Ultrafiltration Fractionation

Dialysis and ultrafiltration Fractionation based on molecular size. Used for fractionation of WSE for further analysis. Potential limitations include rejection of hydrophobic peptides by UF membranes and aggregation of small peptides Kuchroo and Fox (1982b, 1983b), Visser et al. (1983), O Sullivan and Fox (1990), and Fox (1989)... 

Ultimately, the usefulness of tracer methods will partially depend upon how readily they can be incorporated into a field study. Methods that can be applied to filtered water samples are less labor intensive than those requiring some type of fractionation, such as the use of small-volume XAD-8 columns or ultrafiltration. However, column or ultrafiltration fractionation can be streamlined to make them practical for field studies, and the better resolution of DOM chemistry may make the extra effort worthwhile. If fulvic acid or high molecular weight fractions are isolated in a study, these can be saved for potential subsequent analysis of trace moieties as motivated by initial results. Finally, the overall question being addressed in a particular experimental or field study will determine which tracer methods, if any, are included. 

Ultrafiltration was applied to examine the size fractionation of Al, Ca, Cu, Fe, K, Na, and Pb in white and red wines [91]. Metal determinations were performed on the unfiltered wine, the 0.45 p,m membrane-filtered wine and each ultrafiltrate fraction. Aluminum was determined by ET-AAS, while FAAS was employed for Cu and Fe. An electroanalytical technique, stripping potentiometry, was selected for Pb measurement, whereas flame photometry was chosen for K and Na quantification. Fractionation patterns were evaluated and discussed. Castineira et al. [92] combined on-line tangential-flow multistage ultrafiltration with a home-built carbon analyzer and ICP-MS for size fractionation of nonvolatile dissolved organic compounds and metal species in three German white wines. The study showed that the major part of the elements investigated (up to 25) were dissolved in the size fraction of < 1 kDa, with the exception of Ba, Pb, and Sr, which also appeared in other fractions. 

Newcombe et al. 536] also concluded that the adsorption of four NOM ultrafiltration fractions on to activated carbon was consistent with the pore volume distributions of the carbons and the hydrodynamic diameters of the fractions. ... 

Elliott (1978) in a brief communication reported that more than 70% of the aluminium in blood was present in the plasma compartment. In normal subjects, they reported that a very small proportion of the plasma aluminium was ultrafiltrable. These findings were not consistent with thos of Lundin et al. (1978). In the study of (1978) the tendency was for the ultrafiltrable fraction to decrease as the total plasma aluminium concentration fell below 200 fiQlL. At high plasma aluminium levels, studies using polyethene glycol and direct ultrafiltration indicated that 60-70% of the aluminium was bound to high-molecular weight proteins, 10-20% was bound to albumin, and 10-30% was ultrafiltrable. 

Graf et al. (1982, 1981) performed in vivo ultrafiltration studies on patients during hemodialysis. Their results revealed an ultrafiltration fraction of about 20% of total plasma aluminium, suggesting that 80% of the aluminium was protein bound. 

In intact or testosterone-stimulated castrated male rats orally administered 40 mg/kg of hexane or water extracts or an ultrafiltration fraction daily for 20 days, the water extract caused a decrease in the weight of seminal vesicles in intact rats, and an increase in the weight of all accessory sexual organs in castrated rats. In intact rats, the ultrafiltration fraction exhibited the same but smaller effects than the water extract, with no effects observed in castrated rats. No effects on sexual organs were observed on rats administered the hexane extract (Hiermann and Bucar 1997). 

Figure 9.17 Size exclusion chromatography of lignosulphonate fractions after ultrafiltration. Fraction number and approximate molecular mass are shown in the figure [92].

Rabbits injected with l//gAs /kg i.p. excreted 60% of the dose in urine and 6% with feces during the first posttreatment day (Bertolero et al. 1981). iAs was found to be the predominant species in urine during the first 2h posttreatment. At later time points, DMA was the major metabolite (80% of urinary As at 6h). MMA was detected in urine as a minor metabolite (<5%). MMA, DMA, and iAs were found in plasma in similar proportions. At 6h after injection, the ultrafiltrable fraction of As in cytosol of lung, liver, and kidney from injected animals consisted of 3.6%, 30.1%, and 48.5% of iAs, respectively the portion as DMA reached 95.3%, 68.5%, and 47.6%. MMA ranged between 1.3% and 3.3%. In liver and kidney cytosol, most of the As (70.3%-86.5% of total As) was bound to proteins at 4 and 6h after injection. 

Ultrafiltration (qv) (uf) is increasingly used to remove water, salts, and other low molecular-weight impurities (21) water may be added to wash out impurities, ie, diafiltration. Ultrafiltration is rarely used to fractionate the proteins because the capacity and yield are too low when significant protein separation is achieved. Various vacuum evaporators are used to remove water to 20—40% dry matter. Spray drying is used if a powdery intermediate product is desired. Tyophilization (freeze-drying) is only used for heat-sensitive and highly priced enzymes. 

Autofiltration The retention of any material at the surface of the membrane gives rise to the possibility of a secondaiy or a dynamic membrane being formed. This is a significant problem for fractionation by ultrafiltration because microsolutes are partially retained by almost all retained macrosolutes. The degree of retention is quite case-specific. As a rule of thumb, higher pressure and more polarization resiilts in more autofiltration. Autofiltration is particularly problematic in attempts to fractionate macromolecules. 

Sodium poly(a-L-glutamate). It was washed with acetone, dried, dissolved in water and ppted with isopropanol at 5°. Impurities and low molecular weight fractions were removed by dialysis of the aqueous solution for 50h, followed by ultrafiltration through a filter impermeable to polymers of molecular weights greater the 10. The polymer was recovered by freeze-drying. [Mori et al. J Chem Soc, Faraday Trans I 2583 1978.]... 

Phosphoribosyl pyrophosphate synthetase (from human erythrocytes, or pigeon or chicken liver) [9015-83-2] Mr 60,000, [EC 2.7.6.1]. Purified 5100-fold by elution from DEAE-cellulose, fractionation with ammonium sulfate, filtration on Sepharose 4B and ultrafiltration. [Fox and Kelley J Biol Chem 246 5739 197h, Flaks Methods Enzymol6 158 1963 Kornberg et al. J Biol Chem 15 389 7955.]... 

Nonselective membranes can assist enantioselective processes, providing essential nonchiral separation characteristics and thus making a chiral separation based on enantioselectivity outside the membrane technically and economically feasible. For this purpose several configurations can be applied (i) liquid-liquid extraction based on hollow-fiber membrane fractionation (ii) liquid- membrane fractionation and (iii) micellar-enhanced ultrafiltration (MEUF). 

In the short term, we do not expect chiral membranes to find large-scale application. Therefore, membrane-assisted enantioselective processes are more likely to be applied. The two processes described in more detail (liquid-membrane fractionation and micellar-enhanced ultrafiltration) rely on established membrane processes and make use of chiral interactions outside the membrane. The major advantages of these... 

Purification of poloxamers has been extensively investigated due to their use in medical applications, the intention often being to remove potentially toxic components. Supercritical fluid fractionation and liquid fractionation have been used successfully to remove low-molecular weight impurities and antioxidants from poloxamers. Gel filtration, high-performance liquid chromatography (HPLC), and ultrafiltration through membranes are among the other techniques examined [5]. 

The alternative large scale recovery method to precipitation is ultrafiltration. For concentration of viscous exopolysaccharides, ultrafiltration is only effective for pseudoplastic polymers (shearing reduces effective viscosity see section 7.7). Thus, pseudoplastic xanthan gum can be concentrated to a viscosity of around 30,000 centipoise by ultrafiltration, whereas other polysaccharides which are less pseudoplastic, are concentrated only to a fraction of this viscosity and have proportionally lower flux rates. Xanthan gum is routinely concentrated 5 to 10-fold by ultrafiltration. 

The eluted luciferase is precipitated with ammonium sulfate. The precipitate is dissolved in 1 mM Tris-HCl, pH 8, containing 0.1 mM EDTA, 3 mM DTT and 0.1 M NaCl, and chromatographed on a column of Sephacryl S-300 (2.6 x 97 cm) using the same buffer. Luciferase is eluted in two peaks, corresponding to the molecular weights of about 420,000 (an aggregate) and 130,000, in a ratio of about 8 1. The fractions of these two peaks are pooled separately the Mr 420,000 luciferase is concentrated by either ultrafiltration or precipitation with ammonium sulfate. 

Fig. 3. QAE-Sephadex gradient separation of the B fruit extract. An 18 mg (uronic acid equivalents) sample of extract was dissolved in 20 ml of 125 mM imidazole-HCl buffer (pH 7.0) and applied to the column. The column was then eluted with 50 ml 125 mM buffer followed by a 125 mM to 1.5 M buffer gradient (500ml), and, finally, 50 ml of 1.5 M buffer. Fractions of 5 ml were collected and assayed for uronic acids. Groups of fractions (26-41, 45-50, 53-75 and 84-100) were pooled, concentrated by ultrafiltration and analyzed by HPLC.

Buesseler KO, Bauer JE, Chen RF, Eglinton TI, Gustafsson O, Landing W, Mopper K, Moran SB, Santschi PH, Vernon Clark R, Wells ML (1996) An intercomparison of cross-flow filtration techniques used for sampling marine colloids overview and organic carbon results. Marine Chem 55 1-31 Buffle J, Perret D, Newman M (1992) The use of filtration and ultrafiltration for size fractionation of aquatic particles, colloids, and macromolecules. In Enviroiunental particles. Buffle J, van Leeuwen HP (eds) Lewis Publishers, Boca Raton FL, pl71-230... 

Influence of U colloidal transport in organic-poor surface waters has been far less studied. Riotte et al. (2003) reported U losses from 0 to 70% during ultrafiltration experiments for surface waters of Mount Cameroon without nearly any DOC. Even in the low concentration waters, U can be significantly fractionated from other soluble elements by the occurrence of a colloidal phase, probably inorganic in origin. However, such fractionations are not systematic because of the occurrence of various colloidal phases, characterised by different physical and chemical properties, and hence different sorption and/or complexation capacities (Section 2.1). 

Presently, the precise determination of the true dissolved Th fraction in water samples remains a challenge. Results from ultrafiltration experiments on organic-rich water samples from the Mengong river tend to demonstrate that Th concentration is less than 15 ng/L in absence of DOC (Table 2 and Viers et al. 1997), and that Th is still controlled by organic carbon in the final filtrate of the ultrafiltration experiments. The latter conclusion is also supported by the results obtained for the Kalix river (Porcelli et al. 2001). These results therefore not only raised the question of the determination of the amount of dissolved Th in water but also of the nature of Th chemical speciation. 

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