Flow injection analysis: Rayleigh light scattering technique for total protein determination

A novel flow injection analysis (FIA) method with Rayleigh light scattering (RLS) detection was developed for the determination of total protein concentrations. This method is based on the weak intensity of RLS of bromothymol blue (BB) (3',3"-dibromothymolsulfonephthalein) which can be enhanced by the addition of protein in weakly acidic solution. A common spectrofluorimeter was used as a detector. It was proved that the application of this method to quantify the total proteins in real samples by using bovine serum albumin was possible. The RLS signal was detected at lambda(ex)= lambda(em)=572 nm. The linear range was 7.0-70.0 microg mL(-1), the detection limit was 3.75 microg mL(-1), the reproducibility was 5.5% (n=7), and the sample throughput was 26 h(-1).     

Electric conductivity in a fibrous porous medium with thin but polarized double layers

The electric conduction in the fibrous medium constructed by a homogeneous array of parallel, identical, charged, circular cylinders having an arbitrary zeta potential filled with the solution of a symmetrically charged electrolyte is analytically examined. The thickness of the electric double layers surrounding the dielectric cylinders is assumed to be small relative to the radius of each cylinder and to the gap width between two neighboring cylinders, but the polarization of the mobile ions in the diffuse layers is allowed. The effect of interactions among individual cylinders is taken into explicit account by employing a unit cell model. The appropriate equations of conservation of electrochemical potential energies of ionic species are solved for each cell, in which a cylinder is envisaged to be surrounded by a coaxial cylindrical shell of the fluid solution. Analytical expressions for the effective electric conductivity are obtained in closed forms as functions of the porosity of the fiber matrix and other characteristics of the porous system. Comparisons of the results of the cell model with different conditions at the outer boundary of the cell are made. Under an otherwise identical condition, the electric conductivity in a porous medium composed of an array of parallel cylinders in the transverse direction is smaller than that of a suspension of spheres. The effect of interactions among the cylinders or spheres on the effective conductivity can be quite significant under appropriate conditions.     

Biosensors based on enzyme field-effect transistors for determination of some substrates and inhibitors

This paper is a review of the authors' publications concerning the development of biosensors based on enzyme field-effect transistors (ENFETs) for direct substrates or inhibitors analysis. Such biosensors were designed by using immobilised enzymes and ion-selective field-effect transistors (ISFETs). Highly specific, sensitive, simple, fast and cheap determination of different substances renders them as promising tools in medicine, biotechnology, environmental control, agriculture and the food industry.The biosensors based on ENFETs and direct enzyme analysis for determination of concentrations of different substrates (glucose, urea, penicillin, formaldehyde, creatinine, etc.) have been developed and their laboratory prototypes were fabricated. Improvement of the analytical characteristics of such biosensors may be achieved by using a differential mode of measurement, working solutions with different buffer concentrations and specific agents, negatively or positively charged additional membranes, or genetically modified enzymes. These approaches allow one to decrease the effect of the buffer capacity influence on the sensor response in an aim to increase the sensitivity of the biosensors and to extend their dynamic ranges.Biosensors for the determination of concentrations of different toxic substances (organophosphorous pesticides, heavy metal ions, hypochlorite, glycoalkaloids, etc.) were designed on the basis of reversible and/or irreversible enzyme inhibition effect(s). The conception of an enzymatic multibiosensor for the determination of different toxic substances based on the enzyme inhibition effect is also described.We will discuss the respective advantages and disadvantages of biosensors based on the ENFETs developed and also demonstrate their practical application.     

Zirconium(IV)-porphyrins as novel ionophores for fluoride-selective polymeric membrane electrodes

The feasibility of using Zr(IV)-porphyrins as novel ionophores for preparing anion-selective polymeric membrane electrodes is examined. Electrodes constructed using o-nitrophenyl octyl ether plasticized poly(vinyl chloride) membranes containing Zr(IV)-octaethylporphyrin (OEP) dichloride (Zr(IV)[OEP]Cl₂) or Zr(IV)-tetraphenylporphyrin (TPP) dichloride (Zr(IV)[TPP]Cl₂) were found to exhibit enhanced potentiometric selectivity toward fluoride compared to electrodes based on a typical anion-exchanger (e.g. tridodecylmethylammonium chloride). At pH 5.5, the electrodes displayed the following selectivity sequences: ClO₄⁻ > SCN⁻ > I⁻ > F⁻ > NO₃⁻ > Br⁻ > NO₂⁻ > Cl⁻ and F⁻ > ClO₄⁻ > SCN⁻ > I⁻ > NO₂⁻ > NO₃⁻ > Br⁻ > Cl⁻ for membranes doped with Zr(IV)[OEP]Cl₂) and Zr(IV)[TPP]Cl₂, respectively. Both ionophores are shown to operate via a charged carrier mechanism, with 10 mol% of lipophilic tetraphenylborate derivative in the membrane phase required to achieve optimal selectivity. Electrodes prepared with both metalloporphyrin species display super-Nernstian response toward fluoride with slopes typically greater than −100 mV per decade. It is shown, via UV-VIS spectroscopy of the membrane phase, that this behavior occurs due to spontaneous formation of hydroxide ion bridged porphyrin dimers in the membrane in the presence of the lipophilic anionic additive. The dimers are easily converted to monomeric species upon increasing the concentration of fluoride in the sample solution. Decreasing the pH of sample buffer background solution (from pH 5.5 to pH 3) decreases the lower detection limit (DL) of the electrode response toward fluoride (by two-order of magnitude) and improves the electrodes’ selectivity.     

Microwave‐Assisted Palladium‐Catalyzed Direct Arylation of 1,4‐Disubstituted 1,2,3‐Triazoles with Aryl Chlorides

Treatment of 1,4‐disubstituted 1,2,3‐triazoles with aryl chlorides in the presence of potassium carbonate under palladium catalysis and microwave irradiation at 250 °C for 15 min leads to arylation of the triazole at the 5‐position. A variety of functional groups, including ester and hydroxy groups, are compatible. The procedure is suitable for the regioselective preparation of trisubstituted triazoles. Microwave irradiation accelerates the reaction, thus allowing the rapid synthesis of trisubstituted triazoles, which are difficult to synthesize selectively.     

Influence of the Size of Aromatic Chelate Ligands on the Structures of Cadmium(II) Tetracarboxylates Polymers: From 2D Layered Network to 3D Metal‐Organic Framework

To determine the influence of the size of the aromatic chelate ligands on the frameworks of metal tretracarboxylate polymers, two new coordination polymers [Cd(btc)_0.5 (2,2′‐bpy)] ( 1 ) and [Cd(btc)_0.5(phen)]·H₂O ( 2 ) (H₄btc = biphenyl‐3,3′,4,4′‐tetracarboxylic acid, 2,2′‐bpy = 2,2′‐bipyridine, phen = 1,10‐phenanthroline) have been synthesized under similar hydrothermal conditions. In complex 1 , the dimeric Cd₂ units are linked by bridging btc^4? ligand to form a 2D layered network, whereas complex 2 possesses a 3D metal‐organic framework consisting of the dimeric Cd₂ units. The differences of two metal‐organic frameworks demonstrate that the size of the rigid aromatic chelate ligands have an important effect on the structures of their complexes. Additionally, the two complexes show strong fluorescence in the solid state at room temperature.     

Simultaneous determination of DTPA,EDTA, and NTA by capillary electrophoresis after complexation with copper

We present a method for simultaneous determination of the aminopolycarboxylic acids DTPA, EDTA and NTA in dishwashing detergents, paper mill waters, and natural waters by capillary electrophoresis (CE). The complexing agents were examined as their copper(II) complexes and separated by conventional CE with reversed polarity of the applied voltage. The optimum separation conditions were established by varying the pH and phosphate and tetradecyltrimethylammonium bromide (TTAB) concentrations in the run buffer. The separations were carried out in a fused-silica capillary (61 cm×75 m i.d.) filled with phosphate buffer (80 mmol L^–1, TTAB concentration 0.5 mmol L^–1, pH 7.1, voltage –20 kV) using direct UV detection at 191 and 254 nm. With this CE method all the peaks in the electropherograms were properly separated, the calibration plots gave good correlation coefficients and all three complexing agents could be detected in less than 4 min. Linear calibration plots were obtained for CuDTPA, CuEDTA and CuNTA; limits of detection were 0.03 mmol L^–1 for all complexing agents and recoveries for all tested samples were within the range 104±7%. Results obtained from dishwashing detergent samples were found to be reliable and comparable with those from HPLC (R²=0.989) and UV–Vis (R²=0.985) methods.     

Multifold curdlan gel formation by dialysis into aqueous solutions of metal salts

We have found that the dialysis of curdlan dissolved in alkaline solution into aqueous solutions of metal salts yielded multifold gel structures. Aqueous sodium chloride and potassium chloride as well as pure water induced isotropic gels. Aqueous calcium salts induced liquid crystalline gel with refractive index gradient/amorphous gel alternative structure. Aqueous salts of trivalent aluminum and ferric cations induced a rigid liquid crystalline gel, which shrank above a threshold concentration of each salt. On the other hand, Liesegang ring-like pattern was observed with aqueous solutions of mixed salts of calcium chloride and magnesium chloride. The patterns have been classified to discuss the mechanism of forming the variety of structures.     

Direct qualitative analysis of triacylglycerols by electrospray mass spectrometry using a linear ion trap

Triacylglycerols (TAGs) isolated from a biological sample provide a challenge for mass spectrometric analysis because of the complexity of naturally occurring TAGs, which may contain different fatty acyl substituents resulting in a large number of molecular species having the identical elemental composition. We have investigated the use of mass spectrometry to obtain unambiguous information as to the individual TAG molecular species present in a complex mixture of triacylglycerols using a linear ion trap mass spectrometer. Ammonium adducts of TAGs, [M+NH4]+, were generated by electrospray ionization, which permitted the molecular weight of each TAG molecular species to be determined. The mechanisms involved in the decomposition of the [M+NH4]+ and subsequent fragment ions were investigated using deuterium labeling, MS/MS, and MS3 experiments. Collision induced decomposition of [M+NH4]+ ions resulted in the neutral loss of NH3 and an acyl side-chain (as a carboxylic acid) to generate a diacyl product ion. MS/MS data were used to identify each acyl group present for a given [M+NH4]+ ion, and this information could be combined with molecular weight data to identify possible TAG molecular species present in a biological extract. Subsequent MS3 experiments on the resultant diacyl product ions, which gave rise to acylium (RCO+) and related ions, enabled unambiguous TAG molecular assignments. These strategies of MS, MS/MS, and MS3 experiments were applied to identify components within a complex mixture of neutral lipids extracted from RAW 264.7 cells.