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PubMed1984

A new system for washing ion-exchange paper disks used in radiochemical enzyme assays.

A new device has been developed for the removal of radioactive substrate from ion-exchange paper disks, leaving ionized product bound to the paper. It has a number of important advantages over older methods of washing the ion-exchange paper: (1) It is much more thorough and efficient; background absorption of unionized material is reduced to relatively low levels. (2) Fragile media such as DEAE-substituted paper (DE81) are not subjected to the erosive effects of stirrers or agitators. (3) The disks remain organized throughout the sample application and washing procedure, eliminating need for time-consuming sorting of wet radioactive disks after washing. A larger number of assays can therefore be processed conveniently at one time. (4) Radioactive nucleosides and other adsorbable substrates are collected in a cartridge, eliminating the handling and sink-disposal of large volumes or excessive quantities of radioactive materials.

Chromatography, Ion Exchange↗
PubMed1995

A pharmacokinetic study of an ion-exchange solvent-detergent-treated high-purity factor VIII concentrate. Haemophilia Directors for Scotland and Northern Ireland.

An ion-exchange chromatography purified factor VIII concentrate (Liberate) that had undergone a solvent-detergent viral inactivation treatment was compared with an intermediate-purity concentrate (Z8), which was terminally heated at 80 degrees C for 72 h, in 15 haemophilia A patients in a blinded crossover pharmacokinetic study. Both products achieved a peak level close to that predicted (100 IU/dl for Liberate and 103 IU/dl for Z8) and there were no significant differences in the recoveries achieved nor of any of the other pharmacokinetic parameters. We conclude that the pharmacokinetic properties of factor VIII:C, following solvent-detergent treatment and ion-exchange chromatography, are equivalent to those of the lower purity terminally heat-treated product (Z8), and it is therefore likely to be a clinically efficacious concentrate.

Adult↗
PubMed1975

[A new possibility of improving the breaking strength of mineral tooth facets by ion exchange ].

A 140% increase in the breaking strength of Keradens facings may be achieved by ion exchange, i.e., by substitution of the small sodium ions in soda feldspar by the larger potassium ions, which is done by melting the soda feldspar with potassium nitrate. The temperature and the time required for optimum exchange conditions are derived from the results of the present study.

Dental Porcelain↗
PubMed1989

Analysis of cancer-associated colonic mucin by ion-exchange chromatography: evidence for a mucin species of lower molecular charge and weight in cancer.

Cancer-associated mucins in the colon are antigenically distinct and glycosylated differently from their normal counterparts. Mucin-rich glycoconjugate preparations were made from nine non-neoplastic colons, seven colon cancers, and two different xenografts from mucin-producing human colon cancer cell lines, and radiolabeled with 3H. The preparation was applied to a DEAE-cellulose ion-exchange column, and eluted with a discontinuous ascending NaCl gradient resulting in seven discrete fractions or 'species'. Over half of the 3H-labeled glycoconjugates from specimens of non-neoplastic colonic epithelium eluted in fraction V (eluted with 0.25 NaCl). Significantly less of the 3H-labeled glycoconjugates from specimens of colon cancer eluted in fraction V (34%, P less than 0.0005), and there were significant increases in glycoconjugates eluted in fractions IV (P less than 0.008), III (P less than 0.0005), and II (P less than 0.028). Additional samples were prepared without the radiolabeling procedures, chromatographed on a DEAE-cellulose ion-exchange column, and analyzed for monosaccharide content. Each of the fractions contained the monosaccharides expected in mucin-type glycoproteins, but only sialic acid was differentially expressed in the seven fractions or 'species', occurring principally in the more charged species. However, differences in sialic acid content were not sufficient to explain the differences in retention on the ion-exchange column, nor were differences in O-acetylation of the mucins. Mucin-type glycoconjugates from colon cancers are relatively less charged than those from the normal colon, and elute at lower ionic strengths. Of interest, cancer-associated mucins appear to be of lower molecular weight than their normal counterparts. Additional studies of oligosaccharide and apomucin structure will be required to explain the molecular basis of these differences in charge.

Adenocarcinoma↗
PubMed1980

A rapid and simple procedure for the preparation of human lactate dehydrogenase-2 from erythrocytes using an ion-exchange mini-column.

We describe a simple ion-exchange chromatography technique for the rapid preparation of lactate dehydrogenase-2 from lysed human erythrocytes obtained either from plasma or serum clots. The procedure appears capable of scale-up to larger columns thus making it ideal as an initial step in a more extensive purification scheme.

Chromatography, Ion Exchange↗
PubMed1968

Rat small-intestinal beta-galactosidases. Separation by ion-exchange chromatography and gel filtration.

1. The chromatography of rat small-intestinal beta-galactosidase activities on gel-filtration and ion-exchange columns has been studied. Five different substrates were used to measure beta-galactosidase activity (lactose, phenyl beta-galactoside, o-nitrophenyl beta-galactoside, p-nitrophenyl beta-galactoside and 6-bromo-2-naphthyl beta-galactoside) and the activity was measured at one acid and one more neutral pH value. 2. By gel filtration one acid beta-galactosidase, hydrolysing lactose and the hetero-beta-galactosides at about the same rate, and one more neutral beta-galactosidase, hydrolysing lactose much more rapidly than the hetero-beta-galactosides, were separated. 3. By ion-exchange chromatography the acid enzyme was fractionated into two components. These may be individual enzymes or different forms of the same enzyme.

Animals↗
PubMed1983

Purification of reduced nicotinamide adenine dinucleotide by ion-exchange and high-performance liquid chromatography.

Combining ion-exchange (AG MP-1) and reversed-phase (C-18) partition chromatography accomplishes a higher degree of purification of NADH than either method can provide alone. Final elution in 95% ethanol, dehydration with anhydrous sodium sulfate, and storage in dry 1,2-propanediol over molecular sieves prevents hydrolysis of the purified dinucleotide.

Chromatography, High Pressure Liquid↗
PubMed2003

Sephadex and sephadex ion-exchange filtration improves the quality and freezability of low-grade buffalo semen ejaculates.

The effect of sephadex and sephadex ion-exchange filtration on the improvement in quality and freezability of low-grade buffalo semen ejaculates was assessed. Two types of filtration columns were used: one containing only sephadex G-10 (FS) and the other sephadex G-10 along with ion-exchangers (diethyl amino ethane-52 (DEAE-52) cellulose and carboxy methyl-52 (CM-52) cellulose; FS+IE). Unfiltered samples served as controls. Semen ejaculates extended in Tris-citric acid (1:4) (n=16; initial motility 40-50%) were filtered at the rate of 1.5 ml/min under negative pressure at room temperature (28-30 degrees C). The mean recovery rate (%) of motile spermatozoa in the FS (85.9+/-1.51) and FS+IE (77.10+/-2.28) filtrates did not differ significantly. Percentages of sperm motility, normal acrosomes, and intact plasma membranes were highest (P<0.05) in FS+IE, intermediate (P<0.05) in FS and lowest (P<0.05) in controls at the three stages of cryopreservation (postfiltration final dilution, after equilibration, and after freezing). Mean sperm abnormalities were lowest (P<0.05) in the filtrates of FS+IE, moderate (P<0.05) in FS and highest in controls at all stages of freezing. Compared to dilution and equilibration, freezing greatly reduced (P<0.05) the overall percent motility, normal acrosomes and intact plasma membranes. The spermatozoa eluted through FS+IE columns proved more resistant (P<0.05) in bearing dilution, equilibration, freezing and thawing stresses than the spermatozoa from FS and control samples. It is concluded that filtration systems containing an FS+IE column can effectively enhance the quality and freezability of extended, low quality buffalo semen.

Acrosome↗
PubMed1999

The use of histological techniques for the demonstration of ion exchange resins.

AIM: To establish the staining characteristics of certain ion exchange resins in histological material, with a view to enabling confident differential identification. METHODS: Various histological staining procedures were applied to selected pathological material and prepared agar blocks containing the cation exchange resin calcium polystyrene sulphonate and the anion exchange resin cholestyramine. RESULTS: Calcium polystyrene sulphonate uniquely stained strongly by a direct Schiff's reagent procedure without any preoxidation and by the Ziehl-Neelsen method. Cholestyramine was negative by the former method but stained strongly with a standard Congo red technique. CONCLUSIONS: These staining results are consistent with the known structure and properties of polystyrene sulphonate and cholestyramine resins. Polystyrene sulphonate resins have the virtually pathognomonic feature of direct Schiff positivity, while morphology, location, and strong non-birefringent Congo red positivity facilitate the identification of cholestyramine. It is possible that the intrinsic staining characteristics of cholestyramine may be lost once it has bound to its target.

Anion Exchange Resins↗
PubMed2005

Latex-coated polymeric monolithic ion-exchange stationary phases. 1. Anion-exchange capillary electrochromatography and in-line sample preconcentration in capillary electrophoresis.

A sulfonated methacrylate monolithic polymer has been synthesized inside fused-silica capillaries of diameters 50-533-microm i.d. and coated with 65-nm-diameter fully functionalized quaternary ammonium latex particles (AS18, Dionex Corp.) to form an anion-exchange stationary phase. This stationary phase was used for ion-exchange capillary electrochromatography of inorganic anions in a 75-microm-i.d. capillary with Tris/perchlorate electrolyte and direct UV detection at 195 nm. Seven inorganic anions (bromide, nitrate, iodide, iodate, bromate, thiocyanate, chromate) could be separated over a period of 90 s, and the elution order indicated that both ion exchange and electrophoresis contributed to the separation mechanism. Separation efficiencies of up to 1.66 x 10(5) plates m(-1) were achieved, and the monoliths were stable under pressures of up to 62 MPa. Another latex-coated monolith in a 250-microm-i.d. capillary was used for in-line preconcentration by coupling it to a separation capillary in which the EOF had been reversed using a coating of either a cationic polymer or cationic latex particles. Several capillary volumes of sample were loaded onto the preconcentration monolith, and the analytes (inorganic anions) were then eluted from the monolith with a transient isotachophoretic gradient before being separated by electrophoresis in the separation capillary. Linear calibration curves were obtained for aqueous mixtures of bromide, nitrite, nitrate, and iodide. Recoveries of all analytes except iodide were reduced significantly when the sample matrix contained high levels of chloride. The preconcentration method was applied to the determination of iodide in open ocean water and provided a limit of detection of 75 pM (9.5 ng/L) calculated at a signal-to-noise ratio of 3. The relative standard deviation for migration time and peak area for iodide were 1.1 and 2.7%, respectively (n = 6). Iodide was eluted as an efficient peak, yielding a separation efficiency of 5.13 x 10(7) plates m(-1). This focusing was reproducible for repeated analyses of seawater.

Journal Article↗
PubMed1991

Ion exchange chromatography of purified colonic mucus glycoproteins in inflammatory bowel disease: absence of a selective subclass defect.

Previous reports of a selective mucin subclass defect in ulcerative colitis have been reassessed using high performance chromatography (Superose 6 and Mono Q) for mucin purification and fractionation coupled with analysis of the fractions obtained using a combination of enzyme linked lectin and mucin antibody assays. Mucin samples purified from snap frozen rectal biopsy specimens obtained from patients with ulcerative colitis (n = 12), Crohn's disease (n = 5), and non-inflammatory bowel disease control subjects (n = 9) were subject to ion exchange chromatography using a continuous 0-0.35 mol/l NaCl salt gradient with a final 2.5 mol/l NaCl step. In all samples the major proportion (mean (SD) 86.7 (8.9)%) of the mucin detectable by wheat germ agglutinin binding eluted between 0.15 and 0.35 mol/l NaCl with no significant difference in elution profile between ulcerative colitis and control subjects. Significant elution of glycoprotein at less than 0.15 mol/l NaCl did occur, however, when a lower molecular weight mucin containing fraction which contained concanavalin A positive (glucose or mannose containing) material was analysed similarly. Similar ion exchange profiles were obtained when (3H)N-acetylglucosamine labelled mucins were studied after tissue culture of rectal biopsy specimens. No significant alteration in the ion exchange profile of purified mucins in ulcerative colitis has been shown in these studies. It is possible that the previously reported relative depletion of mucin subclass IV (eluting with 0.20 mol/l NaCl) may simply have reflected mucin depletion.

Chromatography, Ion Exchange↗
PubMed1986

Contaminating peptides widely present in ion-exchanged water, reagents, experimental implements and natural sample.

Ion-exchanged water, various reagents, experimental implements and natural sample have been found to be widely contaminated by certain kinds of peptides, whose molecular weights are between 1000 and 6000 daltons. An amino acid analysis showed them to contain serine, glycine and alanine as the three major amino acid components, which constitute about 50% of the total amino acid in the fractions of the contaminants. Ornithine is also commonly abundantly present in the contaminants. The significance of the presence of ornithine and the origin of the contaminants are discussed.

Amino Acids↗
PubMed1987

Allowance for "analyte valence" in the retention model of ion-exchange chromatography. Studies of adenosine 5'-phosphates on DEAE cellulose.

Quantitative expressions are derived for various retention models of the ion-exchange behaviour of multivalent analytes. Their effectiveness as descriptions of experimental results is then tested by application to partition equilibrium studies of the effect of phosphate concentration on the interaction between the three adenosine 5'-phosphates and diethylaminoethylcellulose at pH 4.4. A completely general multi-state model has pointed to quantitative deficiencies in the currently accepted cooperative two-state model, which is, however, superior to a multi-state model devoid of cooperative effects. A similar situation pertains to the ion-exchange behaviour of cytochrome c on carboxymethylcellulose at pH 7.0.

Adenosine Diphosphate↗
PubMed1977

Cellulosic ion-exchange membranes for hemodialysis.

The application of cellulosic ion-exchange membranes to hemodialysis was studied in vitro. The membranes were prepared by radiation-grafting methacrylic acid and vinylpyridine to films of DuPont cellophane PD-215 to produce cation-exchange and anion-exchange membranes, respectively. Solutions of urea, creatinine, glucose, and uric acid were studied for their interactions with and diffusion through the membranes. Ultrafiltration rates were also determined. Cuprophane and PD-215 cellophane were studied as controls. Dialysis plots for the membranes revealed a mechanism of "assisted transport." Initially, the solutes were removed from solution by a sorption/adsorption mechanism followed by a steady-state diffusion process. The calculated diffusivities for these later steady-state regions increased linearly with capacity for urea, creatinine, and uric acid, while for glucose the reverse was true. The combined processes involved provided considerably greater mass transport per unit thickness than either DuPont PD-215 cellophane or Cupropane.

Biocompatible Materials↗
PubMed1995

Gram-scale purification of phosphorothioate oligonucleotides using ion-exchange displacement chromatography.

The purification of oligonucleotides by ion-exchange displacement chromatography is demonstrated on the gram-scale. Using a 50 mmD x 100 mmL (203 ml) column operated in the displacement mode, 1.2 g of a 24mer phosphorothioate oligonucleotide was purified. Product yield for this separation was 70% (780 mg) at a purity of 96.4% and the mass balance recovery of all oligonucleotide was 97.5%. The displacement purification of four additional phosphorothioate oligonucleotides ranging in length from 18 to 25 bases is also demonstrated on the semi-preparative (10-50 mg) scale. All of these oligonucleotides were purified using similar displacement conditions and typical results were 60% yield at 96% purity. The displacement portion of these separations required <15 min and total cycle time including equilibration, feed loading and regeneration can be performed in under 30 min. These results seem to indicate that displacement chromatography may be amenable to generalizations in separation protocol that would greatly reduce the effort required to obtain an optimized purification scheme for moderately long oligonucleotides.

Chromatography, High Pressure Liquid↗
PubMed1994

Solid-phase extraction of small drugs on apolar and ion-exchanging silica bonded phases: towards the development of a general strategy.

In connection with the development of a general strategy for solid-phase extraction (SPE) of drugs, the use of the apolar octadecyl silica bonded phase and ion-exchanging phases with a benzene sulphonic acid or quaternary amine bonded functionality is investigated for the SPE of small polar drugs. This investigation was performed on a set of 15 drugs, belonging to varying pharmacological groups and with varying structures. For each analyte, its adsorption on the C18 and the ion-exchanging phase was controlled for an aqueous solution and for a spiked plasma sample. For those analytes retained on the sorbent, different elution solvents were compared. Although SPE methods could successfully be developed for some drugs, no general solution can be proposed and no solution was found for a few drugs. The main problems are that for these few drugs no sufficient retention is obtained on any SPE phase investigated or the selectivity is too low.

Amines↗
PubMed1977

Separation of tissue and serum acid phosphatase isoenzymes by ion-exchange column chromatography.

I describe a simple, rapid ion-exchange column-chromatographic technique for separating the acid phosphatase (EC 3.1.3.2) isoenzymes in human serum and tissue. Extracts of platelets, spleen, liver, erythrocytes, and prostate were used to determine optimum conditions for separating these isoenzymes. Samples layered on mini-colunms of DEAE-Sephadex A-50 were eluted stepwise with sodium chloride (100, 200, and 300 mmol/liter, buffered with tris (hydroxymethyl)aminomethane). Activity in column effluents was measured with p-nitrophenol phosphate as substrate, and their isoenzyme content was assessed by electrophoresis on polyacrylamide gel. Comparision of activity patterns so derived for various tissues revealed prostatic tissue to be a rich source of acid phosphatase isoenzyme 2 activity. Evaluation of sera from six patients with prostatic cancer revealed isoenzyme patterns with prominent amount of isoenzyme 2 (3.8 to 27.6 U/liter). sera from 10 healthy laboratory technicians contained isoenzyme 2 in the range of 0.3-0.5 U/liter. Samples from two patients with abnormally high activity owing to nonprostatic conditions (Gaucher's disease and carcinoma of lung) exhibited less than 2 U of isoenzyme 2 per liter and acid phosphatase isoenzymes 3-5 that were 50- to 100-fold the normal range. Quantification of isoenzyme 2 by DEAE-Sephadex column chromatography as described appears to provide a more sensitive and specific approach to diagnosis of prostatic cancer.

Acid Phosphatase↗
PubMed2005

Two-component protein adsorption kinetics in porous ion exchange media.

This work provides a theoretical analysis of multicomponent adsorption kinetics for conditions typical of protein adsorption in porous ion exchangers as well as experimental results for the adsorption of lysozyme/cytochrome c mixtures in the cation exchanger SP-Sepharose-FF. The theory predicts the formation of overshoots in the intraparticle concentration profiles and in the total amount adsorbed for the more weakly adsorbed component. An analytical solution valid for the case where the isotherms are rectangular is developed and found to be in good agreement with the limiting behavior of the general numerical solution of the model equations. The experimental results show that the two proteins are competitively adsorbed and that an overshoot of adsorbed cytochrome c occurs during simultaneous adsorption. Model predictions based on the assumption that the adsorption isotherms are rectangular and that lysozyme completely displaces cytochrome c are in qualitative and quantitative agreement with the experimental kinetics suggesting that the overshoot phenomena observed with multicomponent systems in these resins can be explained with a diffusion model without the need to account for flux coupling or electrophoretic contributions to transport.

Adsorption↗