Dextran gels

Recovery and Purification. The dalbaheptides are present in both the fermentation broth and the mycelial mass, from which they can be extracted with acetone or methanol, or by raising the pH of the harvested material, eg, to a pH of 10.5—11 for A47934 (16) (44) and A41030 (41) and actaplanin (Table 2) (28). A detailed review on the isolation of dalbaheptides has been written (14). Recovery from aqueous solution is made by ion pair (avoparcin) or butanol (teicoplanin) extraction. The described isolation schemes use ion-exchange matrices such as Dowex and Amberlite IR, acidic alumina, cross-linked polymeric adsorbents such as Diaion HP and Amberlite XAD, cation-exchange dextran gel (Sephadex), and polyamides in various sequences. Reverse-phase hplc, ion-exchange, or affinity resins may be used for further purification (14,89). 

Sephadex. Other carbohydrate matrices such as Sephadex (based on dextran) have more uniform particle sizes. Their advantages over the celluloses include faster and more reproducible flow rates and they can be used directly without removal of fines . Sephadex, which can also be obtained in a variety of ion-exchange forms (see Table 15) consists of beads of a cross-linked dextran gel which swells in water and aqueous salt solutions. The smaller the bead size, the higher the resolution that is possible but the slower the flow rate. Typical applications of Sephadex gels are the fractionation of mixtures of polypeptides, proteins, nucleic acids, polysaccharides and for desalting solutions. 

Dextran gels have been utilized since the late 1950s (1) for the separation of biopolymers. First attempts on Sephadex (2-5) and Sephadex/Sepharose (6-8) systems are documented for hydrolyzed and native starch glucans. Up until now, particularly for the preparative and semipreparative separation of polysaccharides, a range of efficient and mechanically stable Sephacryl gels (9-14) have been developped. 

There are two types of stationary phases commonly used in exclusion chromatography silica gel and micro-reticulated cross-linked polystyrene gels. A third type of exclusion media is comprised of the Dextran gels. Dextran gels are produced by the action of certain bacteria on a sucrose substrate. They consist of framework of glucose units that can form a gel in aqueous solvents that have size exclusion properties. Unfortunately the gels are mechanically weak and thus, cannot tolerate the high pressures necessary for HPLC and, as a consequence, are of very limited use to the analyst. 

Flodin, P. Dextran Gels and Their Applications in Gel Filtration Halmatd, Uppsala, 1962. 

Fractionation Methods. Ultrafiltration and gel filtration are nondestructive methods which, based on limited experience, can be used for fractionation of mineral complexes from digests. In earlier studies mineral absorption on the gel material was a problem. Lonnerdal (30) introduced a method of treating dextran gels with sodium borohydride in order to eliminate the mineral-binding sites on the gel. In preliminary studies we have recovered more than 90 of Ca, Mg, Fe, Zn and P in samples applied to a borohydride-treated gel column (Sephadex G-50, Pharmacia Fine Chemicals, Piscataway, NJ). Recovery of Ca (Table IV) and Mg, Fe and Zn from ultrafiltration was also good. 

Dextran Gels Proteins and nucleotides can be separated by using cross-linked dextran gels available in various types and particle sizes. The molecular weight of dextran-gels vary considerably depending upon the extent of cross-linked nature. 

The timely adoption of the cross-linked dextran gels (i.e., Sephadex) in late-fifties as a packing material for column chromatography opened an altogether new horizon of chromatographic separation whereby substances, in general, undergo separation more or less as per their molecular size. 

Sephacryl (allyldextran cross-linked by N,N -methylenebisacrylamide) are the two types of dextran gel used in separation. Sephadex is mainly used as a glucose pol)rmer and employed for purification of many molecules such as lectins from Helix pomatia and Viciafaba, exoamylase from Pseudomonas stutzeri [14]. 

A good separation of L-rhamnose, 2-acetamido-2-deoxy-D-glucose, D-glucose, and 2-amino-2-deoxy-D-glucose, eluted in that order, was obtained by Zeleznick80 on a small column packed with the highly cross-linked, dextran gel Sephadex G-25, with 62 15 25 (v/v) butyl alcohol-M acetic acid-water as the eluant however, the gel functioned more as a support for partition chromatography than as a molecular sieve. 

P.Flodin, Dextran Gels and Their Applications in Gel Filtration, Pharmacia, Uppsala, Sweden, 1962. C-S.Wu, Handbook of Size Exclusion Chromatography, M.Dekker Inc., NY, 1995. 

Dextran gels used as molecular sieves are formed by crosslinking dextrans with epichlorhydrin to give a semisynthetic polysaccharide with a well-defined pore size (Sephadex ). 

The molecular-sieve dextran gels are widely used in chemistry, biochemistry, and pharmacy for the analytical and preparative separation of metabolites and other biological products. 

Chang (64) used a porous styrene/divinylbenzene gel to determine small amounts of fatty acid dimers in tall oil. Hase and Harva (66) separated the monomeracid methyl esters from dimers and higher oligomers using a modified dextran gel SEPHADEX LH-20. 

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