Concentration methods ultrafiltration

Two general classes of methods can be functionally defined for preparing concentrates of organic substances. Concentration methods involve the removal of water (e.g., lyophilization, freeze concentration, vacuum distillation, reverse osmosis [RO], and ultrafiltration) and result in a more highly concentrated aqueous solution of organic contaminants. Isolation methods are those methods in which the organic substances are physically removed from the aqueous solution, for example, adsorption onto a solid substrate followed by desorption (I). 

Because the application of NMR spectroscopy to environmental samples is relatively new, we focused our studies on the identification and characterization of DOP by 31P FT-NMR spectroscopy. Ultrafiltration and reverse osmosis concentration techniques were employed to increase the dissolved organic phosphorus concentrations to the detection level of 31P FT-NMR techniques (approximately 10-20 mg of P/L). With these concentration methods a DOP concentration factor of up to 2000 is obtainable. This chapter reports the use of 31P FT-NMR spectroscopy in the analysis of DOP. In... 

Concentration. Clarified filtrates, centrates, or column eluates are usually too dilute for use in their specific applications, thus, substantial amounts of water must be removed. This can be achieved by evaporation or by ultrafiltration. Concentration methods used in industrial settings, such as evaporation, which is done under vacuum, and solvent extraction, may or may not be suitable for dewatering proteins because of their potential for thermal or chemical denaturation, and due to high energy costs associated with evaporation. The benefit of evaporation is that nonvolatile compounds that may stabilize the proteins are retained. 

Schratter, P. (1996) Purification and concentration by ultrafiltration, Methods Md. Biol. 59,115-134. 

A concentration process involves removal of a solvent, typically water from a macromolecular solution. Ultrafiltration is the method of choice for large-scale concentration. The selectivity issue involving removal of water from a macromolecular solution using ultrafiltration is trivial. The main challenge in a concentration process is maintaining a high productivity on account of the increased macromolecular concentration in the feed solution. Some of the main applications of macromolecular concentration using ultrafiltration are listed below [2] ... 

When only proteins remaining soluble after crushing the berries have to be studied, the so-called free-run juice can be prepared. To prepare the free run juice, the suspension obtained from squeezing the berries is filtered through cloth and the suspended particles are separated by centrifugation. The soluble proteins can be concentrated by ultrafiltration on membranes with a cut-off lower than 10 kDa (Waters et al., 1998). Proteins can be collected from the (concentrated) solution by the classical methods for protein precipitation, such as ammonium sulphate at 80% saturation, or organic solvents added to a final concentration... 

Humic and fulvic acids as well as humin were isolated from the samples described in Table 1 by standard methods ( ). In short, humic and fulvic acids are extracted with 0.5 N NaOH under N2. Humic acids are protonated on an ion exchange resin, precipitated by acidifying to pH 2, separated by centrifugation, and lyophilyzed. The soluble fulvic acids are concentrated by ultrafiltration and lyophilyzed. Humin, the residue after treatment with NaOH, is treated with concentrated HC1 HF to remove a large portion of the mineral matter and hydrolyzable substances such as proteins and polysaccharides. 

In soybean concentrates and isolates much of the phytate remains associated with the protein in fact, phytate may constitute as much as 2-3% of the weight of a commercial protein isolate (57). A low-phytate soybean protein isolate can be prepared from soybean flour, however, by allowing endogenous phytase to act on the phytate in a 6% suspension of the flour at pH 5 at a temperature of 65°C (58). Hydrolysis of the phytate facilitates its separation from the bulk of the soybean protein which is then concentrated by ultrafiltration using a membrane which is permeable to phytate and its hydrolysis products but impermeable to protein. The product obtained by this method contains over 90% protein and only about 0.3% phosphorus. 

The initial PIA purification method was developed by Mack et al. (3). These authors used a different, two-step chromatography protocol involving size-exclusion and ion exchange chromatography on Sephadex G-200, Q-Sepharose, and S-Sepharose. A similar purification method has been described recently to isolate a PIA-related polysaccharide polymer in E. coli (7). Briefly, E. coli cells were incubated in 50 raM Tris-HCL buffer (pH 8.0), 100 mg lysozyme, and 0.1 M EDTA at room temperature for 2 h. Phenol/chloroform extraction steps were performed to separate protein and debris contamination from the polysaccharide. Samples were concentrated by ultrafiltration devices (10,000 MW cut off) and fractionated on a fast protein liquid chromatography (FPLC) system with a Sephacryl S-2000 column (equilibration and elution buffer 0.1 MPBS, pH 7.4). 

The general concept of the integration of ion exchange, which effectively increases the concentration of the metal ions to be recovered, with the metal deposition process to enhance the efficiency of the metal deposition can be applied with other separation/concentration methods. One of these methods is based on the use of crossflow membrane ultrafiltration [38]. The complexation-ultrafiltration of the dissolved metal ions allows the concentration of the solution prior to electrolysis. 

The method used is governed by the market application of the exopolysaccharide. In general, the food industry has a requirement for a dry powder, whereas for several other applications, such as enhanced oil recovery, a liquid product is required and the ultrafiltration concentrate is preferred. 

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. 

A disadvantage of the ethanol injection method to produce SUV is the need to use a low lipid concentration, resulting in a low encapsulation efficiency of the aqueous phase. The dispersions can be concentrated by ultracentrifugation, ultrafiltration, or removal of water by evaporation. 

In studies of mice, rats, and dogs, diisopropyl methylphosphonate was rapidly absorbed into plasma (Hart 1976). The plasma data indicate that all three species rapidly absorbed diisopropyl methylphosphonate, although the exact rate was species specific. Although no studies were located regarding human absorption, diisopropyl methylphosphonate is also likely to be absorbed rapidly into the plasma of humans. The ability of porous polymeric sorbents, activated carbon, and dialysis to remove diisopropyl methylphosphonate from human plasma has been studied (McPhillips 1983). The grafted butyl-XAD-4 was found to be the most efficient sorbent for the removal of diisopropyl methylphosphonate from human plasma. Hemoperfusion of plasma over synthetic XAD-4 or butyl-XAD-4 sorbent resin was more efficient than dialysis/ultrafiltration for the removal of diisopropyl methylphosphonate from human plasma the smaller surface of the packed resins provided less area to minimize damage to molecular constituents of the plasma. These methods are useful in reducing diisopropyl methylphosphonate concentrations in the plasma. However, since diisopropyl methylphosphonate and its metabolites are not retained by the body, the need for methods to reduce body burden is uncertain. 

Dissolve the purified SPDP-modified dendrimer of step 5 in 50 mM sodium phosphate, 0.15M NaCl, pH 7.5, or in DMSO at a concentration of at least lOmg/ml. Add a 10-20 X molar excess of an amine-reactive fluorescent molecule (i.e., NHS-rhodamine or a hydrophilic NHS-Cy5 derivative see section on fluorescent probes). React with mixing for 1 hour at room temperature. Purify the fluorescently labeled SPDP-modified dendrimer using gel filtration or ultrafiltration. Follow the method of either step 7 or 8 to conjugate the dendrimer to another protein or molecule. 

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