A novel lysosome-located fluorescent probe for highly selective determination of hydrogen polysulfides based on a naphthalimide derivative.

Hydrogen polysulfides (H2Sn, n > 1) belongs to sulfane sulfur in the reactive sulfur species (RSS) family and plays a significant regulatory role in organisms. Highly selective and lysosome-located probes for detecting hydrogen polysulfides are rare. Thus, it is important to develop a technique to detect the changes of H2Sn level in lysosomes. In this work, a lysosome-targeting fluorescent probe for H2Sn was designed and developed based on a naphthalimide derivative. 4-Hydroxynaphthalimide was selected as the fluorescent group and 2-chloro-5-nitrobenzoate group was used as a specific recognition unit for H2Sn. A morpholine unit was chosen as a lysosome-located group. In the absence of H2Sn, the fluorescent probe exhibited almost no fluorescence. In the presence of H2Sn, the fluorescent probe showed strong fluorescence owing to H2Sn-mediated aromatic substitution-cyclization reactions. The fluorescence emission intensity at 548 nm of the probe showed a good linear relationship toward H2Sn in the range of 2.0 × 10-7 - 9.0 × 10-5 mol·L-1, and the detection limit was found to be 1.5 × 10-7 mol·L-1. The probe possessed a wide work range of pH, including the pH of physiological environment, and high selectivity for H2Sn. There are almost no cytotoxicity and the ability of detecting endogenous and exogenous H2Sn in lysosomes. These results indicate that the fluorescent probe can provide a good tool for intracellular and extracellular detection of H2Sn.

A Highly Selective Fluorescent Probe for Hypochlorous Acid in Living Cells Based on a Naphthalene Derivative.

Hypochlorous acid (HOCl) was crucial for maintaining the homeostasis in cells and plays vital roles in many physiological and pathological processes. In this work, a highly selective fluorescent probe for hypochlorous acid in living cells was constructed and prepared based on a naphthalene derivative. A naphthalene derivative was utilized as the fluorescent group, and N,N-dimethylthiocarbamate was applied as the selective recognition site for HOCl. Before adding HOCl, the fluorescent probe exhibited weak fluorescence. Upon adding HOCl, the fluorescent probe displayed remarkable fluorescence enhancement. The fluorescence intensity at 502 nm showed a linear response to the concentration of HOCl from 3.0 × 10-7 to 1.0 × 10-5 mol·L-1. The detection limit was estimated to be 1.5 × 10-7 mol·L-1 for HOCl. The fluorescent probe showed fast response and outstanding selectivity toward HOCl. It owned good biocompatibility and had also been successfully applied in the confocal imaging of exogenous and endogenous HOCl in living cells.

Ratiometric Determination of Nitroxyl Utilizing a Novel Fluorescence Resonance Energy Transfer-Based Fluorescent Probe Based on a Coumarin-Rhodol Derivative.

Nitroxyl (HNO) is a member of the reactive nitrogen species, and how to detect it quickly and accurately is a challenging task. In this work, we designed and prepared a fluorescent ratiometric probe based on the fluorescence resonance energy transfer (FRET) mechanism, which can detect HNO with high selectivity. The coumarin derivative was used as an energy donor, the rhodol derivative was applied as an energy receptor, and 2-(diphenylphosphine)benzoate was utilized as the recognition group to detect nitroxyl. In the absence of HNO, the rhodol derivative exists in a non-fluorescent spironolactone state, and the FRET process is inhibited. Upon adding HNO, the closed spironolactone form is transformed into a conjugated xanthene structure and the FRET process occurs. This probe could specifically recognize nitroxyl, showing high sensitivity and selectivity. When the HNO concentration was changed from 3.0 × 10-7 to 2.0 × 10-5 mol·L-1, I 543nm/I 470nm exhibited a satisfactory linear correlation with the concentration of HNO. A detection limit of 7.0 × 10-8 mol·L-1 was obtained. In addition, almost no cell toxicity had been verified for the probe. The probe had been successfully applied to the ratiometric fluorescence imaging of HNO in HepG2 cells.

Novel Mitochondria-Targeting and Naphthalimide-based Fluorescent Probe for Detecting HClO in Living Cells.

As a key reactive oxygen species (ROS), hypochlorous acid (HClO) plays an important role in many physiological and pathological processes. The mitochondria-targeting probes for the highly sensitive detection of HClO are desirable. In present work, we designed and synthesized an original mitochondria-localizing and turn-on fluorescent probe for detecting HClO. 4-Aminonaphthalimide was employed as the fluorescent section, the (2-aminoethyl)-thiourea unit was utilized as a typical sensing unit, and the quaternized pyridinium moiety was used as a mitochondria-targeted localization group. When HClO was absent, the probe showed weak fluorescence. In the existence of HClO, the probe revealed a blue fluorescence. Moreover, the turn-on fluorescent probe was able to function in a broad pH scope. There was an excellent linearity between the fluorescence emission intensity at 488 nm and the concentrations of HClO in the range of 5.0 × 10-7 to 2.5 × 10-6 mol·L-1. Additionally, the probe had almost no cell toxicity and possessed an excellent mitochondria-localizing capability. Furthermore, the probe was able to image HClO in mitochondria of living PC-12 cells. The above remarkable properties illustrated that the probe was able to determine HClO in mitochondria of living cells.

A highly selective fluorescent probe for hydrogen polysulfides in living cells based on a naphthalene derivative.

Hydrogen polysulfides (H2Sn, n > 1) are members of reactive sulfur species (RSS) and signaling molecules derived from hydrogen sulfide (H2S). Recently, the functions of H2Sn in physiological and pathological processes have been increasingly recognized. However, their biological effects and detailed mechanisms of action are still little known. Therefore, there is an urgent need to develop highly selective and sensitive techniques for monitoring hydrogen polysulfides (H2Sn) in living cells. In this study, we designed and synthesized a fluorescent probe based on a naphthalene derivative for the detection of hydrogen polysulfides. A naphthalene derivative was applied as the fluorescent main structure and the 2-fluoro-5-nitrobenzoate group was chosen as the recognition unit. In the absence of hydrogen polysulfides, the fluorescent probe displayed almost no fluorescence. In the presence of hydrogen polysulfides, the fluorescent probe exhibited strong fluorescence. The sensing mechanism was based on H2Sn-mediated aromatic substitution-cyclization reactions. The linear range of the response concentration of the probe to hydrogen polysulfide was acquired in a concentration range of H2Sn from 7.5 × 10-7 to 2.5 × 10-5 mol L-1. The detection limit was evaluated to be 5.0 × 10-7 mol L-1 for H2Sn. The fluorescent probe can applied in a wide pH range including physiological condition pH. The fluorescent probe showed high specificity for H2Sn over other reactive sulfur species (RSS). Moreover, the fluorescent probe has been successfully applied to confocal imaging of hydrogen polysulfides in HepG2 cells without cell cytotoxicity. All of such good qualities indicated that it could be used to detect H2Sn in living cells.

A Highly Selective Fluorescent Probe for Hg2+ Based on a 1,8-Naphthalimide Derivative.

Hg2+ has a significant hazardous impact on the environment and ecosystem. There is a great demand for new methods with high selectivity and sensitivity to determine mercury in life systems and environments. In this paper, a novel turn-on Hg2+ fluorescent probe has been reported with a naphthalimide group. The Hg2+ fluorescent probe was designed by the inspiration of the well-known specific Hg2+-triggered thioacetal deprotection reaction. A 1,2-dithioalkyl group was chosen as the specific recognition site of Hg2+. The probe showed weak fluorescence without Hg2+, and the color of the solution was light yellow. In the presence of Hg2+, the probe reacted specifically with the mercury ion to produce an aldehyde and emitted strong fluorescence, and the color of the solution also turned light green, thus realizing the monitoring of the mercury ion. The Hg2+ fluorescent probe showed outstanding sensitivity and selectivity toward Hg2+. Furthermore, the Hg2+ fluorescent probe could work in a wide pH range. The linear relationship between the fluorescence intensity at 510 nm and the concentration of Hg2+ was obtained in a range of Hg2+ concentration from 2.5 × 10-7 to 1.0 × 10-5 M. The detection limit was found to be 4.0 × 10-8 M for Hg2+. Furthermore, with little cell toxicity, the probe was successfully applied to the confocal image of Hg2+ in PC-12 cells.

A FRET-based ratiometric fluorescent probe for highly selective detection of hydrogen polysulfides based on a coumarin-rhodol derivative.

In modern biology, hydrogen polysulfides (H2Sn, n > 1) are members of reactive sulfur species (RSS), with anti-oxidation, cell protection and redox signals in tissues and organs. Therefore, it is crucial to develop a method to monitor the changes of H2Sn level in organisms. We designed and synthesized a ratiometric fluorescent probe for highly selective detection of H2Sn based on the fluorescence resonance energy transfer (FRET) process. In this work, a coumarin derivative was chosen as an energy donor, a rhodol derivative was used as an energy acceptor and a 2-fluoro-5-nitrobenzoate group was applied as a recognition unit for H2Sn. In the absence of H2Sn, the rhodol receptor existed in the non-fluorescent spirolactone state and FRET process was disabled. In the presence of H2Sn, the closed spirolactone form was converted to a conjugated fluorescent xanthenes form to invoke the occurrence of FRET which resulted in a 77 nm red-shift of fluorescence emission from 460 nm to 537 nm. The ratio value of the fluorescence intensity between 537 nm and 460 nm (I537nm/I460nm) of the probe exhibited a good linear relationship toward H2Sn in the range of 3.0 × 10-6-1.0 × 10-4 mol·L-1, and the detection limit was estimated to be 8.0 × 10-7 mol·L-1. In addition, the ratiometric fluorescent probe showed high specificity for H2Sn over other biologically related species. Moreover, the probe displayed little cell toxicity and had been successfully used to the confocal imaging of H2Sn in HepG2 cells by dual emission channels.