Chromatographic system, dynamics

Mixing the additive in the eluent used as a mobile phase can also modify the chromatographic system (dynamic modification), but the use of modified adsorbents has led to an improvement of resolution. Example works include that by Armstrong and Zhou [11], who used a macrocyclic antibiotic as the chiral selector for enantiomeric separations of acids, racemic drugs, and dansyl amino acid on biphenyl-bonded silica. 

So far the plate theory has been used to examine first-order effects in chromatography. However, it can also be used in a number of other interesting ways to investigate second-order effects in both the chromatographic system itself and in ancillary apparatus such as the detector. The plate theory will now be used to examine the temperature effects that result from solute distribution between two phases. This theoretical treatment not only provides information on the thermal effects that occur in a column per se, but also gives further examples of the use of the plate theory to examine dynamic distribution systems and the different ways that it can be employed. 

The theory that results from the investigation of the dynamics of solute distribution between the two phases of a chromatographic system and which allows the different dispersion processes to be qualitatively and quantitatively specified has been designated the Rate Theory. However, historically, the Rate Theory was never developed as such, but evolved over more than a decade from the work of a number of physical chemists and chemical engineers, such as those mentioned in chapter 1. 

As known, SEC separates molecules and particles according to their hydro-dynamic volume in solution. In an ideal case, the SEC separation is based solely on entropy changes and is not accompanied with any enthalpic processes. In real systems, however, enthalpic interactions among components of the chromatographic system often play a nonnegligible role and affect the corresponding retention volumes (Vr) of samples. This is clearly evident from the elution behavior of small molecules, which depends rather strongly on their chemical nature and on the properties of eluent used. This is the case even for... 

In effect, the composition of the mobile phase, and thus the selectivity of the chromatographic system, has been changed. As mentioned in the text, dynamic FAB operates effectively with lower concentrations of matrix than static FAB and although its effect may be minimal it should always be considered. Post-column addition of matrix overcomes potential problems of this nature. 

Klatte, S.J. and Beck, T.L., Molecular-dynamics of tethered alkanes—temperature-dependent behavior in a high-density chromatographic system, 7. Phys. Chem., 91, 5727, 1993. 

Fortunately, the effects of most mobile-phase characteristics such as the nature and concentration of organic solvent or ionic additives the temperature, the pH, or the bioactivity and the relative retentiveness of a particular polypeptide or protein can be ascertained very readily from very small-scale batch test tube pilot experiments. Similarly, the influence of some sorbent variables, such as the effect of ligand composition, particle sizes, or pore diameter distribution can be ascertained from small-scale batch experiments. However, it is clear that the isothermal binding behavior of many polypeptides or proteins in static batch systems can vary significantly from what is observed in dynamic systems as usually practiced in a packed or expanded bed in column chromatographic systems. This behavior is not only related to issues of different accessibility of the polypeptides or proteins to the stationary phase surface area and hence different loading capacities, but also involves the complex relationships between diffusion kinetics and adsorption kinetics in the overall mass transport phenomenon. Thus, the more subtle effects associated with the influence of feedstock loading concentration on the... 

The extract is collected by depressurization on a column packed with a solid sorbent, in a vessel containing the appropriate solvent, in a collection device connected to a chromatograph, or on combined solid phase-solvent traps [92]. For extraction of volatile compounds, such solvents as acetone, CH2CI2, methanol, or liquid nitrogen are used. Silica gel columns are the most popular way of trapping solids. In this case, the selectivity of the process can be improved by selective elution of the sorbent [88, 92]. SFE can be conducted in a static mode in which sample and solvent are mixed and kept for a user-specified time at a constant pressure and temperature, or in a dynamic mode where the solvent flows through the sample in a continuous manner [56]. The extracted analytes can be collected into an off-line device or transferred to an on-line chromatographic system for direct analysis. 

Column efficiency is mainly dependent on the kinetic factors of the chromatographic system such as molecular diffusion, mass-flow dynamics, properties of the column packing bed, flow rate, and so on. The smaller the particles and the more uniform their packing in the column, the higher the efficiency. The faster the flow rate, the less time analyte molecules have for diffusive band-broadening. At the same time, the faster the flow rate, the further analyte molecules are from the thermodynamic equilibrium with the stationary phase. This shows that there should be an optimum flow rate that allows achievement of an optimum efficiency for a given column. Detailed discussions of the... 

The process of analyte retention in high-performance liquid chromatography (HPLC) involves many different aspects of molecular behavior and interactions in condensed media in a dynamic interfacial system. Molecular diffusion in the eluent flow with complex flow dynamics in a bimodal porous space is only one of many complex processes responsible for broadening of the chromatographic zone. Dynamic transfer of the analyte molecules between mobile phase and adsorbent surface in the presence of secondary equilibria effects is also only part of the processes responsible for the analyte retention on the column. These processes just outline a complex picture that chromatographic theory should be able to describe. 

While dynamic distribution of the analyte between the mobile phase and adsorbent surface is a primary process, there are many secondary processes in the chromatographic system that significantly alter the overall analyte retention and selectivity. Detailed theoretical discussion of the influence of secondary equilibria on the chromatographic retention is also given in Chapter 2. 

Bulk property detectors generally have neither the sensitivity nor the linear dynamic range of solute property detectors and, as a consequence, are less frequently used in modem LC analyses. Furthermore, none can be used with gradient elution, flow programming or temperature programming and so they place considerable restrictions on the choice of chromatographic system. They do, however, have certain unique areas of application, some of which have already been mentioned, but their use probably represents less than 5% of all LC analyses. 

The elution profile of an ideal chromatogram depends only on the thermodynamic behavior of the chromatographic system. In a real chromatogram additional mass transfer and fluid dynamic factors have to be taken into account. 

Physical properties and espedally the isotherms depend on temperature as well as eluent composition. Feed and eluent composition influence the viscosity and therefore the fluid dynamics. However, these effects have already to be taken into account when selecting the chromatographic system (Chapter 4). The operating temperature for preparative processes is commonly selected to be to close room temperature for... 

The whole chromatographic system is run for at least 30 min under the isocratic working conditions. For the determination of the dynamic noise the detector signal is registered during 10 min or longer. The noise is determined as shown in Figure 25.4 by... 

The most common extraction techniques for semivolatile and nonvolatile compounds from solid samples that can be coupled on-line with chromatography are liquid-solid extractions enhanced by microwaves, ultrasound sonication or with elevated temperature and pressures, and extraction with supercritical fluid. Elevated temperatures and the associated high mass-transfer rates are often essential when the goal is quantitative and reproducible extraction. In the case of volatile compounds, the sample pretreatment is typically easier, and solvent-free extraction methods, such as head-space extraction and thermal desorption/extraction cmi be applied. In on-line systems, the extraction can be performed in either static or dynamic mode, as long as the extraction system allows the on-line transfer of the extract to the chromatographic system. Most applications utilize dynamic extraction. However, dynamic extraction is advantageous in many respects, since the analytes are removed as soon as they are transferred from the sample to the extractant (solvent, fluid or gas) and the sample is continuously exposed to fresh solvent favouring further transfer of analytes from the sample matrix to the solvent. 

A chromatograph can be considered as a system and subdivided into subsystems. The subdivision of a liquid chromatograph into the first rank subsystem is shown in Fig. 3. The dynamics of the chromatographic system and its subsystems are discussed briefly in this chapter. 

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