Binding of azobenzene and p-diaminoazobenzene to the human voltage-gated sodium channel Nav1.4.

The activity of voltage-gated ion channels can be controlled by the binding of photoswitches inside their internal cavity and subsequent light irradiation. We investigated the binding of azobenzene and p-diaminoazobenzene to the human Nav1.4 channel in the inactivated state by means of Gaussian accelerated molecular dynamics simulations and free-energy computations. Three stable binding pockets were identified for each of the two photoswitches. In all the cases, the binding is controlled by the balance between the favorable hydrophobic interactions of the ligands with the nonpolar residues of the protein and the unfavorable polar solvation energy. In addition, electrostatic interactions between the ligand and the polar aminoacids are also relevant for p-diaminoazobenzene due to the presence of the amino groups on the benzene moieties. These groups participate in hydrogen bonding in the most favorable binding pocket and in long-range electrostatic interactions in the other pockets. The thermodinamically preferred binding sites found for both photoswitches are close to the selectivity filter of the channel. Therefore, it is very likely that the binding of these ligands will induce alterations in the ion conduction through the channel.

Associations of Water-Soluble Macrocyclic Hosts with 4-Aminoazobenzene: Impact of pH.

An investigation of the pH effect on the inclusion complexes of ß-cyclodextrins and calixarenesulfonates with 4-aminoazobenzene was conducted both by experiments and molecular simulations. The whole thermodynamic characterizations of the association between hosts and 4-aminoazobenzene ( K, Δr G0, Δr H0, and TΔr S0) were determined by UV-visible spectroscopy. ß-Cyclodextrin inclusion complexes are not affected by pH change unlike those obtained with calixarenes. All the studied systems were enthalpically favored. Nevertheless, the entropic behavior is different depending on the host. In order to interpret these experimental results, molecular simulations were used to calculate the number of atoms inserted into the cage-like host compounds and the number of water molecules expelled from the cavity.

Interactions of 4,4'-diaminoazobenzene derivatives with telomeric G-quadruplex DNA.

The development of small molecules to stabilize the G-quadruplex structure has garnered significant attention for anticancer drug discovery. Herein, we report the synthesis of several 4,4'-diaminoazobenzene derivatives containing different substituent groups and their ability to bind and stabilize telomeric G-quadruplex DNA. Circular dichroism (CD) spectroscopy was performed to characterize the quadruplex topologies, measure stabilization effects, and evaluate their capabilities for conformational photoregulation. 4,4'-Diaminoazobenzene derivatives were found to moderately stabilize quadruplex structures but not affect conformational photoregulation. This work further develops the design and general understanding of the stabilization effects of small molecules with telomeric G-quadruplex DNA.

A highly facile and selective Chemo-Paper-Sensor (CPS) for detection of strontium.

Chemosensors have attracted increasing attention for their usefulness on-site detection and monitoring. In this study, we elucidated a novel, facile, and highly selective Chemo-Paper-Sensor (CPS) for detection and monitoring of strontium (Sr(2+)) ions, which means a potent colorimetric sensor based on a Chrysoidine G (CG)-coated paper strip. The CPS for highly selective colorimetric detection of strontium ion was handily analyzed to determine the red-green-blue (RGB) value using portable devices such as desktop digital scanner and mobile phone camera, quantitatively. Interestingly, an orange to dark orange color transition was observed when the aqueous and solid paper colorimetric sensor was introduced to Sr(2+) ion, respectively. It was demonstrated that the value of the signal has a linear relationship with concentrations of the strontium in the 500 ppm to 100 ppb range with a detection limit of 200 ppb. We believe that a newly developed Chemo-Paper-Sensor will be useful in a wide range of sensing applications.

Synergistic action of thermoresponsive and hygroresponsive elements elicits rapid and directional response of a bilayer actuator.

A bilayer actuator composed of thermoresponsive and thermo/hygroresponsive elements is developed, which undergoes fast, directional and autonomous curling with a speed of up to 0.7 m s(-1) and recovers its shape by hydration. In situ tensile testing of the thermal response of individual layers provided insights into the mechanism of actuation of thermo/hygromorphic bilayers.

CdTe quantum dots@luminol as signal amplification system for chrysoidine with chemiluminescence-chitosan/graphene oxide-magnetite-molecularly imprinting sensor.

A sensitive chemiluminescence (CL) sensor based on chemiluminescence resonance energy transfer (CRET) in CdTe quantum dots@luminol (CdTe QDs@luminol) nanomaterials combined with chitosan/graphene oxide-magnetite-molecularly imprinted polymer (Cs/GM-MIP) for sensing chrysoidine was developed. CdTe QDs@luminol was designed to not only amplify the signal of CL but also reduce luminol consumption in the detection of chrysoidine. On the basis of the abundant hydroxy and amino, Cs and graphene oxide were introduced into the GM-MIP to improve the adsorption ability. The adsorption capacities of chrysoidine by both Cs/GM-MIP and non-imprinted polymer (Cs/GM-NIP) were investigated, and the CdTe QDs@luminol and Cs/GM-MIP were characterized by UV-vis, FTIR, SEM and TEM. The proposed sensor can detect chrysoidine within a linear range of 1.0×10(-7) - 1.0×10(-5) mol/L with a detection limit of 3.2×10(-8) mol/L (3δ) due to considerable chemiluminescence signal enhancement of the CdTe quantum dots@luminol detector and the high selectivity of the Cs/GM-MIP system. Under the optimal conditions of CL, the CdTe QDs@luminol-Cs/GM-MIP-CL sensor was used for chrysoidine determination in samples with satisfactory recoveries in the range of 90-107%.

Toxic effects of chrysoidine on human serum albumin: isothermal titration calorimetry and spectroscopic investigations.

Chrysoidine is widely used in industry as a type of azo dye, and is sometimes used illegally as a food additive despite its potential toxicity. Human serum albumin (HSA) is one of the most important proteins in blood plasma and possesses major physiological functions. In the present study, the conformational and functional effects of chrysoidine on HSA were investigated by isothermal titration calorimetry (ITC), multiple spectroscopic methods, a molecular docking study and an esterase activity assay. Based on the ITC results, the binding stoichiometry of chrysoidine to HSA was estimated to be 1.5:1, and was a spontaneous process via a single hydrogen bond. The binding of chrysoidine to HSA induced dynamic quenching in fluorescence, and changes in secondary structure and in the microenvironment of the Trp-214 residue. In addition, the hydrogen bond (1.80 Å) formed between the chrysoidine molecule and the Gln-211 residue. The esterase activity of HSA decreased following the addition chrysoidine due to the change in protein structure. This study details the direct interaction between chrysoidine and HSA at the molecular level and the mechanism for toxicity as a result of the functional changes induced by HSA structural variation upon binding to chrysoidine in vitro. This study provides useful information towards detailing the transportation mechanism and toxicity of chrysoidine in vivo.

SiO2@Au nanoshells-based SERS method for detection of sunset yellow and chrysoidine.

A novel surface-enhanced Raman scattering (SERS) substrate made up of SiO2@Au nanoshells has been developed for detecting sunset yellow and chrysoidine. It exhibits an excellent enrichment SERS effect on these two colorant molecules. In this work, density functional theory (DFT) calculations have been carried out to determine the molecular structure and theoretical Raman spectra. This provided a good description of the characteristic peaks of the molecules. In addition, the appropriate thicknesses of the shell and environment pH have been derived to obtain improved SERS signals. The lowest concentration is 1 ppm and 0.5 ppm for sunset yellow and chrysoidine, respectively. Under optimal detection condition, it has proved possible to distinguish each colorant by its characteristic peaks in the SERS spectra of a mixture of the two colorants.

Azo dye/cyclodextrin: new findings of identical nanorods through 2:2 inclusion complexes.

Inclusion complexation behavior of 4-aminoazobenzene (AAB) and 4-amino-2,3'-dimethyl azobenzene (GBC, fast grant GBC) with α- and ß-cyclodextrins (α-CD, ß-CD) is analyzed by scanning electron microscope, transmission electron microscope, Fourier transform infrared spectroscopy, differential scanning calorimetry, powder X-ray diffraction, and proton nuclear magnetic resonance spectroscopy techniques. Transmission electron microscope analysis suggests that identical nanorods formed in AAB/CD inclusion complexes while different dimension nanostructures were observed in GBC/CD inclusion complexes. The nanostructures confirmed that the ratio of 2:2 (guest:host) inclusion complex has been developed to a miniature nanorod. Nanosecond time-resolved fluorescence studies indicated that AAB/GBC have fast life time in water, whereas slow life time in CDs corresponds to a higher-order structure of 2:2 complexes. Thermodynamic parameters and binding affinity of the inclusion complex formation were determined and discussed. van der Waals interactions are mostly responsible for enthalpy-driven complex formation of AAB and GBC with cyclodextrins.

Binding of chrysoidine to catalase: spectroscopy, isothermal titration calorimetry and molecular docking studies.

Chrysoidine is an industrial azo dye and the presence of chrysoidine in water and food has become an environmental concern due to its negative effects on human beings. In this work, the interactions between chrysoidine and bovine liver catalase (BLC) were explored. Obvious loss in catalytic activity was observed after incubation of BLC with chrysoidine, and the inhibition effect of BLC was found to be of the non-competitive type. No profound conformational change of BLC occurs in the presence of chrysoidine as revealed by UV-vis absorption, circular dichroism and fluorescence spectroscopy studies. Isothermal titration calorimetry results indicate that catalase has two sets of binding sites for chrysoidine. Further, molecular docking simulations show that chrysoidine is located within the bottleneck in the main channel of the substrate to the active site of BLC, which explain the activity inhibition of BLC by chrysoidine.

Determination method for p-Phenylazoaniline and 2-methyl-4-(2-tolylazo)aniline in workplace air by high-performance liquid chromatography.

OBJECTIVES: The purpose of this research was to develop a method for the simultaneous determination of p-Phenylazoaniline (also called 4-aminoazobenzene, AAB) and 2-methyl-4-(2-tolylazo)aniline (also called o-aminoazotoluene, AAT) in workplace air for risk assessment. METHODS: The characteristics of the proposed method, such as recovery, limit of quantitation, reproducibility and storage stability of the samples were examined. RESULTS: An air sampling cassette containing two sulfuric acid-treated glass fiber filters was chosen as the sampler. The AAB and AAT were extracted from the sampler filters by methanol and then analyzed by a high-performance liquid chromatograph equipped with a photo-diode array detector. The overall recoveries from spiked samplers were 77-98 and 85-98% for AAB and AAT, respectively. The recovery after 5 days of storage in a refrigerator exceeded 96%. The overall limits of quantitation were 5.00 and 2.50 µg/sample for AAB and AAT, respectively. The relative standard deviations, which represent the overall reproducibility defined as precision, were 0.6-1.8 and 0.5-2.2% for AAB and AAT, respectively. CONCLUSIONS: The proposed method enables 4-h personal exposure monitoring of AAB and AAT at concentrations of 21 to 2,000 µg/m3 for AAB and 10 to 2,000 µg/m3 for AAT, respectively. The proposed method is useful for estimating worker exposure to AAB and AAT.

Higuchi's equation and beyond: overview of the formulation and application of a generalized model of drug release from polymeric matrices.

An account is presented of modeling and experimental work, which complements, in many useful ways, the excellent coverage, afforded by a recent dedicated issue edited by Siepmann and Peppas (Int. J. Pharm. 2011, 418(1)), of the theoretical background and many ramifications, as well as practical applications, of the Higuchi equation. The main notable feature of the said modeling work is formulation and successful practical application, of an ab initio generalized approach, based on solving the fundamental transport equations applicable to the operation of typical matrix-controlled release (MCR) devices. This approach (i) reduces to the Higuchi equation under the pertinent restrictive conditions but can duly handle most of its more complex ramifications and (ii) can be parameterized (by the use of appropriate data from the literature or from independent experiments) to adapt itself to the physics of the particular matrix-solvent-solute system under consideration; as demonstrated here with examples from our laboratory or from the literature. A distinctive feature of the experimental work is the extensive use of simpler "model" MCR systems, chosen so as to promote better understanding of the effect of various physicochemical parameters and mechanisms on drug MCR rate and kinetics. A concept of MCR device efficiency is also discussed.

Ionic liquid-based liquid phase microextraction with direct injection for capillary electrophoresis.

Liquid-liquid microextraction using the water immiscible ionic liquid, 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, EMIM NtfO2, for the concentration and cleanup of basic compounds for analysis by CE has been investigated. Using an electrolyte comprising 1 mol/L alanine and 3 mol/L acetic acid, EMIM NtfO2 could be directly injected into the capillary after liquid phase extraction. Using the basic dye chryisoidine, sensitivity enhancements approaching 1000-fold were obtained by mixing 20 µL of EMIM NtfO2 with 1500 µL of aqueous sample, leaving only 5 µL of the undissolved ionic liquid which was used for injection into the CE. Lower more repeatable enhancement factors of 200-fold were obtained with slightly larger initial 25 µL volumes of EMIM NtfO2 due to the larger residual volume of ionic liquid which made handling easier. This could be extended to basic pharmaceuticals, and the extraction of clozapine and its two active metabolites, nor-clozapine and clozapine-N-oxide, was demonstrated from urine with enrichment factors greater than 100 obtained. Handling of potentially more dangerous samples, such as serum, through in-vial extraction of clozapine and its metabolites and direct injection of the ionic liquid layer was also demonstrated with enhancements in sensitivity of 80. Limits of detection from 3 to 11 µg/L and 6 to 55 µg/L were obtained from urine and serum, respectively, which are sufficiently low to be useful for the determination of these pharmaceuticals clinically for therapeutic drug monitoring and for forensic toxicology.

Azonium-ammonium tautomerism and inclusion complexation of 1-(2,4-diamino phenylazo) naphthalene and 4-aminoazobenzene.

Spectral characteristics of 1-(2,4-diamino phenylazo) naphthalene (FBRR, fat brown RR) and 4-aminoazobenzene (AAB) have been studied in various solvents, varying hydrogen ion concentrations and in ß-cyclodextrin (ß-CD). The inclusion complex of FBRR and AAB with ß-CD were analysed by UV-visible, fluorometry and CAche-DFT methods. Solvent study reveals that only azo tautomer is present in both compounds and the large red shifted absorption and emission maxima of FBRR indicate naphthalene ring effectively increases the π-π* transition. Unusual red shift is observed in acid solutions suggests azonium-ammonium tautomer is present in both molecules. In ß-CD solutions, the large hypsochromic shift is observed in S(0) and S(1) states indicates ortho amino group of FBRR molecule is entrapped in the ß-CD cavity and the large bathochromic shift for AAB in the S(1) state indicates 1:1 inclusion complex is formed.

Photoinduced surface relief grating formation for a single crystal of 4-aminoazobenzene.

Photoinduced surface relief grating (SRG) formation for a single crystal of 4-aminoazobenzene was investigated. It was found that SRG could be inscribed on the (001) surface of the crystal, which might suggest that the photoinduced SRG formation is a general phenomenon observed for single crystals of azobenzene-based molecules as well as for azobenzene-based amorphous systems. In addition, the dependences of the SRG formation upon the orientation of the sample crystal and upon the polarization of the writing beams were found to be different from those observed for previously reported crystalline systems.

Removal and recovery of Chrysoidine Y from aqueous solutions by waste materials.

This article describes the use of bottom ash [a power plant waste] and de-oiled soya [an agricultural waste] as effective adsorbents for the removal of a hazardous azo dye [Chrysoidine Y] from its aqueous solutions. This paper presents an experimental study and discussion of the adsorption characteristics of this dye on the two adsorbents. The adsorbents have been characterized, and also the effects of time, temperature, concentration, pH, and sieve size on the extent of adsorption have been evaluated. Batch adsorption measurements, kinetic studies, and column operations have been performed to elucidate the dye uptake capacity of the adsorbents. The monolayer adsorption capacities at 30 degrees C have been found from Langmuir analysis to be 7.27x10(-5) mol g(-1) and 3.35x10(-5) mol g(-1) for bottom ash and de-oiled soya, respectively. Adsorption kinetics experimental data are indicative of pseudo-second order kinetics during these processes. Column experiments indicate practical utility of the adsorbents for eradicating hazardous dyes from effluents. The recovery of the adsorbed dye from bottom ash and de-oiled soya, have been found to be 85% and 99%, respectively.

Rapid method for the confirmatory analysis of chrysoidine in aquaculture products by ultra-performance liquid chromatography-tandem mass spectrometry.

A sensitive and fast method for the quantification of the illegal dye chrysoidine in aquaculture products with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) is presented. Muscle tissues were made alkaline with sodium hydroxide and extracted with ethyl acetate. After evaporation and subsequent defatting with n-hexane, extracts were directly injected onto the UPLC-column. Chromatography was performed on a C(18) column using 0.1% formic acid in water and an acetonitrile gradient within 6 min. Mass spectrometric analysis was performed in the positive electrospray MS/MS mode. The limit of quantification was 0.25 ng/g, which was 30 times lower than the only previously published method with gas chromatographic detection. A complete validation according to the scientific literature and as defined by the European Union was performed. The applicability of the method was shown in the analysis of more than 50 unknown samples in the framework of a monitoring program.

Investigation on the toxic interaction of chrysoidine hydrochloride-CTMAB combined contamination with calf thymus DNA.

The toxic interaction of the azo dye-chrysoidine hydrochloride combined with cetyltrimethyl ammonium bromide (CTMAB) in living tissue was studied in vitro. The absorption spectrum, resonance light scattering (RLS), circular dichroism (CD) and transmission electron microscopy (TEM) results showed that the toxicity of chrysoidine hydrochloride itself to calf thymus DNA (ct-DNA) is weak, while the chrysoidine hydrochloride-CTMAB combined pollution showed obvious toxic interaction with ct-DNA. The chrysoidine hydrochloride-CTMAB combined contamination can interact with ct-DNA to form an ion-associated complex through electrostatic and hydrophobic forces. The conformation of DNA was changed in the interaction process to show toxic. The experimental results showed that the combination of chrysoidine hydrochloride-CTMAB has higher toxicity to ct-DNA than either chrysoidine hydrochloride or CTMAB individually, and the combined pollution showed a strong toxic co-effect at a dose of 3.0x10(-4) mol L(-1) chrysoidine hydrochloride and 1.6x10(-5) mol L(-1) CTMAB.

High-sensitivity aminoazobenzene chemisorbed monolayers for photoalignment of liquid crystals.

We describe a new type of optically controlled liquid crystal alignment layer that demonstrates unprecedented performance. It consists of an aminoazobenzene-type material with a very simple molecular structure, which is derived from methyl red by a one-step synthesis. We have devised a method of forming covalently attached monolayers of this material on glass by an amine-assisted condensation reaction involving the triethoxysilane end of the molecule. A nematic liquid crystal (LC) cell made with the monolayer and a rubbed polymer layer was switched from a uniform state to a twisted state with a polarized 450 nm control beam having a dose of 5.5 mJ/cm(2). This is equivalent to an average of only one absorbed photon per azobenzene group. Through atomic force microscopy, absorption spectroscopy, spectroscopic ellipsometry, and second harmonic generation experiments, we have confirmed that layers of this type are smooth and uniform with a surface coverage consistent with a monolayer and that the azobenzene groups are tilted, on average, 55 degrees with respect to the surface normal. These characteristics lead to a large interaction energy density between the layer and LC. The monolayer's rapid response in developing anisotropy in this property can be attributed to a large absorption cross section, as well as the favorable tilt angle, which allows for sufficient photoisomerization free volume in a dense layer.

Cloud point extraction and flame atomic absorption spectrometry combination for copper(II) ion in environmental and biological samples.

A cloud point extraction procedure was presented for the preconcentration of copper(II) ion in various samples. After complexation by 4-(phenyl diazenyl) benzene-1,3-diamine (PDBDM) (chrysoidine), copper(II) ions were quantitatively recovered in Triton X-114 after centrifugation. 0.5ml of methanol acidified with 1.0molL(-1) HNO3 was added to the surfactant-rich phase prior to its analysis by flame atomic absorption spectrometry (FAAS). The influence of analytical parameters including ligand, Triton X-114 and HNO3 concentrations, bath temperature, heating time, centrifuge rate and time were optimized. The effect of the matrix ions on the recovery of copper(II) ions was investigated. The detection limit (3S.D.b/m, n=10) of 0.6ngmL(-1) along with preconcentration factor of 30 and enrichment factor of 41.1 with R.S.D. of 1.0% for Cu was achieved. The proposed procedure was applied to the analysis of various environmental and biological samples.