Detection of menadione sodium bisulfite (vitamin K-3) by reversed-phase high performance liquid chromatography with series dual-electrode amperometric detector

A reversed-phase high-performance liquid chromatography with series dual glassy carbon electrodes for the amperometric detection of water-soluble menadione is described. The complex post-column derivatization reaction and the high background currents were avoided. The menadione sodium bisulfite was reduced at -0.3 V vs. SCE at the upstream (generator) electrode and oxidized at +0.2V vs. SCE at the downstream (collector) electrode. The mobile phase was 0.2moll(-1) HAc-NaAc aqueous buffer (pH 5.50) and 40% (v/v) methanol. The linear response was in the range of 35 ng to 15 mu g, with a detection Limit of 15 ng (S/N=3). The correlation coefficient was 0.9997 (n=6). The electrochemical detection with series dual electrodes has a higher selectivity for menadione (vitamin K-3) compound than with UV detection.

Determination of alcohols by high-performance liquid chromatography with fluorimetric detection after pre-column derivatisation with carbazole-9-N-acetic acid and chromatographic behaviour of alcoholic derivatives

Alcohols were derivatised to their carbazole-9-N-acetic acid (CRA) esters with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC . HCl) as the dehydrating agent. Studies on derivatisation conditions indicated that the coupling reaction proceeded rapidly and smoothly in the presence of a base catalyst in acetonitrile to give the corresponding sensitively fluorescent derivatives. The retention behaviour of alcohol derivatives was investigated by varying mobile phase compositions (ACN-water and MeOH-water). The parameters from the equation log k'=A-BX were evaluated by retention data of derivatives using an isocratic elution with different mobile phases. The results indicated that the parameters derived allowed computation of retention factors in good agreement with experiments. At the same time, a general equation was derived that makes possible predictions of partition coefficient in binary mobile phases with different proportions of organic solvent to water based on some simple regression analysis. The LC separation for the derivatised alcohols containing higher carbon alcohols showed good reproducibility on a reversed-phase C-18 column with gradient elution. The detection limits (excitation at 335 nm, emission at 360 nm) for derivatised alcohols (signal-to-noise ratio=3:1) were in the range of 0.1-0.4 pg per injection. (C) 2001 Elsevier Science B.V. All rights reserved.

Simple derivatization method for sensitive determination of fatty acids with fluorescence detection by high-performance liquid chromatography using 9-(2-hydroxyethyl)-carbazole as derivatization reagent

A simple and sensitive method for the determination of short and long-chain fatty acids using high-performance liquid chromatography with fluorimetric detection has been developed. The fatty acids were derivatized to their corresponding esters with 9-(2-hydroxyethyl)-carbazole (HEC) in acetonitrile at 60 degreesC with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride as a coupling agent in the presence of 4-dimethylaminopyridine (DMAP). A mixture of esters of C-1-C-20 fatty acids was completely separated within 38 min in conjunction with a gradient elution on a reversed-phase C-18 column. The maximum fluorescence emission for the derivatized fatty acids is at 365 nm (lambda (ex) 335 nm). Studies on derivatization conditions indicate that fatty acids react proceeded rapidly and smoothly with HEC in the presence of EDC and DMAP in acetonitrile to give the corresponding sensitively fluorescent derivatives. The application of this method to the analysis of long chain fatty acids in plasma is also investigated. The LC separation shows good selectivity and reproducibility for fatty acids derivatives. The R.S.D. (n = 6) for each fatty acid derivative are <4%. The detection limits are at 45-68 fmol levels for C-14-C-20 fatty acids and even lower levels for

Determination of Proper Processing Conditions of Material Surface Heat Treatment Through Finite Element Method

A two-dimensional model has been developed based on the experimental results of stainless steel remelting with the laminar plasma technology to investigate the transient thermo-physical characteristics of the melt pool liquids. The influence of the temperature field, temperature gradient, solidification rate and cooling rate on the processing conditions has been investigated numerically. Not only have the appropriate processing conditions been determined according to the calculations, but also they have been predicted with a criterion established based on the concept of equivalent temperature area density (ETAD) that is actually a function of the processing parameters and material properties. The comparison between the resulting conditions shows that the ETAD method can better predict the optimum condition.

Determination Ofinterfacial Mechanical Parameters For An Al/Epoxy/Al2O3 System Byusing Peel Test Simulations

Peel test measurements and simulations of the interfacial mechanical parameters for the Al/Epoxy/Al2O3 system are performed in the present investigation. A series of Al film thicknesses between 20 and 250 microns and three peel angles of 90, 135 and 180 degrees are considered. Two types of epoxy adhesives are adopted to obtain both strong and weak interface adhesions. A finite element model with cohesive zone elements is used to identify the interfacial parameters and simulate the peel test process. By simulating and recording normal stress near the crack tip, the separation strength is obtained. Furthermore, the cohesive energy is identified by comparing the simulated steady-state peel force and the experimental result. It is found from the research that both the cohesive energy and the separation strength can be taken as the intrinsic interfacial parameters which are dependent on the thickness of the adhesive layer and independent of the film thickness and peel angle.

Nanoindentation of thin-film-substrate system: Determination of film hardness and Young's modulus

In the present paper, the hardness and Young's modulus of film-substrate systems are determined by means of nanoindentation experiments and modified models. Aluminum film and two kinds of substrates; i.e. glass and silicon, are studied. Nanoindentation XP II and continuous stiffness mode are used during the experiments. In order to avoid the influence of the Oliver and Pharr method used in the experiments, the experiment data are analyzed with the constant Young's modulus assumption and the equal hardness assumption. The volume fraction model (CZ model) proposed by Fabes et al. (1992) is used and modified to analyze the measured hardness. The method proposed by Doerner and Nix (DN formula) (1986) is modified to analyze the measured Young's modulus. Two kinds of modified empirical formula are used to predict the present experiment results and those in the literature, which include the results of two kinds of systems, i.e., a soft film on a hard substrate and a hard film on a soft substrate. In the modified CZ model, the indentation influence angle, phi, is considered as a relevant physical parameter, which embodies the effects of the indenter tip radius, pile-up or sink-in phenomena and deformation of film and substrate.

Determination of interface properties between micron-thick metal film and ceramic substrate using peel test

Peel test measurements have been performed to estimate both the interface toughness and the separation strength between copper thin film and Al2O3 substrate with film thicknesses ranging between 1 and 15 mu m. An inverse analysis based on the artificial neural network method is adopted to determine the interface parameters. The interface parameters are characterized by the cohesive zone (CZ) model. The results of finite element simulations based on the strain gradient plasticity theory are used to train the artificial neural network. Using both the trained neural network and the experimental measurements for one test result, both the interface toughness and the separation strength are determined. Finally, the finite element predictions adopting the determined interface parameters are performed for the other film thickness cases, and are in agreement with the experimental results.

Simple phenomenological determination of contact stiffness and elastic modulus of Ce-based bulk metallic glasses through nanoindentation

The viscoelastic deformation of Ce-based bulk metallic glasses (BMGs) with low glass transition temperature is investigated at room temperature. Contact stiffness and elastic modulus of Ce-based BMGs cannot be derived using the conventional Oliver-Pharr method [W. C. Oliver and G. M. Pharr, J. Mater. Res. 7, 1564 (1992)]. The present work shows that the time dependent displacement of unloading segments can be described well by a generalized Kelvin model. Thus, a modified Oliver-Pharr method is proposed to evaluate the contact stiffness and elastic modulus, which does, in fact, reproduce the values obtained via uniaxial compression tests. (c) 2007 American Institute of Physics.

Statistical formulation and experimental determination of growth rate of micrometre cracks under impact loading

A new statistical formulation and a relevant experimental approach to determine the growth rate of microcracks were proposed. The method consists of experimental measurements and a statistical analysis' on the basis of the conservation law of number density of microcracks in phase space. As a practical example of the method, the growth rate of microcracks appearing in an aluminium alloy subjected to planar impact loading was determined to be ca. 10 mu m/mu s under a tensile stress of 1470 MPa and load duration between 0.26 mu s and 0.80 mu s.

Determination of plastic properties by instrumented spherical indentation: Expanding cavity model and similarity solution approach

The present paper aims to develop a robust spherical indentation-based method to extract material plastic properties. For this purpose, a new consideration of-piling-up effect is incorporated into the expanding cavity model; an extensive numerical study on the similarity Solution has also been performed. As a consequence, two semi-theoretical relations between the indentation response and material plastic properties are derived, with which plastic properties of materials can be identified from a single instrumented spherical indentation curve, the advantage being that this approach no longer needs estimations of contact radius with given elastic modulus. Moreover, the inconvenience in using multiple indenters with different tip angles can be avoided. Comprehensive sensitivity analyses show that the present algorithm is reliable. Also, by experimental verification performed oil three typical materials, good agreement of the material properties between those obtained from the reverse algorithm and experimental data is obtained.

Determination Of Interfacial Properties Between Metal Film And Ceramic Substrate With An Adhesive Layer

Peel test measurements and inverse analysis to determine the interfacial mechanical parameters for the metal film/ceramic system are performed, considering that there exist an epoxy interface layer between film and ceramic. In the present investigation, Al films with a series of thicknesses between 20 and 250 mu m and three peel angles of 90, 135 and 180 degrees are considered. A finite element model with the cohesive zone elements is used to simulate the peel test process. The finite element results are taken as the training data of a neural network in the inverse analysis. The interfacial cohesive energy and the separation strength can be determined based on the inverse analysis and peel experimental result. (C) 2008 Elsevier Ltd. All rights reserved.

Determination Of Plastic Properties By Instrumented Spherical Indentation: Expanding Cavity Model And Similarity Solution Approach

The present paper aims to develop a robust spherical indentation-based method to extract material plastic properties. For this purpose, a new consideration of-piling-up effect is incorporated into the expanding cavity model; an extensive numerical study on the similarity Solution has also been performed. As a consequence, two semi-theoretical relations between the indentation response and material plastic properties are derived, with which plastic properties of materials can be identified from a single instrumented spherical indentation curve, the advantage being that this approach no longer needs estimations of contact radius with given elastic modulus. Moreover, the inconvenience in using multiple indenters with different tip angles can be avoided. Comprehensive sensitivity analyses show that the present algorithm is reliable. Also, by experimental verification performed oil three typical materials, good agreement of the material properties between those obtained from the reverse algorithm and experimental data is obtained.

Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity

One of existing strategies to engineer active antibody is to link VH and VL domains via a linker peptide. How the composition, length, and conformation of the linker affect antibody activity, however, remains poorly understood. In this study, a dual approach that coordinates molecule modeling, biological measurements, and affinity evaluation was developed to quantify the binding activity of a novel stable miniaturized anti-CD20 antibody or singlechain fragment variable (scFv) with a linker peptide. Upon computer-guided homology modeling, distance geometry analysis, and molecular superimposition and optimization, three new linker peptides PT1, PT2, and PT3 with respective 7, 10, and 15 residues were proposed and three engineered antibodies were then constructed by linking the cloned VH and VL domains and fusing to a derivative of human IgG1. The binding stability and activity of scFv-Fc chimera to CD20 antigen was quantified using a micropipette adhesion frequency assay and a Scatchard analysis. Our data indicated that the binding affinity was similar for the chimera with PT2 or PT3 and ~24-fold higher than that for the chimera with PT1, supporting theoretical predictions in molecular modeling. These results further the understanding in the impact of linker peptide on antibody structure and activity.

Accurate determination of the absolute phase and temporal-pulse phase of few-cycle laser pulses

A Fourier analysis method is used to accurately determine not only the absolute phase but also the temporal-pulse phase of an isolated few-cycle (chirped) laser pulse. This method is independent of the pulse shape and can fully characterize the light wave even though only a few samples per optical cycle are available. It paves the way for investigating the absolute phase-dependent extreme nonlinear optics, and the evolutions of the absolute phase and the temporal-pulse phase of few-cycle laser pulses.