coefficient of determination (R 2 ) of 0.9947 and a mean squared error (MSE) of 0.0104. The proposed POCT platform, integrating a transparent flexible SERS substrate, a portable Raman spectrometer and a 1D CNN model, provides an efficient strategy for rapid identification and quantitative analysis of MG in fish., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

SERS measurements for monitoring bactericides in dairy products are highly desired for food safety problems. However, the complicated preparation process of SERS substrates greatly impedes the promotion of SERS. Here, we propose acoustofluidic one-step synthesis of Ag nanoparticles on paper substrates for SERS detection. Our method is economical, fast, simple, and eco-friendly. We adopted laser cutting to cut out appropriate paper shapes, and aldehydes were simultaneously produced at the cutting edge in the pyrolysis of cellulose by laser which were leveraged as the reducing reagent. In the synthesis, only 5 μL of Ag precursor was added to complete the reaction, and no reducing agent was used. Our recently developed acoustofluidic device was employed to intensely mix Ag + ions and aldehydes and spread the reduced Ag nanoparticles over the substrate. The SERS substrate was fabricated in 1 step and 3 min. The standard R6G solution measurement demonstrated the excellent signal and prominent uniformity of the fabricated SERS substrates. SERS detection of the safe concentration of three bactericides, including tetracycline hydrochloride, thiabendazole, and malachite green, from food samples can be achieved using fabricated substrates. We take the least cost, time, reagents, and steps to fabricate the SERS substrate with satisfying performance. Our work has an extraodinary meaning for the green preparation and large-scale application of SERS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

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-NH 2 @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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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&#95;0-F, where F&#95;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.

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 R 2  = 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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

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 (MnO 2 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, MnO 2 NS can be recovered by thiocholine to Mn 2+ , 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., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

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.

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 T 1 line, blue T 2 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.

An ingenious nanoscale fluorescent sensor derived from Eu 3+ -postfunctionalized MIL-53 (Al) (Eu 3+ @MIL-53 [Al]) was fabricated though a simple and effective approach. Malachite green (MG) effectively turned off the luminescence of Eu 3+ @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 Eu 3+ @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., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

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.

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.

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.

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, V H and V L gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against LMG, and then thoroughly by database-assisted sequence analysis. Finally, the productive V H and V L 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 (IC 50 ) 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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

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: 2.23×10 8 ). 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.

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

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.

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 (r 2 ) 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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A rapid and simple immunochromatographic strip test assay based on competitive format was developed for leucomalachite green (LMG) detection. LMG-bovine serum albumin and rabbit anti-sheep IgG were immobilized on nitrocellulose membrane for the test line and control line, respectively. Anti-LMG-colloidal gold conjugate was immobilized onto the conjugate pad. For qualitative detection, the cut-off limit of the strip test was determined at 2 µg/L by the naked eye. For quantitative analysis, the working range of the LMG detection was 0.7-2 µg/L with LOD at 0.28 µg/L. A one-step immunochromatographic strip test for LMG detection can be completed within 5 min without any incubation, washing and blocking steps. Analysis results of LMG in aquatic animals obtained from the immunochromatographic strip test were in good agreement with those realized from enzyme-link immunosorbent assay. The developed the immunochromatographic strip test offered rapid detection as a simple (one-step), cost-effective, instrument-free assay and no need for handling reagents., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

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.

Saffron is a highly adulterated spice due to its limited production and high costs. Non-saffron plant material is coloured with synthetic dyes to produce counterfeit saffron. Continuous monitoring of the synthetic dyes used in counterfeit saffron is essential because some dyes are not safe for human consumption. In the present study, 104 commercial saffron samples from 16 countries were screened, and 20 samples were found to contain dyes. Thin-Layer Chromatography (TLC) analysis showed adulteration with magenta- and pink- coloured dyes (18 and 2 samples, respectively), which did not correspond to any of the dyes reported earlier. Mass spectrometry analysis identified the magenta-coloured dye as magenta III or new fuchsin and the pink-coloured dye as rhodamine B. Magenta III is regarded as possibly carcinogenic, and rhodamine B was reported to cause local sarcomas. Both of these synthetic dyes are reported as adulterants in saffron for the first time., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Enriching and locating target analytes into specific "hot spots" are vital for ultrasensitive molecular identification and detection using plasmonic-based techniques. Inspired by mass transportation in lamp wicks, we develop an effective enrichment strategy for highly diluted analytes in which analytes and Au nanoparticles are transported via a solution microflow under the capillarity driving force of glass fiber papers to a heated region. After evaporation, a large volume of a solution containing analytes and Au nanoparticles is condensed into a very limited area, and thus, analyte molecules are effectively enriched and located into surface-enhanced Raman scattering (SERS) hot spots. Using this enrichment strategy, the sensitivity and detection limits of SERS are remarkably improved. Detection levels of crystal violet and anthracene are down to 10 -16 and 10 -10 M, respectively. This enrichment strategy is very robust and easy to implement, and it can potentially be exploited in various plasmonic-based molecular detection and identification techniques.

A fluorescence probe was delicately designed for the detection of malachite green (MG) in water and fish samples. Through the electrostatic self-assembly of CdTe QDs on the surface of polystyrene (PS) microspheres, the fluorescence signal was amplified. After grafting molecularly imprinted film, the fluorescence probe of MIP@PS@CdTe was fabricated and applied to the detection of MG based on fluorescence quenching. The linear range of MG detection was 0.01-20 μmol L - 1 , and the detection limit was 4.7 nmol L - 1 (3σ, n = 9) which was much lower than those of the previous reports. The recoveries of MG in aquaculture water and fish samples ranging from 87.6% to 105.4% illustrated that the detection by MIP@PS@CdTe probe was accurate and reliable., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019. Published by Elsevier Ltd.)

Violation of the Food Sanitation Act regarding detection of Patent Blue V, which is one of the non-permitted dyes for food in Japan, in imported food occurs every year. With respect to the identification of dyes of Patent Blue group, in some cases, each dye has several different names, and in other cases, different dyes have the same name. Thus, there is a risk that the detected dye is misidentified with other dyes of Patent Blue group. In this study, nine commercial available dyes of Patent Blue group, including a reagent with unclear product information, were analyzed by TLC, HPLC and LC-MS/MS. The result showed that with all three methods, the dyes could be clearly identified into one of four types of blue dyes, i.e. Patent Blue V, Azure Blue VX, Isosulfan Blue and Alphazurine A. Unification of nomenclature would reduce the risk of misidentification of dyes of Patent Blue group.

Compared with small model plants like Arabidopsis containing ovules with few cell layers, embryo sac and embryo development of model crop plants such as maize and other grasses are difficult to image. Multiple layers of tissue usually surround the deeply embedded embryo sac and developing embryo. Moreover, reliable cell biological marker lines labeling, for example, nuclei, plasma membrane, cell walls, or cells of a specific identity are often not available. The introduction of markers to study mutants is difficult and time-consuming and may require several generations of backcrosses. In this chapter, we therefore present an easy protocol to image maize ovaries and developing embryo sacs before and after fertilization allowing also high-throughput mutant analysis. The laborious embedding of samples and preparation of thin sections are omitted in this fixing-Feulgen staining-clearing (FFC) method. Optical sectioning through multiple layers of tissue is possible allowing 3D reconstructions of the whole embryo sac if necessary. The advantage of staining cell nuclei using the FFC method described here compared, for example, with DAPI staining is a wide range of Schiff's type reagents available for the Feulgen reaction. Depending on the reagent of choice, various conditions such as different excitation/emission filters or even white light can be applied for imaging. Moreover, in order to better visualize cell division, nuclei polarity as well as cell extent and integrity, periodic acid staining (PAS) of cell walls can be combined with Feulgen staining.

Objective: A rapid quantitative technique had been developed for the screening of malachite green(MG) in aquaculture water using paper spray ionization mass spectrometry(PSI-MS)., Methods: To rapid extract MG, chloroform and deep eutectic solvents(DESs) were used as extraction agent and co-solvent respectively. malachite green D&#95;5(MG-D&#95;5) was used as an internal standard to quantitative determination MG, C&#95;(18) chromatographic column(4. 6 mm×250 mm, 5 μm)., Results: The method showed a good linearity(R~2=0. 999) in the concentration range of 0. 1-10 μg/mL with the detection limit of 0. 03 μg/mL and the quantitative limit of 0. 1 μg/mL. The intra-and inter-day precisions were 3. 5% and 6. 2%, and the recovery ranged from 98. 9% to 106. 9%(n=7). The detection result by PSI-MS were in agreement with those by liquid chromatography-tandem mass spectrometry(LC-MS/MS), but the run time was only 1 min each sample., Conclusion: The method is accurate and reliable. And compared with the conventional LC-MS/MS technology, it is simple, rapid and low-cost.

Rising concern about the use of non-permitted colourants, in common food items such as dals and green vegetables sold in Indian markets, have led to a demand for low-cost point-of-use chemical analysis tools. Conventional food-analysis techniques involving tedious sample preparation protocols are not suited for in-field applications. Surface Enhanced Raman Spectroscopy (SERS) is an analytical technique that is well-suited for point-of-use chemical analysis with molecular level detection capability, which can also serve as a quality assurance tool for businesses. Effective and rapid signal collection from a large-area sample within a field-setting using disposable, low-cost SERS substrates is a key challenge in implementing such a solution. Herein, we demonstrate the use of inkjet-printed thin films comprising of robust nanostructured-silver as flexible, paper-based SERS (P-SERS) swabs for the direct detection of Metanil Yellow (MY) from toor dal (yellow split pigeon peas) samples and Malachite Green (MG) from green peas and green chillies. The macroscopic uniformity of these thin-films in combination with a portable Raman spectrometer equipped with orbital raster scanning (ORS™) technology for signal collection results in an unprecedented precision (RSD ∼ 1.6%) upon characterizing samples saturated with Rhodamine-6G (R6G), a standard Raman probe. As several food-cleansing products have appeared in the marketplace, the adulterant removal efficacy of some commercially available 'washes' as well as products such as 'ozoniser', which was determined by SERS characterization of swabs before and after use, is also reported., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A "detect and destroy" strategy is reported for the spectroscopic determination and photocatalytic degradation of Malachite Green (MG) in aqueous solutions. The intensity of the reflection peak maxima from the TiO 2 -coated 2D-photonic crystal (PhC) at 633 nm wavelength undergoes a gradual decrease with increasing concentrations of MG. The determination of MG was readily achieved in the nanomolar range due to the quenching of the reflection intensity of the peak, measured using a fiber optic probe. The assay works in the 1.0 nM to 10 μM MG concentration range with a detection limit of 1.3 nM. The same TiO 2 -coated 2D-PhC surface can photocatalytically degrade MG in aqueous solutions under UV irradiation. The photocatalytic degradation in the presence of TiO 2 -coated 2D-PhC becomes evident as the blue color of MG changes to colorless with increasing irradiation time. The decrease in absorption is detected at 617 nm. It was found that the photocatalytic efficiency of TiO 2 was synergistically enhanced in the presence of 2D-PhCs. It is concluded that each component of the TiO 2 -coated 2D-PhC system plays a key role in the detection and degradation of MG. Graphical abstractSchematic representation for reflectometric detection and photocatalytic degradation of hazardous Malachite Green dye using TiO 2 -coated two-dimensional photonic crystals.

The aim of the study was to initiate an exhaustive strategy of control by implementing both targeted and non-targeted metabolomics approaches. A LC-MS/MS method including an oxidative step for most of dyes was developed and validated to target the analysis of 14 residues belonging to different families of dyes. The method was suitable for the quantitative confirmation of 13 dyes at low ppb levels. The metabolomics approach objective was to compare fingerprints between farmed fish treated with malachite green and farmed fish treated with victoria pure blue bo. Analytical information on modifications in the metabolome of muscle, liver and plasma was exploited by HRMS following by multivariate statistics and revealed some direct or endogenous metabolites among relevant mass features contributing to the constructed models. These two approaches, either appropriate biomarkers either enlarged targeted dyes are explored concomitantly to help improving the strategy for tracking new illegal practices in aquaculture., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Food safety issues, especially those related to drug residues, have caused concern in recent years. Malachite green (MG), which is frequently used as ectoparasiticide, fungicide, and antiseptic in fish farming, is poisonous and carcinogenic. Several technologies and methods have been developed to accurately detect MG, but they can be time-consuming and expensive. Herein, we prepared a glass fiber paper as a surface-enhanced Raman scattering substrate for rapid and convenient detection of MG residues in fish. Uniform silver nanoparticles were deposited on the glass fiber paper by two in situ reduction steps of silver nitrate. The Raman signal of MG was detected and recorded using a portable Raman spectrometer in 10 s. There was a good linear relationship between the surface-enhanced Raman scattering signal intensity and the concentration of MG between 1 × 10 -7 and 1 × 10 -5 mol/L and the limit of detection was 5 × 10 -10 mol/L. Residual MG in fish samples was extracted and detected with a recovery rate range of 98.9-119.4%. This method provides a novel way to detect MG in fish., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A novel hydrophobic-SERS substrate platform based on silver nanoparticles (AgNPs) functionalized commercially available filter paper was reported. Compared with conventional substrates, not only could this novel SERS substrate meet the requirements of simple and large-scale preparation, but also realized direct droplet-detection with reusable property. In this work, the fabrication, physical characterization, and SERS sensitivity of the substrates to spot-on food determination were studied. The experimental results show the method exhibited high reproducibility with less than 9% spot-to-spot variation in Raman intensity. Furthermore, the method was successfully applied to detect melamine in diluted milk with the instrument detection limit (IDL) down to 1 ppm in a linear correlation (1 ppm-1000 ppm). In addition, this SERS substrate could also be applied successfully in other fields, such as the determination of pesticide (thiram) and dye (malachite green) in real environment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Effective monitoring of the content of malachite green (MG) in aquaculture is of great importance for food safety. Traditional methods for MG assay, such as chromatography and spectroscopy, have been criticized for expensive instrumentation and complicated pretreatments. An MG RNA aptamer (MGA) is a powerful tool for immediate and rapid detection of MG. However, RNA is easily degraded by nucleases and is unstable in the environment, making accurate and reliable detection of MG difficult. In order to address the problems, an innovative levo (l)-MGA with excellent stability is designed to perform the specific recognition function. Interestingly, the gel electrophoresis and fluorescence measurement results indicate that this unnaturally occurring l-aptamer is resistant to nuclease degradation and it can be kept intact in the standard buffer solution under room temperature for quite a long time. A label-free, simple, and efficient method has been developed for sensitive detection of MG in fish tissue, which holds promising potential in food analysis and environmental monitoring.

Semiconductor materials have become competitive candidates for surface-enhanced Raman scattering (SERS) substrates; however, their limited SERS sensitivity hinders the practical applications of semiconductors. Here, we develop a hybrid substrate by integrating anatase/rutile TiO 2 heterostructure with dense plasmonic hotspots of Ag nanoparticle (AgNPs) for efficient photoinduced enhanced Raman spectroscopy (PIERS). The PIERS mechanism is systematically investigated by means of a portable Raman instrument. When ultraviolet (UV) light irradiates the substrate, the TiO 2 -Ag hybrid arrays produce remarkable charge-transfer enhancement, which can be ascribed to the highly efficient charge separation driven by heterojunction and transfer from TiO 2 heterostructure to AgNPs. This platform allows for the rapid detection of multifold organic species, including malachite green (MG), crystal violet (CV), rhodamine 6G (R6G), thiram, and acephate, and as high as 27.8-fold enhancement over the normal SERS is achieved, representing the highest PIERS magnification up to the present time. The intensive PIERS enhancement makes it ultrasensitively detect analyte concentration of an order of magnitude lower than that of SERS method. The improved sensitivity and resolution can be readily realized by simple UV irradiation, which represents a major advantage of our PIERS methodology. Besides, the integration of uniform TiO 2 heterostructure arrays with AgNPs generates superior signal reproducibility with relative standard deviation (RSD) value of less than 14%. In addition, the detected molecules on the substrate can be eliminated by photocatalytic degradation after PIERS measurements by using UV irradiation, which makes the substrate reusable for 15 cycles. The ultrahigh sensitivity, superior reproducibility, and excellent recyclability displayed by our platform may provide new opportunities in field detection analysis coupled with a portable Raman instrument.

Pararosaniline hydrochloride (CPR) is a dye used for colouring paper, leather and natural and artificial fibres. It is also used in analytical and microbiological laboratories. It is a carcinogenic substance of category 1B. In analytical chemistry, it is used for detecting the following among others: bromates, formaldehyde, ozone, sulphite and sulfur dioxide. CPR is a dye commonly used in microbiology for staining preparations, for staining bacteria, antibodies or other organisms. In Poland, about 800 employees were exposed to this substance. The lack of methods for the determination of pararosaniline hydrochloride in workplace air makes it impossible to assess the occupational exposure of workers to this substance. For this reason, a determination method has been developed, which allows for the determination of pararosaniline hydrochloride in the air. This method makes it possible to determine the concentration of CPR in the air at the workplace within the range from 0.002 to 0.04 mg/m 3 (for an air sample of 120 L). The method is based on the adsorption of pararosaniline hydrochloride present in the workplace air on a polypropylene filter, eluting the substance deposited on the filter with methanol and analysing the solution thus obtained using high-performance liquid chromatography with a diode array detector (wavelength of 544 nm). Using an Ultra C18 (250 mm length) chromatographic column at a temperature of 23 °C and the mobile phase of methanol:0.1% phosphoric acid(V) (95:5, v/v) at flow rate of 0.6 mL/min makes it possible to determine the content of pararosaniline hydrochloride in the presence of aniline, nitrobenzene and 4-tolylamine. Limit of detection and limit of quantification were 0.17 ng/mL and 0.51 ng/mL, respectively.

A high-throughput PRiME (process, robustness, improvements, matrix effects, ease of use) sample purification procedure was developed to simplify the multiple steps of traditional SPE in extracting the malachite green and leucomalachite green in Chinese softshell turtle (Pelodiscus sinensis). The sample loading volume, extracting solvent type, and pH value of the employed PRiME hydrophilic-lipophilic balance cartridge for sample purification were optimized to be 3 mL, acetonitrile, and pH 5, respectively. In comparison with traditional SPE, the PRiME process is cost-effective, solvent-saving, and simple to operate, which only consists of a passing through step without traditional sorbent conditioning and impurity washing. Afterward, eluate was analyzed by ultra-performance liquid chromatography-tandem mass spectrometry, and the proposed method was validated for linearity (R 2 > 0.9992), intraday precision (2.44-3.22%), interday precision (3.28-6.58%), sensitivity (LOD ≤ 0.18 μg/kg and, LOQ ≤ 0.60 μg/kg), and recovery (88.7-94.1%, RSD < 6.79%). The results indicated that the PRiME technique can simplify the sample preparation procedure by avoiding the tedious steps, such as conditioning, washing, etc. It would be of significant interest for environmental and food safety applications in the market of Chinese softshell turtle and related products., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A specific and sensitive colorimetric aptasensor is described for the determination of Malachite Green (MG). It is exploiting the inhibition of the peroxidase-like activity of gold nanoparticles (AuNPs). The AuNPs act as enzyme mimics that catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by H 2 O 2 to yield a dark blue solution. The catalytic activity is inhibited by hexadecyl trimethyl ammonium ion, specifically by cetyltrimethylammonium bromide (CTAB), which causes the aggregation of AuNPs. If a (negatively charged) RNA-aptamer against MG is added, it binds to the positively charged CTAB and prevents aggregation. This enhances the enzyme mimicking activity of the AuNPs and leads to the formation of a dark blue solution. However, in the presence of MG, the aptamer binds to MG, and leads to the aggregation of AuNPs again. The aggregated AuNPs possess a light blue color. A colorimetric method (best performed at 650 nm) was work out that can detect MG in a concentration range from 10 to 500 nmol L -1 . The detection limit based on 3σ/k criterion is 1.8 nmol L -1 . The assay is highly specific and accurate. Recoveries from spiked real samples (aquaculture water) ranged from 80% to 120%. Graphical abstract Based on the inhibition of cetyltrimethyal ammonium ion and the enhancement of RNA-aptamer, the differences of the peroxidase-like activities of AuNPs can be greatly enlarged with and without MG, by which a colorimetric aptasensor can be constructed for the detection of Malachite Green (MG).

Collecting highly diluted target analytes into specific hot spot regions is vital for ultrasensitive surface-enhanced Raman spectroscopy (SERS) applications. In this work, a hydrophobic slippery platform was employed as a concentrator to construct colloidal SERS-active substrates regardless of the diffusion limits during droplet evaporation. Within only 140 s, sufficient absorption between the analytes and colloidal Au nanoparticles (Au NPs) was observed by fluorescence imaging. This effect resulted in excellent SERS sensitivity and stability. Compared with the common metal colloid-based SERS substrates, e.g., drying on a silicon wafer or detection in colloidal solutions, this preconcentrated method showed lower detection limits and the lowest detection concentration of crystal violet molecule down to 10 -12 M with a portable Raman spectrometer. Such high signal enhancement was mainly ascribed to the condensation effect of Au colloids/analytes on the hydrophobic slippery substrate, by which almost all probe molecules were guided into the "hot spot" regions of aggregated Au NPs. Using the SERS platform, various illegal additives in realistic food and health-care products, for example, malachite green (1 ppb) added in fish and morphine (0.1 ppm) added in a chafing dish, could be sensitively detected. Therefore, our protocol is a general SERS platform that may provide a simple, fast, and cost-effective approach for trace molecular sensing.

Proteomic analysis by combining PAGE separation, gel slicing and slice-by-slice in-gel digestion and LC-MS/MS has been frequently reported in recent years. Since the MS analysis would provide identities and quantities of the proteins along the whole lane, gel visualization by dye staining could be skipped to save time and labor. In this work, we examined the effects of CBB R-250 staining on the performance of the method and the results showed actually better results were obtained with CBB staining than without, both in nondenaturing PAGE and SDS-PAGE. A primary examination was firstly performed with excised gel bands of purified proteins and the LC-MS/MS results showed that almost all the proteins were detected with higher sequence coverages and quantities from the stained bands than from the unstained. Then a proteomic sample of rat heart soluble proteins was examined for the complete workflow. The sample was separated by both nondenaturing PAGE and SDS-PAGE and the gels were divided to halves for CBB staining and fixation without staining, respectively. Multi-blade cutters were used to simultaneously cut lanes and then slice each lane into about forty squares of the same size. All the gel pieces were analyzed in standard procedures of in-gel digestion, peptide extraction and label-free quantitative LC-MS/MS. The results showed more proteins, about 40% in nondenaturing PAGE and 18% in SDS-PAGE, were detected from the CBB-stained lanes than from the unstained ones. Examination on the detected quantities and square numbers of individual proteins also confirmed about the better detection with CBB staining. As the data showed the detection of proteins with lower molecular masses (e.g. <30 kDa) were more benefited by the staining, we speculate the dye binding might help retaining of the proteins in the gel matrix., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Malachite green is used in aquaculture and fisheries as a fungicide and antiseptic and it is also used in industry as a dye. However, malachite green is carcinogenic and highly toxic for humans and animals. In this study, a spectrophotometric method was developed to detect malachite green. The method was based on the surface plasmon resonance property of gold nanoparticles and interaction between malachite green and gold nanoparticles., Results: Malachite green-gold nanoparticles were rapidly aggregated in the acidic medium; as a result, a color change from red to blue was observed, which was easily detectable by the naked eye. The absorption ratio (A623/A520) of the gold nanoparticles in an optimized system exhibited a linear correlation with malachite green concentration. The method detection limit and linear range were 3 and 50-350 ng mL -1 , respectively. The method was applied successfully to detect malachite green in different samples., Conclusion: The method was simple and rapid to detect malachite green. The most important advantages of the method are the possibility of malachite green determination with very good accuracy and sensitivity using a simple UV-visible spectrometer without any expensive or sophisticated instrumentation and also the versatility of real samples. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Polymethylmethacrylate matrix has been developed as an analytical device for visual and spectrophotometric determination of malachite green. Based on the studied effects of different pH levels, temperature, contact time and diverse ions, optimum analysis conditions have been established. For malachite green, the calibration graph was linear in the range of 0.5-30 mg·L -1 in the samples with the detection limit of 1 mg·kg -1 . The proposed simple method makes it possible to determine malachite green visually in the concentration range 8-30 mg·L -1 using a colour scale. The method has been implemented for determination of malachite green in various sea fish samples., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was validated and used to quantify crystal violet (CV), leucocrystal violet (LCV), malachite green (MG), leucomalachite green (LMG), and brilliant green (BG) residues in frozen fish (121 samples) from various countries, in order to detect the use of prohibited antibiotic dyes in fish for human consumption. The microbial quality of the fish was also assessed along with the effectiveness of a simple treatment with whole fat milk to reduce the levels of CV and LCV contamination. CV and LCV were the only two residues detected. They were found in farmed Pangasius (0.362 to 41.34 μg/kg and 0.178 to 10.58 μg/kg, respectively) and Tilapia (1.24 to 9.48 μg/kg and 1.29 to 2.81 μg/kg). Based on aerobic plate count (APC), 74%, 59%, and 55% of the samples of Tilapia fillets (from China) and Pangasius fillets (United Arab Emirates and Vietnam), and 100% and 50% of the skin samples of Hake (Argentina and U.S.A.) were of unacceptable microbial quality (APC > 10 7 cfu/g). Human pathogens, namely Escherichia coli, Staphylococcus aureus, and Vibrio spp., were detected in most fish. A significant reduction in CV and LCV concentrations by more than a third was achieved after immersing Pangasius and Tilapia fillets in whole fat milk for 120 minutes. These findings support the necessity of regular inspections and monitoring of CV and other antibiotic dye residues in fish, along with routine assessments of fish microbial quality, in order to protect public health. PRACTICAL APPLICATION: The described LC-MS/MS method can be used to rapidly and simultaneously quantify antibiotic dye residues in frozen fish. CV and LCV were detected in farmed Pangasius and Tilapia fillets and their concentrations was reduced by more than one third after immersing the fillets in whole milk for 120 min, a treatment which is not intended to replace safe fish farming practices upstream to artificially lower the level of banned dyes in fish. The findings support the necessity of regular inspections and monitoring of CV and other antibiotic dye residues in fish, along with assessments of fish microbial quality, to protect public health., (© 2019 Institute of Food Technologists®.)

With the advanced development of miniaturized Raman spectroscopy, surface-enhanced Raman spectroscopy (SERS) has extended its applications into the field of point-of-care testing (POCT) and demonstrated its great significance by virtue of its noninvasive property and capability of fingerprint identification. In the SERS-based analysis and/or sensing system, the preparation of a low-cost, high-performance SERS substrate is critically important. In this manuscript, vacuum filtration is utilized to fabricate the silver nanoparticles (AgNPs)-embedded nylon filter membrane (ANFM) as flexible paper-based SERS chips. By characterizing the typical analytes with a miniaturized smartphone-based Raman analyzer, the proposed SERS chips have successfully demonstrated good sensitivity, repeatability, and stability. The lowest concentration as detected can approach 1 pmol for rhodamine 6G (RH6G) and 10 pmol for both crystal violet (CV) and malachite green (MG), respectively. With the help of the microporous structure of the membrane, the ANFM-based SERS chips can implement the separation of small molecules from a complex mixture and can achieve "purified" SERS signals of targeted molecules. Besides, with the function of antifriction resistance and flexibility, the ANFM can serve as SERS papers to preconcentrate the contaminates by multiple swapping and further enhance the SERS signals for point-of-care analysis. Therefore, we demonstrate multifunctions of the flexible ANFM-based SERS chips, which provide a promising solution for the POCT analysis with the SERS technique on account of their flexibility and low fabrication cost.

Malachite green (MG) has been widely used in the aquaculture industry as a fungicide and parasiticide because of its high efficiency and low cost, and it is commonly found in aquatic products and environmental water. However, MG and its primary metabolite, leuco-malachite green (LMG), are also toxic inorganic contaminants that are hazardous to the health of humans and other organisms. A variety of methods have been proposed in recent years for detecting and monitoring MG and LMG. This article was compiled as a general review of the methods proposed for MG and LMG detection, and several important detection parameters, such as the limit of detection, recovery and relative standard deviation, were tabulated. The analytical methods for the determination of MG and LMG in various matrices include high-performance liquid chromatography separation-based methods, liquid chromatography tandem mass spectrometry, surface-enhanced Raman spectroscopy, electrochemical methods, immunological assays, spectrophotometry and fluorescent methods which were described in detail in this article. In addition, some sample preparation techniques were also described. This review can provide expert guidance to the reader on the advantages, disadvantages and applicability of the different methodologies. This review also discussed challenges and several perspectives on the future trends in the determination of MG and LMG.

It is urgent to develop a rapid, reliable, and in-site determination method to detect or monitor trace amounts of toxic substances in the field. Here, we report an alternative surface-enhanced Raman scattering (SERS) method coupled with a portable Raman device on a plasmonic three-dimension (3D) hot spot sensing surface. Plasmonic Ag nanoparticles (AgNPs) were uniformly deposited on 3D TiO 2 nanopore arrays as a sensitive SERS substrate, and further coated with graphene oxide (GO). We demonstrate the plasmon-induced SERS enhancement (5.8-fold) and the improvement of catalytic activity by incorporation of plasmonic AgNPs into the 3D TiO 2 nanopore arrays. The modification of GO on the TiO 2 -Ag nanopore array further increases by a 6.2-fold Raman enhancement compared to TiO 2 -Ag while maintaining good uniformity (RSD < 10%). The optimized TiO 2 -Ag-GO substrate shows powerful quantitative detection potential for drug residues in fish scales via a simple scrubbing method, and the limit of detection (LOD) for crystal violet (CV) was 10 -8 M. The SERS substrate also showed detection practicability of pesticide residues in banana peel with an LOD of 10 -7 M. In addition, our TiO 2 -Ag-GO substrate exhibits excellent SERS self-monitoring performance for catalytic reduction of multiple organics in NaBH 4 solution, and the substrate shows good recyclability of 6 cycles. Such a 3D TiO 2 -Ag-GO substrate is a promising SERS substrate with good sensitivity, uniformity, and reusability, and may be utilized for further miniaturization for point of analytical applications.