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Biomedical subjects

Andreas Seidel-Morgenstern

Publications and source records attributed to Andreas Seidel-Morgenstern.

At least 19 recordsLinked to original sources

Pore-scale dispersion in electrokinetic flow through a random sphere packing.

The three-dimensional velocity field and corresponding hydrodynamic dispersion in electrokinetic flow through a random bulk packing of impermeable, nonconducting spheres are studied by quantitative numerical analysis. First, a fixed bed with interparticle porosity of 0.38 is generated using a parallel collective-rearrangement algorithm. Then, the interparticle velocity field is calculated using the lattice-Boltzmann (LB) method, and a random-walk particle-tracking method is finally employed to model advection-diffusion of an inert tracer in the LB velocity field. We demonstrate that the pore-scale velocity profile for electroosmotic flow (EOF) is nonuniform even under most ideal conditions, including a negligible thickness of the electrical double layer compared to the mean pore size, a uniform distribution of the electrokinetic potential at the solid-liquid interface, and the absence of applied pressure gradients. This EOF dynamics is caused by a nonuniform distribution of the local electrical field strength in the sphere packing and engenders significant hydrodynamic dispersion compared to pluglike EOF through a single straight channel. Both transient and asymptotic dispersion behaviors are analyzed for EOF in the context of packing microstructure and are compared to pressure-driven flow in dependence of the average velocity through the bed. A better hydrodynamic performance of EOF originates in a still much smaller amplitude of velocity fluctuations on a mesoscopic scale (covering several particle diameters), as well as on the microscopic scale of an individual pore.

Journal Article↗

Discontinuous and continuous separation of the monomeric and dimeric forms of human bone morphogenetic protein-2 from renaturation batches.

Bone morphogenetic protein-2 (BMP-2) is one of the most interesting of the approximately 14 BMPs which belong to the transforming-growth-factor-beta (TGF-beta) superfamily. BMP-2 induces bone formation and thus plays an important role as a pharmaceutical protein. Recently, rhBMP-2 has been produced in form of inactive inclusion bodies in Escherichia coli. After solubilization and renaturation the biologically active dimeric form of rhBMP-2 can be generated. However, inactive monomers of BMP-2 are also formed during the renaturation process which must be separated from the active dimeric BMP-2. The purpose of this paper is to present: (a) results of an experimental study of a chromatographic separation of the monomeric and dimeric forms; and (b) a concept for a continuous counter-current simulated moving bed (SMB) process. The capacity of heparin as stationary phase was estimated for different salt concentrations in the mobile phase. A simulation study of a three-zone SMB process was performed applying a two step salt gradient. The results reveal the potential of the process for the purification of the dimeric BMP-2.

Bone Morphogenetic Protein 2↗

Theoretical study of multicomponent continuous countercurrent chromatography based on connected 4-zone units.

Continuous countercurrent or simulated moving bed (SMB) chromatography is a well-established separation technology. Conventional processes are based on four zones which fulfil distinct functions in order to split a feed into two fractions. Frequently there is an interest in isolating a target component out of a feed mixture containing more than two components. Modifications of the classical SMB process are required to solve this task. In the last years several concepts exploiting more than four zones have been suggested. To analyse these concepts the equilibrium theory has been frequently applied, neglecting all kinetic effects. It is the purpose of this paper to apply an equilibrium stage model in order to describe the performance of a combination of two or three 4-zone true moving bed units which are connected in series or integrated into 8- or 12-zone true moving bed units. The performance of such units is evaluated with respect to their potential to continuously separate ternary or quaternary mixtures. The analysis is based on the assumption of linear adsorption isotherms. An important aspect is the introduction of additional purge streams required for a successful operation of integrated 8- and 12-zone units.

Computer Simulation↗

Adsorption behavior of the (+/-)-Tröger's base enantiomers in the phase system of a silica-based packing coated with amylose tri(3,5-dimethyl carbamate) and 2-propanol and molecular modeling interpretation.

The binary adsorption isotherms of the enantiomers of Tröger's base in the phase system made of Chiral Technologies ChiralPak AD [a silica-based packing coated with amylose tri(3,5-dimethyl carbamate)] as the chiral stationary phase (CSP) and 2-propanol as the mobile phase were measured by the perturbation method. The more retained enantiomer exhibits a S-shaped adsorption isotherm with a clear inflection point, the concentration of the less retained enantiomer having practically no competitive influence on this isotherm: In the entire range of concentrations studied, dq2/dC1 approximately 0. By contrast, the less retained enantiomer has a Langmuir adsorption isotherm when pure. At constant mobile phase concentrations, however, its equilibrium concentration in the adsorbed phase increases with increasing concentration of the more retained enantiomer and dq1/dC2 > 0. This cooperative adsorption behavior, opposed to the classical competitive behavior, is exceedingly rare but was clearly demonstrated in this case. Two adsorption isotherm equations that account for these physical observations were derived. They are based on the formation of an adsorbed multi-layer, as suggested by the isotherm data. The excellent agreement between the experimental overloaded elution profiles of binary mixtures and the profiles calculated with the equilibrium-dispersive model validates this binary isotherm model. The adsorption energies calculated by molecular mechanics (MM) and by molecular dynamics (MD) indicate that the chiral recognition arising from the different interactions between the functional groups of the CSP and the molecules of the Tröger's base enantiomers are mainly driven by their Van der Waals interactions. The MD data suggest that the interactions of the (-)-Tröger's base with the CSP are more favored by 8+/-(5) kJ/mol than those of (+)-Tröger's base. This difference seems to be a contributing factor to the increased retention of the - enantiomer on this chromatographic system. The modeling of the data also indicates that both enantiomers can form high stoichiometry complexes while binding onto the stationary phase, in agreement with the results of the equilibrium isotherm studies.

2-Propanol↗

Breakthrough curves and elution profiles of single solutes in case of adsorption isotherms with two inflection points.

The shape of breakthrough curves and elution profiles depends strongly on the course of the specific equilibrium functions characterizing the chromatographic system. For a highly efficient system the equilibrium theory provides a methodology how to predict the band profiles. The concept is frequently applied to analyze single component systems characterized by isotherms possessing simple shapes (Langmuir or anti-Langmuir behaviour). However, adsorption isotherms often possess more complicated shapes and have inflection points in their courses. This leads to the development of composite concentration waves and results in complex shapes of breakthrough curves and elution profiles. In this paper, the equilibrium theory is used to predict breakthrough curves for a chromatographic system characterized by an adsorption isotherm with two inflection points. The results obtained are validated by comparing with numerical solutions of the equilibrium dispersive model.

Adsorption↗

Application of preferential crystallization to resolve racemic compounds in a hybrid process.

The application of preferential crystallization is at present limited to conglomerate forming systems, which cover only a minor part of chiral substances. In this paper, a hybrid process is proposed that extends the applicability of the preferential crystallization principle to the more common racemic compound forming systems. It comprises a preliminary (e.g., chromatographic) enantiomeric enrichment step and preferential crystallization to finally produce the desired pure enantiomer(s). The applicability of preferential crystallization to racemic compounds is demonstrated on the example of mandelic acid as a model system. Direct monitoring of the separation progress is performed using combined online polarimetry and online density measurements. A cyclic crystallization process, which provides alternating the pure mandelic acid enantiomer and the racemic compound, is feasible and allows the resolution of rac-mandelic acid as part of the proposed hybrid approach.

Crystallization↗

Chromatographic enantioseparation of amino acids using a new chiral stationary phase based on a macrocyclic glycopeptide antibiotic.

The separation of the enantiomers of several a-amino acids was studied on a new chiral stationary phase (CSP) which is based on the macrocyclic glycopeptide antibiotic eremomycin attached to silica particles. Retention and separation factors were determined under analytical conditions at ambient temperature for different mobile phase compositions. In order to evaluate the potential with respect to preparative separations the adsorption isotherms of D- and L-methionine were determined for one mobile phase composition applying the elution by characteristic point method. The isotherms were validated by comparing experimentally determined elution profiles with predictions based on the equilibrium dispersive model. Finally, the performance of the eremomycin CSP was compared with a commercially available CSP based on the macrocyclic antibiotic teicoplanin. After determining the isotherms of D- and L-methionine also for the teicoplanin phase, the equilibrium dispersive model was used for both CSP to identify optimal operating conditions. For the separation and conditions considered the new eremomycin CSP revealed a better performance compared to the teicoplanin CSP.

Adsorption↗

[Application of adjustable moving overlapping separation range map in multisegment stepwise gradient elution condition optimization of high performance liquid chromatography].

Computer-assisted optimization of high performance liquid chromatographic separation conditions can be used to obtain optimized experimental conditions in relatively short time and low cost, and has been widely applied to the separation of complex samples. Based on "moving overlapping separation range map" method, a novel method of optimizing multi-segment stepwise gradient elution conditions, "adjustable moving overlapping separation range map" method, was developed. By predicting the retention time, peak width and resolution of each solute under different mobile phase conditions, the overlapping separation range map of the solutes in the sample is plotted. When the mobile phase composition on the current step is optimized, the overlapping separation range map for solutes eluted in next one or two steps is replotted simultaneously by considering the influence of elution conditions for current step on the retention of uneluted solutes. It is helpful to consider the separation of each solute in the sample simultaneously and find optimized separation conditions close to the global optimum by investigating the effect of mobile phase conditions for current step on the separation of solutes eluted in the current and next one or two steps. By slightly adjusting the optimized separation conditions using grid search method, the separation performance can be further improved. The application of "adjustable moving overlapping separation range map" method was illustrated by using experimental data from literature. The advantage of this method in high performance liquid chromatography was verified in binary mobile phase system.

English Abstract↗

Optimization of gradient elution conditions in multicomponent preparative liquid chromatography.

Gradient elution is widely applied in analytical chromatography to reduce the separation time and/or to improve the selectivity. Increasingly the potential of modulating the solvent strength during gradient operation is exploited in preparative liquid chromatography. The purpose of this paper is to investigate theoretically the effect of optimizing free parameters available in gradient chromatography (extents and shapes of gradients) on the productivity of isolating a target component in a multicomponent mixture. An equilibrium stage model was used to quantify and compare different modes of operation (isocratic and various variants of gradient elution). By combining experimental design and artificial neural network concepts, optimal conditions were identified for the production of the second eluting component in a ternary mixture. The strong impact of the shape of gradients on process performance is elucidated.

Chromatography, Liquid↗

Electrokinetic effects on the transport of charged analytes in biporous media with discrete ion-permselective regions.

The influence of external electrical fields on local concentration distributions and the mass transport of ionic background (buffer) species, as well as eluting co- and counterionic tracer molecules, was investigated in a fixed bed of native glass beads by confocal laser scanning microscopy and numerical simulations. Due to the negative surface charge of the porous glass beads and significant electrical double layer overlap, the intraparticle mesopore space becomes ion-permselective. This cation selectivity and the externally superimposed electrical fields induce concentration polarization in the bulk electrolyte solution adjacent to the particles. At the anodic hemisphere of a bead, the actual interplay of convection, diffusion, and electromigration leads to the formation of a convective-diffusion boundary layer with reduced ion concentrations relative to the bulk solution. At the opposite, cathodic hemisphere where counterions leave a bead in the direction of the applied field, electrolyte concentrations increase generating an enriched concentration polarization zone. Complementary data from quantitative confocal laser scanning microscopy and numerical simulations provide insight into the spatial variations of chemical and electrical potential gradients in the hierarchically structured material, including molar flux densities of the background ionic species, and reveal the elution dynamics of co- and counterionic analytes. These results demonstrate that concentration polarization in the external fluid domain, as well as the magnitude and sign of electrophoretic with respect to electroosmotic mobility in the ion-permselective domain, are major local contributions to coupled mass and charge transport, reflecting analyte retention, migration, and dispersion on a macroscopic scale.

Journal Article↗

Numerical analysis of electroosmotic flow in dense regular and random arrays of impermeable, nonconducting spheres.

We present a numerical scheme for analyzing steady-state isothermal electroosmotic flow (EOF) in three-dimensional random porous media, involving solution of the coupled Poisson, Nernst-Planck, and Navier-Stokes equations. While traditional finite-difference methods were used to resolve the Poisson-Nernst-Planck problem, the (electro)hydrodynamics has been addressed with high efficiency using the lattice-Boltzmann method. The developed model allows simulation of electrokinetic transport under most general conditions, including arbitrary value and distribution of electrokinetic potential at the solid-liquid interface, electrolyte composition, and pore space morphology. The approach provides quantitative information on a spatial distribution of simulated velocities. This feature was utilized to characterize EOF fields in regular and random, confined and bulk packings of hard (i.e., impermeable, nonconducting) spheres. Important aspects of pore space morphology (sphere size distribution), surface heterogeneity (mismatch in electrokinetic potentials at confining wall and sphere surface), and fluid phase properties (electrical double layer thickness) were investigated with respect to their influence on the EOF dynamics over microscopic and macroscopic spatial domains. Most important is the observation of a generally nonuniform pore-level EOF velocity profile in the sphere packings (even in the thin double layer limit) which is caused by pore space morphology and which is in contrast to the pluglike velocity distribution in a single, straight capillary under the same conditions.

Journal Article↗

Shortcut method for evaluation and design of a hybrid process for enantioseparations.

Hybrid processes for enantioseparations have a considerable potential for reducing investment and operational costs. An example is the combination of simulated moving bed (SMB) chromatography and selective crystallisation. However, the design of integrated processes is a difficult task. A shortcut method is presented that can serve as a tool for design and estimation of the potential of such processes. The approach requires only limited experimental data and thus allows for systematic parameter studies. The method is based on the determination of the purity-performance characteristic of the SMB process and rigorous application of mass balances. The use of relative mass fluxes allows derivation of simple algebraic expressions for essential process parameters. The significant potential of combining SMB and crystallisation is demonstrated for the example of the separation of mandelic acid enantiomers.

Chromatography↗

Analysis of applying different solvents for the mobile phase and for sample injection.

Overloading a chromatographic column with a compound possessing low solubility in the mobile phase has been investigated. In order to increase the concentration of injection a strong solvent for dissolving the feed was used. The injection of such concentrated samples brings the risk of triggering undesired crystallisation processes. A model system has been investigated with ethanol-water as the mobile phase and DL-threonine as the sample dissolved in pure water. Under extreme overloaded conditions band splitting was observed. Measurements of the adsorption isotherms and systematic solubility studies were carried out. For the process analysis a simplified mathematical model was applied. The simulations of the band profiles were compared with the experimental data.

Adsorption↗

Experimental study and simplified mathematical description of preferential crystallization.

The purpose of this paper is to investigate the concept of preferential crystallization, with a focus on aspects of quantification and application from an engineering point of view. The amino acid threonine was used as a model system in the experiments performed. Application of on-line polarimetry in combination with measurements of an on-line density meter proved the general applicability of this method in order to monitor directly the resolution progress, to recognize the region of "safe" resolution, and to gain information about the crystallization kinetics. For evaluation of the crystal growth kinetics of the desired enantiomers, the method of moments was applied to analyze isothermal single-step crystallization experiments. A population balance model is used in order to simulate the time changes of liquid-phase composition during the preferential crystallization process. Finally, preliminary experimental results regarding the cyclic operation mode are presented.

Journal Article↗

Influence of moderate Joule heating on electroosmotic flow velocity, retention, and efficiency in capillary electrochromatography.

The influence of Joule heating on electroosmotic flow velocity, the retention factor of neutral analytes, and separation efficiency in capillary electrochromatography was investigated theoretically and experimentally. A plot of electrical current against the applied electrical field strength was used to evaluate the Joule heating effect. When the mobile phase concentration of Tris buffer exceeded 5.0 mM in the studied capillary electrochromatography systems using particulate and monolithic columns (with an accompanying power level of heat dissipation higher than 0.35 W/m), the Joule heating effect became clearly noticeable. Theoretical models for describing the variation of electroosmotic flow velocity with increasing applied field strength and the change of retention factors for neutral analytes with electrical field strength at higher Tris buffer concentrations were analyzed to explain consequences of Joule heating in capillary electrochromatography. Qualitative agreement between experimental data and implications of the theoretical model analysis was observed. The decrease of separation efficiency in capillary electrochromatography with macroporous octadecylsilica particles at high buffer concentration can be also attributed to Joule heating mainly via the increased axial diffusion of the analyte molecules and dispersion of solute bands by a nonuniform electroosmotic flow profile over the column cross-section. However, within a moderate temperature range, the contribution of the macroscopic velocity profile in the column arising from radial temperature gradients is insignificant.

Chromatography, Micellar Electrokinetic Capillary↗

Analysis of the isolation of a target component using multicomponent isocratic preparative elution chromatography.

The separation of a certain target component from a multicomponent mixture using isocratic preparative elution chromatography was studied theoretically. In particular, the important and most complicated case was considered that the target component does not elute in the first or last position. To specify the productivity of collecting this component different options are suggested to identify suitable times for fractionation. Using a conventional Craig model, capable to quantify chromatographic processes, the impact of several essential parameters (e.g. threshold concentration, desired purity, injection volume, separation factor between neighboring components, composition of the mixture) is evaluated for a ternary system based on parametric calculations. The paper provides simple tools to evaluate and optimize the productivity and other objective functions relevant in multicomponent preparative chromatography.

Chromatography, Liquid↗

Experimental determination of single solute and competitive adsorption isotherms.

In order to design and to optimise preparative liquid chromatography, the knowledge of the underlying thermodynamic functions, i.e. the adsorption isotherms, is of large importance. Usually these functions can not be predicted and various techniques have been suggested to determine them experimentally. In this paper, several important methods to measure adsorption equilibrium data are discussed and evaluated. The main focus is set on dynamic methods analysing concentration profiles that could be detected at the outlet of fixed-beds packed with the stationary phase of interest. The theoretical background of the different methods is explained using classical equilibrium theory and the equilibrium dispersion model. Each method is illustrated based on experimental data collected in our laboratory. Based on these personal experiences recommendations are given regarding the potential and the applicability of the methods discussed.

Adsorption↗

Online monitoring of preferential crystallization of enantiomers.

Polarimetry is used for continuous online monitoring of optical resolution by preferential crystallization. In combination with refractometry the liquid phase composition is determined, allowing one to follow the resolution progress quantitatively. The measurement techniques were calibrated up to relatively high solution concentrations and combined with the crystallizer. The resolution of DL-threonine was performed by preferential crystallization experiments in aqueous solution varying several process parameters like supersaturation, seed amount, initial enantiomeric excess, and scale. The resolution progress can be conveniently described by profiles of the optical rotation (polarimetric signal) and the crystallization pathway in the corresponding ternary phase diagram. The method outlined is applicable for dynamic process optimization and control purposes in "quasi-continuous" chiral separation processes.

Calibration↗