Detection Of Anions Research Articles

Review on Synthesis of 2-(2-Hydroxyaryl) Benzothiazoles (HBT) for Excited-State Intra-molecular Proton Transfer (ESIPT)-Based Detection of Ions and Biomolecules.

In this review, we present a systematic and comprehensive summary of the recent developments in the synthetic strategies of 2-(2-hydroxyarylsubstituted)-benzothiazole (HBT) framework along with incorporation of various substituents on phenolic and benzothiazole rings which affect the emission process. The literature, spanning the years 2015-2024, on excited-state intramolecular proton transfer (ESIPT)-based studies of HBT derivatives comprising the effects of solvent polarity, substituents, and extended conjugation on fluorophores has been searched. ESIPT, intramolecular charge transfer, and aggregation-induced emissions enable these fluorescent probes to specifically interact with analytes, thereby altering their luminescence characteristics to achieve analyte detection. These fluorescent probes exhibit large Stokes shifts, high quantum yields, and excellent color transitions. Finally, the applications of HBTs as ESIPT-based fluorescent probes for the detection of cations, anions, and biomolecules have been summarized. We anticipate that this review will provide a comprehensive overview of the current state of research in this field and encourage researchers to develop novel ESIPT-based fluorophores with new applications.

Fluorescent sensors for detecting anions

Fluorescent sensors for detecting anions Nicola Edwards, Associate Professor of Chemistry at the University of St. Joseph, is conducting research on the development of fluorescent sensors for anion detection. Fluorescent (chemo)sensors are molecules that register changes in their “glow” (fluorescence ) when bound to an analyte (the substance of interest that is being studied), thus reporting the presence of that analyte. These changes can be detected with the naked eye. and/or instrumentation. Therefore, the presence of this invisible substance can now be detected. Fluorescent sensors are generally more sensitive and have lower limits of detection for a particular analyte compared to colorimetric sensors. Anions—chemical entities with an overall negative charge—are found everywhere and are involved in numerous chemical, biological, and industrial processes. As an example, cavity-fighting kinds of toothpaste and gels contain anion and fluoride. Additionally, chemical compounds containing anions, nitrate and phosphate are used in fertilizers. In this piece, Dr. Nicola Edwards discusses her ongoing work on developing fluorescent sensors for anions.

Detection of sulphur(II) of carbon dots synthesized from Gardenia residue.

The detection of anions using carbon dots (CDs) has received less attention compared to cations. Therefore, the present study aimed to develop a fluorescence sensor based on carbon dots (CDs) capable of detecting S2- in real water samples. The CDs were successfully prepared from the residues of a traditional Chinese herb, Gardenia, which emitted green photoluminescence (PL) under ultraviolet light irradiation. The as-prepared CDs were quasi-spherical in shape and ranged in size from 10 to 30 nm. Different detailed analyses proved that the CDs had good morphology, various functional groups, high water solubility, great optical features, and excellent stability under diverse environmental conditions. The ion detection showed that only Ag+ had the strongest fluorescence quenching effect on the CDs, however, the addition of S2- could recover their fluorescence. Based on these results, an "off-on" fluorescence sensor was achieved to selectively detect the concentration of S2- in real water samples with a limit of detection (LOD) of 39 μM, which further expanded the application of residues from traditional Chinese herbal medicine.

Precise counter anion modulation of the self-assembly behavior-endowed ultrasensitive and specific dual-mode visualization of nitrate

Pt(II) polypyridine complex-based probe exhibits promising performance in anion detection by the change of the absorption and emission properties based on supramolecular self-assembly. However, whether one can develop a modulation strategy of the counter anion to boost the detection sensitivity and anti-interference capability of the Pt(II) complex-based probe remains a big challenge. Here, an effective modulation strategy was proposed by precisely regulating the interaction energy through adjusting the type of the counter anions, and a series of probes have been synthesized by counter anion (X = Cl-, ClO4-, PF6-) exchange in [Pt(tpy)Cl]·X (tpy=2,2′:6′,2′’-terpyridine), and thus the colorimetric-luminescence dual-mode detection toward nitrate was achieved. The optimal [Pt(tpy)Cl]·Cl probe shows superior nitrate detection performance including a limit of detection (LOD) (8.68 nM), rapid response (<0.5 s), an excellent selectivity and anti-interference capability even facing 14 common anions. Moreover, a polyvinyl alcohol (PVA) sponge-based sensing chip loaded with the probe enables the ultra-sensitive detection of nitrate particles with an ultralow detection limit of 7.6 pg, and it was further integrated into a detection pen for the accurate recognition of nitrate particles in real scenarios. The proposed counter-anion modulation strategy is expected to start a new frontier for the exploration of novel Pt(II) complex-based probes.

A Highly Stable Multifunctional Bi-Based MOF for Rapid Visual Detection of S2- and H2S Gas with High Proton Conductivity.

Metal organic frameworks (MOFs) constructed with bismuth metal have not been widely reported, especially multifunctional Bi-MOFs. Therefore, developing multifunctional MOFs is of great significance due to the increasing requirements of materials. In this work, a 3D Bi-MOF (Bi-TCPE) with multifunctionality was successfully constructed, demonstrating high thermal stability, water stability, a porous structure, and strong blue fluorescence emission. We evaluated the properties of Bi-TCPE in detecting anions (S2-, Cr2O72-, and CrO42-) in aqueous solution, along with the rapid visual detection of H2S gas and proton conduction. In terms of anion detection, Bi-TCPE achieved the rapid detection of trace S2- in aqueous solutions, while the Ksv value was 1.224 × 104 M-1 with a limit of detection (LOD) value of 1.93 μM through titration experiments. Furthermore, Bi-TCPE could sensitively detect Cr2O72- and CrO42-, with Ksv values of 1.144 × 104 and 1.066 × 104 M-1, respectively, while LOD reached 2.07 and 2.18 μM. Subsequently, we conducted H2S gas detection experiments, and the results indicated that Bi-TCPE could selectively detect H2S gas at extremely low concentrations (2.08 ppm) and with a fast response time (<10 s). We also observed significant color changes under both UV light and sunlight. Therefore, we developed a H2S detection test paper for the rapid visual detection of H2S gas. Finally, we evaluated the proton conductivity of Bi-TCPE, and the experimental results showed that the proton conductivity of Bi-TCPE reached 4.77 × 10-2 S·cm-1 at 98% RH and 90 °C, achieving an excellent value for unmodified and encapsulated MOFs. In addition, Bi-TCPE showed high stability in proton conduction experiments (it remained stable after 21 consecutive days of testing and 12 cycles of testing), demonstrating relatively high application value. These results indicate that Bi-TCPE is a multifunctional MOF material with great application potential.

A supramolecular fluorescence sensor array for the differentiation of multiple anions and prediction of iodine in artificial urine using machine learning.

The simultaneous discrimination and detection of multiple anions in an aqueous solution has been a major challenge due to their structural similarity and low charge radii. In this study, we have constructed a supramolecular fluorescence sensor array based on three host-guest complexes to distinguish five anions (F-, Cl-, Br-, I-, and ClO-) in an aqueous solution using anionic-induced fluorescence quenching combined with linear discriminant analysis. Due to the different affinities of the three host-guest complexes for each anion the anion quenching efficiency for each host-guest complex was likewise different, and the five anions were well recognized. The fluorescence sensor array not only distinguished anions at different concentrations (0.5, 10, and 50 µM) with 100% accuracy but also showed good linearity within a certain concentration range. The limit of detection (LOD) was < 0.5 µM. Our interference study showed that the developed sensor array had good anti-interference ability. The practicability of the developed sensor array was also verified by the identification and differentiation of toothpaste brands with different fluoride content and the prediction of the iodine concentration in urine combined with machine learning.

Pyrene Appendant Triazole-based Chemosensors for Sensing Applications.

Over the last two decades, the design and development of fluorescent chemosensors for the targeted detection of Heavy Transition-metal (HTM) ions, anions, and biological analytes, have drawn much interest. Since the introduction of click chemistry in 2001, triazole moieties have become an increasingly prominent theme in chemosensors. Triazoles generated via click reactions are crucial for sensing various ions and biological analytes. Recently, the number of studies in the field of pyrene appendant triazole moieties has risen dramatically, with more sophisticated and reliable triazole-containing chemosensors for various analytes of interest described. This tutorial review provides a general overview of pyrene appendant-triazole-based chemosensors that can detect a variety of metal cations, anions, and neutral analytes by using modular click-derived triazoles.

Bis-butoxyphenyl dipyridine cation with high affinity for PF6−: Enhancement of solid fluorescence and inhibits Escherichia coli and Staphylococcus aureus activity through anion-π+ interactions

The contamination of anions and the overuse of antibiotics pose significant hazards to human health. Excess anions can cause serious damage to organisms and the natural environment, while the misuse of antibiotics has led to the emergence of multi-drug resistant bacteria. Therefore, it is important to develop multifunctional fluorescent sensing devices that integrate the rapid and sensitive detection of specific anions and that exhibit excellent antimicrobial activity against multi-resistant bacteria. This study describes the synthesis of three water-soluble cationic fluorescent molecules, DPPT-R (R = 5C, 6C, 7C). Their structures were characterized using 1H NMR spectroscopy, HRMS, and single crystal X-ray diffraction. Spectroscopic properties of the probes were recorded using UV–visible absorption and fluorescence spectroscopy. The recognition mode and mechanism of the fluorescent probes towards PF6− were investigated. Under aqueous solution conditions involves the PF6− induced aggregation of the probe via strong electrostatic interactions, leading to fluorescence quenching. Adjusting the alkyl chain length of the probe can enhance the sensitivity for the detection of the target anions. The detection limits are 0.892 μM, 0.786 μM and 0.444 μM for R = 5C, 6C, 7C, respectively. In addition, DPPT-R and DPPT-R@PF6 exhibit good inhibitory activity and fluorescence imaging capability against methicillin-resistant Staphylococcus aureus and multidrug-resistant Escherichia coli. Among them, the side chain of DPPT-R@PF6 with an almost 90° bend may be more advantageous for membrane insertion, thus enhancing its antimicrobial activity. In particular, after DPPT-R and PF6− form the DPPT-R@PF6 complex, the complex enhances its solid-state fluorescence, intensity of bacterial fluorescence imaging, and antibacterial activity through anion-π+ interactions. Therefore, this study can provide a new strategy and theoretical basis for the design and application of multifunctional fluorescent sensing materials and antibacterial molecules based on electrostatic interactions and anion-π+ interactions.

Electrochemical Sensor Based on a MXene Nanosheet–Gold Nanourchin Hybrid as a Superoxide Dismutase Mimic for Real-Time Detection of Superoxide Anions Released from Living Cells

Electrochemical Sensor Based on a MXene Nanosheet–Gold Nanourchin Hybrid as a Superoxide Dismutase Mimic for Real-Time Detection of Superoxide Anions Released from Living Cells

Voltammetric Sensing of Chloride Based on a Redox-Active Complex: A Terpyridine-Co(II)-Dipyrromethene Functionalized Anion Receptor Deposited on a Gold Electrode.

A redox-active complex containing Co(II) connected to a terpyridine (TPY) and dipyrromethene functionalized anion receptor (DPM-AR) was created on a gold electrode surface. This host-guest supramolecular system based on a redox-active layer was used for voltammetric detection of chloride anions in aqueous solutions. The sensing mechanism was based on the changes in the redox activity of the complex observed upon binding of the anion to the receptor. The electron transfer coefficient (α) and electron transfer rate constant (k0) for the modified gold electrodes were calculated based on Cyclic Voltammetry (CV) experiments results. On the other hand, the sensing abilities were examined using Square Wave Voltammetry (SWV). More importantly, the anion receptor was selective to chloride, resulting in the highest change in Co(II) current intensity and allowing to distinguish chloride, sulfate and bromide. The proposed system displayed the highest sensitivity to Cl- with a limit of detection of 0.50 fM. The order of selectivity was: Cl- > SO42- > Br-, which was confirmed by the binding constants (K) and reaction coupling efficiencies (RCE).

Advancements in Small Molecule Fluorescent Probes for Superoxide Anion Detection: A Review.

Superoxide anion (O2•-), a significant reactive oxygen species (ROS) within biological systems, plays a widespread role in cellular function regulation and is closely linked to the onset and progression of numerous diseases. To unveil the pathological implications of O2•- in these diseases, the development of effective monitoring techniques within biological systems is imperative. Small molecule fluorescent probes have garnered considerable attention due to their advantages: simplicity in operation, heightened sensitivity, exceptional selectivity, and direct applicability in monitoring living cells, tissues, and animals. In the past few years, few reports have focused on small molecule fluorescence probes for the detection of O2•-. In this small review, we systematically summarize the design and application of O2•- responsive small molecule fluorescent probes. In addition, we present the limitations of the current detection of O2•- and suggest the construction of new fluorescent imaging probes to indicate O2•- in living cells and in vivo.

Synthesis of Cd-Cu nanoparticles supported on HOPG for use as electrocatalytic material for nitrate reduction

Bimetallic nanoparticles application has attracted great attention in the field of electrocatalysis because they can enhance the catalytic properties toward certain reactions of technological interest, such as nitrate reduction. For this purpose, Cd-Cu bimetallic nanoparticles were prepared on highly oriented pyrolytic graphite substrates by sequential electrodeposition of both metals. The process was studied by conventional electrochemical techniques, and the generated deposits were characterized by scanning electron microscopy − energy dispersive X-ray spectroscopy analysis. Initially, Cu nanoparticles were prepared by a simple potentiostatic pulse applied to the carbonaceous substrate. Scanning electron microscopy images showed high density of mainly hemispherical-shaped Cu crystals distributed preferably on step edges and surface defects of the highly oriented pyrolytic graphite electrode. Subsequently, Cd particles were prepared in the same way by immersing the Cu nanoparticles / highly oriented pyrolytic graphite modified substrate in an electrolyte solution containing Cd2+ ions, exploiting the phenomenon of underpotential deposition. It was evidenced that the existing crystals grew in size, most of them showing the typical shapes of those for Cd nanoparticles. The application of the Cd-Cu nanoparticles / highly oriented pyrolytic graphite electrode as a feasible electrocatalyst material for nitrate reduction was investigated by cyclic voltammetry for different nitrate concentrations, also considering the possibility of using this procedure as a method for the anion detection.

Development of a fluorescent scaffold by utilizing quercetin template for selective detection of Hg2+: Experimental and theoretical studies along with live cell imaging

Quercetin is an important antioxidant with high bioactivity and it has been used as SARS-CoV-2 inhibitor significantly. Quercetin, one of the most abundant flavonoids in nature, has been in the spot of numerous experimental and theoretical studies in the past decade due to its great biological and medicinal importance. But there have been limited instances of employing quercetin and its derivatives as a fluorescent framework for specific detection of various cations and anions in the chemosensing field. Therefore, we have developed a novel chemosensor based on quercetin coupled benzyl ethers (QBE) for selective detection of Hg2+ with “naked-eye” colorimetric and “turn-on” fluorometric response. Initially QBE itself exhibited very weak fluorescence with low quantum yield (Φ = 0.009) due to operating photoinduced electron transfer (PET) and inhibition of excited state intramolecular proton transfer (ESIPT) as well as intramolecular charge transfer (ICT) within the molecule. But in presence of Hg2+, QBE showed a sharp increase in fluorescence intensity by 18-fold at wavelength 444 nm with high quantum yield (Φ = 0.159) for the chelation-enhanced fluorescence (CHEF) with coordination of Hg2+, which hampers PET within the molecule. The strong binding affinity of QBE towards Hg2+ has been proved by lower detection limit at 8.47 µM and high binding constant value as 2 × 104 M−1. The binding mechanism has been verified by DFT study, Cyclic voltammograms and Jobs plot analysis. For the practical application, the binding selectivity of QBE with Hg2+ has been capitalized in physiological medium to detect intracellular Hg2+ levels in living plant tissue by using green gram seeds. Thus, employing QBE as a fluorescent chemosensor for the specific identification of Hg2+ will pave the way for a novel approach to simplifying the creation of various chemosensors based on quercetin backbone for the precise detection of various biologically significant analytes.

Phenolate-thiazole based reversible “turn-on” chemosensor for the selective detection of carbonate anion: X-ray crystallography, DFT/TDFT, and cell study

A new phenolate-thiazole derivative (L) has been synthesized and structurally characterized.The chemo-sensing activity of L is detected by the naked eye for the aqueous carbonate anion in the pH range of 4 to 8. The selective ‘turn-on’ fluorescence occurs through the formation of a stable intermediate L∙CO32–(1) following the PET mechanism. The limit of detection (LOD) is found 0.18 µM based on the absorbance-based assay.The quinonoid form of bromophenol unit binds strongly with CO32– through thiazole nitrogen and hydrazinic nitrogen. Further, the selective holding of CO32– anion over other planar tetranuclear anions (e.g., SO32–, NO3–) happens with several intra and intermolecular hydrogen bonds as envisaged by the DFT/TDFT study. The formation mechanism of L∙CO32– is proposed based on experimental and theoretical studies. The biological experiments (MTT and cell imaging)reveal the non-cytotoxicity nature of L and the biocompatible uptake of L mostly in the cytoplasm at physiological pH.

Nonequilibrium Anion Detection in Solid-Contact Ion-Selective Electrodes.

Low-cost and portable nitrate and phosphate sensors are needed to improve farming efficiency and reduce environmental and economic impact arising from the release of these nutrients into waterways. Ion selective electrodes (ISEs) could provide a convenient platform for detecting nitrate and phosphate, but existing ionophore-based nitrate and phosphate selective membrane layers used in ISEs are high cost, and ISEs using these membrane layers suffer from long equilibration time, reference potential drift, and poor selectivity. In this work, we demonstrate that constant current operation overcomes these shortcomings for ionophore-based anion-selective ISEs through a qualitatively different response mechanism arising from differences in ion mobility rather than differences in ion binding thermodynamics. We develop a theoretical treatment of phase boundary potential and ion diffusion that allows for quantitative prediction of electrode response under applied current. We also demonstrate that under pulsed current operation, we can create functional solid-contact ISEs using lower-cost molecularly imprinted polymers (MIPs). MIP-based nitrate sensors provide comparable selectivity against chloride to costlier ionophore-based sensors and exhibit >100,000 times higher selectivity against perchlorate. Likewise, MIP-based solid contact ion-selective electrode phosphate sensors operated under pulsed current provide competitive selectivity against chloride, nitrate, perchlorate, and carbonate anions. The theoretical treatment and conceptual demonstration of pulsed-current ISE operation we report will inform the development of new materials for membrane layers in ISEs based on differences in ion mobility and will allow for improved ISE sensor designs.

Alternative Methods for the Detection of Superoxide Anion Generation in Platelets.

Reactive oxygen species (ROS) are highly unstable oxygen-containing molecules. Their chemical instability makes them extremely reactive and gives them the ability to react with important biological molecules such as proteins, nucleic acids, and lipids. Superoxide anions are important ROS generated by the reduction of molecular oxygen reduction (i.e., acquisition of one electron). Despite their initial implication exclusively in aging, degenerative, and pathogenic processes, their participation in important physiological responses has recently become apparent. In the vascular system, superoxide anions have been shown to modulate the differentiation and function of vascular smooth muscle cells, the proliferation and migration of vascular endothelial cells in angiogenesis, the immune response, and the activation of platelets in hemostasis. The role of superoxide anions is particularly important in the dysregulation of platelets and the cardiovascular complications associated with a plethora of conditions, including cancer, infection, inflammation, diabetes, and obesity. It has, therefore, become extremely relevant in cardiovascular research to be able to effectively measure the generation of superoxide anions byhuman platelets,understand the redox-dependent mechanisms regulating the balance between hemostasis and thrombosis and, eventually,identify novel pharmacological tools for the modulation of platelet responses leading to thrombosis and cardiovascular complications. This study presents three experimental protocols successfully adopted for the detection of superoxide anions in platelets and the study of the redox-dependent mechanisms regulating hemostasis and thrombosis: 1) dihydroethidium (DHE)-based superoxide anion detection by flow cytometry; 2) DHE-based superoxide anion visualization and analysis by single platelet imaging; and 3) spin probe-based quantification of superoxide anion output in platelets by electron paramagnetic resonance (EPR).

Electrochemical biosensing platform based on AuNWs/rGO-CMC-PEDOT:PSS composite for the detection of superoxide anion released from living cells

Detection of superoxide anion (O2·-) levels holds significant importance for the diagnosis and even clinical treatments of oxidative stress-related diseases. Herein, we prepared a composite electrode material to encapsulate copper-zinc superoxide dismutase (SOD1) for biosensing of O2·-. The sensing material consists of gold nanowires (AuNWs), reduced graphene oxide (rGO), carboxymethyl cellulose (CMC) and PEDOT:PSS. CMC provides abundant -COOH to bind SOD1, with a high adsorption coverage of 1.499 × 10−9 mol cm−2 on the sensor surface. rGO and PEDOT endow the composite with significant conductivity, whereas PSS has antifouling capability. Moreover, AuNWs exhibit excellent electrical conductivity and a high aspect ratio, which promotes electron transfer, and ultimately enhances the catalytic performance of the enzyme. Meanwhile, SOD1(Cu2+) catalyzes the dismutation of O2·- to O2 and H2O2, and H2O2 is then electrochemically oxidized to generate amperometric signals for determination of O2·-. The sensor demonstrates outstanding detection performance for O2·- with a low detection limit of 2.52 nM, and two dynamic ranges (14.30 nM–1.34 μM and 1.34 μM–42.97 μM) with corresponding sensitivity of 0.479 and 0.052 μA μM−1cm−2, respectively. Additionally, the calculated apparent Michaelis constant (Kmapp) of 1.804 μM for SOD1 demonstrates the outstanding catalytic activity and the surface-immobilized enzyme's substrate affinity. Furthermore, the sensor shows the capability to dynamically detect the level of O2·- released from living HepG2 cells. This study provides an inovative design to obtain a biocompatible electrochemical sensing platform with plenty of immobilization sites for biomolecules, large surface area, high conductivity and flexibility.

A Novel Bisquaternary Ammonium Compound as an Anion Sensor-ESI-MS and Fluorescence Study.

Electrospray ionization mass spectrometry (ESI-MS) analysis is frequently associated with noncovalent adduct formation, both in positive and negative modes. Anion binding and sensing by mass spectrometry, notably more challenging compared to cation binding, will have major research potential with the development of appropriate sensors. Here, we demonstrated identification of stable bisquaternary dication adducts with trifluoroacetate (TFA-), Cl- and HSO4- in positive-mode ESI-MS analysis. The observed adducts were stable in MS/MS mode, leading to the formation of characteristic fragment ions containing a covalently bound anion, which requires bond reorganization. This phenomenon was confirmed by computational methods. Furthermore, given that anion detection and anion sensor chemistry have gained significant prominence in chemistry, we conducted an analysis of the fluorescent properties of bisquaternary ammonium compound as a potential anion sensor.

Theoretical Investigation of Bimolecular Carbon Chain Growth Reactions in the Interstellar Media.

Detection of molecular anions in interstellar media implies that negatively charged species play a prominent role in astrochemical reactions. Among the observed species, carbon chain anions are important, as they can be precursors for the production of complex organic molecules. These anions can form either via direct electron attachment to the corresponding neutral species or through chain growth reactions of smaller anions, resulting in longer chains. In a recent study, crossed beam experiments coupled with velocity map imaging techniques were used to investigate the carbon chain growth reaction C2n- + C2H2 → C2n+2Hm- + H2-m (n = 1-3, m = 0,1). Products and branching ratios were established from experimental data. In the present work, electronic structure calculations and on-the-fly direct dynamics simulations were used to study these reactions. Energy profiles were investigated by using different density functional methods. Direct trajectory simulations were performed at the experimental collision energies using the B3LYP/6-31+G* level of theory. Trajectory analysis showed a variety of reaction pathways, and detailed atomic-level reaction mechanisms are presented.

A Enhanced Fluorescent Probe for Simultaneous Detection and Discrimination of Hydrogen Bisulfite Anions and Glutathione.

Bisulfite (HSO3-) and biological thiols molecules, such as glutathione (GSH), cysteine (Cys), and homocysteine (Hcy), play important roles in organisms. Developing a fluorescent probe that can simultaneously detect and distinguish HSO3- and biological thiols is of great significance. In this study, ethyl(2E,4Z)-5-chloro-2-cyano-5-(7-(diethylamino)-2-oxo-2H-chromen-3-yl)penta-2,4-dienoate (CCO) as a novel enhanced fluorescence probe was synthesized by integrating coumarin derivatives and ethyl cyanoacetate, which can simultaneous detection and discrimination of hydrogen bisulfite anions and glutathione. The sensing mechanism was elucidated through spectral analysis and some control experiments. In weakly alkaline environments, the probe not only has good selectivity for HSO3- and GSH, but also has a lower detection limits of 0.0179 µM and 0.2034 µM. The probe exhibited fuorescent turn-on for distinguishing with 296 and 28 fold the fluorescent intensity increase at 486 and 505nm, respectively, through diferent excitation wavelengths. This provides a new method for simultaneous detection and discrimination of HSO3- and biological thiol cell levels and further applications.