The effect of hydrolysis time on amino acid analysis

Determining the amino acid content of a protein involves the hydrolysis of that protein, usually in acid, until the protein-bound amino acids are released and made available for detection. Both the variability in the ease of peptide bond cleavage and differences in the acid stability of certain amino acids can significantly affect determination of a protein's amino acid content. By using multiple hydrolysis intervals, a greater degree of accuracy can be obtained in amino acid analysis. Correction factors derived by linear extrapolation of serial hydrolysis data are currently used. Compartmental modeling of the simultaneous hydrolysis (yield) and degradation (decay) of amino acids by nonlinear multiple regression of serial hydrolysis data has also been validated and applied to determine the amino acid composition of various biological samples, including egg-white lysozyme, human milk protein, and hair. Implicit in the routine application of serial hydrolysis in amino acid analysis, however, is an understanding that correction factors, derived either linearly or through the more accurate nonlinear multiple regression approach, need to be determined for individual proteins rather than be applied uniformly across all protein types.     

Immobilisation of electroactive macrocyclic complexes within titania films

The 4-carboxyphenyl-appended macrocyclic ligand trans-6,13-dimethyl-6-((4-carboxybenzyl)amino)-1,4,8,11-tetraazacyclotetradecane-6-amine (HL10) has been synthesised and complexed with Co(III). The mononuclear complexes [Co(HL10)(CN)]2+ and [CoL10(OH)]+ have been prepared and the crystal structures of their perchlorate salts are presented, where the ligand is bound in a pentadentate mode in each case while the 4-carboxybenzyl-substituted pendent amine remains free from the metal. The cyano-bridged dinuclear complex [CoL10-mu-NC-Fe(CN)5]2- was also prepared and chemisorbed on titania-coated ITO conducting glass. The adsorbed complex is electrochemically active and cyclic voltammetry of the modified ITO working electrode in both water and MeCN solution was undertaken with simultaneous optical spectroscopy. This experiment demonstrates that reversible electrochemical oxidation of the Fe(II) centre is coupled with rapid changes in the optical absorbance of the film.     

Numerical simulation of DNA sample preconcentration in microdevice electrophoresis

A numerical model is presented for the accurate and efficient prediction of preconcentration and transport of DNA during sample introduction and injection in microcapillary electrophoresis. The model incorporates conservation laws for the different buffer ions, salt ions, and DNA sample, coupled through a Gaussian electric field to account for the field modifications that cause electromigration. The accuracy and efficiency required to capture the physics associated with such a complex transient problem are realized by the use of the finite element-flux corrected transport (FE-FCT) algorithm in two dimensions. The model has been employed for the prediction of DNA sample preconcentration and transport during electrophoresis in a double-T injector microdevice. To test its validity, the numerical results have been compared with the corresponding experimental data under similar conditions, and excellent agreement has been found. Finally, detailed results from a simulation of DNA sample preconcentration in electrophoretic microdevices are presented using as parameters the electric field strength and the other species concentrations. The effect of the Tris concentration on sample stacking is also investigated. These results demonstrate the great potential offered by the model for future optimization of such microchip devices with respect to significantly enhanced speed and resolution of sample separation.     

A computational study of the reactions of a beta-diketiminatoaluminium(I) complex with the hydrogen atom and the electron

A computational study has been performed to examine the reactions of a model beta-diketiminatoaluminium (I) complex with the hydrogen atom and with the electron. It was found that the hydrogen atom adds to the metal centre exothermically (DeltaH(rxn)=-202 kJ mol(-1)), and the spin density in the resulting radical resides entirely on the beta-diketiminato ligand. The spin density of the corresponding radical anion is very similar to the H-adduct.     

Anisotropic hydrogel thickness gradient films derivatized to yield three-dimensional composite materials

We report the preparation of in-plane density gradients of amino-terminated molecules and gold particles through derivatization of laterally varying thickness gradients of poly(acrylic acid) (PAA) or poly(acrylamide) (PAAm) films. PAA and PAAm gradients were formed by Zn(II)-catalyzed electropolymerization of acrylic acid (AA) or acrylamide (AAm) in the presence of an in-plane electrochemical potential gradient applied to Au or indium-tin-oxide (ITO) working electrodes. PAA thickness gradients were converted into density gradients of fluorocarbons or biocompatible groups by derivatizing with NH(2)CH(2)(CF(2))(6)CF(3) or an Arg-Gly-Asp (RGD)-containing peptide, respectively. X-ray photoelectron spectroscopy (XPS) and XPS imaging were used to characterize the modified PAA gradients. Transition regions as narrow as 104 mum were achieved for fluorocarbon gradients. PAAm gradients were treated with gold particles to form a density gradient of gold particles. Surface plasmon resonance imaging and scanning electron microscopy (SEM) as well as UV-visible absorption measurements were used to characterize the gold particle density gradients. It is likely that the gold particles were attached both on the surface and inside the PAAm film.     

Multilevel microfluidics via single-exposure photolithography

This communication reports a new method to form multilevel features in a single layer of SU-8 photoresist to facilitate the generation of 3D microfluidic chips. The method utilizes the spatial dependence of diffracted light intensity to selectively overexpose masked regions of photoresist and requires only a UV light source and a single transparency mask. 3D structures are formed within microfluidic channels using this selective overexposure method, with feature sizes being determined by the exposure dose and mask feature sizes. The dimensions of the internal features and the microfluidic channels can be varied independently according to these parameters, and any number of different heights can be obtained in a single exposure step. The method provides a simple means of forming 3D microfluidic structures with integrated features, including mixing structures, flow stabilization ridges, and separation weirs to increase the capabilities of microfluidic chips in a variety of microchemical applications.     

Ionic Association and Mobility IV. Ionophores in Dimethylsulfoxide at 25°C

The results of conductance measurements on pyridinium picrate, tetraphenylo-sonium picrate, potassium picrate, tetraphenylantimony picrate, tetrapropylam-monium, tetrafluoroborate, tetramethylammonium hexafluorophosphate ion association noncoulombic interaction in dimethyl sulfoxide (DMSO) at 25°C in the concentration range 1–15×10^–4 M are reported. The data were analyzed by the Justice modification of the Fuoss–Hsia equation. Except for pyridinium picrate all salts studied were found to be associated.Application of the Justice Barthel–Bjerrum model of ion association permitted calculation of the noncoulombic portion of the potential of mean force, W _±. Ionic limiting conductances were calculated for six ions using known values of previously determined transport numbers. A table of most current limiting ionic conductances for a variety of ions in DMSO at 25°C has been established.     

Syntheses and Structural Aspects of Cyclodextrin/Dialkylamine Inclusion Compounds

We report the syntheses and structural aspects of cyclodextrin host–guest inclusion compounds containing linear secondary alkylamines (dipropyl, dibutyl, dipentyl, dihexyl, and dioctyl) at 25 °C. Elemental analysis, ¹³C CP-MAS NMR spectroscopy, and powder X-ray diffraction analysis confirm the inclusion process. The basic host structure of the products is similar to that of typical cyclodextrin inclusion systems. ¹³C MAS NMR experiments show a different resonance pattern for the confined guest molecules with respect to the amine in the liquid phase. The presence of different resonance signals for the homologous carbon atoms of both dialkylamine branches is evidence for the non-symmetric location of the amine in the cyclodextrin channels.     

Square Planar (SP-4) and Octahedral (OC-6) Complexes of Platinum(II) and -(IV) with Predetermined Chirality at the Metal Center

cis-Bis-homoleptic platinum(II) complexes, with predetermined helical chirality at the metal center, can be obtained by using strongly sterically interacting ligands. With this aim, two new ligands, (8R,10R)-2-(2'-thienyl)-4,5-pinenopyridine, th4,5ppy (2), and (8R,10R)-2-(2'-thienyl)-5,6-pinenopyridine, th5,6ppy (4), were synthesized and coordinated to platinum. The structures of the resulting complexes, Pt(th4,5ppy)(2) (5) and Pt(th5,6ppy)(2) (6), were determined by X-ray diffraction, and it was found that they both crystallize with a Delta-cis configuration. Thermal oxidative additions (TOA) of alkyl halides were performed with both complexes leading, in the case of 5, to a mixture of isomers and, in the case of 6, to isomerically pure products. The predetermination of chirality at the metal center is therefore preserved in the octahedral (OC-6) platinum(IV) complexes. Crystals of Pt(th4,5ppy)(2) (5) are orthorhombic, of space group P2(1)2(1)2(1), with a = 12.973(1) ?, b = 13.619(2) ?, c = 17.665(2) ?, alpha = beta = gamma = 90 degrees, and Z = 4. Final R = 0.0268 and R(w) = 0.0424 for 3101 observed reflections. Crystals of Pt(th5,6ppy)(2) (6) are hexagonal, of space group P6(1), with a = 11.5465(4) ?, b = 11.5465(4) ?, c = 35.356(3) ?, alpha = beta = 90 degrees, gamma = 120 degrees, and Z = 6. Final R = 0.0424 and R(w) = 0.0845 for 2660 observed reflections. Neither molecule possesses a crystallographic C(2) symmetry.     

Cationic Lanthanide Luminescent Copolymer: Design,Synthesis and Interaction with DNA

Polymerizable rare earth complex Eu(AA)₃Phen was synthesized by complexion of europium ion, acrylic acid (AA), and 1,10-phenanthroline (Phen). The structure and fluorescence properties of the complex were studied by elemental analysis, ¹H-NMR spectroscopy, and fluorescence spectroscopy. Eu-containing copolymer poly(PEGMA-co-MMA-co-METAC-co-Eu(AA)₃Phen) (PPMMEu) was then synthesized by free radical copolymerization of Eu(AA)₃Phen and other functional monomers including poly(ethylene glycol) methyl ether methacrylate (PEGMA) and [2-(Methacryloyloxy) ethyl] trimethylammonium chloride (METAC). ¹H-NMR spectroscopy and fluorescence spectroscopy were used to characterize the copolymer and the interactions between the copolymer and DNA was investigated by TEM, fluorescence spectroscopy, and agarose gel electrophoresis. The desired luminescent cationic copolymer was successfully obtained. The copolymer can form micelles in water solution and can efficiently bind to DNA molecules through electrostatic interaction. The results suggest the potential use of PPMMEu in bioprobes and gene vectors.