Electrophoresis adsorption

Fig. 40. Influence of adsorption in zone and two-dimensional electrophoresis. (A) In zone electrophoresis adsorption causes contamination of the fractions. (B) In two-dimensional electrophoresis adsorption has no influence on the purity of the fractions.

Behavior. Diffusion, Brownian motion, electrophoresis, osmosis, rheology, mechanics, and optical and electrical properties are among the general physical properties and phenomena that are primarily important in coUoidal systems (21,24—27). Of course, chemical reactivity and adsorption often play important, if not dominant, roles. Any physical and chemical feature may ultimately govern a specific industrial process and determine final product characteristics. 

A. W. K. Tiselius (Uppsala) electrophoresis and adsorption analysis, especially for discoveries concerning the complex nature of the serum proteins. 

Discrimination between the enantiomers of a racemic mixture is a complex task in analytical sciences. Because enantiomers differ only in their structural orientation, and not in their physico-chemical properties, separation can only be achieved within an environment which is unichiral. Unichiral means that a counterpart of the race-mate to be separated consists of a pure enantiomeric form, or shows at least enrichment in one isomeric form. Discrimination or separation can be performed by a wide variety of adsorption techniques, e.g. chromatography in different modes and electrophoresis. As explained above, the enantioseparation of a racemate requires a non-racemic counterpart, and this can be presented in three different ways ... 

The form of the effective mobility tensor remains unchanged as in Eq. (125), which imphes that the fluid flow does not affect the mobility terms. This is reasonable for an uncharged medium, where there is no interaction between the electric field and the convective flow field. However, the hydrodynamic term, Eq. (128), is affected by the electric field, since electroconvective flux at the boundary between the two phases causes solute to transport from one phase to the other, which can change the mean effective velocity through the system. One can also note that even if no electric field is applied, the mean velocity is affected by the diffusive transport into the stationary phase. Paine et al. [285] developed expressions to show that reversible adsorption and heterogeneous reaction affected the effective dispersion terms for flow in a capillary tube the present problem shows how partitioning, driven both by electrophoresis and diffusion, into the second phase will affect the overall dispersion and mean velocity terms. 

In addition to chromatography based on adsorption, ion pair chromatography (IP-HPLC) and capillary electrophoresis (CE) or capillary zone electrophoresis (CZE) are new methods that became popular and are sufficiently accurate for these types of investigations. Other methods involving electrochemical responses include differential pulse polarography, adsorptive and derived voltammetry, and more recently, electrochemical sensors. 

Conventional colloid chemistry and elaitrochemistry have always been clo ly related with each other, the keywords electrophoresis, double layer theory, and specific adsorption describing typical asp ts of this relationship. In more ro nt times, new aspects have arisen which again bring colloid chemistry into contact with modem developments in electrcolloidal particles as catalysts for electron transfer reactions and as photocatalysts. In fact, the similarity between the reactions that occur on colloidal particles and on compact electrodes has often been emphasized by calling the small particles microelectrodes . 

Chemical surface modifications The first surface modification for the purpose of eliminating EOF and protein adsorption was recommended by Hjerten.28 The attachment of vinyl silanes allowed the polymerization of a variety of molecules to the surface. Most of the chemical modifications used for preparing capillaries for electrophoresis originated from the experience acquired over the years preparing GC and LC stationary phases. Chemical modification should conform to certain requirements, including the prevention of adsorption, the provision of stable and constant EOF over a wide pH range, chemical stability, ease of preparation, and reproduciblity of preparation. The effects of silanization of the inner surface of capillaries on electrophoretic separations have been extensively studied.26-29... 

Gassner, B., Friedl, W., and Kenndler, E., Wall adsorption of small anions in capillary zone electrophoresis induced by cationic trace constituents of the buffer, /. Ckromatogr., 680, 25, 1994. 

Hjerten, S., High-performance electrophoresis. Elimination of electroendos-mosis and solute adsorption,. Ckromatogr., 347, 191, 1985. 

Green, J. S. and Jorgenson, J. W., Minimizing adsorption of proteins on fused silica in capillary zone electrophoresis by the addition of alkali metal salts to the buffers, /. Chromatogr., 478, 63, 1989. 

Three bacterial species (E. coli, P. putida, and S. rubidae) were separated on isoelectric focusing in methylcellulose coated capillaries, and three bacterial species (P. fluorescens, E. aerogenes, and M. luteus) and the yeast S. cerevisae, were separated by capillary electrophoresis in the presence of polyethylene oxide.101 The polymers served to minimize adsorption to the walls without causing cellular lysis. 

Various methods have been used to examine the composition of proteins adsorbed to SAMs. Overall adsorption patterns can be examined with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) [50, 76, 77]. Absorbed proteins are eluted from the surface with surfactant (SDS), and then separated by electrophoresis. The proteins of interest are examined by western blotting [50, 76, 77]. Protein-specific antibodies can be used to detect proteins of... 

TABLE 1 Electrophoresis of BSA adsorbed on glass at various ionic strengths. Initial adsorption (at 0.25% w/v, BSA) at 1.96 yg/cm2, and electrophoretic mobility U. 

Influence of the Surface Concentration of BSA. Compared to the corrected moving boundary electrophoretic mobility of BSA in solution, the mobility of BSA adsorbed onto glass is considerably faster at all ionic strengths at 1.96 pg/cm2 and somewhat faster at lower ionic strengths 1.38 pg/cm2. However, at lower adsorption densities (1.05 and 0.64 pg/cm2), the adsorbed BSA moves more slowly in the applied electric field than BSA in moving boundary electrophoresis under otherwise identical conditions, and at the lowest surface adsorption (0.64 pg/cm2) the mobility of the adsorbed BSA are even somewhat slower than in cellulose acetate gel at all conditions of ionic strength investigated. 

Figure 6.2 Electrostatic adsorption mechanism of Brunelle [1] (a) surface polarization as a function of pH (b) measurement of PZC of some oxides (equivalent to isoelectric point) by electrophoresis.

Paper chromatography Ion exchange chromatography Electrophoresis Gas chromatography Adsorption chromatography Gel permeation chromatography... 

Adsorption chromatography Size exclusion chromatogaphy Thin-layer chromatography Gas chromatography Electrophoresis... 

Zonal techniques are the most frequently used form of electrophoresis and involve the application of a sample as a small zone to a relatively large area of inert supporting medium which enables the subsequent detection of the separated sample zones. A wide range of supporting media have been developed either to eliminate difficulties caused by some media (e.g. the adsorptive effects of paper) or to offer additional features (e.g. the molecular sieving effects of polyacrylamide gel). 

---