Run-to-run sample separation reproducibility has been compared on two commercial high performance capillary electrophoresis units which differ in the mode by which the capillary temperature is thermostatted. Three standard analytes, differing dramatically in molecular character and size, were used for the analysis: benzoic acid, a 14 amino acid peptide from human chorionic gonadotropin, and ribonuclease A represent, respectively, small stable organic molecules, small peptides with little or no secondary structure, and proteins with secondary structure. These standards were evaluated with regard to reproducibility of migration time, peak area, and peak height. The analyses, performed in buffers of optimum pH for the separations, demonstrated that the liquid and forced air convection thermostatted systems both performed extremely well. The reproducibility, as judged by the percent coefficient of variance (% CV) of replicate analyses, was generally found to be less than 1 % (migration time); the reproducibility decreased in the order migration time > peak height > peak area. Whereas the absolute % CV values for MTrel (migration relative to a standard) observed with the liquid thermostatted system were 2- to 4-fold lower than those observed with the forced air convection thermostatted system, there was little statistically significant difference between the two. As expected, the data indicated a reduction in reproducibility as the complexity of the analyte increased, perhaps as the result of an increased potential for wall interactions. Comparing separations in which low (≈?1 watt/meter [W/m] of capillary) and high (>5 W/m) Joule heat was generated by altering the sodium chloride content of the buffer revealed few statistically significant differences in the reproducibility obtained from the two systems. With these particular standard analytes and their respective buffer systems, there appears to be little difference between forced air convection and liquid thermostatting of the capillary. 相似文献
This work utilizes on-column ligand synthesis and affinity capillary electrophoresis (ACE) to determine binding constants (Kb) of 9-flourenylmethyloxy carbonyl (Fmoc)-amino acid derivatives to the glycopeptide antibiotics ristocetin (Rist) and teicoplanin (Teic). In this technique, two separate plugs of sample are injected on to the capillary column and electrophoresed. The initial sample plug contains a d-Ala-d-Ala terminus peptide and either one or two non-interacting standard(s). The second plug contains a Fmoc-amino acid-N-hydroxysuccinimide (NHS) ester. The electrophoresis is then carried out with an increasing concentration of Rist or Teic in the running buffer. Upon electrophoresis the initial d-Ala-d-Ala peptide reacts with the Fmoc-amino acid yielding a new Fmoc-amino acid-d-Ala-d-Ala peptide derivative. Continued electrophoresis results in the binding of Rist or Teic to the Fmoc-amino acid-d-Ala-d-Ala peptide derivatives. Analysis of the change in the relative migration time ratio (RMTR) or electrophoretic mobility () of the Fmoc-amino acid-d-Ala-d-Ala peptide derivatives relative to the non-interacting standards, as a function of the concentration of Rist and Teic, yields a value for
Kb. These findings demonstrate the advantage of coupling on-column ligand synthesis to ACE for estimating binding parameters between antibiotics and ligands.Abbreviations Rist
Ristocetin
- Teic
Teicoplanin
- ACE
Affinity capillary electrophoresis
- RMTR
Relative migration time ratio 相似文献
Multiple-injection affinity capillary electrophoresis (MIACE) is used to determine binding constants (Kb) between receptors and ligands using as model systems vancomycin and teicoplanin from Streptomyces orientalis and Actinoplanes teichomyceticus, respectively, and their binding to D-Ala-D-Ala peptides and carbonic anhydrase B (CAB. EC 4.2.1.1) and the binding of the latter to arylsulfonamides. A sample plug
containing a non-interacting standard is first injected followed by multiple plugs of sample containing the receptor and then
a final injection of sample containing a second standard. Between each injection of sample, a small plug of buffer is injected
which contains an increasing concentration of ligand to effect separation between the multiple injections of sample. Electrophoresis
is then carried out in an increasing concentration of ligand in the running buffer. Continued electrophoresis results in a
shift in the migration time of the receptor in the sample plugs upon binding to their respective ligand. Analysis of the change
in the relative migration time ratio (RMTR) or electrophoretic mobility (μ) of the resultant receptor–ligand complex relative to the non-interacting standards, as a function of the concentration of
ligand yields a value for Kb. The MIACE technique is a modification in the ACE method that allows for the estimation of binding affinities between biological
interactions on a timescale faster than that found for standard ACE. In addition sample volume requirements for the technique
are reduced compared to traditional ACE assays. These findings demonstrate the advantage of using MIACE to estimate binding
parameters between receptors and ligands. 相似文献
Liquid and solid cyclic peroxides derived from aliphatic ketones are explosive materials so their enthalpies of vaporization
and other thermodynamic or condensed-phase properties cannot be measured directly. In this work the enthalpies of vaporization
of peroxides at 298.15 K were estimated simply from gas chromatographic retention times measured at different temperatures.
The technique correlates changes in the retention times of compounds whose enthalpies of vaporization are known (called the
reference series), with those of the compounds of interest. If tR′ is the adjusted retention time (retention time of each compound minus the retention time of unretained diethyl ether, used
as solvent) a plot of ln tR′ against 1/T for each compound (reference compounds and cyclic peroxides) results in a straight line (r2 > 0.99 for all compounds). The enthalpy of transfer from solution to the vapor state (ΔsolgHm) can be obtained by multiplying the slope by the gas constant (R). A second plot correlates the enthalpies of transfer from
solution to the vapor state (ΔsolgHm), as measured by gas–liquid chromatography (GLC), with enthalpies of vaporization of reference materials (ΔvapHm at 298.15 K) available in the literature. C9–C15 fatty acid methyl esters and hydrocarbons were used as reference compounds. The enthalpies of vaporization of the cyclic
organic peroxides were calculated from the equation of the line obtained in this second correlation, the slope of which was
ΔvapHm (at 298.15 K)/ΔgsolHm. The experiments were performed under isothermal conditions with a DB-5 capillary column, flame-ionization detection (FID),
and nitrogen as carrier gas. The column temperature was varied over a range of at least 30–70 K between 403 and 473 K, with
chromatograms being acquired at 10 K intervals. Enthalpies of vaporization of cyclic organic peroxides are not available in
the literature, and the values given in this paper, obtained by gas chromatography, are the first to be reported. 相似文献
The transition states of intramolecular 1,4 and 1,5 H-atom transfers, from/to primary (p), secondary (s) or tertiary alkyl (t) and primary (pa), secondary (sa) or tertiary (ta) allyl carbon atoms, have been studied at the level of the semiempirical quantum-chemical method AM1 with the UHF approximation. The activation and reaction enthalpies were calculated and compared with experimental data available in the literature and the calculated data obtained for analogous reactions in alkyl radicals. Correlations were found between the activation enthalpies and the dissociation enthalpies of the bonds broken and formed. 相似文献
A CZE method was developed and validated for the analysis of Olmesartan medoxomil (OLMD) in tablets. The influences of pH, buffer concentration, applied voltage and capillary temperature on the migration time of OLMD were investigated. About 50 mM pH 6.5 phosphate buffer were used as background electrolyte. The optimum instrument parameters were found to be 30 °C temperature with 30 kV applied voltage and diode array detection was carried out at 210 nm. OLMD was hydrodynamically injected (Pinj = 50 mbar, tinj = 3 s) and an internal standard, diflunisal (IS), was used to improve the precision and repeatability. Under these conditions, the migration time of OLMD was 2.32 min and the total analysis time was shorter than 5 min. Linearity range for the developed method was found to be 2.0–50.0 μg mL−1 and the limit of detection was 0.5 μg mL−1. The developed method was applied for the analysis of OLMD in pharmaceutical tablet formulations.
This article presents an analytical approach for identification problems related to ion transport problems. In the first part
of the study, relationship between the flux jL : = (D(x)ux(0, t)x=0{\varphi_L := (D(x)u_x(0, t)_{x=0}} and the current response I(t){{\mathcal I}(t)} is analyzed for various models. It is shown that in pure diffusive linear model case the flux is proportional to the classical
Cottrelian IC(t){{\mathcal I}_C(t)}. Similar relationship is derived in the case of nonlinear model including diffusion and migration. These results suggest
acceptability of the flux data as a measured output data in ion transport problems, instead of nonlocal additional condition
in the form an integral of concentration function. In pure diffusive and diffusive-convective linear models cases, explicit
analytical formulas between inputs (diffusion or/and convection coefficients) and output (measured flux data) are derived.
The proposed analytical approach permits one to determine the unknown diffusion coefficient from a single flux data given
at a fixed time t1 > 0, and unknown convection coefficient from a single flux data given at a fixed time t2 > t1 > 0. Linearized model of the nonlinear ion transport problem with variable diffusion and convection coefficients is analyzed.
It is shown that the measured output (flux) data can not be given arbitrarily. 相似文献
This article describes a method for preparation of low concentration gas standard mixtures of biogenic volatile organic compounds (BVOCs) emitted primarily by plants. A set of 10 plant volatiles including α-pinene, β-pinene, 3-carene, linalool, methyl salicylate, α-cedrene, β-caryophyllene, β-farnesene, aromadendrene and α-humulene was used in the study. Gas standard mixture of these compounds was generated using a capillary diffusion system (CDS). Diffusion coefficient (D) and saturation vapour pressure (ps) data of these compounds were calculated from experimentally determined gas chromatographic retention indices (RI) and empirical relationships between D and ps versus RI. A comparison of the calculated and measured concentrations of investigated compounds has proved that designed CDS can be successfully used for the proper quantification of BVOCs. 相似文献
A rapid and universal capillary zone electrophoresis (CZE) method was developed to determine the dissociation constants (pKa) of the 20 standard proteogenic amino acids. Since some amino acids are poorly detected by UV, capacitively coupled contactless
conductivity detection (C4D) was used as an additional detection mode. The C4D coupling proved to be very successful on a conventional CE-UV instrument, neither inducing supplementary analyses nor instrument
modification. In order to reduce the analysis time for pKa determination, two strategies were applied: (i) a short-end injection to reduce the effective length, and (ii) a dynamic
coating procedure to generate a large electroosmotic flow (EOF), even at pH values as low as 1.5. As a result, the analysis
time per amino acid was less than 2 h, using 22 optimized buffers covering a pH range from 1.5 to 12.0 at a constant ionic
strength of 50 mM. pKa values were calculated using an appropriate mathematical model describing the relationship between effective mobility and
pH. The obtained pKa values were in accordance with the literature.
Figure
a UV (1) and C4D (2) detectors placed on-line on the CE capillary. b Curve of effective mobility as a function of pH for histidine 相似文献
The above article (DOI: 10.1002/cem.1112) was published online on 14 February 2008. An error was subsequently identified: the captions for Figures 1 and 2 were omitted; they should read as follows: Figure 1. Orthogonality criterion (θA) for the octane data as a function of number of components (A) calculated using the standard PLS algorithm and SIMPLS. Figure 2. Orthogonality criterion (θA) for the wines data as a function of number of components (A) calculated using the standard PLS algorithm and SIMPLS. 相似文献
Energy stored in vibrational level ν = 1 of several individual dipolar diatomic molecules AB which are trapped in a rare gas matrix M is automatically accumulated in a higher level ν > 1 of a single molecule AB. This remarkable cascade of energy upwards competes with a cascade of energy downwards. the radiative decay. The interplay of both cascades, first observed by Dubost and Charneau, is explained a simple model. The model incorporates three processes into a master equation for the relative populations Pν(t) of levels ν: (a) migration of single quanta by resonance energy transfer, AB(1) + AB(0) ? AB(0) + AB(1); (b) phonon assisted excitation of upper levels, AB(1) + AB(ν) → AB(0) + AB(ν+1); and (c) radiative decay, AB(ν) → AB(ν-1). The model assumes that there is only one isotopic species AB which has a small but nonzero vibrational anharmonicity, that the temperature is low, T → 0 K, the concentration ratio ?M/?AB is large and that, initially, at time t = 0, a small fraction p1 of molecules AB is excited to level ν = 1. The master equation has only two parameters, the radiative lifetime trad and k 2/[?AB?1k(1,1 → 0,2)trad], where k(1,1 → 0,2) is the reference rate constant of process (b). The master equation is solved in closed form for the Pν(t). For trad = 14 ms and k = 0.2, very satisfactory qualitative agreement is found for the theoretical Pν(t) and the experimental time evolution of the relative population of vibrational levels of 12C16O in an argon matrix, for ?M/?AB = 2000 at T = 9 K. In agreement with experimental results it is concluded that the risetime of the fluorescence signals decreases whereas population inversion increases for decreasing values of ?M/?AB. At long times, t > trad, any population inversion should disappear. 相似文献
The power–time curves of the oscillating extraction system were determined at different temperatures for the extraction of
hydrochloric acid and acetic acid with primary amine N1923 (R–CH(NH2)–R1), R, R1 represent alkyl of C9–11 in chloroform using the titration microcalorimetric method. The apparent activation energy was calculated from the induction
period (tin), the first oscillation period (tp.1) and the second oscillation period (tp.2).
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
The correct (event weighted) average of kt, 〈kt〉, has been calculated for pseudostationary laser-induced polymerization for a kinetic scheme with chain-length dependent termination and compared to the average k̄t obtained by formally solving for kt the expression for the second moment of the chain-length distribution valid for chain-length independent termination (represented by the product of rate of polymerization νp and weight average degree of polymerization P̄w). It is shown that there is a fair agreement between the two quantities. This may be used to recover experimentally the power-law governing the dependence of kt on chain-length, especially its exponent. 相似文献
The migration of 237Np in an undisturbed Chinese loess column was investigated by direct γ-ray method. The column was taken from a field test site and installed in a laboratory simulation hall. Radionuclide 237Np in the form of neptunium nitrate, mixed with quartz, was introduced into the column and covered with loess. Artificial
rainfall was applied to the column for about 3 years and, the counting rates of 237Np in the column from 56 to 616 days at different vertical positions were detected with a γ-ray detection system. Based on the counting rates of 237Np in the simulation column at different vertical positions and the distance from the source layer, the relationship of the
mass center of 237Np in the column at different experimental periods to the experimental time was established, Cm = 0.36 log(t)-2.75. Here Cm is the mass center of 237Np in the column, cm, and t is the experimental time in days. Based on this relationship, the mass center of 237Np for the 1,073-day experiment was predicted and compared to that obtained with the final destructive method. The good agreement
between the prediction and the experimental values indicates that the direct γ-ray method could be used to predict the migration of strongly adsorbed radionuclides such as 237Np in environmental media with the help of laboratory simulation columns. 相似文献
Contributions to the Chemistry of Phosphorus. 87. 1,2-Di-tert-butyl-3-iso-propyl-cylclotriphosphane, a Stable Mixed-substituted Cyclotriphosphane The first kinetically stable mixed-substituted cyclotriphosphane, 1,2-di-tert-butyl-3-iso-propyl-cyclotriphosphane, (PBut)2(PPri) ( 1 ), was synthesized by [2+1]-cyclocondensation of K(But)P–P(But)K with PriPCl2 in n-pentane. Mainly (PBut)4 as well as mixed-substituted cyclotetra- and cyclopentaphosphanes are formed as by-products. 1 could be isolated in a pure state by high vacuum distillation and was thoroughly characterized. It forms two diastereomers, the more stable of which with a cis-standing tert-butyl and iso-propyl group can be stored without decomposition under inert conditions at room temperature for several days. Through thermolysis of 1 beside other alkylcyclophosphanes the mixed-substituted cyclotetraphosphanes (PBut)2(PPri)2 ( 2 ) and (PBut)3(PPri) ( 3 ) are formed and their 31P NMR parameters are reported. 相似文献
A simple, rapid, and accurate method for the separation and determination of honokiol and magnolol in Magnoliaofficinalis and related herbal medicines was developed by combination of flow injection (FI) and capillary zone electrophoresis (CZE). The analysis was carried out using an unmodified fused-silica capillary (50-μm I.D.; total length 7.5 cm; effective length 4.5 cm). A series of optimization steps afforded the following conditions: the sample solvent consisted of 150 mM NaOH and a running buffer composed of 10 mM sodium tetraborate/10 mM sodium dihydrogenphosphate (NaH2PO4) at pH 12 was applied for the separation of the analytes. The separation could be achieved within 5 min with a sample throughput rate of up to 28 h−1. The repeatability (defined as the relative standard deviation, RSD) for honokiol and magnolol was 2.0% and 1.6% with peak area evaluation, 3.6% and 2.0% with peak height evaluation, and 2.0% and 1.4% with migration time evaluation, respectively. Regression equations revealed linear relationships (r = 0.9991–0.9998) between the peak area of each analyte and the concentration. 相似文献
The study and modelling peak shape of in capillary electrochromatography (CEC), obtained using butyl acrylate (BA)-based monolithic columns, is described in this article. A modified-Gaussian model, which is a function of conventional experimental parameters: retention time (tR), peak height (H0) and standard deviation (σ0) at the peak maximum, and left (A) and right (B) halfwidths, was used to describe the peaks of thiourea and several polyaromatic hydrocarbons compounds, which were eluted under several applied voltages. A mean relative error below 2% in the prediction of peak shape is obtained. Based on peak parameters, an easy and reliable estimation of global chromatographic performance, efficiency and peak capacity of BA-based monoliths was also considered. A comparison in terms of chromatographic performance of BA-based monoliths in CEC with CE mode and RPLC microparticulate columns was also performed. 相似文献
Summary: The sol–gel transition of two thermoreversible gelling mixtures made of xanthan gum and locust‐bean gum has been studied by using in situ, time‐resolved dynamic light scattering (DLS) and in situ rheology. A critical dynamical behavior was observed near the sol–gel transition, which was characterized by the presence of power‐law spectra over three and four decades in the time‐intensity correlation function g2(t) − 1 ∼ t−μ and over four and three decades in the oscillatory shear experiment G′(ω) ∼ G″(ω) ∼ ωn. A comparison of the critical exponents obtained (μ1 ≈ 0.36, μ2 ≈ 0.32 and n1 ≈ 0.62, n2 ≈ 0.67) was made as a function of the dependence of the two mixing ratios according to the theory by Doi and Onuki. New experiments were also performed to compare the critical exponents on such a thermoreversible system.
Double‐logarithmic plot of the time‐intensity correlation functions g2(t) − 1 versus the delay time, t, at a 90° scattering angle and at several temperatures of the mixture 1. 相似文献