Preparation and ion chromatographic properties of a new core-shell chromatographic support Al2O3/SiO2-10

A new stationary phase Al₂O₃/SiO₂-10 was prepared and characterized by XPS, XRD, SEM and surface analysis. The anion exchanger properties of this new stationary phase were investigated by the separation of inorganic anions in ion chromatography (IC). pH of the mobile phase, concentration and strength of the Lewis base of the elute, and the organic modifier of the mobile phase strongly affect the separation of inorganic anions, and anion exchange selectivities of the analyte on the new support are significantly different from quaternary ammonium styrene based anion exchangers. The result of separation of inorganic anions shows that the new stationary phase provides excellent column efficiency, well-defined chromatographic peaks and favorable retention times.     

Ion-exchange high-performance liquid chromatography of nucleotides and polypeptides on new types of ion-exchange sorbents, based on polystyrene-coated silicas.

A novel type of ion exchanger was prepared by multipoint covalent binding of polystyrene chains onto the surface of porous silica followed by polymer-analogous modification of the bonded layer. Both anion and cation exchangers were synthesized and examined in the separation of nucleotides and proteins. Rapid and efficient separation of basic polypeptides on strong anion exchangers and that of acidic polypeptides on strong cation exchangers could be achieved. For the separation of complete mixtures of polypeptides the application of zwitter-ionic ion exchangers can be recommended.     

Physicochemical basis of the ion-exchange separation of gold cyanide complexes

The mechanism of the separation of gold cyanide complexes is discussed, along with ion exchanger selection, selectivity, elution, and industrial applications. The ion-exchange mechanism for the sorption of gold cyanide complexes is established, and a criterion is suggested for selecting the anion exchanger for their extraction (specifically, the pK _a of the anion exchanger). The selectivity of the sorption of gold cyanide complexes by anion exchangers with rarely distributed ionogenic groups is demonstrated. A procedure for the elution of gold cyanide complexes using alkaline solutions is developed.     

Ion exchange extraction of platinum(IV) and palladium(II) from hydrochloric acid solutions

Sorption concentration of platinum(II, IV) and palladium(II) from freshly prepared and aged two-yearold hydrochloric acid solutions by a series of anion exchangers with different functional groups and of different physical structure of Purolite and CYBBER grades was studied. The high sorption ability of the ion exchangers in relation to the extracted chlorocomplexes of noble metals is shown. It was demonstrated that palladium(II) from all tested ion exchangers can be completely desorbed with thiourea solutions acidified with hydrochloric acid, while complete desorption of platinum is achieved only from Purolite S 985 anion exchanger of the complexforming type and Purolite A 111 weak base anion exchanger.     

Organic Ion Exchangers. Synthesis and Their Behaviour in the Retention of Some Metal Ions

Summary: Three pyridine strong base anion exchangers as beads were obtained by quaternization reactions of a 4-vinylpyridine : 8% divinylbenzene copolymer of gel type. These resins possess methyl / ethyl / butyl radicals as substituents on N⁺ atoms and have exchange capacities of 4.80 mEq/g and 2.10 mEq/mL. For pyridine strong base anion exchangers, the behaviours in the retention processes of Cr(VI) as oxyanions and Ga(III) as [GaCl₄]⁻ complex anion were evaluated with the bath method. All the resins exhibited retention properties, but the retained amounts of the metal cations are different as a function of the alkyl length as substituent on N⁺ atoms and the complex anion nature. Thus, Cr(VI) oxyanions are best retained by the resin with  CH₃ as substituent on N⁺ atoms while [GaCl₄]⁻ complex anion by the resin with  C₄H₉ as substituent on N⁺ atoms. By aminolysis reaction of an ethylacrylate : acrylonitrile : divinylbenzene copolymer as beads of macroporous type with NH₂OH · HCl in the presence of C₂H₅OH a new chelating ion exchanger was performed which contains both amidoxime and hydroxamic acid functional groups. This ion exchanger has the retention property for different metal cations but its retention capacities values are strongly dependent of the nature of metal cation and the counterion as well as pH of the solution. Thus, in the static conditions Zn(II) cation with NOequation/tex2gif-stack-1.gif anion as counterion is retained with the best result at pH = 5. As an example, for the aqueous metal cation solution of 10⁻² M concentration for Zn(NO₃)₂ the resin possess at equilibrium a retention capacity of 6.70 mmol Zn/g dry resin and for Cu(II) from Cu(NO₃)₂ solution of same concentration, the retention capacity is 0.22 mmol Cu/g dry resin and Fe(III) from Fe(NO₃)₃ solution is not retained.     

Speciation of aluminum in soil extracts using cation and anion exchangers followed by a flow-injection system with fluorescence detection using lumogallion.

Flow-injection analysis (FIA) with fluorescence detection of aluminum using lumogallion was applied to the chemical speciation of aluminum in soil extracts after the separation of aluminum species with ionic exchangers. Aluminum complexes with organic substances (anion species) can be specified from other species by using a strongly acidic cation exchanger in the pH range of 3 to 5. Furthermore, aluminum can be separated into three categories, namely, (i) the Al3+ and Al-OH complex, (ii) aluminum organic complexes (cation species), and (iii) its anion species by using a strongly acidic and a weakly acidic cation exchanger at around pH 5. A considerable percentage of water-soluble aluminum in soils was found to be complexes with humic substances.     

Anion-exchange chromatography of DNA restriction fragments.

The abilities of several high-performance liquid chromatography (HPLC) anion-exchange packings to separate DNA restriction fragments, ranging in size from 50 to 23,000 base pairs, were studied. The ion exchangers investigated include the porous packings Protein-Pak DEAE-5PW, Nucleogen-DEAE 4000-7, Poros-Q and BakerBond WP-PEI, and the non-porous packings TSK Gel DEAE-NPR, Gen-Pak FAX, and ProPac PA1. The results indicated that the non-porous packings could separate all 18 fragments (less than 600 base pairs) in a pBR322 DNA-HaeIII digest, while of the porous packings, only Nucleogen-DEAE 4000-7 could resolve DNA fragments in this size range. Only Gen-Pak FAX and TSK Gel DEAE-NPR could significantly resolve the very large DNA fragments (125-23,000 base pairs) of a lambda DNA-HindIII digest. The chromatographic parameters governing this separation by Gen-Pak FAX were optimized so that six of eight fragments were resolved. Split-peak phenomena were observed at low flow-rates when employing non-poros packings, but were eliminated by the incorporation of organic modifiers or surfactants, suggesting that, under certain conditions, hydrophobicity may play a significant role in separations on this packing. Gen-Pak FAX also separated 21 of 23 fragments in a 1000-base pair DNA ladder, a performance which, in addition to the quantitative capabilities of HPLC, makes anion-exchange chromatography a powerful method complementary to slab-gel electrophoresis, and perhaps preferable over agarose gel electrophoresis for applications such as the confirmation of plasmid integrity.     

DNA binding during expanded bed adsorption and factors affecting adsorbent aggregation

DNA-induced aggregation and contraction of expanded bed adsorption chromatography beds have been examined using strong anion exchanger Q HyperZ and calf thymus DNA in buffers containing added NaCl. Two batches of adsorbent with different ionic capacities were used allowing the effects of different ligand densities to be examined. Very high dynamic binding capacities at 10% breakthrough were found in the absence of added salt. However, the highest binding capacities ( approximately 10 and approximately 19mgDNAml(-1) gel) were found in buffers containing added salt at concentrations of either 0.25 or 0.35M, for the low and high ligand density adsorbents, respectively. Bed contraction was observed, but did not correlate with dynamic binding capacity or with the amount of DNA loaded. No differences in bed contraction were seen by varying the concentration of DNA loaded in the range of 20-80mugml(-1) even though the dynamic binding capacity was reduced as DNA concentration was increased. The extent of bed contraction during DNA loading was found to be a function of added salt concentration and ligand density of the adsorbent. The results imply that ligand density significantly affects the salt tolerance of anion exchangers when binding DNA. However, more importantly, with the adsorbents examined here, attempts to reduce bed aggregation by feedstock conditioning with added salt may increase DNA binding leading to a reduction in expanded bed adsorption performance compromising protein capture in real feedstocks.     

A kinetics of hydrolysis of ring phosphorus oxoanion in ion exchanger phase

Ring phosphorus oxoanion, hexapotassium dodecaoxohexaphosphate(III), (DOHP), was hydrolyzed in anion exchangers in order to elucidate the physicochemical state in the solid phase. The hydrolysis of DOHP obeys pseudo first order kinetics as is the case in aqueous solution. The hydrolysis rate constants k were determined as a function of the external solution pH or proton concentration in an anion exchanger. It was estimated from the Donnan equation and from the spectral change of an acid base indicator in the anion exchanger. The difference in k in aqueous and in the anion exchanger phase is discussed.     

Liquid chromatography on unmodified and modified spheron gels I. Nucleosides,bases and related compounds

Summary The ethylene glycol methacrylate gel Spheron and ion exchangers produced by the chemical modification of this gel (such as the cation exchanger Spheron S and anion exchanger Spheron DEAE) are compared with octadecylsilica as column packing materials for reversedphase chromatography of nucleic acid constituents and related compounds. The different separation selectivities of the individual materials can be utilized for the chromatographic separation of these compounds.     

A potential application for large diameter pellicular supports in the reversed phase study of ultra high molecular mass polystyrenes

Summary Ultra high molecular mass (> 7 million dalton) polystyrenes are prone to degradation in size exclusion chromatography. In gradient elution reversed phase HPLC they do not elute visibly on small particle size porous supports. However, large diameter C18 pellicular particles were successfully employed for reversed phase study of ultra high molecular mass (15 million dalton) polystyrenes without polymer degradation during elution. Although retention for the lower molecular mass polystyrenes was lower than on small diameter porous particles, the medium high molecular mass polystyrenes (0.5–1 million dalton) showed similar retention. The addition of small diameter porous particles in small quantities, to the large diameter pellicular particles, increased the amount of retention of the low molecular mass polystyrenes without affecting the higher molecular mass polystyrenes.     

Structure–affinity relationships for the binding of actinomycin D to DNA

Molecular models of the complexes between actinomycin D and 14 different DNA hexamers were built based on the X-ray crystal structure of the actinomycin–d(GAAGCTTC)2 complex. The DNA sequences included the canonical GpC binding step flanked by different base pairs, nonclassical binding sites such as GpG and GpT, and sites containing 2,6-diamino- purine. A good correlation was found between the intermolecular interaction energies calculated for the refined complexes and the relative preferences of actinomycin binding to standard and modified DNA. A detailed energy decomposition into van der Waals and electrostatic components for the interactions between the DNA base pairs and either the chromophore or the peptidic part of the antibiotic was performed for each complex. The resulting energy matrix was then subjected to principal component analysis, which showed that actinomycin D discriminates among different DNA sequences by an interplay of hydrogen bonding and stacking interactions. The structure–affinity relationships for this important antitumor drug are thus rationalized and may be used to advantage in the design of novel sequence-specific DNA-binding agents.     

Polystyrene immobilized ionenes as novel stationary phase for ion chromatography

Several aliphatic ionenes (2-6-, 6-6-, 10-6-ionene) have been prepared as ion exchangers for the development of novel high-performance stationary phases for anion chromatography (IC). A macroporous polystyrene/divinylbenzene (PS/DVB) resin with adjusted cation exchange capacity was used as support. Therefore the immobilization of ionenes to polystyrene carriers with remaining positive surface charge became possible for the first time. Strong ion-exchange interactions, resulting in high retention times, between the stationary phase and inorganic as well as organic anionic analytes have been observed. The influence of different ionenes on the retention behaviour during the ion chromatographic separation was investigated. Additionally, partly aromatic and polar ionene backbones were prepared and their retention behaviour as anion exchanger was investigated. The highest number of theoretical plates obtained was about 90.000 per meter. The signal asymmetries were generally lower than obtained for surface functionalized anion exchangers.     

The preparation of nano‐scope chitosan‐oligomer copper complexes and their interaction with DNA

Water‐soluble low molecular weight chitosan of nanometer level and its copper complexes were prepared, and characterized by IR spectra, elemental analysis and gel permeation chromatography (GPC). The modes and mechanism of these copper complexes interaction with DNA were studied by a fluorescent probe method and electrophoresis analysis. It is suggested that there are electrostatic and intercalation modes of copper complexes interacting with DNA. At first, the cationic complex electrostaticly binds to the negatively charged phosphate backbone of DNA, and then a portion of the complex intercalates between the base pairs on the DNA duplex strand. Copyright © 2005 John Wiley & Sons, Ltd.     

A mass spectrometric investigation of non-covalent interactions between ruthenium complexes and DNA

Electrospray ionisation mass spectrometry was used to investigate reactions between six ruthenium compounds and three different non self-complementary duplex oligonucleotides containing 16 base pairs. Each of the compounds studied formed non-covalent complexes containing between one and five ruthenium molecules bound to DNA. Competition experiments involving duplex 16mers and pairs of ruthenium compounds were used to determine the order of relative binding affinities of the metal compounds. Other competition experiments involving ruthenium compounds, and the organic DNA binding agents daunomycin and distamycin, provided information about the sites and modes of DNA binding of the ruthenium compounds.     

DNA-histidine complex formation in isoelectric histidine buffers.

The free solution electrophoretic mobility of two DNA molecules of different molecular masses, 18 base pairs and 2686 base pairs, has been measured in isoelectric histidine buffers with and without added low-molecular-mass electrolytes. Extensive DNA-histidine complex formation is observed in isoelectric histidine buffer, as evidenced by distortion and splitting of the peaks in the electropherograms. Peak distortion and splitting can be decreased or eliminated by adding low-molecular-mass neutral salts to the solution, suggesting that the DNA-histidine complexes are stabilized by electrostatic interactions. The ability of various neutral salts to disrupt the DNA-histidine complexes depends on the molecular mass of the DNA and the concentration and type of added salt.     

Protein adsorption on novel acrylamido-based polymeric ion exchangers. II. Adsorption rates and column behavior

Uptake kinetics and breakthrough behavior were determined for bovine serum albumin (BSA) and alpha-chymotrypsinogen (alphaCHY) in new polymeric ion-exchange media based on acrylamido monomers. Two anion exchangers and a cation exchanger were investigated. As shown in Part I of this work, the two anion exchangers have different morphologies. The first one, BRX-Q, comprises a low-density gel with a matrix of denser polymeric aggregates. While this material has a very low size-exclusion limit for neutral probes, it exhibits an extremely high binding capacity for BSA. The second anion exchanger, BRX-QP, comprises large open pores but has a very low binding capacity. The cation exchanger, BRX-S, also comprises large open pores but exhibits an intermediate capacity; likely as a result of the presence of smaller pores. Dynamic protein uptake experiments showed that the highest mass transfer rates are obtained with BRX-Q. The apparent diffusivity is also highest for this material and increases substantially as the protein concentration is reduced. For these particles, the external film resistance is dominant at very low protein concentrations. Much lower rates and apparent diffusivities are obtained for BRX-QP. Finally intermediate rates and apparent diffusivities are found with BRX-S. The concentration dependence of the apparent pore diffusivity is much less pronounced in this case. The apparently paradoxical result that mass transfer rates are highest for the material with the smallest neutral-probe size-exclusion limit can be explained in terms of a general conceptual model where parallel pore and adsorbed-phase diffusion paths exist in these particles. In the first case, adsorbed phase diffusion in gel pores is dominant, while in the second transport is dominated by diffusion in a macroporous network. In the third case, both contributions are important. The conceptual model provides an accurate prediction of the breakthrough behavior of columns packed with these media using independently determined rate parameters. Dynamic binding capacities of 80-140 mg/ml were observed for BSA on BRX-Q in ca. 1.5 cm columns operated at 300-900 cm/h in agreement with theoretical predictions.     

Polyelectrolyte Sorbents for Ion Chromatography

A method for the synthesis of new anion exchangers for ion chromatography has been proposed. The method is based on the fact that interaction of water-soluble anionic polymers with materials containing negatively charged sulfo groups on the surface gives rise to polyelectrolyte complexes. It is demonstrated that replacement of functional groups in molecules of the polymer affects the selectivity of the ion-chromatographic determination of ions. Recommendations are given for selecting a polymer for the determination of mixtures of inorganic anions and anionic complexes of transition metals in ethylenediaminetetraacetic acid.     

Methods for Preparing High Performance Stationary Phases for Anion-Exchange Chromatography

The development of new high-performance anion-exchange stationary phases is one of the priority tasks of modern ion chromatography (IC). Most of the research in this field is aimed at improving the selectivity of anion exchangers and increasing the sensitivity, efficiency, and rapidity of the analysis. The selectivity of the stationary phases is largely determined by the chemistry of the adsorbent surface; therefore, it is particularly important to understand the relationship between the structure of the functional layer of the anion exchanger and its chromatographic properties. This review is devoted to methods for modifying polymer matrices in order to obtain highly efficient anion exchangers for the separation of inorganic and organic anions.     

The equilibrium sorption of sulfuric acid by weakly basic polyacrylic anion exchangers synthesized on the basis of polyamines

The equilibrium ion-exchange sorption of sulfuric acid by the free base form of weakly basic polyacrylic anion exchangers with the gel (Relite MG1) and porous (Relite MG1/P) structures was studied. It was proved that the bisulfate variety of the anion exchangers did not form. Solutions of resinates were found to be nonideal. Their nonideality was analyzed in terms of the theory of exchange equilibria to suggest a model of the exchange of doubly-charged anions on fixed polyamine exchange centers of Relite MG1 and Relite MG1/P anion exchangers. The type of the ion was found to have no effect on sorption parameters.