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PubMed1998

Purification of alpha 1 proteinase inhibitor from human plasma fraction IV-1 by ion exchange chromatography.

BACKGROUND AND OBJECTIVES: Alpha-proteinase inhibitor (PI) protects the lungs from proteolytic damage caused by elastase and can be used to treat congenital emphysema. We describe an improved method of purification of alpha 1 PI from redissolved fraction IV-1 paste. MATERIALS AND METHODS: The process used dimethylaminoethyl anion exchange chromatography, sulfopropyl cation exchange chromatography, virus inactivation by dry heat, and tri-n-butyl-phosphate/cholate treatment, followed by a second strong cation exchange chromatography. Optimizations of loading conditions for ion exchange chromatography at small scale (20-60 ml of suspension) are described. Virus inactivation was adjusted to provide the best yield of alpha 1 PI consistent with effective inactivation. The process has been effectively scaled up. RESULTS: The final product was approximately 90% pure by SDS-PAGE, with a 60-70% yield from starting fraction IV-1 paste. The process has been characterized by methods including nonreduced SDS-PAGE, alpha 1 PI inhibition assay, and biuret protein assay. CONCLUSION: The method described is an effective way of preparing large quantities of alpha 1 PI from fractionated plasma.

Anions↗
PubMed2000

[Purification of prothrombin in Nitschmann fraction III by membrane radial column ion-exchange liquid chromatography].

A method is described for the purification of human prothrombin complex concentrate (PCC) from Nitschmann fraction III by membrane radial column ion-exchange chromatography, which allows large sample volumes to be processed at low operation pressure. The Nitschmann fraction III (15 g) was mixed with 1,000 mL 0.06 mol/L Tris-HCl(pH 7.5). The centrifuged supernatant(10,000 r/min, 15 min, 20 degrees C) was applied onto a DEAE ion-exchange liquid chromatographic column(XK-16 DEAE fast flow Sepharose and DEAE membrane radical column chromatography) with almost the same excellent separation efficiency. The parameters of sample flow rate, elution flow rate and sample capacity were optimized.

Blood Coagulation Factors↗
PubMed1984

Glycated hemoglobin in uremic patients as measured by affinity and ion-exchange chromatography.

We estimated glycated ("glycosylated") hemoglobin in erythrocytes of hemodialyzed uremic patients by the aminophenylboronic acid affinity-chromatographic method and by an ion-exchange chromatographic method. As expected, apparent HbA1 concentrations were above normal in the uremic patients, owing to interference by the carbamylated hemoglobin. However, there was no significant difference between values for uremic and normal subjects when glycated hemoglobin was measured by the affinity method. Evidently it is unaffected by the presence of hemoglobin species modified by reactants not displaying a cis-1,2-diol group, such as carbamylated hemoglobin. For this reason it is preferable to the ion-exchange chromatographic method for accurate measurement of glycated hemoglobin.

Adult↗
PubMed2000

Behavior of basic compounds in ion-exchange capillary electrochromatography with low-pH carrier electrolytes.

This work describes the separation of basic aromatic compounds by capillary electrochromatography employing acidic carrier electrolytes and bare silica as well as strong cation-exchange stationary phases. A mixed-mode separation mechanism was involved, comprising chromatographic interactions (adsorption, ion-exchange) as well as electrophoretic migration. The influence of ion-exchange on the retention/migration of the solutes could be manipulated according to procedures commonly employed in ion chromatography. These include variations of the eluting strength and/or the concentration of the competing ion present in the background electrolyte. Using this approach, separation times could be shortened and changes in selectivity could be achieved for a number of analytes.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed2004

Development of urethane acrylate composite ion-exchange membranes and their electrochemical characterization.

With the objective of introducing antifouling characteristics into interpolymer types of cation and anion exchange membranes, the surface of these membranes was coated with a 12-microm-thick urethane acrylate layer and was cured by UV radiation of wavelengths 308 and 172 nm under a complete inert atmosphere. Different urethane acrylate composite ion exchange membranes developed were characterized in NaCl solution by measuring their ion-exchange capacity, volume fraction of water, contact angle with water, membrane conductance, and membrane potential. It was found that the electrochemical transport properties of urethane acrylate composite cation-exchange membranes were increased due to resonance stabilization of the urethane group, which acts as a weak acid and dissociates as a negatively charged urethane ion and a positively charged proton. This contributes toward the net charge density of the membrane matrix responsible for enhanced selectivity and conductivity, while for urethane acrylate composite anion-exchange membranes reduction in net charge density was responsible for reduction in electrochemical transport properties. Counterion transport number, permselectivity, and counterion diffusion coefficient values for these membranes were also estimated. Experiments were also carried out in higher homologs of sodium carboxylate solutions in order to observe the fouling tendencies of these membranes. It was concluded that it is possible to obtain antifouling characteristics of ion-exchange membranes by coating and curing thin hydrophilic layers of urethane acrylate on their surfaces without sacrificing their electrochemical transport properties.

Journal Article↗
PubMed2000

[Determination of ibandronate by high performance ion exchange chromatography].

A method for the determination of Ibandronate by high performance ion exchange chromatography is reported. Ibandronate can form a complex compound with the copper ion added in the mobile phase. The complex can be detected at UV 240 nm. The effects of pH of mobile phase and concentration of methanol to the retention time were studied. Concentration of 10% methanol and pH 2.5 were suitable for the detection in less than 5 min. The flow rate was 1.0 mL/min. The relative standard deviation of peak area was 0.8%. The linear correlation was observed from 300 mg/L to 1,500 mg/L of Ibandronate (r = 0.9989). The lowest detectable concentration of Ibandronate was 6 mg/L. The method is simple, rapid and reproducible.

Bone Resorption↗
PubMed2004

Removal of ammonium from human urine through ion exchange with clinoptilolite and its recovery for further reuse.

Ammonium, from separately collected human urine, had been removed through transfer onto the ammonium selective natural zeolite, clinoptilolite, through ion exchange. In the subsequent treatment steps of washing with tap water, ammonium removed from urine was eluted from the surface of the clinoptilolite to be recovered for further reuse. Different quantities of clinoptilolite were used for a survey of the capacity of the zeolite for the process and to identify removal efficiencies based on initial ammonium loads. The highest surface concentration attained under experimental conditions employed was 15.44 mg ammonium per gram of clinoptilolite for an initial concentration of 110 mg ammonia per litre, and the highest removal was 98%, obtained for a loading of 1 mg ammonium per gram clinoptilolite. In the subsequent elution process, better removals were observed as pH was increased and the highest removal was attained at pH 13. The recovery was calculated as 9.73 mg ammonium per gram of clinoptilolite, corresponding to an efficiency of 63% only through washing with tap water. The results have given positive indications for the possibility of using ion exchange with clinoptilolite for the removal of ammonium from human urine and an incentive for improving methods of elution for its recovery for further reuse.

Ammonia↗
PubMed2005

T dependence of vibrational dynamics of water in ion-exchanged zeolites A: a detailed Fourier transform infrared attenuated total reflection study.

In order to explore the influence of cation substitution on the vibrational dynamics of water molecules in zeolites, the evolution of structural properties of the O-H stretching band of water in fully hydrated Na-A and Mg-exchanged A zeolites has been studied, for different percentages of induced ion exchange, by Fourier transform infrared attenuated total reflection spectroscopy as a function of temperature. The differences revealed in the O-H stretching band shapes have been accounted by fitting the spectra as a sum of four components, corresponding to water molecules exhibiting different types of hydrogen bonding. The dependencies of the relative intensities, peak wave numbers, and bandwidths of the resolved components on temperature and Mg2+ content have been discussed. Evidence of the "structure-maker" role played by a zeolitic surface on physisorbed water, systematically enhanced by increasing the percentage of induced ion exchange, is given in the whole explored temperature range.

Journal Article↗
PubMed1994

Two-dimensional high-performance liquid chromatography using monodisperse polymer beads containing segregated chemistries prepared by pore size specific functionalization. Single-column combinations of size exclusion or ion exchange with reversed-phase chromatography.

Separation media for the complete separation of complex samples that require a combination of size exclusion or ion-exchange with reversed-phase chromatographic modes in a single column have been prepared from size monodisperse 10 microns poly(glycidyl methacrylate-co-ethylene dimethacrylate) beads using a pore size specific functionalization process. To achieve the first combination of chromatographic modes, the large pores of the beads were selectively hydrolyzed to diols using aqueous poly(styrenesulfonic acid), while highly hydrophobic octadecyl groups were introduced into the small pores by reaction of the remaining epoxide groups with octadecylamine. These beads provide excellent protein recoveries and may be used for the direct injection separation of samples containing both hydrophilic proteins and hydrophobic drugs. Beads containing diethylamino groups in the large pores and octadecyl functionalities in the small pores were also prepared by size-selective modification. A plot of log k' against ionic strength of the mobile phase for these beads shows the absence of hydrophobic interactions and documents the clean ion-exchange mechanism of protein separation. Examination of the small pores in both types of separation media confirmed that their hydrophobicity was sufficient to allow the separations of small molecules in reversed-phase mode. Column packed with these dual-chemistry beads exhibited high efficiencies and were used successfully for the separations of proteins and alkylbenzenes or drugs.

Animals↗
PubMed1970

Chloride and potassium activities in luminal fluid of turtle thyroid follicles as determined by selective ion-exchanger microelectrodes.

Cl(-) and K(+) activities in the follicular lumen and the intraluminal potential of the same lumen were measured simultaneously with specific liquid ion-exchanger and 3 M KCl microelectrodes, respectively, in turtle thyroid glands. The Cl(-) equilibrium potential between the thyroid interstitium and the lumen calculated from the measured Cl(-) activities was higher than the directly measured intraluminal potential. These data indicate that Cl(-) is actively transported out of the follicular lumen in turtle thyroid gland. On the other hand, the calculated K(+) equilibrium potential was not different from the directly measured potential, which indicates that K(+) is probably distributed passively according to the electrochemical gradients between the interstitial and the luminal compartments in turtle thyroid gland. Results obtained from the specific liquid ion-exchanger microelectrode studies correlate well with those obtained from previous radioautographic observations and chemical determinations.

Animals↗
PubMed2001

Ion-exchange resins: carrying drug delivery forward.

Ion-exchange resins (IER), or ionic polymer networks, have received considerable attention from pharmaceutical scientists because of their versatile properties as drug-delivery vehicles. In the past few years, IER have been extensively studied in the development of novel drug-delivery systems (DDSs) and other biomedical applications. Some of the DDSs containing IER have been introduced into the market. In this review, the applications of IER in drug delivery research are discussed.

Journal Article↗
PubMed1984

Isolation of a proform of porcine secretin by ion-exchange and reversed-phase high-performance liquid chromatography.

A polypeptide with secretin-like bioactivity has been isolated from upper small intestinal porcine tissue by ion-exchange and reversed-phase high-performance liquid chromatography (HPLC). The purification was followed by determination of biological activity. Its elution position in the ion-exchange HPLC indicated that it was less basic than secretin. Amino acid analysis showed that it contained an additional glycine residue as compared to secretin. Digestions by trypsin and subtilisin established that the polypeptide was a variant form of secretin in which the previously known secretin is extended C-terminally by a glycine residue.

Amino Acid Sequence↗
PubMed2002

Adsorption and isotope effects by ion exchange with aqueous-2-aminomethyl-18-crown-6 bonded merrifield resin.

Magnesium isotopes effects were investigated by chemical ion exchange using synthesized 2-aminomethyl-18-crown-6 (AM18C6) bonded Merrifield peptide resin. It was found that separation factors larger those reported previously were obtained, and the hydration and isotope mass effects are more significant than that of the complexation. The capacity of the crown ion exchanger was 2.3 meq/g dry resin. The adsorption capacity of the resin for magnesium ion was 26.8 mg/g dry resin at pH 7. The heavier isotopes of magnesium were enriched in the solution phase, while the lighter isotopes were enriched in the resin phase. The separation factors of (24)Mg-(25)Mg, (24)Mg-(26)Mg, and (25)Mg-(26)Mg were 1.0085, 1.0162, and 1.0081, respectively.

Journal Article↗
PubMed2005

Controlled oxidative protein refolding using an ion-exchange column.

Column-based refolding of complex and highly disulfide-bonded proteins simplifies protein renaturation at both preparative and process scale by integrating and automating a number of operations commonly used in dilution refolding. Bovine serum albumin (BSA) was used as a model protein for refolding and oxido-shuffling on an ion-exchange column to give a refolding yield of 55% after 40 h incubation. Successful on-column refolding was conducted at protein concentrations of up to 10 mg/ml and refolded protein, purified from misfolded forms, was eluted directly from the column at a concentration of 3 mg/ml. This technique integrates the dithiothreitol removal, refolding, concentration and purification steps, achieving a high level of process simplification and automation, and a significant saving in reagent costs when scaled. Importantly, the current result suggests that it is possible to controllably refold disulfide-bonded proteins using common and inexpensive matrices, and that it is not always necessary to control protein-surface interactions using affinity tags and expensive chromatographic matrices. Moreover, it is possible to strictly control the oxidative refolding environment once denatured protein is bound to the ion-exchange column, thus allowing precisely controlled oxido-shuffling.

Chromatography, High Pressure Liquid↗
PubMed2006

Extraction of inhibitor-free metagenomic DNA from polluted sediments, compatible with molecular diversity analysis using adsorption and ion-exchange treatments.

PCR inhibitor-free metagenomic DNA of high quality and high yield was extracted from highly polluted sediments using a simple remediation strategy of adsorption and ion-exchange chromatography. Extraction procedure was optimized with series of steps, which involved gentle mechanical lysis, treatment with powdered activated charcoal (PAC) and ion-exchange chromatography with amberlite resin. Quality of the extracted DNA for molecular diversity analysis was tested by amplifying bacterial 16S rDNA (16S rRNA gene) with eubacterial specific universal primers (8f and 1492r), cloning of the amplified 16S rDNA and ARDRA (amplified rDNA restriction analysis) of the 16S rDNA clones. The presence of discrete differences in ARDRA banding profiles provided evidence for expediency of the DNA extraction protocol in molecular diversity studies. A comparison of the optimized protocol with commercial Ultraclean Soil DNA isolation kit suggested that method described in this report would be more efficient in removing metallic and organic inhibitors, from polluted sediment samples.

Adsorption↗
PubMed1992

Reversed-phase chromatography of phenylthiocarbamyl amino acid derivatives of physiological amino acids: an evaluation and a comparison with analysis by ion-exchange chromatography.

Reversed-phase chromatography of phenylthiocarbamyl (PTC) amino acid derivatives of physiological amino acids was evaluated and compared with the traditional method of ion exchange. The PTC amino acid derivatives were stable for at least 32 h at ambient temperature before injection. The relationship of detector response to concentration for the PTC derivatives was linear from 39 to 1250 pmol. With few exceptions, the within- and between-run precisions of plasma amino acid retention times were less than 0.2 and 0.3%, respectively; the within- and between-run precisions of their concentrations were less than 4.0 and 5.0%, respectively. Twenty-four plasma samples were quantitated by both reversed-phase and ion-exchange chromatography; fifteen of the twenty amino acids determined had correlation coefficients in the range 0.81-1.00. Nine non-standard amino acids and ten therapeutic drugs were added to plasma; D-glucosaminic acid and alpha-amino-beta-guanidinopropionic acid co-eluted with alpha-aminoadipic acid and threonine, respectively. Of the ten drugs added, only metronidazole and theophylline co-eluted with beta-alanine and histidine, respectively. The precision, stability, and sensitivity of the method render it ideal for the quantitation of plasma amino acids.

Amino Acids↗