Ratiometric Fluorescence Signal Research Articles

NIR‐II Ratiometric Fluorescent Nanoplatform for Real‐Time Monitoring and Evaluating Cancer Sonodynamic Therapy Efficacy

AbstractReal‐time monitoring the therapeutic process of sonodynamic therapy (SDT) is essential to optimize the treatment course in time and eventually improve the efficacy. The generation of singlet oxygen (1O2) is a quintessential characteristic of SDT, which permits non‐invasive monitoring of SDT by real‐time imaging of 1O2 inside the tumor. Nonetheless, the majority of probes are unable to measure 1O2 in real time because of its short half‐life and strong oxidative capacity. Here, the study constructs a ratiometric nanoplatform (DTPI) utilizing two fluorescent probes and the sonosensitizer TiO2. The poisonous 1O2 generated by DTPI following ultrasonic (US) radiation efficiently destroys tumor cells. The structural disruption of fluorescent dye IR‐1061 by 1O2 leads to a reduction in the DTPI fluorescence signal at 1100 nm, while US radiation has no impact on the fluorescence signal at 1550 nm. Thus, DTPI provides a precise and consistent reflection of the treatment efficacy at the tumor site, leveraging the ratiometric fluorescence signal and variations in oxygen content throughout the treatment process. This ratiometric‐fluorescence‐based reflection strategy establishes an effective and dependable platform for the real‐time monitoring and assessment of the cancer therapeutic effect through ratiometric probes.

Determination of Acetylcholinesterase Activity Based on Ratiometric Fluorescence Signal Sensing.

Acetylcholinesterase (AChE) plays an important role in the treatment of human diseases, environmental security and global food supply. In this study, the simple fluorescent indicators and MnO2 nanosheets were developed and integrated to establish a ratiometric fluorescence sensing system for the detection of AChE activity. Two fluorescence signals could be recorded independently at the same excitation wavelength, which extended the detection range and enhanced the visibility of results. Fluorescence of F-PDA was quenched by MnO2 nanosheets on account of inner filtering effect. Meanwhile, the nonfluorescent OPD was catalytically oxidized to 2,3-diaminophenazine by MnO2 nanosheets. The acetylcholine (ATCh) was catalytically hydrolyzed by AChE to enzymatic thiocholine, which decomposed MnO2 to Mn2+, recovered the fluorescence of F-PDA and reduced the emission of ox-OPD. Utilizing the fluorescence intensity ratio F468/F558 as the signal readout, the ratiometric fluorescence method was established to detect AChE activity. Under the excitation wavelength of 410nm, the ratio F460/F558 against the AChE concentration demonstrated two linear relationships in the range 0.05 -1.0 and 1.0-50 U·L- 1 with a limit of detection (LOD) of 0.073 U·L- 1. The method was applied to the detection of AChE activity and the analysis of the inhibitor Huperzine-A. Due to the advantages of high sensitivity and favorable selectivity, the method possesses an application prospect in the activity deteceion of AChE and the screening of inhibitors.

NIR-II Ratiometric Fluorescence Probes Enable Precise Determination of the Metastatic Status of Sentinel Lymph Nodes.

Accurately determining the metastatic status of sentinel lymph nodes (SLNs) through noninvasive imaging with high imaging resolution and sensitivity is crucial for cancer therapy. Herein, we report a dual-tracer-based NIR-II ratiometric fluorescence nanoplatform combining targeted and nontargeted moieties to determine the metastatic status of SLNs through the recording of ratio signals. Ratiometric fluorescence imaging revealed approximately 2-fold increases in signals in tumor-draining SLNs compared to inflamed and normal SLNs. Additionally, inflamed SLNs were diagnosed by combining the ratio value with the enlarged size outputted by NIR-II fluorescence imaging. The metastatic status diagnostic results obtained through NIR-II ratiometric fluorescence signals were further confirmed by standard H&E staining, indicating that the ratiometric fluorescence strategy could achieve distant metastases detection. Furthermore, the superior imaging quality of ratiometric probes enables visualization of the detailed change in the lymphatic network accompanying tumor growth. Compared to clinically available and state-of-the-art NIR contrast agents, our dual-tracer-based NIR-II ratiometric fluorescence probes provide significantly improved performance, allowing for the quick assessment of lymphatic function and guiding the removal of tumor-infiltrating SLNs during cancer surgery.

New Tritopic Terpyridine‐based Fluorescent Probe for the Efficient Determination of Th4+, Zn2+ and F− Ions

AbstractThe present work demonstrates the determination of thorium (Th4+), zinc (Zn2+) and fluoride (F−) ions by employing a substituted tripodal terpyridine (TP) as a fluorescence sensor. The distinct fluorescence intensities of the free TP and TP‐M complexes (M=Zn2+or Th4+) with the same emission wavelength at 468 nm were observed in steady‐state photoluminescence (PL) spectra at pH 7.0 with a 1 : 3 ratio of metal to the ligand. A ratiometric dual fluorescence signals for Zn2+ and Th4+ were displayed by the probe TP at varied concentrations. Furthermore, the quantitative determination of F− ions was achieved through the addition of F− to the tripod‐terpyridine‐zinc complex (TP‐Zn2+) where the fluorescence intensity decreases gradually. Notably, the present work facilitates the LOD (limit of detection) as 1.1×10−10 M, 1.01×10−8 M and 3.18×10−10 M and further LOQ (limit of quantification) as 0.37×10−9 M, 0.33×10−7 M and 5.26×10−9 M for Zn2+, Th4+ and F− ions, respectively. The predicted absorption spectra of TP show HOMO to LUMO+2 electronic excitation at 358 nm whereas the complex TP‐Th4+ exhibits HOMO to LUMO+1 electronic excitation at 568 nm.

A ratiometric dual-fluorescent paper-based synthetic biosensor for visual detection of tetracycline on-site

The excessive use of tetracycline poses a threat to human health, making it essential to monitor and regulate its usage. While whole-cell biosensors offer a simple and cost-effective method, their utility is constrained by limitations in sensitivity, portability, and robustness, hindering real-time measurements within complex environmental contexts. In this study, a ratiometric i/cTetR synthetic biosensing test strip with an engineered modified dual-fluorescence reporting was developed for detecting Tet antibiotics in water and food. First, the standardized unidirectional promoter PtetR by tailoring and screening TetR transcription factor binding sites and verified by molecular docking, shortening the detection time. Secondly, decoupling the sensing and reporting modules enhances the biosensor's performance, eliminating genetic background leakage and tripling the output signal. Thirdly, a ratiometric dual fluorescence signal i/cTetR biosensing test strip was designed. Under the light box LED/UV light source, the dual signal output method significantly reduced false negative results and enhanced the anti-interference capability of the biosensor. The i/cTetR strips can detect Tet in tap water (5–1280 μg/mL) and milk (50–3200 μg/kg) within 45 min in high volume on-site without separation and purification. This study provides a standardized and universal sensing method for the field detection of antibiotic contaminants.

An enzyme-free and label-free ratiometric fluorescence signal amplification biosensor based on DNA-silver nanoclusters and catalytic hairpin assembly for tetracycline detection

Sensitive and accurate in antibiotic detection is crucial for food security, environmental conservation, public health, and environmental protection. In this study, we developed an enzyme-free and label-free ratiometric fluorescence biosensor for highly sensitive detection of tetracycline (TET). This biosensor utilized a combination of target-triggered catalytic hairpin assembly (CHA) and DNA-silver nanocluster (DNA-AgNCs) dual-color fluorescent signal probes. Upon TET binding to the aptamer, it initiated the leakage of CHA initiator, resulting in the formation of numerous double-stranded DNA complexes. This process exposed the foothold sequence of hairpin-structured DNA-AgNCs (H3-AgNCs) that exhibit yellow fluorescence. Subsequently, the conformational changes in H3-AgNCs significantly reduced the yellow fluorescence intensity (FI) and reveal the auxiliary DNA-AgNCs (a-DNA-AgNCs) linkage sequence. The binding of a-DNA-AgNCs brought two dark AgNCs close, forming nanocluster dimers emitting red fluorescence. The results demonstrate that the variation in ratiometric signal is linear with TET concentration between 0.1 and 50 ng/mL, with a detection limit as low as 0.086 ng/mL. Additionally, this approach exhibited good recovery results with spiked milk samples. This enzyme-free and label-free combined signal amplification method with proportional signal output based on DNA-AgNCs provides valuable empirical reference for creating low-cost, stable, and highly-sensitive biosensors.

Ratiometric luminescent sensor based on BSA-coated gold/silver nanoclusters for the selective determination and spatiotemporal imaging of gallic acid in plants.

Anewfluorescence sensing strategyhas been developed. Four bimetallic nanoclusters, gold/silver, gold/copper, gold/molybdenum and gold/cobalt, were prepared using bovine serum albumin (BSA) as a reducing and stabilizing agent. The fluorescence properties of four nanoclusters were explored by solid-state UV and XPS. The gold/silver nanoclusters (BSA-Au/Ag NCs) with the best ratiometric fluorescence properties for gallic acid (GA) in plants were selected to realize the sensitive detection of GA. GA affected the conformation of BSA, thereby disrupting the luminescent environment of the nanoclusters, resulting in a pronounced fluorescence quenching at 566nm. The ratiometric fluorescence signal (I566/I453) was used for trace detection of GAin plants. It has a wide response range of 1.25-40.0μM and a low detection limit of 45.27nM. GA was detected at 19.49μM in the plant extract, and the spiked recoveries ranged from 96.09 to 104.6%. In addition, due to the non-toxic and biocompatible properties of BSA, BSA-Au/Ag NCs have also been validated for fluorescence imaging of plant tissues. It realized the comparison of GA content in different parts of plants and the difference of GA content in plants after abiotic stress. Therefore, the developed strategy offers potential application for the analytical study of active substances in plants.

Covalent Coupling Assisted Hydrophilic Perovskite Spheres for Ratiometric Fluorescent Visual Multichannel Immunoassay.

Quantitative fluorescence immunoassay is essential for the construction of biosensing mechanisms and the quantification of trace markers. But the interference problems caused by low fluorescence efficiency and broad fluorescence spectrum of fluorescent probes have hindered the continued development of ratiometric fluorescence sensing in biosensing. Perovskite materials, with ultra-high color purity (FWHM < 30nm) and photoluminescence quantum yield (PLQY) (close to 100%), are expected to be next-generation fluorescent probes. However, poor water stability and biocompatibility are still non-negligible in biosensor applications. In this work, hyperstatic perovskite fluorescent microspheres prepared by swelling-shrinking method can be used as ratiometric fluorescence signals and biological immunoassay platforms. Meanwhile, inspired by p-aminophenol (AP) controlled synthesis and the catalytic reaction of 4-aminophenol phosphate (APP) triggered by alkaline phosphatase (ALP), a strategy to prepare fluorescent nanoparticles as fluorescence signals for ALP detection is proposed. Most importantly, it is proposed for the first time to combine this enzymatic fluorescence with perovskite materials using covalent linkage to create a novel cascade immunoassay and use it for quantitative and visualization determination of hepatitis B surface antigen (HBsAg)for application verification. These results indicate the biosensing potential of perovskite materials and provide a pathway for high sensitivity enzyme detection and enzyme triggered immune detection.

Precisely Constructing Renal-Clearable and LAP-Activatable Ratiometric Molecular Probes for Early Diagnosis of Acute and Chronic Kidney Injury Via Optimizing Asymmetric DPP Dyes.

Fluorescence analysis is an increasingly important contributor to the early diagnosis of kidney diseases. To achieve precise visualization of the kidneys and early diagnosis of related diseases, an asymmetric pyrrolopyrrolidone (DPP) dye platform with C-aromatic substituents and N-lipophilic/hydrophilic modification was constructed. Based on these, we developed the renal-clearable, water-soluble, and kidney injury biomarker leucine aminopeptidase (LAP) activated ratiometric fluorescent probe DPP-S-L. In the mouse model of cisplatin-induced acute kidney injury and during the development of type 2 diabetes to diabetic kidney disease, we visualized for the first time the upregulation of LAP in the kidney and urine by dual-channel ratiometric fluorescence signal and diagnosed the kidney injury earlier and more sensitively than blood/urine enzyme detection and tissue analysis. This study showcases an excellent asymmetric DPP dye platform and renal-clearable ratiometric fluorescent probe design strategy that is extended to determination and visualization of other biomarkers for early disease diagnosis.

A novel dual-mode sensing strategy for the determination of 1,4-dithiothreitol based on tannic acid stabilized copper nanocluster

In this paper, a novel sensing strategy with both ratiometric fluorescence and colorimetric responses towards 1,4-dithiothreitol (DTT) was developed based on tannic acid-templated copper nanoclusters (TA-CuNCs). The one-step reduction approach was utilized to obtain TA-CuNCs with the fluorescence emission peak located at 436 nm. Through the catalytic oxidation of Cu2+, o-phenylenediamine (OPD) could be converted to 2,3-diaminophenazine (DAP) with an obvious fluorescence signal at 557 nm. Additionally, the obtained yellow DAP showed a clear absorption peak at 420 nm, DAP could cause a significant fluorescence quenching of TA-CuNCs on account of inner-filter effect (IFE). The oxidation process between Cu2+ and OPD could be largely prevented by 1,4-dithiothreitol (DTT), whose sulfhydryl groups bound to Cu2+, and the fluorescence intensity of DAP was diminished, whereas the fluorescence of the TA-CuNCs was restored. The ratiometric fluorescence signal and absorbance intensity changed with the concentration of DTT in the linear ranges of 5–45 μM and 5–40 μM, respectively. And the limits of detection (LOD) for two modes were found to be 2.98 and 2.03 μM, respectively. Furthermore, the dual-mode assay method was used for the determination of DTT accurately and reliably in human serum samples.

In Situ Generated Dye@MOF/COF Heterostructure for Fluorescence Detection of Chloroquine Phosphate and Folic Acid via Different Luminescent Channels.

Metal-organic framework (MOF) and covalent-organic framework (COF) hybrid materials can combine the unique properties of MOF and COF components, and their applications in fluorescence sensing have attracted more and more attention. Herein, ZIF-90 is grown on 3D-COF by a simple in situ growing method in which the 7-amino-4-methylcoumarin (AMC) is encapsulated in ZIF-90 to construct a fluorescent sensor. Chloroquine phosphate (CQP) can coordinate with Zn2+ to decompose the ZIF-90 and release AMC. At 365 nm excitation, the ratiometric fluorescence signal AMC/3D-COF (I430/I598) increases linearly with CQP in a linear range of 4 × 10-5 to 4 × 10-4 M in urine. Under 340 nm excitation, quantitative analysis of CQP in the serum (3 × 10-6 to 4 × 10-5 M) is based on the fluorescence intensity of Zn-CQP/3D-COF (I384/I598). In addition, AMC@ZIF-90/3D-COF (1) exhibits high anti-interference and selectivity in sensing of FA with a "turn off" mode, with a correlation range of 1 × 10-5 to 1 × 10-3 M. The fluorescence color changes triggered by CQP under different excitation conditions, and the different fluorescence responses caused by CQP make it a highly secure anticounterfeiting platform. The synthesized dye@MOF/COF hybrids not only provide a new way to integrate multiple emission to design fluorescent probes for differentiation detection but also offer ideas for the design of anticounterfeiting platforms.

Intelligent quantitative recognition of sulfide using machine learning-based ratiometric fluorescence probe of metal-organic framework UiO-66-NH2/Ppix

Sulfides possess either high toxicity or play crucial physiological role such as gas transmitter dependent upon dosage, hence the significant for their rapid sensitive and selective concentration determination. Herein, a machine learning enhanced ratiometric fluorescence sensor was engineered for sulfide determination by incorporating the nanometal-organic framework (UiO-66-NH2) along with protoporphyrin IX (Ppix). The blue fluorescence at 431 nm originated from the moiety of UiO-66-NH2 by 365 nm excitation serves as an internal calibration reference signal, while the red fluorescence at 629 nm from the moiety of Ppix serves as the analytical signal, and the intensity is correlated to the amount of sulfides. The fluorescence color of the sensor gradually varies from blue to red upon sequential addition of copper and sulfide ions, resulting in RGB (Red, Green, Blue) feature values for corresponding sulfide concentrations, which facilities the advanced data processing techniques using machine learning algorithms. On the basis of fluorescence image fingerprint extraction and machine learning algorithms, an online data analysis model was developed to improve the precision and accuracy of sulfide determination. The established model employed Linear Discriminant Analysis (LDA) and was subjected to rigorous cross-validation to ensure its robustness. By analyzing the correlation between RGB feature values and sulfide concentrations, the study highlighted a significant positive relationship between the red feature values and sulfide concentrations. The application of machine learning techniques on the ratiometric fluorescence signal of the UiO-66-NH2/Ppix probe demonstrated its potential for intelligent quantitative determination of sulfides, offering a valuable and efficient tool for pollution detection and real-time rapid environmental monitoring.

Ratiometric Fluorescence and Afterglow Lifetime Dual-Channel Nanoprobe for Simultaneous Imaging of HOCl and Temperature in Arthritis.

Arthritis is a joint disorder that potentially causes permanent joint damage and eventual disability without effective treatment. Clinical detection methods, including in vitro blood tests and anatomical imaging, still have limitations in achieving real-time in situ early detection of arthritis. In this work, a dual-channel luminescence nanoprobe (AGNPs-Cy7) is reported, which combines a cyanine dye and a photochemical reaction-based afterglow system for real-time in vivo imaging of arthritis. AGNPs-Cy7 simultaneously detect hypochlorous acid (HOCl) and temperature, two important indicators associated with the early development of arthritis, by monitoring the respective changes in independent ratiometric fluorescence and afterglow lifetime signals. The anti-interference properties of both the ratiometric fluorescence signal and afterglow lifetime signal enhance sensing accuracy compared to the single luminescence intensity. The developed probe successfully reveals the simultaneous increase in HOCl concentration and temperature in an arthritis mouse model.

A luminescent MOF-based nonenzymatic probe for colorimetric/photothermal/fluorescence triple-mode assay of uric acid in body fluids

Monitoring the levels of uric acid (UA) in body fluids is of great significance in the clinical diagnosis and therapy of related diseases. Herein, a novel nanocomposite R6G@Fe-MOF based nonenzymatic probe is presented to provide a ratiometric fluorescent, colorimetric, and photothermal triple read-out signal for the visual, sensitive, and convenient assay of UA. The framework structure of the in situ encapsulated R6G@Fe-MOF is found to decompose upon the addition of UA, resulting in the reduction of Fe3+ to Fe2+. This reduction will lead to a rapid increase in fluorescence emission (FL) at 430 nm. Simultaneously, the FL at 573 nm will decrease remarkably due to the inner filter effect (IFE) between UA and R6G@Fe-MOF. Furthermore, the reaction of the generated Fe2+ with potassium ferricyanide (K3 [Fe(CN)6]) can in situ generate Prussian blue (PBNPs) with outstanding color and photothermal properties, which allow for easy colorimetric and photothermal signal readout. The detection limits (LOD) for the colorimetric, fluorometric and photothermal detection are low at 1.68 μM, 0.236 μM, and 1.32 μM respectively. Ultimately, it is successfully employed to determine UA in urine, serum, and saliva, yielding satisfactory results. The constructed R6G@Fe-MOF sensor provides a simple, sensitive, and accurate determination of UA that can be tailored to meet the needs of various applications, and also provides new perspectives for the design and development of versatile sensors for diverse uses.

A Readily Available Red‐Emitting Methylthio‐Substituted Salicylaldehyde Azine with AIE Feature for Ratiometric Detection of HClO

AbstractDeveloping a simple, selective, and rapid reliable method to monitor hypochlorous acid (HClO) in vivo is very meaningful due to its important physiological and pathological functions. In this research, a novel AIE based fluorescent probe for ratiometric detection of HClO was fabricated by a simple one‐step synthesis, in which the methyl sulfide group and salicylaldehyde azine serve as the recognition unit and fluorophore, respectively. This probe was visible‐light‐excitable, and showed a palpable aggregation‐induced emission into the red region with high stability to Cu2+ and pH. Oxidation of the methyl sulfide group can be achieved rapidly and specifically by HClO in physiological condition, which enables it to display a rapid, ratiometric fluorescent response to HClO with high selectivity over other biologically pertinent species. By exploiting the probe‐based tool, we successfully validated its practical utility in selectively recognizing HClO in living cells via ratiometric fluorescence signals, thereby providing a potential method for investigating the relevant functions of HClO in biosystems.

Competitive ratiometric fluorescent lateral flow immunoassay based on dual emission signal for sensitive detection of chlorothalonil

In this study, we develop a competitive ratiometric fluorescent lateral flow immunoassay (CRF-LFIA) based on dual emission fluorescence signal, which has great advantage in visual and quantitative detection of Chlorothalonil (CTN). Red-emitted fluorescent magnetic nanobeads (FMNBs) and green-emitted aggregation-induced emission fluorescent microsphere (AIEFM) are synthesized and conjugated to antibodies and antigens respectively, resulting in competitive binding with the analyte. The ratiometric fluorescence signal which comes from the overlap of these two fluorescence emissions. FMNBs probes also provide immunomagnetic separation (IMS) to enrich the analysts and resist complex matrix effects. This strip generates a visually discernible yellow-to-green fluorescence color change in the presence of CTN (2 ng/mL), which could be incisively observed by naked eye. Moreover, the limit of detection (LOD) reached 0.152 ng/mL by measurement of color (Red-Green-Blue, RGB) signals. Method validation shows a good correlation between CRF-LFIA and LC-MS/MS.

Construction of a nanoscale metal-organic framework aptasensor for fluorescence ratiometric sensing of AFB1 in real samples.

Aflatoxins B1 (AFB1) that could contaminate agricultural products has received sustained attention due to its high toxicity and wide distribution. Therefore, sensitive and facile detection method for AFB1 is significant for food safety and control. In this work, a ratiometric fluorescence NMOFs-Aptasensor was developed based on the combination of Cy3-modified aptamer and zirconium-based nanoscale metal-organic frameworks (NMOFs). NMOFs served as energy donors, and Cy3 labeled on the AFB1 aptamer was used as an acceptor. An energy donor-acceptor pair was fabricated in the NMOFs-Aptasensor. With AFB1 selectively caught by the AFB1 aptamer, the fluorescence of the NMOFs-Aptasensor changed via fluorescence resonance energy transfer (FRET), and the fluorescence spectra changed accordingly. The ratiometric fluorescence signal was utilized to quantitatively measure AFB1. The reported NMOFs-Aptasensor presented great detection performance from 0 to 3.33ngmL-1, with an LOD of 0.08ngmL-1. Moreover, the fluorescence sensor was successfully applied to detect AFB1 in real samples.

Cell-Membrane-Localizing Fluorescence Probes for Aminopeptidase N.

Aminopeptidase N (APN), a transmembrane ectoenzyme, plays multifunctional roles in cell survival and migration, angiogenesis, blood pressure regulation, and viral uptake. Abnormally high levels of the enzyme can be found in some tumors and injured liver and kidney. Therefore, noninvasive detection methods for APN are in demand for diagnosing and studying the associated diseases, leading to two dozen activatable small-molecule probes reported up to date. All of the known probes, however, analyze the enzyme activity by monitoring fluorescent molecules inside cells, despite the enzymatic reaction taking place on the outer cell membrane. In this case, different cell permeability and enzyme kinetics can cause false signal data. To address this critical issue, we have developed two cell-membrane-localizing APN probes whose enzymatic products also localize the outer cell membrane. The probes selectively respond to APN with ratiometric fluorescence signal changes. A selected probe, which has two-photon imaging capability, allowed us to determine the relative APN levels in various organ tissues for the first time: 4.3 (intestine), 2.1 (kidney), 2.7 (liver), 3.2 (lung), and 1.0 (stomach). Also, a higher APN level was observed from a HepG2-xenograft mouse tissue in comparison with the normal tissue. Furthermore, we observed a significant APN level increase in the mouse liver of a drug (acetaminophen)-induced liver injury model. The probe thus offers a reliable means for studying APN-associated biology including drug-induced hepatotoxicity simply by ratiometric imaging.

Construction of a laccase mimic enzyme with fluorescence properties for kanamycin multi-mode analysis

A copper-based dual-ligand biomimetic enzyme (Cu-BH) inspired by natural laccase was constructed, which had promising laccase-like activity and excellent fluorescence properties. More remarkably, Cu-BH can oxidize not only traditional phenolic substrates, but also catecholamines and amine substrates. As expected, OPD could be catalyzed by Cu-BH to produce the yellow fluorescent product DAP with absorbance and emission peaks at 426 nm and 566 nm, respectively. The generated DAP could quench the Cu-BH fluorescence at 440 nm by fluorescence internal filtration effect (IFE), thus constructing a ratiometric fluorescence signal(F566/F440). Kanamycin further promoted the catalytic activity of Cu-BH by accelerating electron transfer, resulting in a gradual increase in absorbance at 426 nm and fluorescence signal amplification at 566 nm. Based on this, a sensitive dual-mode sensing strategy was established to analyze kanamycin with the LOD of 0.033 and 0.26 nM, which was successfully applied to the monitoring of kanamycin in tap water, milk and honey. The visualization analysis was also achieved by RGB reading from a smartphone. In conclusion, this novel biomimetic enzyme (Cu-BH) had provided a new application path in bioanalysis and environmental monitoring.

A ratiometric fluorescent probe for simultaneous detection of L-ascorbic acid and alkaline phosphatase activity based on red carbon dots/polydopamine nanocomposite

Herein, efficient red carbon dots (R-CDs) were synthesized by one-step hydrothermal treatment of N-(4-amino phenyl) acetamide and (2,3-difluoro phenyl) boronic acid. The optimal emission peak of R-CDs was at 602 nm (under 520 nm excitation) and the absolute fluorescence quantum yield of R-CDs was 12.9%. Polydopamine, which was formed by the self-polymerization and cyclization of dopamine in alkaline condition, emitted characteristic fluorescence with peak position of 517 nm (under 420 nm excitation) and affected the fluorescence intensity of R-CDs through inner filter effect. L-Ascorbic acid (AA), which was the hydrolysis product of L-ascorbic acid-2-phosphate trisodium salt under the catalytic reaction of alkaline phosphatase (ALP), effectively prevented the polymerization of dopamine. Combined with the ALP-mediated AA production and the AA-mediated polydopamine generation, the ratiometric fluorescence signal of polydopamine with R-CDs was correlated closely with the concentration of both AA and ALP. Under optimal conditions, the detection limits of AA and ALP were 0.28 μM during linear range of 0.5–30 μM and 0.044 U/L with linear range of 0.05–8 U/L, respectively. This ratiometric fluorescence detection platform can efficiently shield the background interference of sophisticated samples by introducing a self-calibration as reference signal in a multi-excitation mode, which can detect AA and ALP in human serum samples with satisfactory results. Such R-CDs/polydopamine nanocomposite provides a steadfast quantitative information and makes R-CDs be excellent candidate for biosensors via combining target recognition strategy.