Anti-cancer drug axitinib: a unique tautomerism-induced dual-emissive probe for protein analysis.

A commercial anti-cancer drug, axitinib, exhibits very stable dual emissions for discrimination of human serum albumin.

Harnessing a simple ratiometric fluorescent probe for albumin recognition and beyond.

A commercially available naphthalene fluorophore serves as a ratiometric indicator for albumin, showcasing its applications in albumin-based supramolecular recognition.

Ratiometric determination of etomidate based on an albumin-based indicator displacement assay (IDA).

The first fluorescent sensor based on the indicator displacement assay (IDA) for on-site determination of etomidate.

A novel dual-color fluorescent sensor with two pKas for on-site detection of pH in food.

Tracking pH fluctuations in food samples is important for ensuring food freshness. Fluorescent probes have been widely applied as promising tools for the on-site detection of pH changes; however, most of them can be applied only at either lower or higher pH ranges because their response structures commonly have a single acid dissociation constant (pKa). To address this problem, we designed a fluorescent sensor, called HMB, containing a methylpiperazine group with two pKa values, which exhibited a unique dual-color response to pH changes over a wide pH range. Furthermore, the HMB-based test strips are easily prepared and used as portable labels for the visual monitoring of food spoilage that results in microbial and anaerobic glycolytic pathways in real food (such as cheese and shrimp). To the best of our knowledge, this is the first fluorescent pH sensor with two pKa values, and we expect that this work will inspire more sensor designs for food quality control.

Rational design of a FA1-targeting anti-interference fluorescent probe for the point-of-care testing of albuminuria.

Albuminuria is a crucial urine biomarker of human unhealthy events such as kidney diseases, cardiovascular diseases, and diabetes. However, the accurate diagnosis of albuminuria poses a significant challenge owing to the severe interference from urine fluorescence and urine drugs. Here, we report a novel flavone-based fluorescent probe, DMC, by incorporating the FA1-targeting methylquinazoline group into a flavone skeleton with the extend π-conjugation. DMC exhibited a rapid response time, high sensitivity, and selectivity towards human serum albumin (HSA) in urine. Moreover, the red-shifted fluorescence and the FA1-targeted HSA-binding of DMC efficiently mitigated the interference from both urine fluorescence and urine drug metabolites. Furthermore, the establishment of a portable testing system highlighted the potential for point-of-care testing, offering a user-friendly and accurate approach to diagnose A2-level and A3-level albuminuria. We expect that the success of this DMC-based diagnostic platform in real urine samples can signify a significant advancement in early clinical diagnosis of albuminuria and its associated diseases.

Fast, portable, selective, and ratiometric determination of ochratoxin A (OTA) by a fluorescent supramolecular sensor.

Ochratoxin A (OTA), a mycotoxin found in various food items, possesses significant health risks due to its carcinogenic and toxic properties. Thus, detecting OTA is crucial to ensure food safety. Among the reported analytical methods, there has yet to be one that achieves fast, selective, and portable detection of OTA. In this study, we explore a novel supramolecular sensor, DOCE@ALB, utilizing human serum albumin as the host and a flavonoid fluorescent indicator as the guest. On the basis of indicator displacement assay, this sensor boasts an ultra-fast response time of just 5 s, high sensitivity with a limit of detection at 0.39 ppb, exceptional selectivity, and a noticeable ratiometric fluorescence response to OTA. This discernible color change and portability of the sensor make it suitable for on-site OTA detection in real food samples, including flour, beer, and wine, simply using a smartphone. In comparison to previously reported methods, our approach has showcased notable advantages in both response time and portability, addressing a critical need for food safety and regulatory compliance.

Intermolecular proton transfer from flavonol to human serum albumin triggers a red-shifted ratiometric fluorescence response.

Intermolecular proton transfer from a flavonol-based probe to the arginine (Arg222) in drug site 1 of human serum albumin triggers an unusual red-shifted ratiometric fluorescence response, which can be applied in the point-to-care diagnosis of hypoalbuminemia.

A portable paper-based testing device for fast and on-site determination of nitroxynil in food.

Nitroxynil (NTX) is a common anthelmintic veterinary drug for the management of fascioliasis in food-producing sheep and cattle. Since excessive NTX residue in food can lead to several adverse side effects, such as allergic skin reaction and respiratory irritation, it is of great importance to develop an efficient analytical method for NTX determination. Herein, we report a simple fluorescent detection method based on a novel supramolecular probe capable of detecting NTX with a fast response (5 s), high sensitivity (107 nM), high selectivity, and acceptable anti-interference property. Moreover, the portable paper-based test strips were facilely prepared and successfully realized on-site determination of NTX in real edible animal products simply with the aid of a smartphone. To the best of our knowledge, this is the very first report on the portable detection of NTX. This study also provides a promising strategy for the fast and portable detection of analyte based on the host-guest system, which will lead to improved fluorescent probe design for food analysis.

The first fluorescent sensor for the detection of closantel in meat.

Closantel is widely used in the management of parasitic infestation in livestock, but is contraindicated in humans due to its high toxic to human retina. Thus, development of a fast and selective method for the detection of closantel residues in animal products is highly needed yet still challenging. In the present study, we report a supramolecular fluorescent sensor for closantel detection through a two-step screening process. The fluorescent sensor can detect closantel with a fast response (<10 s), high sensitivity, and high selectivity. The limit of detection is 0.29 ppm, which is much lower than the maximum residue level set by government. Moreover, the applicability of this sensor has been demonstrated in commercial drugs tablets, injection fluids, and real edible animal products (muscle, kidney, and liver). This work provides the first fluorescence analytical tool for accurate and selective determination of closantel, and may inspire more sensor design for food analysis.

Dual-mode supramolecular fluorescent probe for rapid and on-site detection of chlorpyrifos in the environment.

Chlorpyrifos (CPF) as a classic organophosphorus pesticide has been widely used in agricultural applications to control insects and worms. CPF in the environment can cause deaths of diverse kinds of aquatic organism and bring a high risk to human health. Therefore, the development of effective analytical method for CPF is of great importance. In this work, a novel dual-mode albumin (ALB)-based supramolecular probe FD@ALB was designed and prepared for rapid detection of CPF in the environment. The limit of detection is 0.57 µM (∼ 0.2 ppm) with a wider detection range up to 200 µM, which is satisfactory for application. The sensing mechanism can be ascribed to CPF-induced phosphorylation of ALB, thus leading to a change in the binding microenvironment of FD dye. Moreover, the paper-based test strips were used in conjunction with the FD@ALB, realizing the portable detection of CPF. This method was demonstrated to be suitable for on-site detection of CPF in various kinds of environmental samples, including water, soil, and food samples, with the aid of a smartphone. To the best of our knowledge, this is the first analytical method achieving a combination of the rapid and ratiometric detection of CPF in the environment.

Smartphone-based on-site detection of hydrogen peroxide in milk by using a portable ratiometric fluorescent probe.

The on-site detection of hydrogen peroxide (H2O2) is important for maintaining food safety as the ingestion of H2O2 can lead to serious pathological conditions. However, most reported fluorescent probes require a fluorometer to ensure readable signal output and reliable detection result. Consequently, the fluorescent detection of H2O2 can be realized only within a standard laboratory setting. Herein, we report a novel supramolecular strategy that can successfully convert the typical off-on response to H2O2 into a ratiometric response, which allows the on-site detection of H2O2 when used in conjunction with a smartphone-based 3D-printed miniaturized testing system. This method has acceptable sensitivity, good anti-interference ability, and desirable accuracy compared to a standard detection method. More importantly, this portable ratiometric method can be used to detect H2O2 residue in commercial milk samples with the simple testing apparatuses.

Naphthalene-based fluorescent probe for on-site detection of hydrazine in the environment.

The widespread occurrence of hydrazine residues in the environment, including in water, soil, and organisms, is a potential health threat to humans. Therefore, the development of an efficient method for the detection of hydrazine in environmental samples is highly desirable although it poses a significant challenge. In this study, we designed and synthesized a series of naphthalene-based fluorescent dyes through structural engineering and developed a novel probe for hydrazine detection. The probe could provide a distinct fluorescence response toward hydrazine in aqueous solution with high sensitivity and selectivity. Moreover, paper-based test strips can be easily fabricated using this probe, enabling the portable on-site detection of hydrazine with the aid of a smartphone. Furthermore, we demonstrated that this probe is capable of recognizing hydrazine in various environmental samples, including water, soil, plants, and zebrafish embryos. This research provides a promising tool for the detection of hydrazine in the environment.

General Method for Pesticide Recognition Using Albumin-Based Host-Guest Ensembles.

The massive use of pesticides nowadays has led to serious consequences for the environment and public health. Fluorescence analytical methods for pesticides are particularly advantageous with respect to simplicity and portability; however, currently available fluorescence methods (enzyme-based assays and indicator displacement assays) with poor universality are only able to detect few specific pesticides (e.g., organophosphorus). Making use of the multiple flexible and asymmetrical binding sites in albumin, we herein report a set of multicolor albumin-based host-guest ensembles. These ensembles exhibit a universal but distinctive fluorescent response to most of the common pesticides and allow array-based identification of pesticides with high accuracy. Furthermore, the simplicity, portability, and visualization of this method enable on-site determination of pesticides in a practical setting. This albumin host strategy largely expands the toolbox of traditional indicator displacement assays (synthetic macrocycles as hosts), and we expect it to inspire a series of sensor designs for pesticide detection.

Rapid and ratiometric fluorescent detection of phosgene by a red-emissive ESIPT-based-benzoquinolone probe.

Phosgene is a highly toxic gas that poses a serious threat to human health and public safety. Therefore, it is of great importance to develop an available detection method enabling on-the-spot measurement of phosgene. In this paper, we report a novel ESIPT fluorescent probe for phosgene detection based on quinolone fluorophore. This probe exhibits rapid response (in 10 s), stable signal output (last for 10 min), high sensitivity (LOD âˆ¼ 6.7 nM), and distinct emission color change (red to green) towards phosgene. The sensing mechanism was investigated by using 1H NMR, HRMS and fluorescence lifetime techniques, confirming that the amidation reaction between phosgene and quinolone effectively suppressed the ESIPT process of probe. Eventually, this probe was fabricated into polymer nanofibers by electrospinning and successfully employed to monitor gaseous phosgene with high specificity. This work provided a promising analytical tool for rapid and ratiometric detection of phosgene both in solution and in the gas phase.

A surfactant-assisted approach enables the fluorescence tracking of benfluralin in plants.

Developing an effective detection method for benfluralin (BFA) is of great significance, since BFA as most widely used herbicides can be bioaccumulated by aquatic organisms in environment, possessing potential risks to human health. Owing to aggregation-caused quenching effect, most fluorescent detection methods based on donor-acceptor organic fluorophores suffered from very low sensitivity towards BFA in water system, hampering the bioimaging application in plants. In this work, we reported a novel surfactant-assisted fluorescent probe enabling detection of BFA in water with a high sensitivity. The involvement of specific surfactant Triton X100 (TX100) could amplify the response signal of probe more than 100-fold. The detection limit for BFA was determined to be 80 nM, satisfying the environmental protection requirements. Moreover, we demonstrated applications of this strategy for the fluorescent imaging of BFA in plant. The absorbance of BFA into roots of Arabidopsis thaliana and castor seedlings was successfully observed based on this method.

A near-infrared dicyanoisophorone-based fluorescent probe for discriminating HSA from BSA.

Despite the rapid development of fluorescent probe techniques for the detection of human serum albumin (HSA), a probe that discriminates between HSA and bovine serum albumin (BSA) is still a challenging task, since their similar chemical structures. As a continuation of our work, herein, a dicyanoisophorone-based fluorescent probe DCO2 is systematically studied for discrimination of HSA from BSA. The photophysical and sensing performances of DCO2, including basic spectroscopic properties, sensing sensitivity, and selectivity, exhibits that DCO2 could selectively bind with HSA and display remarkable fluorescence enhancement (∼254-fold) at 685 nm. The gap of the fluorescent response of DCO2 between HSA and BSA is an obvious increase from 21% to 73% compared to the previous probe DCO1. The sensing mechanism was elucidated by Job's plot, displacement experiment, and molecular docking, suggesting that the specific response to HSA originated from the rigid donor structure and steric hindrance. DCO2 could be buried in the DS1 pocket of HSA, and only partly wedged into the DS1 pocket of BSA with exposing twisted N,N-diethylamino group outside. Application studies indicated that DCO2 has well detective behavior for HSA in the biological fluids. This work could provide a new approach to design HSA-specific near-infrared fluorescence probes.

Modulating donor of dicyanoisophorone-based fluorophores to detect human serum albumin with NIR fluorescence.

It is urgently needed to develop NIR-fluorescent probe for detection of human serum albumin (HSA) since the interference of short-wavelength-fluorescence from endogenous species in real serum and urine. However, most previous reports were located in the short-wavelength region (<600 nm). In this work, a series of dicyanoisophorone (DCO)-based fluorophores 1-4 with different donor groups have been designed and investigated. A systematic study of their photophysical properties has been carried out. Among these probes, 4 exhibited NIR emission with the highest fluorescence brightness and the most sensitive signal response to HSA. Further studies demonstrated that 4 could strongly bind into the DS1 pocket of HSA with a 1:1 ratio. Importantly, the method based on 4 has been proven to be capable of sensing HSA in real serum and urine samples.

Fluorescence discrimination of HSA from BSA: A close look at the albumin-induced restricted intramolecular rotation of flavonoid probe.

Discrimination of human serum albumin (HSA) from bovine serum albumin (BSA) based on the fluorescence probe technique is still challenging due to similar chemical structures. In this work, a novel flavonoid-based fluorescent probe AF is reported for successful discrimination of HSA from BSA. The sensing performances of probe, including sensing dynamic, sensitivity and selectivity, have been carefully studied. Moreover, sensing mechanism was elucidated by Job's plot, displacement experiment, and molecular docking, suggesting that the specific response to HSA originated from the albumin-induced restricted intramolecular rotation (RIR) of probe. This work may provide a simple way for designing of novel probes for HSA with high selectivity.

A fluorescent probe with dual acrylate sites for discrimination of different concentration ranges of cysteine in living cells.

Monitoring of cysteine (Cys) is of significant importance for studying Cys-involved biological functions and clinically diagnosing Cys-related diseases. Recently, few fluorescent probes with two different reacting sites were reported to be capable of sensing different concentration ranges of Cys with distinct fluorescence signals, particularly suiting for bioimaging. However, due to relative sophisticated synthesis and moderate selectivity, the applications of these probes were still severely restricted. In this work, we proposed a novel probe design strategy by utilizing two same reacting groups, instead of two different reacting groups, to simplify the synthesis route and minimize the interference from competing species. Same reacting groups in a probe with different steric hindrances could exhibit different reactivities to Cys. This probe showed distinguishable fluorescence peak wavelengths towards low and high concentration ranges of Cys, giving green and blue emissions, respectively. Moreover, this probe was successfully applied for monitoring of Cys concentration in living cells. We believe this work provided a simpler strategy for dual-site fluorescent probes to sense difference concentration ranges of Cys, which may inspire more probe design in future.

Nile-Red-Based Fluorescence Probe for Selective Detection of Biothiols, Computational Study, and Application in Cell Imaging.

A new colorimetric and fluorescence probe NRSH based on Nile-red chromophore for the detection of biothiols has been developed, exhibiting high selectivity towards biothiols over other interfering species. NRSH shows a blue shift in absorption peak upon reacting with biothiols, from 587 nm to 567 nm, which induces an obvious color change from blue to pink and exhibits a 35-fold fluorescence enhancement at 645 nm in red emission range. NRSH displays rapid (<1 min) response for H2S, which is faster than other biothiols (>5 min). The detection limits of probe NRSH towards biothiols are very low (22.05 nM for H2S, 34.04 nM for Cys, 107.28 nM for GSH and 113.65 nM for Hcy). Furthermore, NRSH is low cytotoxic and can be successfully applied as a bioimaging tool for real-time monitoring biothiols in HeLa cells. In addition, fluorescence mechanism of probe NRSH is further understood by theoretical calculations.