A Dual Emission Fluorescence Probe Based on Silicon Nanoparticles and Rhodamine B for Ratiometric Detection of Kaempferol.

In this paper, blue fluorescent silicon nanoparticles (SiNPs) with outstanding optical properties and robust stability were synthesized by a simple one-step hydrothermal method. By introducing red emissive rhodamine B (RhB) into SiNPs solution, a dual emission nanoprobe (SiNPs@RhB) was constructed, which showed excellent pH stability, salt resistance and photobleaching resistance. The SiNPs@RhB probe could emit two peaks at 444 nm and 583 nm under 365 nm excitation. It was found that the fluorescence intensity of the two emission peaks decreased in different degrees with the addition of different concentrations of kaempferol (Kae). According to this phenomenon, a novel ratiometric fluorescence method was established for the detection of Kae via utilizing SiNPs@RhB as nanoprobe. The detection range and limit of detection (LOD) were 0.5 ~ 150 µM and 0.24 µM, respectively. The ratiometric fluorescence method exhibited the superiority of rapid detection, excellent stability, wide linear range and high sensitivity. The detection mechanism was studied by ultraviolet visible absorption spectra, fluorescence spectra and fluorescence lifetime. Furthermore, the method was applied to the detection of Kae in real samples (kaempferia powder, sea buckthorn granules and sea buckthorn dry emulsion).

Metal-Organic Framework Based on Ratiometric dual-Fluorescent Sensor Using for Accurate Quantification and on-Site Visual Detection of Ascorbic Acid.

Ascorbic acid is very important to the metabolic process of the body, but excessive intake can lead to diarrhea, kidney calculi and stomach cramps. However, complicated production procedures and harsh experimental settings limit many detection methods, and a simpler and more accurate measurement method is needed. In this study, a smartphone-assisted ratiometric fluorescence sensor was developed for the portable analysis of ascorbic acid. Leveraging the catalytic properties of MIL-53(Fe) to expedite the conversion of H2O2 into hydroxyl radicals, thereby facilitating the oxidation of o-phenylenediamine and terephthalic acid bridging ligand. The sensor showcased exceptional sensitivity in detecting ascorbic acid within a linear range of 0.3-100 µM, boasting an impressive limit of detection at 0.15 µM. Furthermore, through the utilization of color extraction RGB values captured by smartphones, accurate detection of ascorbic acid was achieved with a detection limit of 0.4 µM. Real fruit samples exhibited robust spiked recovery rates ranging from 91 to 119%, accompanied by relative standard deviations ≤ 4.7%. The MIL-53(Fe) nanozyme-based smartphone-assisted ratiometric fluorescence sensor offers an ascorbic acid fluorescence detection device that is visible, accurate, sensitive, and reasonably priced.

Eu3+-functionalized covalent organic framework for ratiometric fluorescence detection and adsorption of tetracycline and information steganography.

Functional materials with organic/inorganic composites as the main matrix and rare earth ion complexes as the guest have shown a very broad application prospect for antibiotic sensors. However, Eu3+-complex often relies on a single fluorescence response signal, which is susceptible to changes in the detection environment and cannot simultaneously detect and remove tetracycline (TC). Herein, green fluorescent covalent two-dimensional organic framework (COF-TD) is synthesized, followed by modification of Eu3+ to synthesize COF-TD@Eu3+. In the ratiometric sensor, Eu3+ serves as the recognition site and specific response probe for TC, while COF-TD is the fluorescence reference and carrier for Eu3+. Due to the antenna effect, TC enhances the red fluorescence of Eu3+, while the green fluorescence of COF-TD remains almost stable. Based on the change of fluorescence intensity and fluorescence color from green to red, the efficient ratiometric sensing can be finished in 1 min. The developed method shows high sensitivity with a detection limit of 0.3 µM and high selectivity to TC which makes the method applicable to detect TC in traditional Chinese medicine preparations. In addition, due to the high specific surface area of COFs and specific adsorption sites, COF-TD@Eu3+ also shows good performance for TC removal. The findings show that the maximum adsorption capacity is 137.3 mg g-1 and the adsorption equilibrium is reached in 30 min. Smartphone assisted COF-TD@Eu3+ for both ratiometric fluorescence detection and detecting the absorption of TC is proposed for the first time. The molecular cryptosteganography that transforms the selective response of COF-TD@Eu3+ to binary strings is anticipated to advance utilization of nanomaterials in logic sensing and information safety.

A reliable fluorescence "turn-on" aptasensor based on dual-emitting europium metal-organic frameworks for ultrasensitive and selective detection of sulfamethazine.

A high-performance fluorescent "turn-on" aptasensor (Eu-MOFs@SMZ-Apt) for sulfamethazine (SMZ) determination was designed using dual-emitting europium metal-organic frameworks (Eu-MOFs) as a signal transducer and an amplifier. Eu-MOFs featuring dual emission peaks (430 nm and 620 nm) were first prepared via a facile self-assembly strategy employing Eu (III) ions and 2-aminoterephthalic acid as precursors. The high-affinity aptamer was bonded with Eu-MOFs to form Eu-MOFs@SMZ-Apt through the amidation reaction. Benefiting from the integration of inherent virtues from Eu-MOFs and aptamer, the Eu-MOFs@SMZ-Apt-based sensor allowed sensitive and selective determination of SMZ with good linear relationships in a range of 1.4-40 ng mL-1 and a low detection line (0.379 ng mL-1). This sensor was successfully applied to the determination of trace SMZ in real samples with satisfactory recoveries (86.47-113.52%) and a relative standard deviation (<6.51). Consequently, the Eu-MOFs@SMZ-Apt ratiometric fluorescence sensor furnishes new possibilities for the accurate detection of various pollutants in food.

Rapid and reliable ratiometric fluorescence detection of isoquercitrin based on a high-nuclearity Zn(II)-Nd(III) nanomolecular sensor.

Rapid and quantitative detection of isoquercitrin (Isq) has been attracting much attention due to its outstanding pharmacological and physiological activities. Herein, an interesting 48-metal Zn(II)-Nd(III) nanocluster (1, molecular sizes 1.3 × 2.8 × 3.1 nm) with salen-type Schiff base ligand was constructed as molecular sensor for the luminescence detection of Isq. 1 exhibits visible ligand-centered emission and NIR luminescence of Nd(III), and shows ratiometric fluorescence response to Isq with high sensitivity even in the presence of other interferences. The fluorescence sensing behavior can be expressed by a second-order equation I1060nm/I480nm = A*[Isq]2 + B*[Isq] + C, which is used to quantitatively analyze the Isq concentrations in DMF and FCS. The LODs to Isq for the ligand-centered and lanthanide emissions of 1 in DMF are 0.21 µM and 0.11 nM, respectively. The quenching of the ligand-centered emission of 1 caused by Isq is attributed to the competitive absorption of light energy and "inner effect", while, the luminescence enhancement is due to the "antenna effect".

Construction of MOF@COF-derived composites for ratiometric fluorescence detection of water with ultralow background.

A ratiometric sensor with ultralow background is highly desired due to its low environmental influence and high sensitivity. Herein, inspired by the solubility difference of carboxylate in aqueous and organic solvents, we prepared a core-shell structure porous zirconia-covalent organic framework (COF) composite through thermal hydrolysis of UiO-66-COF precursors in organic alkali solution. The ligand 2-aminoterephthalic acids (H2BDC-NH2) of UiO-66 were transformed into 2-aminoterephthalate salts (ATA salts) that existed in zirconium-oxo clusters building units. The composites emitted only yellow emission (597 nm) from the COF in organic solvent due to the insolubility of ATA salts that induce aggregation-caused quenching (ACQ) and the protection of the COF shell. Contrarily, when water was added into mixture, the ATA salts were released into solution and its fluorescence recovered at 446 nm, while the fluorescence of COF was quenched due to the blockage of the intramolecular charge transfer (ICT) process by water. Thus, a high-sensitivity ratiometric fluorescence method is obtained with ultralow background signal and fast response (less than 1 min) for sensing water in organic solvent. We believe that the proposed ratiometric fluorescence sensor based on the zirconia-COF composite will provide the guidance for detection with wide applications.

Dual-emission carbon dots for ratiometric fluorescence sensing of thiabendazole in fruits.

Quantitative determination of pesticides in fruits and vegetables is essential for human healths. Herein, a new dual-emission carbon dots with high fluorescence stability at a pH range of 4-10 and a temperature range of 0-60 °C was synthesized. And a novel ratiometric fluorescence probe was proposed to detect thiabendazole (TBZ) residue with a wide linear range (0-1000 µM) and low detection limit (0.15 µM). The emission at 512 nm exhibited a special "turn-off" fluorescence sensing of TBZ due to internal filter effect, while that at 361 nm barely changed and worked as reference. Furthermore, the ratiometric fluorescence strategy was successfully applied for determining TBZ in fruits with good recoveries (96.73%-111.17 %, 93.29%-120.78 % and 96.28%-100.57 %, respectively). Notably, the constructed ratiometric fluorescence probe had comparable accuracy to HPLC in detecting unknown concentrations of TBZ in pear juice, demonstrating dual-emission carbon dots possess wide and promising applicability for fluorescence sensing pesticides in the future.

Highly Sensitive Ratiometric Fluorescent Flexible Sensor Based on the RhB@ZIF-8@PVDF Mixed-Matrix Membrane for Broad-Spectrum Antibiotic Detection.

Antibiotics play an essential role in the treatment of various diseases. However, the overuse of antibiotics has led to the pollution of water bodies and food safety, affecting human health. Herein, we report a dual-emission MOF-based flexible sensor for the detection of antibiotics in water, which was prepared by first encapsulating rhodamine B (RhB) by a zeolite imidazolium ester skeleton (ZIF-8) and then blending it with polyvinylidene difluoride (PVDF). The luminescent properties, structural tunability, and flexible porosity of the MOF-based composites were combined with the processability and flexibility of polymers to prepare luminescent membranes. The sensor is capable of dual-emission ratiometric fluorescence sensing of nitrofurantoin (NFT) and oxytetracycline (OTC), exhibiting sensitive detection of fluorescence burst and fluorescence enhancement, respectively, with detection limits of 0.012 µM and 8.9 nM. With the advantages of visual detection, high sensitivity, short detection time, and simplicity, the highly sensitive ratiometric fluorescent flexible sensor has great potential for detecting antibiotics in an aqueous environment. It will further stimulate interest in luminescent MOF-based mixed matrix membranes and their sensing applications.

Construction of nanohybrid Tb@CDs/GSH-CuNCs as a ratiometric probe to detect phosphate anion based on aggregation-induced emission and FRET mechanism.

The simple preparation of a nanohybrid of terbium-doped carbon dots/glutathione-capped copper nanoclusters (Tb@CDs/GSH-CuNCs) was for the first time developed for ratiometric detection of phosphate anion (Pi). Blue-emission of Tb@CDs can trigger non-luminescence of GSH-CuNCs for aggregation-induced emission (AIE) performance due to the strong reserved coordination capacity of Tb3+. Thus, Tb@CDs/GSH-CuNCs rapidly generated dual-emission signals at 630 nm and 545 nm by directly mixing the two individual materials via the AIE effect, alongside fluorescence resonance energy transfer (FRET) process. However, by the introduction of Pi, both AIE and FRET processes were blocked because of the stronger binding affinity of Tb3+ and Pi than that of Tb3+ and -COOH on Tb@CDs, thus realizing successful ratiometric detection of Pi. The linear concentration range was 0-16 µM, with the limit of detection (LOD) of 0.32 µM. The proposed method provided new ideas for designing nanohybrid of CDs and nanoclusters (MNCs) as ratiometric fluorescent probes for analytical applications.

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 3.33 ng mL-1, with an LOD of 0.08 ng mL-1. Moreover, the fluorescence sensor was successfully applied to detect AFB1 in real samples.

Bimetallic metal-organic frameworks-based ratiometric fluorescence sensor for the quantitative detection of thiram in fruits samples.

The identification of residual thiram (Tr) in foods is vital in view of its harmful effects on human health. Herein, a ratiometric fluorescence sensor (I435/I590) based on rhodamine B/NH2-MIL-53(Al0.75Fe0.25) was constructed for the detection of Tr. Interestingly, the probe RhB/NH2-MIL-53(Bim) assisted by Cu2+ could rapidly and sensitively recognize Tr with a low detection limit of 0.11 µg/mL in 10 min. The fluorescence sensing mechanism was investigated using fluorescence spectra, UV-Vis absorption spectra, the fluorescence lifetime and quantum yield. The results showed that the excellent sensing performance was attributed to fluorescence resonance energy transfer, electrostatic interaction, and photoinduced electron transfer. In addition, the practical application of this platform showed acceptable relative recoveries for Tr (84.03-107.81 %), and precisions were also achieved (relative standard deviation ≤ 8.69 %, n = 3). These results show that the presented herein can be applied to monitor the Tr content in real fruit samples.

A ratiometric fluorescent biosensing platform for ultrasensitive detection of Salmonella typhimurium via CRISPR/Cas12a and silver nanoclusters.

Rapid, sensitive and specific detection of bacteria is of great importance. Herein, we developed a versatile biosensing platform for ultrasensitive detection of pathogenic bacteria, termed as SCENT-Cas (Silver nanoCluster Empowered Nucleic acids Test using CRISPR/Cas12a). Simply, the species-specific invA gene of Salmonella typhimurium (S. typhi) was isothermally amplified using LAMP, which subsequently triggered the trans-cleavage of CRISPR/Cas12a. The trans-cleavage degraded any single-stranded DNA (ssDNA) non-specifically. A DNA-templated AgNCs probe was then employed, in which green fluorescence emissive AgNCs effectively converted to red fluorescence emissive AgNCs when placed in close vicinity to a pre-designed converter ssDNA. As such, the trans-cleavage was utilized for shredding converter ssDNA, enabling the green-to-red fluorescent change to form a ratiometric biosensing platform. With this strategy, target nucleic acid was dexterously converted into ratiometric fluorescence that was recorded to detect as low as 1 CFU/mL S. typhi with a dynamic range from 1 to 108 CFU/mL. To our knowledge, this is the first report regarding the use of ratiometric fluorescence in CRISPR/Cas-based detection, which minimizes interference and improves reliability. Lastly, this proposed strategy was challenged by detecting S. typhi contamination in real food samples. Our work enriches CRISPR/Cas toolbox in biosensing by providing a desirable method for bacterial detection.

Paper-based device for the selective determination of doxycycline antibiotic based on the turn-on fluorescence of bovine serum albumin-coated copper nanoclusters.

An enhanced ratiometric fluorescence sensor was built for on-site visual detection of doxycycline (DOX) through the interaction with bovine serum albumin on the surface of red emissive copper nanoclusters. Upon the addition of weakly fluorescent DOX, the red fluorescence from copper nanoclusters gradually decreased through the inner-filter effect (IFE), while a green fluorescence appears and significantly increases, forming an interesting fluorescent isosbestic point, which was assigned to DOX due to sensitization effect of bovine serum albumin. On the basis of this ratiometric fluorescence, the system possessed good limit of detection (LOD) of 45 nM and excellent selectivity for DOX over other tetracyclines. Based on these findings, a paper-based sensor has been fabricated for distinct visual detection of trace DOX and combined with smartphone color recognizer for quantitative detection of DOX (LOD = 83 nM). This method shows broad application prospects in environmental monitoring and food safety.

Synthesis of intrinsic dual-emission type N,S-doped carbon dots for ratiometric fluorescence detection of Cr (VI) and application in cellular imaging.

In this paper, intrinsic dual-emission fluorescent carbon dots (CDs) doped with N and S atoms have been firstly fabricated. The characterization results show that CDs are successfully synthesized with two separate fluorescence emissions at 468 nm and 628 nm, respectively. The strong and selective interaction of Cr (VI) ions with CDs lead to obvious fluorescence decrease of CDs at 468 nm, which is caused by a mixed quenching mechanism. At the same time, the fluorescence at 628 nm increase. Interestingly, the CDs solution show obvious color change under the daylight and UV light, so visualization detection of Cr (VI) can be realized in water samples. Based on the data of the emission intensity ratios of F468/F628, Cr (VI) can be detected from 3.8 to 38.9 µM combined with the linear correlation coefficient of 0.998, and the lowest detection concentration is 47.2 nM. The platform is satisfactorily applied to the detection of Cr (VI) ions in water samples. In addition, the CDs could be applied as fluorescent probes for cell imaging with dual fluorescent emission.

Determination of cyanide based on a dual-emission ratiometric nanoprobe using silver sulfide quantum dots and silicon nanoparticles.

A novel ratiometric fluorescent nanoprobe was designed for the sensitive determination of cyanide anion (CN-) by the electrostatic attraction between positively charged silicon nanoparticles (Si NPs) and negatively charged silver sulfide quantum dots (Ag2S QDs). The nanoprobe exhibited two well-resolved emission peaks at 446 nm and 540 nm under a single excitation wavelength (360 nm). In the presence of CN-, the fluorescence of Ag2S QDs at 540 nm was remarkably quenched, while the fluorescence of the Si NPs at 446 nm remained constant, establishing the desired conditions for ratiometric fluorescence detection. Under optimal conditions, the ratiometric fluorescence assay showed good linearity (R2 = 0.9921) within the range 0.05-15 µM, and the limit of detection was calculated to be 56 nM (at an S/N ratio of 3). The proposed Ag2S QD/Si NP nanoprobe has been successfully used to determine CN- in water and sprouting potato samples with satisfactory recoveries in the range 97-110.5%.

A selectivity-enhanced ratiometric fluorescence imprinted sensor based on synergistic effect of covalent and non-covalent recognition units for ultrasensitive detection of ribavirin.

Development of methods with high selectivity and sensitivity for detection of trace ribavirin (RBV) is of great importance for environmental protection and food safety. Herein, we proposed a simple yet valid strategy to construct the highly selective ratiometric fluorescence sensing platform (BA-LMOFs@MIP) for analysis of RBV based on boric acid-functionalized lanthanide metal-organic framework (BA-LMOFs) coupled with molecularly imprinted polymer (MIP). In this strategy, BA-LMOFs featured with dual-emission and pH-responsive behavior were first synthesized as supporter. Benefiting from boric acid group of BA-LMOFs, RBV was easily immobilized onto its surface, taking advantage of template immobilization-based surface imprinting means to fabricate BA-LMOFs@MIP with dual recognition sites for the first time. The synergistic effect of covalent boronate affinity-based recognition unit and non-covalent imprinting sites enabled BA-LMOFs@MIP to exhibit superior selectivity and binding efficiency to RBV. BA-LMOFs as signal tag endowed BA-LMOFs@MIP with desirable sensitivity, photostability and hydrophilicity. More importantly, BA-LMOFs@MIP-based sensor displayed a wide linear range for RBV from 25 to 1200 ng mL-1 with a detection limit down to 7.62 ng mL-1. The sensor was finally applied to RBV determination in real samples, and the obtained results revealed that BA-LMOFs@MIP would be a promising candidate for monitoring of RBV in complex systems.

Ratiometric fluorescence determination of chlortetracycline based on the aggregation of copper nanoclusters triggered by aluminum ion.

The aggregation-induced emission (AIE) characteristic of copper nanoclusters (CuNC) was for the first time used to construct a ratiometric fluorescence probe (CuNC-Al3+) for detection of chlortetracycline (CTC). Aluminum ion (Al3+) can aggregate free CuNC and make it emit a bright and stable red fluorescence. A slight excess of Al3+ in CuNC-Al3+ solution can form a CTC-Al3+ complex to limit the conformational rotation of CTC molecule and enhance CTC fluorescence. So, the red fluorescence of CuNC-Al3+ probe and the enhanced CTC fluorescence are used as a reference signal and a response signal to detect CTC, respectively. The method developed shows a good linear relationship between the CTC concentration and the fluorescence intensity ratio (I495/I575) in the range 0.1-3.0 µM, and the detection limit is 25.3 nM (S/N = 3). In addition, the fluorescent color of CuNC-Al3+ probe changes from red to yellow-green as the concentration of CTC increases. Based on this observation, a fluorescent test paper has also been fabricated. Schematic illustration of Al3+ inducing CuNC to produce AIE performance and detecting CTC.

Aspartic acid assisted one-step synthesis of stable CsPbX3@Asp-Cs4PbX6 by in situ growth in NH2-MIL-53 for ratiometric fluorescence detection of 4-bromophenoxybenzene.

A molecularly imprinted ratiometric fluorescent sensor was synthesized for the detection of 4-bromophenoxybenzene (BDE-3) based on perovskite quantum dots and metal organic framework. First, aspartic acid (Asp) was introduced during the synthesis of perovskite CsPbX3 for the formation of a core-shell structure of CsPbX3@Asp-Cs4PbX6. Due to the protection of the shell layer Cs4PbX6, the stability of the core CsPbX3 was improved significantly. Compared to CsPb(BrI)3, the ultraviolet and thermal stabilities of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 were increased by 26 times and 32 times, respectively, and, compared to CsPbBr3, these stabilities of CsPbBr3@Asp-Cs4PbBr6 were increased by 3 times and 13 times, respectively. The water stabilities of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 and CsPbBr3@Asp-Cs4PbBr6 were greatly improved too. Then, a ratiometric fluorescence sensor was constructed by in situ growth of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 in metal organic framework (NH2-MIL-53) for the detection of BDE-3, in which the orange fluorescence of CsPb(BrI)3@Asp-Cs4Pb(BrI)6 (614 nm) was regarded as the reference signal and the cyan fluorescence of NH2-MIL-53 (494 nm) was used as the fluorescence response signal. To improve the selectivity of the sensor, the molecular imprinting polymer (MIP) was modified on the NH2-MIL-53 and an imprinting factor of 3.17 was obtained. Under 365 nm light excitation, the fluorescent response signal at 494 nm was quenched gradually by BDE-3 in the range 11.4 to 68.5 nmol/L, while the reference signal at 614 nm remained unchanged. The limit of detection and limit of quantification were 3.35 and 11.2 nmol/L, respectively, and the fluorescent color of the sensor changed gradually from cyan to green to orange, which illustrated that the developed sensor has an ability to recognize BDE-3 specifically, a good anti-interference ability, and a sensitively visual detection ability. Moreover, the sensor was successfully applied to the BDE-3 detection in polyethylene terephthalate plastic bottle, polyvinyl chloride plastic bag, and circuit board with satisfactory recoveries (96.3-108.1%) and low relative standard deviations (5%). The preparation processes of NH2-MIL-53, NH2-MIL-53-CsPb(BrI)3@Asp-Cs4Pb(BrI)6, and the MIP-NH2-MIL-53-CsPb(BrI)3@Asp-Cs4Pb(BrI)6 composites.

A ratiometric fluorescence probe based on graphene quantum dots and o-phenylenediamine for highly sensitive detection of acetylcholinesterase activity.

By using graphene quantum dots (GQDs) and o-phenylenediamine (OPD), a ratiometric fluorescence probe was designed for the highly sensitive and selective detection of AChE. GQDs with strong fluorescence were synthesized by the one-step hydrothermal method. The optimal emission wavelength of GQDs was 450 nm at the excitation wavelength of 375 nm. MnO2 nanosheets with a wide absorption band of 300-600 nm were prepared at room temperature. Because of the extensive overlap between the absorption spectrum of MnO2 nanosheets and the excitation and emission spectra of GQDs, the fluorescence of GQDs at 450 nm was efficiently quenched by the inner-filter effect. Meanwhile, due to the peroxidase-like activity of MnO2 nanosheets, OPD was catalytically oxidized to 2,3-diaminophenazine (oxOPD), a yellow fluorescent substance with a new emission peak at 572 nm. When AChE was present, the substrate acetylthiocholine (ATCh) was hydrolyzed to thiocholine (TCh) that is capable of decomposing MnO2 nanosheets. Therefore, the quench of GQDs and the oxidation of OPD by MnO2 nanosheets were suppressed, resulting in the fluorescence recovery of GQDs at 450 nm, while the fluorescence decrease of oxOPD at 572 nm. Utilizing the fluorescence intensity ratio F450/F572 as the signal readout, the ratiometric fluorescence method was established to detect AChE activity. The ratio F450/F572 against the AChE concentration demonstrated two linear relationships in the range 0.1-2.0 and 2.0-4.5 mU mL-1 with a detection limit of 0.09 mU mL-1. The method was applied to the detection of positive human serum samples and the analysis of the inhibitor neostigmine. Due to the advantages of high sensitivity, favorable selectivity, and strong anti-interference, the method possesses an application prospect in clinical diagnosis of AChE and the screening of inhibitors. Graphical abstract Schematic presentation of a ratiometric fluorescence method for the detection of acetylcholinesterase (AChE). The fluorescence of graphene quantum dots (GQDs) is quenched and o-phenylenediamine (OPD) is oxidized to generate fluorescent product 2,3-diaminophenazine (oxOPD) by MnO2 nanosheets. When AChE is present, acetylthiocholine iodide (ATCh) is hydrolyzed to thiocholine (TCh) with reducibility for decomposing MnO2 nanosheets. Due to the decomposition of MnO2 nanosheets, the quenching of GQDs and oxidation of OPD are suppressed. The fluorescence of GQDs at 450 nm is enhanced, while the fluorescence of oxOPD at 572 nm is reduced. The fluorescence intensity ratio F450/F572 is used to establish the ratiometric fluorescence method for AChE activity.

Determination of pathogenic bacteria-Bacillus anthrax spores in environmental samples by ratiometric fluorescence and test paper based on dual-emission fluorescent silicon nanoparticles.

Many lanthanide ions-based probes have been widely used for detecting anthrax spores biomarker-dipicolinic acid (DPA). However, little work has realized detection of bacillus anthrax spores in real environmental samples. In this work, a novel ratiometric fluorescent nanoprobe based on europium (Eu)-doped silicon nanoparticles (Eu@SiNPs) was fabricated for the first time by one-pot method without post-modification for determination of the DPA in bacillus subtilis spores (simulant bacillus anthrax spores). Based on Eu(III) in the Eu@SiNPs could be sensitized by DPA to emit intrinsic fluorescence and the fluorescence intensity of SiNPs in the Eu@SiNPs almost remained stable, a new ratiometric fluorescent method for determination of micro DPA in bacillus subtilis spores and bacillus subtilis spores in real environmental samples, such as Yellow river water, tap water and soil was established. Under the optimum conditions, the limit of detection (LOD) of the method toward bacillus subtilis spores was as low as 2.38×104 spore/mL. Simple, fast and visual DPA and bacillus subtilis spores determination was also achieved by the Eu@SiNPs-based test paper. Therefore, the newly established method was expected to be a powerful tool for efficiently determination of bacillus anthrax spores to avoid anthrax threats.