A simple ion chromatography-mass spectrometry method for amino acid analysis in beer.

The amino acid footprint of different beer samples was analyzed using ion chromatography coupled with electrospray ionization mass spectrometry. A tailor-made polymer-based cation-exchange resin was operated with a mass spectrometry-compatible eluent under isocratic conditions on a standard high-performance liquid chromatography system coupled to a single quadrupole mass spectrometer using formic acid as a volatile eluent ion source. The partially separated peaks of the isomeric pair isoleucine/leucine were processed according to their area response ratio using vertical peak splitting or Gaussian fit. Additionally, the chromatographic resolution of the isomers was optimized with an adjusted, solely aqueous mobile phase from 0.85 to 2.92. Ion suppression in the electrospray ion source was investigated for the derivatization-free method and found to be insignificant (recovery value 100 ± 15%) for 15 out of the 20 analytes. Quantitative results for various beer and mixed-beer beverages were found to be in high agreement with existing methods. Simultaneous photometric detection demonstrated the method's ability to successfully remove most of the interfering matrix compounds.

Sample displacement chromatography of monoclonal antibody charge variants and aggregates.

The rise of biosimilar monoclonal antibodies has renewed the interest in monoclonal antibody (mAb) charge variants composition and separation. The sample displacement chromatography (SDC) has the potential to overcome the low separation efficiency and productivity associated with bind-elute separation of mAb charge variants. SDC in combination with weak cation exchanging macroporous monolithic chromatographic column was successfully implemented for a separation of charge variants and aggregates of monoclonal IgG under overloading conditions. The charge variants composition was at-line monitored by a newly developed, simple and fast analytical method, based on weak cation exchange chromatography. It was proven that basic charge variants acted as displacers of IgG molecules with lower pI, when the loading was performed 1 to 1.5 pH unit below the pI of acidic charge variants. The efficiency of the SDC process is flow rate independent due to a convection-based mass transfer on the macroporous monolith. The productivity of the process at optimal conditions is 35 mg of purified IgG fraction per milliliters of monolithic support with 75-80% recovery. As such, an SDC approach surpasses the standard bind-elute separation in the productivity for a factor of 3, when performed on the same column. The applicability of the SDC approach was confirmed for porous particle-based column as well, but with 1.5 lower productivity compared to the monoliths.

Design and synthesis of naphthalene-based chiral strong cation exchangers and their application for chiral separation of basic drugs.

In continuation of our efforts to synthesize a highly dedicated strong cation exchanger, we introduce four chiral stationary phases based on a laterally substituted naphthalene core featuring chiral 2-aminocyclohexansulfonic acid as the chiral cation-exchange site. The selectors were modified with two different terminal units, which enabled immobilization to the silica support by thiol-ene radical reaction or azide-yne click chemistry. The chromatographic parameters of these chiral stationary phases were determined using a set of chiral amines, mainly from the family of ß-blocker pharmaceuticals. The chiral stationary phases immobilized by means of click chemistry were found to be superior to those possessing the sulfide linker to the silica support. The chromatographic results and visualization of density functional theory-calculated conformations of the selectors hint at a combination of a steric and electronic effect of the triazole ring in the course of chiral resolution of the target analytes.

Nanosized cation exchanger for the electrophoretic separation and preconcentration of catecholamines and amino acids.

The first example of application of nanosized polystyrene-based cation exchanger (NSCE) with sulfo groups as a dynamic coating of capillary walls was demonstrated. The conditions of dynamic coating formation were optimized and ensured the long-term stability of the coating. Capillary-to-capillary and day-to-day repeatabilities were 4% and 3%, correspondingly. The NSCE coating stability at various pH and influence of pH on the EOF mobility were investigated. The developed NSCE-modified coated capillaries provided improved resolution (Rs = 0.9-3.2 for catecholamines and Rs = 1.7-2.8 for amino acids) and efficiencies (330-520 ×103 t.p./m) of basic analytes, which are 1.5 times higher compared to untreated capillary. The optimized conditions were as follows: 50 mM phosphate buffer solution at pH 2.2 with 5 µM NSCE. The effect of the NSCE concentration in BGE on the electrophoretic mobilities of the analytes was investigated. The various online concentration techniques were tested in order to decrease the LODs. The simultaneous application of NSCE capillaries and field-amplified sample stacking provided the lowest LODs of catecholamines and amino acids and allowed to determine these analytes in human urine.

A weak cation exchanger by encapsulating silica with maleic anhydride-modified polyvinyl alcohol.

A novel weak cation exchanger is described by encapsulating silica with a (or multiple) layer(s) of maleic anhydride-modified polyvinyl alcohol coating(s). The preparation method is facile and fast, which is simply performed by dipping silica particles into maleic anhydride-modified polyvinyl alcohol solution as-synthesized, then filtering and curing thermally, and finally generating a thin coating onto silica particles. Multiple layers of maleic anhydride-modified polyvinyl alcohol coatings can be generated by repeating above steps, offering an easy way to manipulate the capacity. The obtained weak cation exchanger demonstrated high separation efficiency and good selectivity toward common inorganic cations, for example, high plate count of 81 000 per meter was obtained for NH4 + . Simultaneous separation of alkali and alkaline earth metals could be achieved within 14 min under isocratic conditions.

Integration of Fenton's reaction based processes and cation exchange processes in textile wastewater treatment as a strategy for water reuse.

The remediation of a real textile wastewater aiming its reuse in the textile industry was carried out by integrating two processes: (i) a chemical or electrochemical advanced oxidation process (AOP or EAOP) based on Fenton's reaction for organics degradation, and (ii) a cation exchange process using marine macroalgae for removal of the iron acting in the Fenton's reaction based processes. Four AOPs/EAOPs at acidic pH 2.8 were tested: Fenton, photo-Fenton with ultraviolet A (UVA) radiation (PF/UVA), electro-Fenton (EF) and photoelectro-Fenton with UVA radiation (PEF/UVA). These processes provided very high color removals. After a running time of 45 min, the color removals were 68-95% for the Fenton process, 76-94% for the EF process, 80-98% for the PF/UVA process and 85-100% for the PEF/UVA process. In contrast, the mineralization was negligible for all the processes, indicating the generation/presence of persistent colorless compounds. The PF process was selected as first treatment stage due to its ability for color removal and related lower costs. A set of six marine macroalgae (Gracilaria caudata, Gracilaria cervicornis, Ascophyllum nodosum, Fucus spiralis, Laminaria hyperborea and Pelvetia canaliculata) were tested for iron uptake. Laminaria hyperborea showed the highest ion exchange capacity and affinity for iron species. Its application allowed the removal of all the iron acting in the PF process (3.4 mg/L). The textile wastewater resulting from the application of PF process followed by cation exchange with Laminaria hyperborea was successfully reused in scouring, bleaching and dyeing processes.

Sequestration of Cr (VI) from water using agar-polyvinyl alcohol based cation exchanger.

Present study focuses on the use of a biodegradable and cost-effective cation exchanger for removal of Cr (VI) metal ions from water sources. Semi-IPN was prepared through grafting of acrylamide onto agar-polyvinyl alcohol backbone in presence of boric acid and ammonium per sulphate as crosslinker-initiator system. Graft copolymer was converted to cation exchanger through phosphorylation. Characterization was done using methods such as FTIR, SEM-EDX and XRD. Semi-IPN exhibited higher thermal resistance. The findings revealed that the optimum conditions for Cr (VI) removal are pH = 4.0; contact time (min) = 360; adsorbent dose (mg) = 125 and metal ion concentration(mg/L) =2. The adsorption kinetics of Cr (VI) ions are best fit by the pseudo second order kinetic with 0.99 R2 and Kf (rate constant) was found to be 0.97 thereby supporting the Freundlich isotherm. The adsorption isotherm models used in this study were consistent with the Freundlich model, but the pseudo second order model was the most accurate description of the adsorption kinetics. The present investigation showed an excellent potential with 85 % adsorption capacity for the removal of Cr (VI). Moreover, reusability studies showed that the cation exchanger can be used effectively up to four cycles.

Transformation of Cu2O into Metallic Copper within Matrix of Carboxylic Cation Exchangers: Synthesis and Thermogravimetric Studies of Novel Composite Materials.

In order to systematize and expand knowledge about copper-containing composite materials as hybrid ion exchangers, in this study, fine metallic copper particles were dispersed within the matrix of a carboxyl cation exchanger (CCE) with a macroporous and gel-type structure thanks to the reduction of Cu2O particles precipitated within the matrix earlier. It was possible to introduce as much as 22.0 wt% Cu0 into a gel-type polymeric carrier (G/H#Cu) when an ascorbic acid solution was used to act as a reducer of Cu2O and a reagent transforming the functional groups from Na+ into the H+ form. The extremely high shrinkage of the porous skeleton containing -COOH groups (in a wet and also dry state) and its limited affinity for water protected the copper from oxidation without the use of special conditions. When macroporous CCE was used as a host material, the composite material (M/H#Cu) contained 18.5 wt% Cu, and copper particles were identified inside the resin beads, but not on their surface where Cu2+ ions appeared during drying. Thermal analysis in an air atmosphere and under N2 showed that dispersing metallic copper within the resin matrix accelerated its decomposition in both media, whereby M/H#Cu decomposed faster than G/H#Cu. It was found that G/H#Cu contained 6.0% bounded water, less than M/H#Cu (7.5%), and that the solid residue after combustion of G/H#Cu and M/H#Cu was CuO (26.28% and 22.80%), while after pyrolysis the solid residue (39.35% and 26.23%) was a mixture of carbon (50%) and metallic copper (50%). The presented composite materials thanks to the antimicrobial, catalytic, reducing, deoxygenating and hydrophobic properties of metallic copper can be used for point-of-use and column water/wastewater treatment systems.

Factors affecting mixed-mode retention properties of cation-exchange stationary phases.

Cation-exchange stationary phases were characterized in different chromatographic modes (HILIC, RPLC, IC) and applied to the separation of non-charged hydrophobic and hydrophilic analytes. The set of columns under investigation included both commercially available cation-exchangers and self-prepared PS/DVB-based columns, the latter consisting of adjustable amounts of carboxylic and sulfonic acid functional groups. The influence of cation-exchange site and polymer substrate on the multimodal properties of cation-exchangers was identified using selectivity parameters, polymer imaging and excess adsorption isotherms. Introducing weakly acidic cation-exchange functional groups to the unmodified PS/DVB-substrate effectively reduced hydrophobic interactions, whilst a low degree of sulfonation (0.09 to 0.27% w/w sulphur) mainly influenced electrostatic interactions. Silica substrate was found to be another important factor for inducing hydrophilic interactions. The presented results demonstrate that cation-exchange resins are suitable for mixed-mode applications and offer versatile selectivity.

Moving adsorption belt system for continuous bioproduct recovery utilizing composite fibrous adsorbents.

A continuous protein recovery and purification system based on the true moving bed concept is presented. A novel adsorbent material, in the form of an elastic and robust woven fabric, served as a moving belt following the general designs observed in known belt conveyors. The composite fibrous material that forms the said woven fabric showed high protein binding capacity, reaching a static binding capacity equal to 107.3 mg/g, as determined via isotherm experiments. Moreover, testing the same cation exchange fibrous material in a packed bed format resulted in excellent dynamic binding capacity values (54.5 mg/g) even when operating at high flow rates (480 cm/h). In a subsequent step, a benchtop prototype was designed, constructed, and tested. Results indicated that the moving belt system could recover a model protein (hen egg white lysozyme) with a productivity up to 0.5 mg/cm2/h. Likewise, a monoclonal antibody was directly recovered from unclarified CHO_K1 cell line culture with high purity, as judged by SDS-PAGE, high purification factor (5.8), and in a single step, confirming the suitability and selectivity of the purification procedure.

Denitrification of nitrate in regeneration waste brine using hybrid cation exchanger supported nanoscale zero-valent iron with/without palladium nanoparticles.

The Sustainable Development Goals require that reducing waste is a priority. This work described the application of an innovative zero-waste hybrid ion exchange nanotechnology that concurrently removed nitrate and induced denitrification to ammonia, with the ability to generate fertilizer for the agriculture sector from the recycled by-products. Herein, hybrid cation exchanger-supported zero-valent iron (Fe0), and bimetallic Fe0/Pd nanoparticles (HCIX-Fe0 and HCIX-Fe0/Pd) were synthesized and successfully validated for denitrification of nitrate in spent waste brine that contained nitrate. The kinetics of nitrate catalysis by both HCIX-Fe0 and HCIX-Fe0/Pd were compared and presented by six kinetic models, namely, zero-order, pseudo first- and second-order reaction, pseudo first- and second-order adsorption, and Elovich. HCIX-Fe0/Pd displayed a higher kinetic value than HCIX-Fe0, with k1 of 0.0019 and 0.0026 min-1, respectively. Nitrate was predominantly catalysed to NH4+ at a ratio of ammonia to other nitrogen compounds of around 80:20. Although HCIX-Fe0/Pd showed slightly better (14%) kinetic results, it was determined as unfavourable for real-life application due to low selectivity toward N2 gas and the need to use H2 gas. Based on practicability, the HCIX-Fe0 was further validated. The effect of salt (using NaCl) and the role of initial pH conditions were optimized and discussed. The recovery of nitrate removal was also calculated, and a recovery range of 91.42-99.14% was obtained for three consecutive runs. The sustainable, novel, zero waste hybrid ion exchange nanotechnology using the combination of two fixed-bed columns containing nitrate-selective resin for nitrate removal and novel HCIX-Fe0 for nitrate reduction to NH4+ may be a promising sustainable solution toward the goal of discharging zero nitrate waste to the environment.

A review of the design of packing materials for ion chromatography.

The development of ion chromatography has made remarkable progress in the past few decades, and it is now widely used for the analysis of common ions and organic compounds. Ion chromatography has many advantages, such as fast, high sensitivity, good selectivity and support for simultaneous analysis of multiple ionic compounds. In order to meet the high requirements of material analysis, new packing materials for ion chromatography with higher sensitivity and selectivity have been developed. In this paper, a lot of knowledge of ion chromatography is reviewed, and the development of ion chromatographic packings in recent years, especially in the last five years, is summarized.

Preparation of a polymer-based weak cation exchanger for ion chromatography via atom transfer radical polymerization.

A novel polymer-based weak cation exchanger (WCX) for ion chromatography has been described. It was prepared by grafting tert-butyl acrylate and maleic anhydride onto the surface of poly(glycidyl methacrylate-divinylbenzene) microspheres via atom transfer radical polymerization, followed by hydrolytic treatment to produce acrylic and maleic acid groups. The obtained WCX showed better separation and higher selectivity for model cations relative to solely acrylic or maleic acid. Simultaneous separation of alkali and alkaline earth cations (including NH4+) was achieved in a single isocratic run under suppressed mode, and ion exchange was found to dominate the separation process. Its utility was demonstrated for determination of several cations in a beer sample and the recovery was ranging from 98.4 to 109.9%.

Copper Rich Composite Materials Based on Carboxylic Cation Exchangers and Their Thermal Transformation.

The effect of a cupric deposit (Cu2+, CuO) on the thermal decomposition of carboxylic cation exchangers (CCEs) is not known, and such studies may have practical significance. CCEs have a very high ion exchange capacity, so an exceptionally large amount of CuO (which is a catalyst) can be precipitated inside them. Two CCEs, macroreticular (Amberlite IRC50) and gel-like (Amberlite IRC86), served as a polymeric support to obtain copper-rich hybrid ion exchangers. Composites with CuO particles inside a polyacrylic matrix (up to 35.0 wt% Cu) were obtained. Thermal analyses under air and under N2 were performed for CCEs in the H+ and Cu2+ form with and without a CuO deposit. The results of sixteen experiments are discussed based on the TG/DTG curves and XRD patterns of the solid residues. Under air, the cupric deposit shifted the particular transformations and the ultimate polymeric matter decomposition (combustion) toward lower temperatures (even about 100-150 °C). Under N2, the reduction of the cupric deposit to metallic copper took place. Unique composite materials enriched in carbonaceous matter were obtained, as the products of polymeric matrix decomposition (free radicals and hydrogen) created an additional amount of carbon char due to the utilization of a certain amount of hydrogen to reduce Cu (II) to Cu0.

Salt-tolerant cation exchanger-containing sulfate groups as a viable alternative for mixed-mode type and heparin-based affinity resins.

Ion-exchange chromatography is still one of the most popular protein separation techniques. Before chromatographic separation, the high salt concentration in various samples necessitates additional steps. Therefore, low salt tolerance of ion-exchange resins is a drawback that needs to be addressed. Herein, the differences in salt tolerance and hydrophobicity of strong cation-exchange TOYOPEARL resins of sulfonium and sulfate-types were investigated. Despite only a minor structural difference, differences in selectivity and salt tolerance between the sulfate and sulfonic groups were detected. In silico calculations were also carried out for model substances representing the sulfonium and sulfate groups, wherein significant differences in hydrophobicity was observed. These experiments confirmed the hypothesis that the salt tolerance, higher affinity, and selectivity for certain vitamin K dependent clotting factors are interrelated and dependent on the presence of the sulfate group. Separation of clotting factor IX from the prothrombin complex concentrate further to confirmed the affinity for these proteins. The results show that the use of only a resin with the sulfate ligand and not with the sulfonic acid ligand allows for a facile and rapid separation of clotting factor IX and other vitamin K dependent clotting factors.

Features of Thermomechanical Stability of Anionic-Cation Exchange Matrix "Polikon AC" on Viscose Non-Woven Materials.

The thermomechanical stability of the anion-cation exchange matrix "Polikon AC" on viscose nonwoven materials is investigated. In this work, a molecular model of a solvation environment for experimentally obtained "Polikon AC" mosaic membranes is refined. Mosaic membranes on a viscose fiber base were fabricated by the method of polycondensation filling. The temperature dependence of deformation was investigated for dry and wet anion and cation exchange membrane components at a constant tensile load of 1.5 N and a heating rate of 8 °C/min. The effect of moisture content on the deformation of anionite and cationite fragments under a constant external tensile load of 1.5 and 3 N in a temperature range up to 100 °C was studied.

[Preparation of difunctional polymer-based cation exchangers and their application in ion chromatography].

Ion chromatography (IC) is one of the most important means for the separation and analysis of cationic compounds. Research on high-performance cation exchangers is of great significance for the advancement of modern IC techniques. Herein we proposed a newly developed modification method for polymer-based particles, based on thiol-radical-mediated polymerization. Specifically, acrylic acid and maleic anhydride were used as monomers and sodium 2-mercaptoethanesulfonate was used as the thiol modifier to prepare novel difunctional cation exchangers, functionalized with both carboxylic and sulfonic groups. With simple strong acid eluents, typical cations were well separated with good resolution. The retention behaviors of metal cations and organic amines on the cation exchangers were investigated using multiple chromatographic models. The good performances of the cation exchangers were also demonstrated by the gradient separation of ten cations within 24 min. The new method was easy to perform with high efficiency. Additionally, the cation exchange capacities of the cation exchangers were easily modulated by adjusting the initial content of the thiol modifier.

Immobilization of inulinase on KU-2 ion-exchange resin matrix.

We develop a technique for the sorption of inulinase from Kluyveromyces marxianus on the KU-2 matrix cation-exchanger. The most appropriate conditions for immobilization are: 25 °C, pH 4.5, incubation time 1.5 h, protein content during immobilization 10 mg/g of carrier. At higher (20-100 mg/g) concentrations, inulinase forms local high-concentration domains on the ion-exchanger surface, the enzyme is adsorbed on the protein, not on the carrier. 62% of the initial catalytic activity of inulinase is immobilized on KU-2 by the adsorption method. Upon the enzyme immobilization on KU-2, the number of unordered structures in the protein is reduced by ~10%, that points to the compaction of the molecule. Hydrogen bonds are formed only between the sulfo groups of carrier and the protein molecule, without affecting other structures of the cation exchanger. The highest activity of the inulinase immobilized on KU-2 was observed at 70 °C (cf. 50 °C for the native enzyme). Heating of the solution for 60 min in the temperature range of 50-80 °C failed to inactivate completely the immobilized enzyme; the complete loss of its ability to hydrolyze inulin was achieved only after more than 1 h incubation at 90 °C.

A Calcium/Cation Exchanger Participates in the Programmed Cell Death and in vitro Virulence of Entamoeba histolytica.

Entamoeba histolytica is the etiologic agent of human amoebiasis, disease that causes 40,000 to 100,000 deaths annually worldwide. The cytopathic activity as well as the growth and differentiation of this microorganism is dependent on both, extracellular and free cytoplasmic calcium. However, few is known about the proteins that regulate the calcium flux in this parasite. In many cells, the calcium extrusion from the cytosol is performed by plasma membrane Ca2+-ATPases and calcium/cation exchangers. The aim of this work was to identify a calcium/cation exchanger of E. histolytica and to analyze its possible role in some cellular processes triggered by calcium flux, such as the programmed cell death and in vitro virulence. By searching putative calcium/cation exchangers in the genome database of E. histolyica we identified a protein belonging to the CCX family (EhCCX). We generated a specific antibody against EhCCX, which showed that this protein was expressed in higher levels in E. histolytica than its orthologous in the non-pathogenic amoeba E. dispar. In addition, the expression of EhCCX was increased in trophozoites incubated with hydrogen peroxide. This E. histolytica exchanger was localized in the plasma membrane and in the membrane of some cytoplasmic vesicles. However, after 10 min of erythrophagocytosis, EhCCX was found predominantly in the plasma membrane of the trophozoites. On the other hand, the parasites that overexpress this exchanger contained higher cytosolic calcium levels than control, but the extrusion of calcium after the addition of hydrogen peroxide was more efficient in EhCCX-overexpressing trophozoites; consequently, the programmed cell death was retarded in these parasites. Interestingly, the overexpression of EhCCX increased the in vitro virulence of trophozoites. These results suggest that EhCCX plays important roles in the programmed cell death and in the in vitro virulence of E. histolytica.

Synthesis and Ion-Exchange Properties of Graphene Th(IV) Phosphate Composite Cation Exchanger: Its Applications in the Selective Separation of Lead Metal Ions.

In this study, graphene Th(IV) phosphate was prepared by sol-gel precipitation method. The ion-exchange behavior of this cation-exchanger was studied by investigating properties like ion-exchange capacity for various metal ions, the effect of eluent concentration, elution behavior, and thermal effect on ion-exchange capacity (IEC). Several physicochemical properties as Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) study, thermal studies, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) studies were also carried out. The material possessed an IEC of 1.56 meq·dry·g-1 of the exchanger and was found to be nano-composite. The selectivity studies showed that the material is selective towards Pb(II) ions. The selectivity of this cation-exchanger was demonstrated in the binary separation of Pb(II) ions from mixture with other metal ions. The recovery was found to be both quantitative and reproducible.