Colorimetric Changes Research Articles

The Caffeine Analysis in Tea Bag and Robusta Coffee Using UHPLC Methods

Caffeine, a natural psychostimulant, enhances arousal by inhibiting the effects of adenosine on the dopamine pathway. This study aimed to analyze and compare caffeine concentrations in seven robusta coffee samples (coded RC) and five black tea bags (coded BT) using both qualitative and quantitative methods. Qualitative tests employed Dragendorff reagents for coffee and Parry reagents for tea, confirming caffeine presence through colorimetric changes. Quantitative analysis utilized reverse-phase UHPLC, with a mobile phase of aquabidestilata and methanol (60:40), a C18 stationary phase at 40°C, and detection at 272 nm. Validation parameters demonstrated strong performance, with an RSD of 0.573%, a linear correlation coefficient of 0.998 for concentrations between 100–250 ppm, a LoD of 1.02 ppm, a LoQ of 3.10 ppm, and 99.88% accuracy. Results showed robusta coffee sample RC3 contained the highest caffeine concentration at 26.00 mg/serving, while black tea sample BTB3 exhibited 2.44 mg/serving. Despite higher caffeine levels in robusta coffee compared to black tea, both remained below the SNI threshold of 50 mg/serving, confirming their safety for consumption.

Multi-Stimuli Responsive Viologen-Imprinted Polyvinyl Alcohol and Tricarboxy Cellulose Nanocomposite Hydrogels

Photochromic inks have shown disadvantages, such as poor durability and high cost. Self-healable hydrogels have shown photostability and durability. Herein, a viologen-based covalent polymer was printed onto a paper surface toward the development of a multi-stimuli responsive chromogenic sheet with thermochromic, photochromic, and vapochromic properties. Viologen polymer was created by polymerizing a dialdehyde-based viologen with a hydroxyl-bearing dihydrazide in an acidic aqueous medium. The viologen polymer was well immobilized as a colorimetric agent into a polyvinyl alcohol (PVA)/tricarboxy cellulose (TCC)-based self-healable hydrogel. The viologen/hydrogel nanocomposite films were applied onto a paper surface. The coloration measurements showed that when exposed to ultraviolet light, the orange layer printed on the paper surface switched to green. The photochromic film was used to develop anti-counterfeiting prints using the organic hydrogel composed of a PVA/TCC composite and a viologen polymer. Reversible photochromism with strong photostability was observed when the printed papers were exposed to UV irradiation. A detection limit was monitored in the range of 0.5–300 ppm for NH3(aq). The exposure to heat (70 °C) was found to reversibly initiate a colorimetric change.

Dual-mode sensing platform for Bisphenol A detection via bifunctional CsPbBr3@Cu-MOF assisted fluorescence and colorimetric analysis

BackgroundBisphenol A (BPA) has been identified as an endocrine disruptor with numerous detrimental effects on human health. There is an urgent need to develop fluorescence/colorimetric dual-mode sensing approaches with expanded detection linear range, increased accuracy, and enhanced application flexibility for BPA detection. The utilization of fluorescence and colorimetric signals in point-of-care applications and real-time sensitive sensing further highlights the significance of developing novel and efficient fluorescence/colorimetric dual-mode sensing platform with high-efficiency probes. ResultsHerein, a fluorescence and colorimetric dual-mode aptasensor was developed by using CsPbBr3@Cu-MOF as the aptamer immobilization matrix and signal generator. CsPbBr3 was functionalized with Cu-MOF using a simple two-step strategy. This strategy involved in-situ formation and modified ligand-assisted precipitation technique with 4,4′-bipyridine (4,4-Bpy) serving as the bifunctional linker. The resulting CsPbBr3@Cu-MOF exhibited improved stability in water and enhanced fluorescence. Additionally, it functioned as peroxidase mimetic to oxidize 3,3′,5,5′-tetramethyl benzidine (TMB), leading to a colorimetric change from colorless to blue. In the presence of BPA, aptamers were removed from CsPbBr3@Cu-MOF. Consequently, the fluorescence and peroxidase-activity of CsPbBr3@Cu-MOF were recovered, resulting in the enhanced fluorescence intensity and color change of TMB. Using this system, the proposed aptasensor demonstrated detection ranges of 1.0–80.0 nM with a LOD of 0.60 nM for the colorimetric method, and a linearity range of 0.1–100 nM with a LOD of 0.02 nM for the fluorescence method. SignificanceThe obtained CsPbBr3@Cu-MOF composites showed excellent fluorescence properties, good peroxidase-like activity, and aqueous stability. Furthermore, the proposed dual-mode aptasensor demonstrated simplicity, cost-effectiveness and good anti-interference abilities. This can be extended to the construction of other dual-mode sensors by changing aptamers and provides novel insights on the potential applications of perovskites in bioanalysis.

A synergetic effect of light and anion: near-infrared colorimetric monitoring of nitric oxide (NO) release from fluoride/cyanide anions and a water responsive ruthenium nitrosyl complex.

Near-infrared (NIR) spectral-responsive multitasking chemical systems are always advantageous in biological and environmental systems. Although ruthenium complexes are highly attractive compounds for many applications, studies on NIR optical responsive sensors are very limited because of their synthetic difficulties. The sensing of fluoride and cyanide ions using ruthenium nitrosyl complexes is not known. In this study, we report the synthesis and characterization of a new terpyridine-based ruthenium nitrosyl complex [Ru(Cl)2NO(terpy-C6H4OH)] 1·Ru-OH. The complex exhibited distinct NIR absorptions at 680 nm, as well as a visible color change with the fluoride ion in DMSO-CH3CN medium. However, when the solvent is changed to DMSO-H2O, it responds only with a cyanide ion with a distinct colorimetric change. Binding of the F- ion leads to deprotonation of 1·Ru-OH; the deprotonated complex is also used for the colorimetric detection of a trace amount of water in DMSO and acetonitrile with a limit of detection (LOD) of 0.034 wt% and 0.007 wt%, respectively. Ruthenium nitrosyl complexes have appeared as promising platforms for light-controlled release of nitric oxide (NO), which can be beneficial for therapeutic application. NO release studies of 1·Ru-OH by UV-vis and FT-IR spectroscopy confirm that it can release NO in a light-controlled manner. In addition, the NO release could be monitored by the naked eye with a color change and spectral change in the NIR region. Importantly, NO release studies revealed that the rate of NO release could be modulated in the presence of the F- ion. Here, the fluoride ion acts as an allosteric regulator. These results demonstrate that 1·Ru-OH is both a promising multitasking colorimetric and NIR sensor and a colorimetric responsive NO-releasing agent.

Novel Colorimetric and Near-Infrared Ratiometric Fluorescent Probe for Sensing Cysteine in Food Samples, Plants, and Living Cells.

Cysteine (Cys) is a crucial biothiol that acts a significant function in food samples and biological systems, including plant roots and living cells. Hence, we developed a novel colorimetric and near-infrared ratiometric fluorescent probe (CT), composed of coumarin and tetrahydroacridine-conjugated indole salt, for the detection of Cys. Upon reaction with Cys, the probe undergoes a specific N-substitution reaction, resulting in a notable colorimetric change and a significant ratiometric fluorescent response in both visible and near-infrared emission channels. These dual-channel ratiometric fluorescence changes are completely independent, enabling the probe to obtain great selectivity, sensitivity, and exceptional detection accuracy. Leveraging these attributes, the probe was employed to provide accurate quantitative analysis of Cys in food samples. Furthermore, confocal imaging demonstrated that the probe could monitor both exogenous and endogenous Cys levels in living cells and track Cys changes in plant roots under heavy metal stress. This work presents a dependable and accurate imaging solution for tracking and identifying Cys of real food, plants, and living cells.

A commercially available dye as a highly versatile colorimetric fluoride sensor

Reported herein is the successful use of the commercially available organic dye bromocresol green (compound 1) for high-performing and practical fluoride detection. This dye responds to fluoride anion with high selectivity (virtually no response to other halide anions), excellent sensitivity (limits of detection as low as 0.22 ppm) and rapid response times, and maintains functionality when it is adsorbed on filter paper. Mechanistic studies determined that the fluoride-induced colorimetric changes were due to intermolecular hydrogen bonding between compound 1 and fluoride. Finally, the broad applicability of this sensor was demonstrated through the colorimetric response of compound 1 – functionalized paper to fluoride in commercial mouthwash. These findings underscore compound 1’s potential as a practical and commercially viable fluoride sensor, in addition to compound 1’s better-known role as a colorimetric pH indicator.

Benzothiazole-quinoline based probe for simultaneous colorimetric detection of CN- and Cu2+ ions, with fluorescence sensing for Cu2+: Mechanistic insights and practical applications in environmental monitoring and cellular analysis

The development and synthesis of notably targeted and colorimetric sensor based on an azomethine compound for the distinct recognition of Cu2+ and CN− ion individually in an aqueous dimethyl formamide solution is performed. In the presence of CN− and Cu2+, the sensor BTZ showed impressive colorimetric changes, going from pale yellow to orange and pale yellow to dark yellow, respectively. In the meantime, FL spectrum (Cu2+) and UV–vis spectroscopy (CN−/Cu2+) were used to assess the sensing features. The plausible binding mechanisms of CN− and Cu2+ ions with sensor BTZ have been studied using the 1H NMR titration, Job's plot and DFT technique. The bathochromic shift produced by the intramolecular charge transfer (ICT) transition may have been the source of the phenomenon. Furthermore, CN− ion in the commercial substance is quickly identified and measured with the naked eye by using sensor BTZ. It was found that the BTZ's LOD for CN− and Cu2+ was 0.280 × 10−7 M and 1.153 × 10−7 M, respectively. Moreover, 1:1 binding ratio for the reaction of CN− and Cu2+ ions with sensor BTZ were demonstrated by Job's plot, which was dependent on analytical data. The findings show that BTZ is an easy-to-use and practical probe for concurrently sensing cyanide and copper ions in environmental samples and living cells that have less cytotoxicity.

Novel förster resonance energy transfer (FRET)-based ratiometric fluorescent probe for detection of cyanides by nucleophilic substitution of aromatic hydrogen (SNArH)

The development of novel fluorescent probes for real-time detection of cyanides (CN−) in environmental and biological systems has become a significant focus in chemical sensing. Particularly, ratiometric fluorescence sensing offers a unique method for precise and quantitative detection of cyanides, even under complex conditions. We report herein the design of a new ratiometric fluorescent probe for cyanides based on modulation of Förster resonance energy transfer (FRET) coupled with novel cyanide-induced nucleophilic substitution of aromatic hydrogen (SNArH). The target probe (R1) is developed by introducing coumarin fluorophores as FRET donors into a 3-nitro-naphthalimide acceptor, which is easily synthesized and exhibits a colorimetric change from colorless to faint yellow and a significant ratiometric fluorescence shift (Δλ = 114 nm) upon cyanide binding. A clear ratiometric signal at I582/I468 was obtained, with a limit of detection of 5.69 μM. The sensing mechanism was confirmed through 1H NMR titration and LC-MS analysis. Additionally, R1-loaded strips were easily prepared, serving as a portable device for detecting CN− with visible color changes. The probe R1 has been successfully utilized for real-time monitoring of cyanide in food materials and water samples. Importantly, fluorescence bioimaging studies in HeLa cells were conducted, demonstrating the probe’s capability for ratiometric detection of exogenous CN− in living systems.

A spectroscopy-based proof-of-concept (POC) for developing loading of pathogen analyzer (LOPA) for dairy products

Detection of bacterial contamination in dairy products of daily use is a challenge worldwide. We have utilized Methylene Blue Reduction Test (MBRT) for quantification of the microbial count in dairy products (milk) and developed a proof-of-concept (POC) based on this for in-filed applications. In this study, we have used pasteurized milk contaminated with model bacteria Escherichia coli, and Staphylococcus aureus for the calibration and validation of the developed POC. The conversion of MB to Leuco-MB i.e., the colorimetric change due to the reduction of MB to Leuco-MB in presence of microbes has been utilized as the tool to detect presence of microbes in milk. The absorbance peak for methylene blue (MB) at 664 nm decreases significantly in presence of microbes and the blue color becomes faded. In our study, we have employed methylene blue (MB) discolouration phenomenon to estimate the microbial count in milk samples using our developed spectroscopy based POC. The limit of detection (LOD) and the limit of quantitation (LOQ) of the POC were found to be 0.32 CFU/mL and 0.97 CFU/mL. The end users of the developed POC are primarily those involved in the production, processing, testing, regulation, and research of dairy products to ensure they meet safety standards and protect public health. These include retailers, dairy farmers, dairy processors, quality control laboratories, regulatory agencies and research institutions. In our experiment, we have observed a significant change in MB absorption in the milk contaminated with microbes. The indigenously developed sensor strips designed for the working of the POC turn to colorless Leuco-MB compared to milk without the microbes. The analysis of the strips has been measured in the developed device.

Colorimetric and Fluorimetric Detection of Fe(III) Using a Rhodamine-Imidazole Hydrazone Based Chemosensor: Photophysical Properties, DFT, TGA, and DSC Studies.

Rhodamine-imidazole hydrazones (RIH-1 & RIH-2) based chemosensors have been synthesized. These are characterised and evaluated by FT-IR spectroscopy, 1H-NMR, 13C-NMR, LCMS, absorption and fluorescence spectroscopy. These chemosensors exhibit enhanced sensitivity and selectivity in detecting the biologically significant Fe3+ metal ion through both colorimetric and fluorescence changes. The optical properties have been investigated using binary acetonitrile-water (7:3 v/v) semi-aqueous solution. The probe RIH-1 can be deployed as a fluorescent and colorimetric probe for the detection of Fe3+ ion. It shows an absorption band at 559nm and an intensity band at 579nm increasing up to 50-fold with the increase in the concentration of Fe3+ with the detection limit as low as 11nM. In the visible light, RIH-1 helps in the detection of Fe3+ ion through the naked eye, while the addition of Fe3+ to the probe RIH-1 results in a colour change from colourless to pink. This is primarily due to the opening of the lactone ring in RIH-1. Notably, RIH-1 probe displays a high quantum yield of 0.51, after binding with Fe3+ ions. Indeed, it has been found that sensor RIH-1 is very effective in sensing Fe3+ ions through both fluorescence based and visual detection methods. Additionally, DFT studies of these chemosensors have been evaluated, TGA and DSC analysis showed good thermal stability.

Towards portable kits for on-site colorimetric detection of aqueous hydrazine using piperidine-thiophene-barbituric acid push-pull probe.

Owing to the carcinogenicity and environmental risks as well as the wide industrial use of hydrazine, we report herein a colorimetric probe for its ratiometric detection in pure water. The developed probe possesses push-pull architecture with 2-(piperidyn-1-yl)thiophene as the donor, N,N'-dibutylbarbituric as the acceptor, and butadiene as the spacer. In contrast to weak solvatochromic behavior in organic solvents, the probe showed distinct optical photophysical properties in water resulting from the formation of nanoscopic aggregates. The probe underwent pronounced spectral changes upon the addition of hydrazine including an 11.5-fold decrease in absorbance and ~2.4-fold fluorescence quenching. The mechanistic investigation revealed the rapid formation of hydrazone upon the interaction of the probe with hydrazine via retro-Knoevenagel reaction as confirmed experimentally and corroborated with DFT calculations. The induced colorimetric and fluorometric changes were utilized in hydrazine sensing with excellent selectivity over other biologically relevant analytes with a detection limit of 0.76 µM in aqueous media. The practical utility of the probe was assessed in real-life natural water samples, while we have also developed a cost-effective portable kit for the on-site hydrazine detection both in the solution and vapor phases.

Solvent-free mechanosynthesis of Schiff bases derived from thiadiazole with potential application in sensing ionic species and NLO applications

In this work, we report the solvent-free mechanosynthesis of three Schiff bases (SBs) derived from 1,3,4-thiadiazole covalently bonded to indole (Th-In), quinoline (Th-Qn) and triphenylamine (Th-TPA) groups. This green chemistry method showed a drastic reduction in reaction time and avoidance of solvents for obtaining SBs, with good reaction yields. The molecular structures of SBs were investigated by computational chemistry calculations, with global reactivity parameters indicative of electron acceptor behavior, estimation of frontier molecular orbitals, and their theoretical band gaps of 3.49 and 3.4 eV for Th-In and Th-Qn, respectively, and the lowest of 299 eV for Th-TPA. To evaluate the possible NLO response, the hyperpolarizabilities (β) were also estimated, which are several orders of magnitude higher than the urea value used as a reference. The absorption and emission spectra were calculated using TD-DFT. These results showed a larger redshift for Th-TPA, with possible charge transfer processes more intense than those obtained for Th-In and Th-Qn. The photophysical properties of the Schiff bases were determined, with band gap values for Th-In, Th-Qn, and Th-TPA of 2.53, 2.66, and 2.39 eV in solution and 2.77, 2.26, and 2.21 eV in film, respectively. When the SBs were evaluated as potential naked-eye colorimetric chemosensors, colorimetric changes were observed in Fe2+, Fe3+, Pb2+, Cu2+, and Ag+ ions, with Ag+ being the most promising. Z-scan was used to evaluate the nonlinear response of the SBs and showed saturable absorber behavior with self-focusing processes. Cyclic voltammetry was used to determine the electrochemical band gaps, which were 2.70, 2.71, and 1.98 eV for Th-In, Th-Qn, and Th-TPA, respectively. Th-TPA exhibits superior optical and electrical properties among the three SBs investigated. In addition, it exhibits photochromism, which can be used in conjunction with its other properties in NLO and sensor applications.

Optical and Catalytic Properties of Nanozymes for Colorimetric Biosensors: Advantages, Limitations, and Perspectives

AbstractDetection of colorimetric signals is commonly used in various analytical methods and for testing in non‐laboratory and resource‐limited settings. The performance of colorimetric assays is largely based on nanoparticles and their unique optical properties. Multifunctional nanoparticles combining optical and enzyme‐like catalytic properties—known as nanozymes—hold great promise for analytical applications as signal‐generating labels. However, the extensive focus on the catalytic properties leaves their unique optical properties overlooked. In this article, the use of the optical and catalytic properties of nanozymes is reviewed for analytical applications relying on the inherent optical properties of nanozymes, the colorimetric detection of a catalytically‐formed product, and colorimetric changes of nanoparticles caused by the catalytically‐formed product. The impact of the extinction coefficient of nanozymes and reaction products, as well as the kinetic parameters of nanozymes on the sensitivity and limit of detection of assays, are quantitatively evaluated. Finally, the existing limitations and prospects of nanozymes for colorimetric biosensors are summarized.

An activatable near-infrared fluorescent probe with large Stokes shift for visualizing peroxynitrite in Alzheimer’s disease models

Alzheimer’s disease (AD), characterized by its incurable nature and prevalence among the elderly, has remained a focal point in medical research. Increasing evidence suggests that peroxynitrite (ONOO−) serves as a crucial biomarker for the diagnosis of AD. In this study, we present a novel, easily available, high-yield, and cost-effective near-infrared (NIR) fluorescent probe, CDCI-ONOO. This probe utilizes a coumarin-dicyanoisophorone conjugate as the fluorophore and diphenylphosphinic chloride as the recognition site, enabling the detection of ONOO− both in vitro and in vivo. Upon interaction with ONOO−, CDCI-ONOO exhibits a distinct maximum emission peak at 715 nm with a substantial Stokes shift of 184 nm. The probe demonstrates excellent selectivity and sensitivity (LOD = 144 nM), along with noticeable colorimetric and fluorescence changes after the reaction. Comprehensive analyses using high-performance liquid chromatography (HPLC), high-resolution mass spectrometry (HRMS), and density functional theory (DFT) calculations confirm that the reaction with ONOO− restores the initially quenched Intramolecular Charge Transfer (ICT), resulting in the formation of CDCI-OH, a product that emitting fluorescence in the near-infrared region. Furthermore, we demonstrated the successful application of CDCI-ONOO for ONOO− detection in neuronal cells and imaging of ONOO− in the brains of mice. These findings underscore the potential of CDCI-ONOO as a near-infrared fluorescent probe for in vivo ONOO− detection, offering a significant avenue for advancing our understanding of AD pathology and diagnosis.

A fluorescently-doped thermoplastic polymer as a proof-of-principle device for the detection and capture of H2S and its HS− anion

Mono and dinuclear Zn(salen-type) complexes 1 and 2 are presented as efficient sensing and adsorption materials for HS-. The addition of HS− provokes colorimetric and fluorometric changes, useful for optical absorption and fluorescence analyses. Moreover, the limit of detection for HS− for both complexes falls in the sub-micromolar range of concentration. The adsorption/desorption process of HS- nanoporous crystalline sulfonated polyphenyleneoxide (NC-sPPO) films doped with Zn(salen-type) complexes and an interesting recycling-potential of this system in the detection of hydrogen sulfide, even over time, was proven. The presented results provide proof-of-principle that a fluorescent complex immobilized in a suitable thermoplastic polymer could be the right material to use in a HS- measuring device that might lead to new developments in the field of optical sensing.

Measuring the capacity of yeast for surface display of cell wall-anchored protein isoforms by using β-lactamase as a reporter enzyme.

Yeast surface display is a promising biotechnological tool that uses genetically modified yeast cell wall proteins as anchors for enzymes of interest, thereby transforming yeast cell wall into a living catalytic material. Here, we present a comprehensive protocol for quantifying surface-displayed β-lactamase on the cell wall of model yeast Saccharomyces cerevisiae. We use β-lactamase as a reporter enzyme, which we tagged to be anchored to the cell wall closer to its N or C terminus, through the portion of the Pir2 or Ccw12 cell wall proteins, respectively. The catalytic activity of surface-displayed β-lactamase is assessed by its ability to hydrolyze nitrocefin, which produces a colorimetric change that is quantitatively measured by spectrophotometric analysis at 482 nm. This system enables precise quantification of the potential of S. cerevisiae strains for surface display, continuous real-time monitoring of enzyme activity, and facilitates the study of enzyme kinetics and interactions with inhibitors within the cell's native environment. In addition, the system provides a platform for high-throughput screening of potential β-lactamase inhibitors and can be adapted for the visualization of other enzymes, making it a versatile tool for drug discovery and bioprocess development.

A New, Rapid, Colorimetric Chemodosimeter, 4-(Pyrrol-1-yl)pyridine, for Nitrite Detection in Aqueous Solution.

With increasing concerns over environmental impact and overall health of both the environment and its people, a need to quantify contaminants is of the utmost importance. Chemosensors with low detection limits and a relative ease of application can address this challenge. Nitrite ions are known to be detrimental to both the environment and human health. A new colorimetric chemodosimeter has been prepared from the homolytic photochemical cleavage of a reaction between pyrrole and pyridine. The product, 4-(pyrrol-1-yl)pyridine, yields a limit of detection of 0.330 (±0.09) ppm for the detection of nitrite in aqueous solution, employing a colorimetric change from yellow to pink. It is also highly selective for nitrite when various competitive anions such as SO3 2-, NO3 -, PO4 3-, SO4 -2, Cl-, F-, I-, Br-, AcO-, and CN- are present in great excess. The molecule's especially high sensitivity to nitrite is apparently the result of a complex supramolecular mechanism, characterized by both dynamic light scattering of the aggregate and the Tyndall effect. Consequently, this new sensor provides a simple, low-cost way to rapidly detect nitrite anions in aqueous solution.

Development of a Gold Nanoparticle-Based Sensor for Authentication of Organic Milk Based on Differential Levels of miRNA.

Dairy production systems significantly impact environmental sustainability, animal welfare, and human health. Intensive farming maximizes output through high-input practices, raising concerns about environmental degradation, animal welfare, and health risks from antibiotic residues. Conversely, organic farming emphasizes sustainable practices, animal welfare, and minimal synthetic inputs, potentially enhancing biodiversity, soil health, and milk quality. MicroRNAs (miRNAs), non-coding RNAs regulating gene expression, are promising biomarkers due to their response to various conditions. In this study, miRNAs bta-miR-103 and bta-miR-155, which are abundant in milk from pasture-fed cows, were selected. Additionally, bta-miR-215, which is abundant in milk fat from intensive systems, was also studied, in order to differentiate dairy production systems. A novel, cost-effective gold nanoparticle (AuNP)-based sensor was developed for miRNA detection, leveraging the unique plasmonic properties of AuNPs for visual detection. The method involves functionalizing AuNPs with complementary RNA probes and detecting miRNA-induced aggregation through colorimetric changes. This rapid, results in 30 min, and sensitive, visual limit of detection of 200 nM, assay requires minimal instrumentation and can be easily interpreted, offering significant advantages for field implementation in characterizing dairy production systems. This study demonstrates the successful application of this sensor in detecting miRNAs in 350 nM miRNA spiked raw milk, highlighting its potential for in situ dairy industry applications.

Event-specific loop-mediated isothermal amplification for living modified cotton MON88701, MON531, MON15985, MON88913, and COT102

Gossypium hirsutum L., commonly known as upland cotton, is cultivated globally for natural fiber, feed, and seed oil. To enhance agricultural productivity and quality, living modified (LM) cotton has been developed and utilized since the late 1990s. Due to environmental concerns, such as biodiversity risks associated with living modified organisms (LMOs), the Korean government, including the Ministry of Environment, has been conducting LMO natural environment monitoring and post-management projects. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay to detect five specific LM cotton events (MON88701, MON531, MON15985, MON88913, and COT102), which were the most abundant volunteers observed from 2019 to 2021 in South Korea. The event-specific LAMP assays for the five LM cotton events were established with a 40-min reaction time using LAMP reaction buffer, Bst DNA polymerase, and event-specific primers. The limit of detection ranged from 0.01 to 1 ng/μL. The assays' specificity and sensitivity were validated through colorimetric changes, fluorescence intensity measurements, and conventional PCR. To demonstrate the practical application of the LAMP assays, we tested 22 LM cotton volunteers collected from the natural environment in 2021. Each LAMP assay event-specifically amplified the respective LM cotton volunteers. These results indicate that the developed LAMP assays are effective tools for the efficient management and detection of LM cotton in field surveys, supporting regulatory compliance and environmental monitoring efforts.

A specific ‘Turn-on’ fluorogenic probe for the detection of phosphate ions

Phosphate (PO43-) ions are an essential trace element in the human body and play a vital role in biological and ecological systems. However, excess consumption of PO43- ions causes various serious health issues and becomes hazardous to public safety. Thus, selective tracking and quantification of PO43- ions is a significant concern to the scientific communities. In the present article, we have introduced a chromo-fluorogenic probe APES, which instantly shows a characteristic color-fluorimetric change towards PO43- ions. APES shows an instant greenish-yellow fluorescence at 530 nm and a yellow to orange naked-eye visible colorimetric change in the presence of PO43- ions. The detection and quantification limit of PO43- ions employing APES is estimated as 1.5 µM and 5.0 µM respectively. The interaction affinity of APES towards PO43- ions is found by the Benesi-Hildebrand equation as 0.42 × 105 M-2 in a 20 % water-DMSO medium. Also, our modeled probe is biologically significant as it can detect and quantify PO43- ions within the range of 50 % water medium. A super handy paper-strips-based detection tool has also been fabricated utilizing APES which could be employed to track trace amounts of PO43- ions. The mechanism involved in the detection of PO43- ions is elucidated through 1H NMR and density functional theoretical analysis. Also to deeply entrust the selectivity with quantification capability and practical usability of our probe APES, a real sample analysis is performed to explore the fruitful real-time analysis by experimenting with different sources of water.