Recent Progress in Folic Acid Detection Based on Fluorescent Carbon Dots as Sensors: A Review.

Folic acid (FA) is a water-soluble vitamin found in diverse natural sources and is crucial for preserving human health. The risk of health issues due to FA deficiency underscores the need for a straightforward and sensitive FA detection methodology. Carbon dots (CDs) have gained significant attention owing to their exceptional fluorescence performance, biocompatibility, and easy accessibility. Consequently, numerous research studies have concentrated on developing advanced CD fluorescent probes to enable swift and precise FA detection. Despite these efforts, there is still a requirement for a thorough overview of the efficient synthesis of CDs and their practical applications in FA detection to further promote the widespread use of CDs. This review paper focuses on the practical applications of CD sensors for FA detection. It begins with an in-depth introduction to FA and CDs. Following that, based on various synthetic approaches, the prepared CDs are classified into diverse detection methods, such as single sensing, visual detection, and electrochemical methods. Furthermore, persistent challenges and potential avenues are highlighted for future research to provide valuable insights into crafting effective CDs and detecting FA.

A Dual-Mode Detection Sensor Based on Nitrogen-Doped Carbon Dots for Visual Detection of Fe(III) and Ascorbic Acid via a Smartphone.

Accurately and promptly detecting Fe3+ and ascorbic acid (AA) is a crucial objective. In this study, nitrogen-doped carbon dots (N-CDs) were synthesized using a one-step hydrothermal synthesis method with 6,9-diamino-2-ethoxyacridine lactate as the precursor. The introduction of Fe3+ and AA resulted in both fluorescence (FL) quenching and enhancement of the synthesized N-CDs. The fluorescent response of the N-CDs probe to Fe3+ was observed in the concentration range of 5-20 µM and 25-50 µM, with a limit of detection (LOD) of 290 nM. Remarkably, the fluorescence of the N-CDs was recovered upon the addition of AA to the N-CDs-Fe3+ system. Using the "off-on" fluorescent N-CDs probe, a linear range of 40-90 µM was achieved with an LOD of 0.69 µM. Additionally, the feasibility of employing a smartphone equipped with an RGB Color Picker was demonstrated for the analysis of Fe3+ and AA concentrations, providing a novel visual detection method. Furthermore, the application of N-CDs in solution demonstrated considerable potential for visually detecting Fe3+ and AA. The proposed dual-mode detection sensor was found to be simple, efficient, and stable, enabling the successful determination of Fe3+ and AA in practical samples with satisfactory results.

Poly-ß-cyclodextrin strengthen Pr6O11 porous oxidase mimic for dual-channel visual recognition of bioactive cysteine and Fe2.

To conveniently monitor bioactive cysteine (Cys) and Fe2+ in practice, a kind of poly-ß-cyclodextrin strengthen praseodymium oxide (Pr6O11) porous oxidase mimic (p-ß-CD@Pr6O11) was constructed by virtue of the strong coordination between nano Pr6O11 and poly-ß-cyclodextrin substrate. After its microstructure and physicochemical property were characterized in detail, it was noted that porous p-ß-CD@Pr6O11 exhibited excellent enzyme-like catalytic activity to accelerate the oxidation of 3,3',5,5,'-tetramethylbanzidine (TMB) and 2,2'-azinobis (3-ethylbenzo-thiazoline-6-sulfonic acid) ammonium salt (ABTS) with significant color-enhancement effect in the air. Based on the signal amplification, trace Cys could exclusively deteriorate the UV-vis absorbance at 653 nm of p-ß-CD@Pr6O11-TMB and Fe2+ alter the one at 729 nm of p-ß-CD@Pr6O11-ABTS with visual color changes. Under the optimized conditions, the proposed p-ß-CD@Pr6O11-TMB and p-ß-CD@Pr6O11-ABTS systems were successfully applied for dual-channel monitoring of Cys in Cys capsules and fetal bovine serum and Fe2+ in agricultural products with quite low detection limits, i.e., 7.8×10-9 mol·L-1 for Cys and 6.93×10-8 mol·L-1 (S/N=3) for Fe2+, respectively. The synergetic-enhancement detection mechanisms to Cys and Fe2+ were also proposed.

Rapid visual detection of Giardia duodenalis in faecal samples using an RPA-CRISPR/Cas12a system.

Giardiasis is a common intestinal infection caused by Giardia duodenalis, which is a major economic and health burden for humans and livestock. Currently, a convenient and effective detection method is urgently needed. CRISPR/Cas12a-based diagnostic methods have been widely used for nucleic acid-based detection of pathogens due to their high efficiency and sensitivity. In this study, a technique combining CRISPR/Cas12a and RPA was established that allows the detection of G. duodenalis in faecal samples by the naked eye with high sensitivity (10-1 copies/µL) and specificity (no cross-reactivity with nine common pathogens). In clinical evaluations, the RPA-CRISPR/Cas12a-based detection assay detected Giardia positivity in 2% (1/50) of human faecal samples and 47% (33/70) of cattle faecal samples, respectively, which was consistent with the results of nested PCR. Our study demonstrated that the RPA-CRISPR/Cas12a technique for G. duodenalis is stable, efficient, sensitive, specific and has low equipment requirements. This technique offers new opportunities for on-site detection in remote and poor areas.

Development of Loop-Mediated Isothermal Amplification Assays for the Rapid and Accurate Diagnosis of Exserohilum turcicum for Field Applications.

Northern corn leaf blight (NCLB), caused by Exserohilum turcicum, is one of the most devastating foliar diseases of maize. Rapid and accurate diagnosis for this disease is urgently needed but still limited. Here, we establish a field-deployable diagnostic method to detect E. turcicum based on loop-mediated isothermal amplification (LAMP) assays. A software application called K-mer Elimination by Cross-reference was used to search for the specific sequences belonging to E. turcicum by comparing the whole genome sequence between E. turcicum and other known maize pathogens. Five LAMP primer sets were designed based on specific and single-copy fragments of E. turcicum. Post-LAMP analyses indicated that only the primer set, Et9468_set1, was the most suitable, producing a ladder-like amplification pattern in the agarose gel electrophoresis and a strong fluorescence signal in the presence of SYBR Green I. The LAMP assay using Et9468_set1 primers demonstrated a high level of specificity in distinguishing E. turcicum from six other common fungal pathogens of maize, as well as 12 more fungal and oomycete strains including the epiphytic fungi from maize leaves and other crop pathogens. Moreover, it exhibited remarkable sensitivity by detecting five copies per reaction, which was approximately 104 times more sensitive compared with conventional PCR. The LAMP assay successfully detected E. turcicum in field maize leaves without DNA extraction, demonstrating its suitability for rapid on-spot detection of NCLB. Our study provides a direct LAMP diagnostic method to detect E. turcicum, which enables on-site pathogen detection in the field and the development of preventive strategies for NCLB management.

A double-emission molecularly imprinted ratiometric fluorescent sensor based on carbon quantum dots and fluorescein isothiocyanate for visual detection of p-nitroaniline.

A novel molecularly imprinted ratiometric fluorescent sensor CQDs@MIP/FITC@SiO2 for the detection of p-nitroaniline (p-NA) was constructed through the mixture of CQDs@MIP and FITC@SiO2 in the ratio of 1:1 (VCQDs@MIP:VFITC@SiO2). The polymers of CQDs@MIP and FITC@SiO2 were prepared by sol-gel method and reversed-phase microemulsion method, respectively. CQDs@MIP was used as the auxiliary response signal and FITC@SiO2 was used as the reference enhancement signal. The signal was measured at excitation/emission wavelengths of 365/438, 512 nm. The sensor showed good linearity in the concentration range 0.14-40.00 µM (R2 = 0.998) with a detection limit of 0.042 µM for p-NA. The color change of "blue-cyan-green" could be observed by the naked eye under 365 nm UV light, thus realizing the visual detection of p-NA. The sensor presented comparable results compared with high-performance liquid chromatography (HPLC) method for the detection of p-NA in hair dye paste and aqueous samples with recoveries of 96.8-103.7% and 95.8-104.4%, respectively. It was demonstrated that the constructed sensor possesses the advantages of simplicity, excellent selectivity, superior sensitivity, and outstanding stability.

An all-inorganic lead-free metal halide double perovskite for the highly selective detection of norfloxacin in aqueous solution.

Lead-based perovskites are highly susceptible to environmental influences, and their application in analytical chemistry, especially in aqueous solution, has been reported rarely. All-inorganic lead-free metal halide perovskites have been considered as a substitute for lead-based perovskites. Herein, a Cs2RbTbCl6 perovskite microcrystal (PMCs), which emits strong yellow-green fluorescence with a maximum emission wavelength at 547 nm, was for the first time  synthesized and characterized. The Cs2RbTbCl6 PMCs could be well dispersed in N,N-dimethylacetamide (DMF), and its fluorescence could be significantly enhanced by the addition of norfloxacin (NOR) in the aqueous solution. We found that the Cs2RbTbCl6 PMCs can be used as fluorescent probes (excitation, 365 nm; emission, 547 nm) to selectively detect NOR in a concentration range from 10.0 to 200.0 µM with the limit of detection (LOD) being 0.04 µM. The Cs2RbTbCl6 PMCs could also be adsorbed on filter paper to fabricate as a fluorescent test paper for visual detection of NOR under 365-nm ultraviolet (UV) lamp irradiation. The proposed method has the potential to establish a new analytical method to visualize the detection of NOR in aqueous environments and also promotes the application of all-inorganic lead-free perovskites for analytical detection in aqueous environments.

Novel biocompatible N-rich AIE fluorescent probe for live cell imaging and visual onsite detection of uranium.

A sensitive and biocompatible N-rich probe for rapid visual uranium detection was constructed by grafting two trianiline groups to 2,6-bis(aminomethyl)pyridine. Possessing excellent aggregation-induced emission (AIE) property and the advantages to form multidentate chelate with U selectively, the probe has been applied successfully to visualize uranium in complex environmental water samples and living cells, demonstrating outstanding anti-interference ability against large equivalent of different ions over a wide effective pH range. A large linear range (1.0 × 10-7-9.0 × 10-7 mol/L) and low detection limit (72.6 nmol/L, 17.28 ppb) were achieved for the visual determination of uranium. The recognition mechanism, photophysical properties, analytical performance and cytotoxicity were systematically investigated, demonstrating high potential for fast risk assessment of uranium pollution in field and in vivo.

Graphene quantum dots on TiO2 nanotubes as a light-assisted peroxidase nanozyme.

Hybrid nanozyme graphene quantum dots (GQDs) deposited TiO2 nanotubes (NTs) on titanium foil (Ti/TiO2 NTs-GQDs) were manufactured by bestowing the hybrid with the advantageous porous morphology, surface valence states, high surface area, and copious active sites. The peroxidase-like activity was investigated through the catalytic oxidation of chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2, which can be visualized by the eyes. TiO2 NTs and GQDs comprising oxygen-containing functional groups can oxidize TMB in the presence of H2O2 by mimicking peroxidase enzymes. The peroxidase-mimicking activity of hybrid nanozyme was significantly escalated by introducing light illumination due to the photosensitive features of the hybrid material. The peroxidase-like activity of Ti/TiO2 NTs-GQDs enabled H2O2 determination over the linear range of 7 to 250 µM, with a LOD of 2.1 µM. The satisfying peroxidase activity is possibly due to the unimpeded access of H2O2 to the catalyst's active sites. The porous morphology provides the easy channeling of reactants and products. The periodic structure of the material also gave rise to acceptable reproducibility. Without material functionalization, the Ti/TiO2 NTs-GQDs can be a promising substitute for peroxidases for H2O2 detection.

Cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of Shigella flexneri.

Pathogen infections including Shigella flexneri have posed a significant threat to human health for numerous years. Although culturing and qPCR were the gold standards for pathogen detection, time-consuming and instrument-dependent restrict their application in rapid diagnosis and economically less-developed regions. Thus, it is urgently needed to develop rapid, simple, sensitive, accurate, and low-cost detection methods for pathogen detection. In this study, an immunomagnetic beads-recombinase polymerase amplification-CRISPR/Cas12a (IMB-RPA-CRISPR/Cas12a) method was built based on a cascaded signal amplification strategy for ultra-specific, ultra-sensitive, and visual detection of S. flexneri in the laboratory. Firstly, S. flexneri was specifically captured and enriched by IMB (Shigella antibody-coated magnetic beads), and the genomic DNA was released and used as the template in the RPA reaction. Then, the RPA products were mixed with the pre-loaded CRISPR/Cas12a for fluorescence visualization. The results were observed by naked eyes under LED blue light, with a sensitivity of 5 CFU/mL in a time of 70 min. With no specialized equipment or complicated technical requirements, the IMB-RPA-CRISPR/Cas12a diagnostic method can be used for visual, rapid, and simple detection of S. flexneri and can be easily adapted to monitoring other pathogens.

Hybrid nanoflower-based electrochemical lateral flow immunoassay for Escherichia coli O157 detection.

An electrochemical biosensor has been developed for detection of Escherichia coli O157 by integrating lateral flow with screen-printed electrodes. The screen-printed electrodes were attached under the lateral flow detection line, and organic-inorganic nanoflowers prepared from E. coli O157-specific antibodies as an organic component were attached to the lateral flow detection line. In the presence of E. coli O157, an organic-inorganic nanoflower-E. coli O157-antimicrobial peptide-labelled ferrocene sandwich structure is formed on the lateral flow detection line. Differential pulse voltammetry is applied using a smartphone-based device to monitor ferrocene on the detection line. The resulting electrochemical biosensor could specifically detect E. coli O157 with a limit of detection of 25 colony-forming units mL-1. Through substitution of antibodies of organic components in organic-inorganic nanoflowers, biosensors have great potential for the detection of other pathogens in biomedical research and clinical diagnosis.

Visual detection of microRNAs using gold nanorod-based lateral flow nucleic acid biosensor and exonuclease III-assisted signal amplification.

An ultrasensitive method for the visual detection of microRNAs (miRNAs) in cell lysates using a gold nanorod-based lateral flow nucleic acid biosensor (GN-LFNAB) and exonuclease III (Exo III)-assisted signal amplification. The Exo III-catalyzed target recycling strategy is employed to generate a large number of single-strand DNA products, which can be detected by GN-LFNAB visually. With the implementation of a unique recycling strategy, we have demonstrated that the miRNA in the concentration as low as 0.5 pM can be detected without the need for instrumentation, providing a detection limit that surpasses previous reports. The new biosensor is ultrasensitive and can be applied to the reliable monitoring of miRNAs in cell lysates with high accuracy. The approach offers a simple and rapid tool for cancer diagnosis and clinical biomedicine, thanks to its flexibility, simplicity, cost-effectiveness, and convenience. This new method has the potential to significantly improve the detection and monitoring of cancer biomarkers, ultimately contributing to more effective cancer diagnosis and treatment.

Fluorescence/visual aptasensor based on Au/MOF nanocomposite for accurate and convenient aflatoxin B1 detection.

A dual-mode fluorescence/visual aptasensor was developed for straightforward and accurate determination of aflatoxin B1 (AFB1) based on an Au/metal-organic framework (Au/MOF) composite. Aptamer-modified Au/Fe3O4 (Apt/Au/Fe3O4) served as the recognition element, and Au/MOF modified with complementary chains and 3,3',5,5'-tetramethylbenzidine (cDNA/TMB/Au/MOF) acted as the fluorescence and visual probes. These components are integrated to form conjugates (Apt/Au/Fe3O4-cDNA/TMB/Au/MOF). Upon the introduction of AFB1, some cDNA/TMB/Au/MOF dissociated from Apt/Au/Fe3O4, enabling the use of detached probes for visual detection. The undecomposed conjugates were isolated magnetically for use in fluorescence detection. As the AFB1 concentration increases, the visual signal intensifies and fluorescence intensity diminishes. Thus, the proposed aptasensor achieves the simultaneous fluorescence and visual determination of AFB1, obviating the need for material and reagent substitutions. The detection limits were established at 0.07 ng mL-1 for the fluorescence mode and 0.08 ng mL-1 for the visual mode. The effectiveness of the aptasensor was further validated by quantifying AFB1 in real samples.

Portable luminescent fiber- and glove-based nanosensor for multicolor visual detection of tetracycline in food samples.

An intelligent fluorescent nanoprobe (lignite-CDs-Eu) was constructed by an effective and facile method based on lignite-derived carbon dots (CDs) and lanthanide europium ions (Eu3+), which exhibited high sensitivity, low detection limit (13.35 nM) and visual color variation (from blue to red) under ultraviolet light towards tetracycline (TC) detection. Significantly, portable and economical sensors were developed using lignite-CDs-Eu immobilized fiber material of filter paper and wearable glove with the aid of color extracting and image processing application (APP) in the smartphone. Facile, fast and real-time visual detection of TC in food samples was realized. Moreover, logic gate circuit was also designed to achieve intelligent and semi-quantitative inspection of TC. To some extent, this study extended the cross-application of intelligent computer software in food analytical science, and provided a certain reference for the development of small portable detection sensors which were suitable for convenience and non-professional use in daily life.

A smartphone-based colorimetric assay using Cu-tannic acid nanosheets (Cu-TA NShs) as a laccase-mimicking nanozyme for visual detection of quercetin in vegetables.

The interaction of Cu-tannic acid nanosheets (Cu-TA NShs) as nanozyme in a surfactant solution of CTAB under relatively acidic conditions is shown to exhibit a catalytic effect on quercetin (Qur). This catalytic property of Cu-TA NShs, which mimics laccase enzyme with many advantages, has been applied to developing a selective colorimetric sensor for the determination of trace amounts of Qur in vegetable samples. This strategy presents a desirable linear relationship between the absorbance signal intensity and the concentrations of Qur from 0.350 to 32.09 µM with a detection limit (LOD) of 0.064 µM (S/N = 3). The feasibility of the proposed portable colorimetric sensor for in situ analysis of the real samples has been validated with the high-performance liquid chromatography (HPLC) method as reference method, and two-tailed test (t test) statistical analysis certifies good agreement between the results. This enzyme-free and sensitive naked-eye sensor with the smartphone-based color map is promising to provide technical support for the rapid and visual detection of Qur in vegetables.

Three in one: coordination-induced emission for inherent fluorescent Al-MOF synthesis combined with inner filter effect@aggregation-induced emission mechanisms for designing color tonality and ratiometric sensing platforms.

Three phenomena, namely coordination-induced emission (CIE), aggregation-induced emission (AIE), and inner filter effect (IFE), were incorporated into the design of a ratiometric and color tonality-based biosensor. Blue fluorescent Al-based metal-organic frameworks (FMIL-96) were prepared from non-emissive ligand and aluminum ions via CIE. Interestingly, the addition of tetracycline (TC) led to ratiometric detection and color tonality, as the blue emission at 380 nm was quenched (when excited at 350 nm) due to IFE, while the green-yellowish emission at 525 nm was enhanced due to AIE. Based on that, an ultra-sensitive visual-based color tonality mode with smartphone assistance was developed for detection of TC. The sensor exhibited a linear relationship within a broad range of 2.0 to 85.0 µM TC with a detection limit of 68.0 nM. TC in milk samples was quantified with high accuracy and precision. This integration of smartphone and visual fluorescence in solution is accurate, reliable, cost-effective, and time-saving, providing an alternative strategy for the semi-quantitative determination of TC on-site.

A low-voltage alternant direct current electroporation chip for ultrafast releasing the genome DNA of Helicobacter pylori bacterium.

Nucleic acid detection, as an important molecular diagnostic method, is widely used in bacterial identification, disease diagnosis. For detecting the nucleic acid of bacteria, the prerequisite is to release nucleic acids inside the bacteria. The common means to release nucleic acids is the chemical method, which involves complex processes, is time-consuming, and remains chemical inhibitors. Compared with chemical methods, electroporation as a physical method has the advantages of easy operation, short-time consumption, and chemical reagents free. However, the current works using electroporation often necessitates high-frequency or high-voltage conditions, entailing bulky power devices. Herein, we propose a low-voltage alternant direct current (LADC) electroporation chip and the corresponding miniature device for ultrafast releasing the genome DNA from Helicobacter pylori (H. pylori) for detection. We connected a micrometer-interdigital electrode in the chip with a 20 V portable battery to make the miniature device. Using this low-voltage device, our chip released genome DNA of H. pylori within only 5 ms, achieving a cell lysis rate of 99.5%. We further combined this chip with a colorimetric loop-mediated isothermal amplification assay to visually detect H. pylori within ~ 25 min at 10 CFU/µL. We detected 11 clinical samples using the chip, and the detection results were consistent with those of the clinical standard. The results indicate that the LADC electroporation chip is useful for ultrafast release of genome DNA from bacteria and is expected to promote the development of nucleic acid detection in POCT and other scenarios.

A Low-Cost Handheld Centrifugal Microfluidic System for Multiplexed Visual Detection Based on Isothermal Amplification.

A low-cost, handheld centrifugal microfluidic system for multiplexed visual detection based on recombinase polymerase amplification (RPA) was developed. A concise centrifugal microfluidic chip featuring four reaction units was developed to run multiplexed RPA amplification in parallel. Additionally, a significantly shrunk-size and cost-effective handheld companion device was developed, incorporating heating, optical, rotation, and sensing modules, to perform multiplexed amplification and visual detection. After one-time sample loading, the metered sample was equally distributed into four separate reactors with high-speed centrifugation. Non-contact heating was adopted for isothermal amplification. A tiny DC motor on top of the chip was used to drive steel beads inside reactors for active mixing. Another small DC motor, which was controlled by an elaborate locking strategy based on magnetic sensing, was adopted for centrifugation and positioning. Visual fluorescence detection was optimized from different sides, including material, surface properties, excitation light, and optical filters. With fluorescence intensity-based visual detection, the detection results could be directly observed through the eyes or with a smartphone. As a proof of concept, the handheld device could detect multiple targets, e.g., different genes of African swine fever virus (ASFV) with the comparable LOD (limit of detection) of 75 copies/test compared to the tube-based RPA.

Comparative Evaluation of PCR-Based, LAMP and RPA-CRISPR/Cas12a Assays for the Rapid Detection of Diaporthe aspalathi.

Southern stem canker (SSC) of soybean, attributable to the fungal pathogen Diaporthe aspalathi, results in considerable losses of soybean in the field and has damaged production in several of the main soybean-producing countries worldwide. Early and precise identification of the causal pathogen is imperative for effective disease management. In this study, we performed an RPA-CRISPR/Cas12a, as well as LAMP, PCR and real-time PCR assays to verify and compare their sensitivity, specificity and simplicity and the practicality of the reactions. We screened crRNAs targeting a specific single-copy gene, and optimized the reagent concentrations, incubation temperatures and times for the conventional PCR, real-time PCR, LAMP, RPA and Cas12a cleavage stages for the detection of D. aspalathi. In comparison with the PCR-based assays, two thermostatic detection technologies, LAMP and RPA-CRISPR/Cas12a, led to higher specificity and sensitivity. The sensitivity of the LAMP assay could reach 0.01 ng µL-1 genomic DNA, and was 10 times more sensitive than real-time PCR (0.1 ng µL-1) and 100 times more sensitive than conventional PCR assay (1.0 ng µL-1); the reaction was completed within 1 h. The sensitivity of the RPA-CRISPR/Cas12a assay reached 0.1 ng µL-1 genomic DNA, and was 10 times more sensitive than conventional PCR (1.0 ng µL-1), with a 30 min reaction time. Furthermore, the feasibility of the two thermostatic methods was validated using infected soybean leaf and seeding samples. The rapid, visual one-pot detection assay developed could be operated by non-expert personnel without specialized equipment. This study provides a valuable diagnostic platform for the on-site detection of SSC or for use in resource-limited areas.

Enhanced Molecularly Imprinted Fluorescent Test Strip for Rapid and Visual Detection of Norfloxacin via a Smartphone.

Paper-based test strips with on-site visual detection have become a hot spot in the field of target detection. Yet, low specific surface area and uneven deposition limit the further application of test strips. Herein, a novel "turn-on" ratio of molecularly imprinted membranes (Eu@CDs-MIMs) was successfully prepared based on a Eu complex-doped polyvinylidene fluoride membrane for the selective, rapid and on-site visual detection of norfloxacin (NOR). The formation of surface-imprinted polymer-containing carbon dots (CDs) improves the roughness and hydrophilicity of Eu@CDs-MIMs. Fluorescence lifetimes and UV absorption spectra verified that the fluorescence enhancement of CDs is based on the synergistic effect of charge transfer and hydrogen bonding between CDs and NOR. The fluorescent test strip showed a linear fluorescent response within the concentration range of 5-50 nM with a limit of detection of 1.35 nM and a short response time of 1 min. In comparison with filter paper-based test strips, Eu@CDs-MIMs exhibit a brighter and more uniform fluorescent color change from red to blue that is visible to the naked eye. Additionally, the applied ratio fluorescent test strip was combined with a smartphone to translate RGB values into concentrations for the visual and quantitative detection of NOR and verified the detection results using high-performance liquid chromatography. The portable fluorescent test strip provides a reliable approach for the rapid, visual, and on-site detection of NOR and quinolones.