Colorimetric chiral fluorescent sensors for Eu3+ and sequential enantioselective sensing of malate anion

Novel phenanthroline Schiff base fluorescent sensors L1 , L2 , and D1 were designed and synthesized. The sensing abilities of the compounds in the presence of metal cations (Li⁺, Na⁺, K⁺, Ag⁺, Mg²⁺, Ba²⁺, Ca²⁺, Mn²⁺, Pb²⁺, Hg²⁺, Ni²⁺, Zn²⁺, Cd²⁺, Co²⁺, Cu²⁺, Cr³⁺, Fe³⁺, Fe²⁺, Al³⁺, and Eu³⁺) were studied by UV‐vis and fluorescent spectroscopy. The compounds L1 , L2 , and D1 could act as Eu³⁺ ion turn‐off fluorescent sensors based on ligand‐to‐metal binding mechanism in DMSO‐H₂O solution (v/v = 1:1, 10 mM Tris, pH = 7.4). Additionally, the L1 –Eu³⁺ and D1 –Eu³⁺ complexes could be applied as turn‐on enantioselective sensors sensing of malate anion isomers with color changes. Furthermore, biological experiments using living PC‐12 cells demonstrated that L1 and D1 had excellent membrane permeability and could be used as effective fluorescent sensors for detecting Eu³⁺ and malate anion in living cells.     

'Naked-eye' enantioselective chemosensors for N-protected amino acid anions bearing thiourea units

Four linear thiourea anion receptors (1-3) derived from simple amino acid have been synthesized and their bonding properties with various chiral N-protected amino acid anions were examined by using UV-vis and fluorescence titration experiments. Receptors 1a, 2, and 3 exhibit excellent enantioselective recognition abilities towards N-Boc-protected alanine anion in the UV-vis spectra, obvious difference in the color of solution indicate that the enantiomers of N-Boc-alanine anion could be distinguished by naked eye directly. Receptor 1a is also found to carry out enantioselective fluorescent recognition of the N-acetyl-glutamate. (1)H NMR experiments suggest that hydrogen-bonding interaction between the host and guest is the main factor in the recognition process.     

A ratiometric fluorescent probe for bisulphite anion,employing intramolecular charge transfer

4‐(1H‐benzimidazol‐2‐yl)benzaldehyde (1) has been developed as a new ratiometric fluorescent probe for bisulphite, based on the modulation of intramolecular charge transfer (ICT). Upon mixing with bisulphite in aqueous ethanol, an aldehyde–bisulphite adduct was formed and the ICT of the probe was switched off, which resulted in a ratiometric fluorescence response with an enhancement of the ratios of emission intensities at 368 and 498 nm. The detection range of the probe for bisulphite is in the 2.0–200 µmol/L concentration range and the detection limit is 0.4 µmol/L. Probe 1 produces a ratiometric fluorescent response to bisulphite with a marked emission wavelength shift (130 nm) and displays high selectivity for bisulphite over other anions. Copyright © 2012 John Wiley & Sons, Ltd.     

Multimode selective detection of mercury by chiroptical fluorescent sensors based on methionine/cysteine

Two multimode Hg(II) sensors, L‐MethBQA and L‐CysBQA, were obtained by fusing methionine or S‐methyl cysteine, into a bis‐quinolyl amine‐based chiral podand scaffold. Quinolyl groups serve as the fluorophore and possess nitrogen lone pairs capable of chelating metal ions. On exposure to Hg²⁺ or Zn²⁺, these sensors show signal enhancement in fluorescence. However, Cu²⁺ quenches their fluorescence in 30:70 acetontrile/water. L‐CysBQA complexes with Hg²⁺, producing an exciton‐coupled circular dichroism spectrum with the opposite sign to the one that is produced by Cu²⁺ or Zn²⁺ complexation. L‐CysBQA binds Hg²⁺ more strongly than Zn²⁺ and is shown to differentiate Hg²⁺ from other metal ions, such as Zn²⁺, Cu²⁺, Ni²⁺, and Pb²⁺, exceptionally well. The synergistic use of relatively soft sulfur, quinoline‐based chiral ligands and chiroptically enhanced fluorescence detection results in high sensitivity and selectivity for Hg²⁺. Chirality, 2011. © 2011 Wiley‐Liss, Inc.     

Chiral single‐walled carbon nanotubes as chiral selectors in multimode enantioselective sensors

Single‐walled carbon nanotube‐(7,6) chirality was used for the design of multimode enantioselective sensors using different carbon matrices such as graphene paste, graphite paste, and carbon nanopowder‐based paste. l ‐ and d ‐malic acids were used as model analytes. The responses of the multimode sensors were evaluated for potentiometric and differential pulse voltammetry (DPV) modes. When carbon nanopowder was used as matrix, the multimode sensor was enantioselective for d ‐malic acid in the concentration range 10^?3 to 10^?15 mol/L for the potentiometric mode and 10^?5 to 10^?8 mol/L for the DPV mode. The graphite paste‐based sensor was enantioselective for l ‐malic acid in the ranges: 10^?10 to 10^?13 for the potentiometric mode and 10^?4 to 10^?7 mol/L for the DPV mode. The sensors based on graphene and chiral single‐walled carbon nanotubes were enantioselective for d ‐malic acid, and a response was obtained only in the DPV mode. Accordingly, the matrix influenced both the enantioselectivity and the sensitivity of the measurements. The application of the sensors was for the enantioanalysis of malic acid in wines and apple juice samples. The proposed method is fast and reliable and allows the quantification of l ‐ and d ‐malic acids using electrochemical methods based on different principles, from the real samples after a buffering of the samples. The enantioanalysis of malic acid in wine and juice samples was performed with high recoveries (higher than 90.00%) and low relative standard deviation (RSD) (%) values (lower than 1.00%).     

Genetically encoded intracellular sensors based on fluorescent proteins

Green fluorescent protein from Aequorea victoria and its many homologs are now widely used in basic and applied research. These genetically encoded fluorescent markers can detect localization of cell proteins and organelles in living cells and also cells and tissues in living organisms. Unique instruments and methods for studies of molecular biology of a cell and high throughput drug screenings are based on fluorescent proteins. This review deals with the most intensively evolving directions in this field, the development of genetically encoded sensors. Changes in their spectral properties are used for monitoring of cell enzyme activities or changes in concentrations of particular molecules.     

Renal basolateral membrane anion transporter characterized by a fluorescent disulfonic stilbene

Summary The fluorescence enhancement of 4,4-dibenzamido-2,2-disulfonic stilbene (DBDS) upon binding to membranes was used to examine proximal tubule stilbene binding sites. Equilibrium binding studies of DBDS to renal brush border (BBMV) and basolateral membrane vesicles (BLMV) were performed using a fluorescence enhancement technique developed for red blood cells (A.S. Verkman, J.A. Dix and A.K. Solomon,J. Gen. Physiol. 81:421–449, 1983). In the absence of transportable anions, DBDS bound reversibly to a single class of sites on BLMV isolated from rabbit (K _d =3.8 m) and rat (3.2 m); 100 m dihydro-4,4-diisothiocyano-2,2-disulfonic stilbene (H₂DIDS) blocked >95% of binding. H₂DIDS inhibitable DBDS binding was not detected using rat or rabbit BBMV. In rabbit BLMV, DBDSK _d doubled with 10mm SO₄, 50mm HCO₃ and 100mm Cl, but was not altered by Na or pH (6–8). In stopped-flow experiments the exponential time constant for DBDS binding slowed with SO₄, HCO₃ and Cl, but was unaffected by Na. These results are consistent with competitive binding of DBDS and anions at an anion transport site. To relate DBDS binding data to anion transport inhibition we used³⁵SO₄ uptake to characterize several modes of rabbit BLM anion transport: H/SO₄ and Na/SO₄ cotransport, and Cl/SO₄ countertransport. Each transport process was electroneutral and was inhibited by H₂DIDS, furosemide, probenecid, chlorothiazide and DBDS. The apparentK _t 's for DBDS (3–20 m) were similar toK _d for DBDS binding. These studies define a class of anion transport sites on the proximal tubule basolateral membrane measureable optically by a fluorescent stilbene.     

Novel spiking methods developed for anion exchange chromatography operating in a continuous process

Many manufacturers of biopharmaceuticals are moving from batch to continuous processing. While this approach offers advantages over batch processing, demonstration of viral clearance for continuous processes is challenging. Fluctuating output from a continuous process chromatography column results in a nonhomogeneous load for the subsequent column and must be considered when designing viral clearance studies. One approach to clearance studies is to downscale the connected unit operations and introduce virus by in-line spiking. This is challenging to be implemented at the contract research organization performing the clearance study given the complexity of systems and level of expertise required. Alternately, each unit operation could be evaluated in traditional batch mode but the spiking and loading conditions be modified to mimic the variance introduced by the transition between two connected columns. Using a standard chromatography system, we evaluated a flow-through anion exchange chromatography step in a monoclonal antibody (mAb) manufacturing process using five different methods to introduce the virus to the column. Our data show that whether the virus or the mAbs were introduced in concentrated peaks, or as a homogeneous batch, the clearance of mouse minute virus was similar. This study introduces an alternative way to evaluate viral clearance in a continuous process and demonstrates the robustness of anion exchange chromatography unit operating in continuous processing.     

Enantioselective anion exchangers based on cinchona alkaloid‐derived carbamates: Influence of C8/C9 stereochemistry on chiral recognition

Four diastereomeric chiral stationary phases (CSPs) based on quinine, quinidine, epiquinine, and epiquinidine tert‐butyl carbamate selectors were synthesized and evaluated under ion exchange HPLC conditions with a set of racemic N‐acylated and N‐oxycarbonylated α‐amino acids as selectands. The enantioseparation potential of quinine‐ and quinidine‐derived CSPs proved to be far superior to that of their C₉‐epimeric congeners. The absolute configuration of C₉ stereogenic center of the cinchonan backbone of these selectors was identified as the structural feature controlling the elution order. Guided by an X‐ray structure of a most favorable selector–selectand complex and the observed chromatographic enantioseparation data, a chiral recognition model was advanced. The contributions of ion‐pairing, π–π donor–acceptor, hydrogen bonding and steric interactions were established as crucial factors. Chirality 11:522–528, 1999. © 1999 Wiley‐Liss, Inc.     

The anion recognition properties of hydrazone derivatives containing anthracene

A series of artificial receptors, hydrazone derivatives containing anthracene, have been designed and synthesized. The interaction of these receptors with biologically important anions was determined by UV–vis, fluorescence and ¹H NMR titration experiments and theoretical investigation. Results indicate that the receptor (1) without NO₂ shows no binding ability for various anions. The other receptors (2 and 3) show the highest binding ability for acetate (AcO⁻) among studied anions (fluoride (F⁻), dihydrogen phosphate (H₂PO₄⁻), chloride (Cl⁻), bromide (Br⁻), iodide (I⁻)); and the binding ability for AcO⁻ is not interfered by the existence of other anions. The additions of AcO⁻, F⁻ and H₂PO₄⁻ can arouse different degrees of fluorescence quenching. ¹H NMR titration shows that the interaction between the receptor 2 and F⁻ firstly depends on the hydrogen-bond formation; later the interacted site NH is deprotonated and the added F⁻ forms hydrogen bond with the near CH in Schiff base. Moreover, visual color changes accompany guest binding, enabling this system to act as colorimetric anion sensors.     

Synthesis and biological effect of lysosome-targeting fluorescent anion transporters with enhanced anionophoric activity

Two lysosome-targeting fluorescent anion transporters derived from coumarins, trifluoromethylated arylsquaramides and morpholines were synthesized, and their specificity and efficiency to target and alkalize lysosomes were investigated. They are able to target lysosomes specifically. Compared with the previous analogue without trifluoromethyl substituents, these two conjugates, in particular the one having a 3,5-bis(trifluoromethyl) substituent, exhibit significantly higher ability to facilitate the transport of chloride anions, alkalize lysosomes and reduce the activity of lysosomal Cathepsin B enzyme. The present finding suggests that improving the anionophoric activity of lysosome-targeting fluorescent anion transporters is favorable to the efficiency to alkalize lysosomes and deactivate lysosomal Cathepsin B enzyme.     

A novel technique for identification of sites of anion transport in intact cells and tissues using a fluorescent probe

Abstract The purported blocker of anion transport 4, 4′ di-isothiocyano-2-2′ stilbene disulfonate (DIDS) has been shown to partially inhibit ³⁶Cl^? influx, ³⁶CIO^?₃ influx and ³⁵SO^2?₄ influx into Pisum salivum L. cv. Feltham First seedlings. This inhibitory effect could be prevented by pretreatment with the respective unlabelled medium. There was no effect of DIDS on ¹⁴C methylamine influx. The results are consistent with the hypothesis that the binding of DIDS to the site of anion-carrier interaction is responsible for its observed inhibitory effects on anion fluxes. The fluorescent properties of DIDS upon binding to membrane proteins was exploited in an attempt to examine the major sites of anion pumping in whole roots. The results show clearly that in the presence of DIDS the epidermal layers became brightly fluorescent, while cortical layers did not fiuoresce. Lycopersicum esculentum cells taken from locular fluid were plasmolysed using sucrose solution, and the patterns of fluorescence in the presence of DIDS showed in an unambiguous way that the fluorescence is associated with cell membranes. The potential usefulness of this technique to probe sites of anion transport in whole plants and tissues is discussed.     

倍他洛尔立体拆分菌株的筛选与鉴定

从土壤中分离到的52株微生物中,经筛选得到一个能立体选择性水解倍他洛尔中间体的菌株。根据个体形态特征和茵落形态特征初步确定其为红酵母,根据16SDNA序列最后确定其为胶红酵母（Rhodotorula mucilaginosa）DQ832198。该菌株可用于化学酶法制备左旋倍他洛尔的研究。     

Ultrasound‐mediated synthesis of camphoric acid‐based chiral salens for the enantioselective trimethylsilylcyanation of aldehydes

New chiral salen ligands were prepared by the ultrasound‐irradiated condensation of optically active (1R, 3S)‐1,2,2‐trimethyl‐1,3‐diaminocyclopentane with aromatic 1‐hydroxyaldehydes. The ultrasound‐mediated process is more convenient due to shorter reaction times, energy economy, and easier isolation of the products. The in situ formed Ti(IV)(salen) complexes, evaluated as catalysts in the enantioselective trimethylsilylcyanation of benzaldehyde, were found to be efficient for this process, originating the corresponding product in high yields (72–99%) and selectivities of up to 79%. The lowest energy transition states were determined by computational studies. These results were in qualitative agreement with the experimentally observed ones. Chirality, 2010. © 2009 Wiley‐Liss, Inc.     

Studies on synthesis and anion recognition properties of (3'-nitrobenzo)[2,3-d]-(3'-nitrobenzo)[9,10-d]-1,4,8,11-tetraazacyclotetradecane-5,7,12,14-tetraone

A novel artificial receptor, (3'-nitrobenzo)[2,3-d]-(3'-nitrobenzo)[9,10-d]-1,4,8,11-tetraazacyclotetradecane-5,7,12,14-tetraone, was designed and synthesized. The interactions of this receptor with different anions were determined by UV-vis and (1)H NMR titration experiments, and their affinity constants to different anions were compared with those of other similar/different systems. The results indicated that this receptor showed a high selective and recognitive ability for F(-) among F(-), Cl(-), Br(-), I(-), AcO(-), OH(-), and H(2)PO(4)(-). Moreover, the low energy configuration of this receptor was further determined by means of theoretical investigations.     

Optimization of bilayer floating tablet containing metoprolol tartrate as a model drug for gastric retention

The purpose of the present study was to develop an optimized gastric floating drug delivery system (GFDDS) containing metoprolol tartrate (MT) as a model drug by the optimization technique. A 2³ factorial design was employed in formulating the GFDDS with total polymer content-to-drug ratio (X₁), polymer-to-polymer ratio (X₂), and different viscosity grades of hydroxypropyl methyl cellulose (HPMC) (X₃) as independent variables. Four dependent variables were considered: percentage of MT release at 8 hours, T_50%, diffusion coefficient, and floating time. The main effect and interaction terms were quantitatively evaluated using a mathematical model. The results indicate that X₁ and X₂ significantly affected the floating time and release properties, but the effect of different viscosity grades of HPMC (K4M and K10M) was nonsignificant. Regression analysis and numerical optimization were performed to identify the best formulation. Fickian release transport was confirmed as the release mechanism from the optimized formulation. The predicted values agreed well with the experimental values, and the results demonstrate the feasibility of the model in the development of GFDDS.     

Cataluminescence‐based sensors: principle,instrument and application

The cataluminescence (CTL)‐based sensor is a new promising type of chemical transducer, and has attracted much attention of researchers for its potential versatile applications in public safety, emission control and environmental protection. In this review, we briefly introduce the development history of CTL‐based sensors and summarize existing explanations of the CTL reaction mechanism as well as three research strategies for mechanism the CTL mechanism. In the following, all the function units of a typical CTL‐based sensor system are described and the investigation of the sensor materials. CTL‐based sensor arrays, are discussed in detail. We classify the recent novel hyphenated techniques based on CTL coupled to other analysis techniques into the preconcentration‐CTL hyphenated technique, nebulization‐CTL hyphenated technique, plasma‐assisted CTL technique and tandem CTL technique according to the type of analysis combined with CTL and provide a detailed account of novel hyphenated techniques. Owing to the appearance of these novel techniques, the application range of CTL has been expanded as well as the sensitivity and selectivity of CTL system has been greatly improved. Finally, the applications of CTL‐based sensor and sensor arrays in the last several years are classified and summarized. Copyright © 2014 John Wiley & Sons, Ltd.     

A new peptidyl fluorescent chemosensors for the selective detection of mercury ions based on tetrapeptide

A novel peptidyl chemosensor (PySO₂-His-Gly-Gly-Lys(PySO₂)-NH₂, 1) was synthesized by incorporation of two pyrene (Py) fluorophores into the tetrapeptide using sulfonamide group. Compound 1 exhibited selective fluorescence response towards Hg(II) over the other metal ions in aqueous buffered solutions. Furthermore, 1 with the potent binding affinity (K_d = 120 nM) for Hg(II) detected Hg(II) without interference of other metal ions such as Ag(I), Cu(II), Cd(II), and Pb(II). The binding mode of 1 with Hg(II) was investigated by UV absorbance spectroscopy, ¹H NMR titration experiment, and pH titration experiment. The addition of Hg(II) induced a significant decrease in both excimer and monomer emissions of the pyrene fluorescence. Hg(II) interacted with the sulfonamide groups and the imidazole group of His in the peptidyl chemosensor and then two pyrene fluorophores were close to each other in the peptide. The decrease of both excimer and monomer emission was mainly due to the excimer/monomer emission change by dimerization of two pyrene fluorophores and a quenching effect of Hg(II).     

A fluorescent chiral chemosensor for the recognition of the two enantiomers of chiral carboxylates

A new 7-nitrobenz-2-oxa-1,3-diazole (NBD)-based fluorescent chiral chemosensor (NBD-1) was prepared and applied to the recognition of the two enantiomers of the tetrabutylammonium salts of N-t-Boc-α-amino acids and chiral carboxylic acids including naproxen. In particular, the chiral recognition by the new fluorescent chiral chemosensor for the two enantiomers of N-t-Boc-threonine (tetrabutylammonium salt) was quite excellent, the Stern-Volmer constant ratio (K(D)/K(L)) for the two enantiomers being as high as 4.89.     

An electrochemical sensor based on cellulose nanocrystal for the enantioselective discrimination of chiral amino acids

A novel electrochemical sensor based on 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized cellulose nanocrystals (TOCNCs) and l-cystines (l-Cys) modified Au electrode (TOCNC/l-Cys/Au) has been fabricated for detection and discrimination of the enantiomers of phenylalanine (Phe), leucine (Leu), and valine (Val). The three amino acids are in connection with metabolism diseases. The TOCNC/l-Cys/Au electrode exhibited obvious peak current difference for the amino acid enantiomers by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The TOCNCs on the electrode surface expressed different interactions with d- and l-amino acids, so the electrochemical recognitions of the three amino acid enantiomers were achieved. TOCNCs were characterized by Fourier transform infrared (FT-IR) and scanning electron microscopy (SEM). The modified electrodes were characterized by SEM and electrochemical techniques. According to DPV, peak currents of the two enantiomers decreased linearly with their concentrations. Furthermore, satisfactory results were obtained when this electrode was applied to measure the d- and l-Phe mixture. The experimental results show that TOCNCs are suitable material for chiral sensor. The contrast of serum sample of healthy people and patients with type 2 diabetes also was proposed, and significant difference was exhibited on the modified electrode. This work is significant for the screening, diagnosis, and treatment of multiple metabolic diseases.