Fractionation methods dialysis

The foremnner of the modern methods of asphalt fractionation was first described in 1916 (50) and the procedure was later modified by use of fuller s earth (attapulgite [1337-76-4]) to remove the resinous components (51). Further modifications and preferences led to the development of a variety of fractionation methods (52—58). Thus, because of the nature and varieties of fractions possible and the large number of precipitants or adsorbents, a great number of methods can be devised to determine the composition of asphalts (5,6,44,45). Fractions have also been separated by thermal diffusion (59), by dialysis (60), by electrolytic methods (61), and by repeated solvent fractionations (62,63). 

Water soluble protein with a relative molecular mass of ca. 32600, which particularly contains copper and zinc bound like chelate (ca. 4 gram atoms) and has superoxide-dismutase-activity. It is isolated from bovine liver or from hemolyzed, plasma free erythrocytes obtained from bovine blood. Purification by manyfold fractionated precipitation and solvolyse methods and definitive separation of the residual foreign proteins by denaturizing heating of the orgotein concentrate in buffer solution to ca. 65-70 C and gel filtration and/or dialysis. 

The successful of recovery of RNase A functional activity by a heat-induced AR method suggested the possibility of recovering RNase A immunoreactivity as well. The immunoreactivity of native RNase A and RNase A that was incubated at a concentration of 4 mg/mL in 10% neutral buffered formalin for 1 day and then freed of formaldehyde by dialysis against PBS was compared using capture enzyme-linked immunosorbent assay (ELISA). Selected fractions that... 

Regardless of their method of fabrication, most liposome preparations need to be further classified and purified before use. To remove excess aqueous components that were not encapsulated during the vesicle formation process, gel filtration using a column of Sephadex G-50 or dialysis can be employed. To fractionate the liposome population according to size, gel filtration using a column of Sepharose 2B or 4B should be done. 

Gel filtration with Sephadex was used by Ghassemi and Christman [426] to make separations by molecular size on water samples concentrated by vacuum evaporation. Fluorescence was also used as one method for following the fractionation. Molecular size was also used by Gjessing [427] but with pressure dialysis as the method of separation. A similar method of concentration and separation was used by Brown [428] to follow the dispersion of these materials as fresh and salt water mixed in the Baltic Sea. 

The first fractionation of urinary ampholytes in this way was carried out by Boulanger et al. (BIO) in 1952 with the use of ion-exchange resins. They had designed this procedure previously for the fractionation of ampholytes in blood serum (B8). According to this method, deproteinized urine was subjected to a double initial procedure aiming at the separation of low-molecular weight substances from macro-molecular ones. One of the methods consisted of the fractionation of urinary constituents by means of dialysis, the second was based on the selective precipitation of urinary ampholytes with cadmium hydroxide, which, as had previously been demonstrated, permits separation of the bulk of amino acids from polypeptides precipitated under these circumstances. Three fractions, i.e., the undialyzable part of urine, the dialyzed fraction, and the so-called cadmium precipitate were analyzed subsequently. 

A constant observation when the MRP were separated by various methods was that antioxidative effect was found in many different fractions. Both the dialysates and the retentates from dialysis were antioxidative to some extent. All the electrophoresis fractions exhibited some antioxidative effect. Attempts to separate the MRP by column chromatography on Sephadex G-50 have resulted in several fractions with some antioxidative effect, and so on. This indicates that several antioxidative products are formed by the Maillard reaction, possibly differing in molecular size and chemical structure, but perhaps with one single antioxidative functional group in common, such as a free radical function. However, it can not be excluded that the MRP contain a few entirely different antioxidants with different modes of action. Various mechanisms have also been suggested. Eichner (6) claimed MRP to inactivate the hydroperoxides formed by the lipid oxidation. There are also reports on the complex binding of metals by MRP (18, 19). 

Techniques can be classified into two main categories those that detect total metal concentrations and those that detect some operationally defined fraction of the total. Methods which detect total concentrations such as inductively coupled plasma spectrometry, neutron activation analysis, atomic absorption spectrometry and atomic emission spectrometry have no inherent speciation capabilities and must be combined with some other separation technique(s) to allow different species to be detected (approach A in Fig. 8.2). Such separation methods normally fractionate a sample on the basis of size, e.g. filtration/ultrafiltration, gel filtration, or a combination of size and charge, e.g. dialysis, ion exchange and solvent extraction (De Vitre et al., 1987 Badey, 1989b Berggren, 1989 1990 Buffle et al., 1992). In all instances the complexes studied must be relatively inert so that their concentrations are not appreciably modified during the fractionation procedure. 

The comparison of a number of dialytic extracts with the parent coals is given in Table 1L These results indicate that the elemental composition of the dialytic extract closely mirrors that of the organic fraction of the coal. Similar conclusions were reached when coal liquids were separated via the dialytic method. The conclusion that dialysis does not concentrate any particular compound type deserves further investigation, since obtaining a representative sample is crucial to the utility of the method. In Table H, it is particularly interesting to note that in each case the "organic sulfur 1 from the classical coal analysis is almost identical to the sulfur content directly determined on the dialytic extract. 

At the close of the dialysis the plasma was centrifuged to remove small amounts of precipitate and the clear plasma resulting was subjected to starch-block preparative zone electrophoresis by the technique of Kunkel and Slater (15). At the close of the electrophoresis 1-cm. segments of the starch block were cut and transferred to sintered glass funnels, and the proteins were quantitatively eluted with five successive 2-ml. aliquots of 0.9% sodium chloride. The filtrates containing the protein fractions were then made up to a constant volume and mixed, and small aliquots were analyzed for total protein content by the method of Lowry et al. (18). 

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