Profile of Separation

There is another factor which helps to make gel electrofocusing simpler than gel electrophoresis. The profile of the separation is not affected by the structure of the gel. However, if the gel has small pores, it may slow down the over-all time of separation, and perhaps stop very large protein molecules altogether. 

In electrophoresis, the gel directly affects the profile of the separation. The protein components of the sample are fractionated by the gel in accordance with the different charges they carry and the size of their 

In gel electrofocusing, however, no matter how each protein component is distributed at the beginning of a run, it always ends up at the point on the gradient where the pH is equal to the isoelectric point of the protein component. It becomes sharply concentrated there and stays there. In other words, the final position of each protein component is independent of the gel pores. This can be strikingly demonstrated by parallel tests, applying the sample at different places on the gel. After electro-focusing, it will be found that the same component gathers in the same place. If this should not be the case, the pores of the gel are too small for the protein molecules to move. 

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Figure 2 Chromatographic profile of separation of proteins in rat serum on a DEAE anion-exchange column at pH 4.5. The conditions of the separation are detailed in Sec. III. Individual fractions were pooled as indicated into eight major pools and analyzed by 2-D PAGE (Fig. 3).

Figure 9.152 Profile of separation of relaxed DNA from supercoiled plasmid pBR329 on a DEAE-NPR column. The reaction was carried out with 1 unit of calf thymus topoisomerase I. Twenty microliters of the reaction mixture was applied on the column and eluted with a linear gradient of 0.5 to 0.65 M NaCl for 30 minutes. DNAs from peaks a and b were collected manually and analyzed by electrophoresis on a 1% agarose gel after ethanol precipitation. Inset O.C., open circular S.C., supercoiled. (From Onishi et al., 1993.)...

Figure 17.3.12 AES depth profiles for carbon and fluorine in 2000-A-thick MgPc films. Carbon profiles of separate samples were normalized to a common curve, (a) MgPc film immersed in aqueous 0.1 M KPF6. (b) MgPc film oxidized in 0.1 M KPF solution.

Fig. 3 Profile of separation methamphetamine enantiomer (A) in precolunm and (B) analytical column.

The resulting concentration profile of separated zones is rectangular in shape, because steady-state concentration in the sample zones is adapted to the concentration of the leading ion according to Kohlrausch s regulating function [33]... 

For those dmgs that are administered as the racemate, each enantiomer needs to be monitored separately yet simultaneously, since metaboHsm, excretion or clearance maybe radically different for the two enantiomers. Further complicating dmg profiles for chiral dmgs is that often the pharmacodynamics and pharmacokinetics of the racemic dmg is not just the sum of the profiles of the individual enantiomers. 

Advantages to Membrane Separation This subsertion covers the commercially important membrane applications. AU except electrodialysis are pressure driven. All except pervaporation involve no phase change. All tend to be inherently low-energy consumers in the-oiy if not in practice. They operate by a different mechanism than do other separation methods, so they have a unique profile of strengths and weaknesses. In some cases they provide unusual sharpness of separation, but in most cases they perform a separation at lower cost, provide more valuable products, and do so with fewer undesirable side effects than older separations methods. The membrane interposes a new phase between feed and product. It controls the transfer of mass between feed and product. It is a kinetic, not an equihbrium process. In a separation, a membrane will be selective because it passes some components much more rapidly than others. Many membranes are veiy selective. Membrane separations are often simpler than the alternatives. 

Quantifying the effect of surface roughness or morphology is difficult, however. Surface preparations that provide different degrees of surface roughness also usually produce surfaces that have different oxide thicknesses and mechanical properties, different compositions, or different contaminant levels. The problem of separation of these variables was circumvented in a recent study [52] by using a modified microtome as a micro milling machine to produce repeatable, well-characterized micron-sized patterns on clad 2024-T3 aluminum adherends. Fig. 2 shows the sawtooth profile created by this process. 

Preswelled Sephacryl S-1000 was prepared in a K26/100 column (88 X 2.6 cm). Equilibration with 0.005 M NaOH containing 0.002% NaN3 at a flow rate of 0.67 ml/min was achieved after 20 hr. Sample solutions were applied with a 5-ml injection loop. The mass and iodine-complexing potential of separated glucan components was determined off-line for each of the subsequently eluted 5-ml fractions. Based on the determined mass of carbohydrate for each of the fractions, elution profiles such as Fig. 16.1 were constructed. 

Reproducibility of separation for a Dextran T-500 sample was tested on a semipreparative Sephacryl system S-500/S-1000 (65 + 95x1.6 cm) over a period of 6 months. The elution profiles of Dextran T-500 could be superimposed with deviations in the elution axis of 3 ml ( 1 fraction), and deviations in carbohydrate content within 5% referring to the maximum value at V,e, = 213 ml (Fig. 16.8). 

Figure 16.15 shows the resulting chromatograms for the three glucan fractions obtained by previous preparative separation on Sephacryl S-200/S-1000 (Fig. 16.14). From the normalized fraction chromatograms, the elution profile of the initial mixture has been reconstructed by mixing 50% fraction 1, 40% fraction 2, and 10% fraction 3. Compared to the chromatogram of the preparative Sephacryl S-200/S-1000 system, separation with the TSK/ Superose system yields improved resolution in the low dp (high V, ) domain.

The flow profiles of electrodriven and pressure driven separations are illustrated in Figure 9.2. Electroosmotic flow, since it originates near the capillary walls, is characterized by a flat flow profile. A laminar profile is observed in pressure-driven systems. In pressure-driven flow systems, the highest velocities are reached in the center of the flow channels, while the lowest velocities are attained near the column walls. Since a zone of analyte-distributing events across the flow conduit has different velocities across a laminar profile, band broadening results as the analyte zone is transferred through the conduit. The flat electroosmotic flow profile created in electrodriven separations is a principal advantage of capillary electrophoretic techniques and results in extremely efficient separations. 

The separation of enantiomers is a very important topic to the pharmaceutical industry. It is well recognized that the biological activities and bioavailabilities of enantiomers often differ [1]. To further complicate matters, the pharmacokinetic profile of the racemate is often not just the sum of the profiles of the individual enantiomers. In many cases, one enantiomer has the desired pharmacological activity, whereas the other enantiomer may be responsible for undesirable side-effects. What often gets lost however is the fact that, in some cases, one enantiomer may be inert and, in many cases, both enantiomers may have therapeutic value, though not for the same disease state. It is also possible for one enantiomer to mediate the harmful effects of the other enantiomer. For instance, in the case of indacrinone, one enantiomer is a diuretic but causes uric acid retention, whereas the other enantiomer causes uric acid elimination. Thus, administration of a mixture of enantiomers, although not necessarily racemic, may have therapeutic value. 

The pharmacological activities of the isomers should be compared in vitro and in vivo in both animals and humans. Separate toxicological evaluation of the enantiomers would not usually be required when the profile of the racemate was relatively benign but unexpected effects - especially if unusual or near-effective doses in animals or near planned human exposure - would warrant further studies with the individual isomers. 

Stationary batteries serve predominantly as an emergency power supply, i.e., they are on continuous standby in order to be discharged for brief periods and sometimes deeply, up to 100 percent of nominal capacity, in the rare case of need. The following profile of requirements for the separator thus arises very low electrical resistance, low acid displacement, no leaching of substances harmful to float-... 

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