Ion-exchange Column Research Articles

Resource Recovery from Abandoned Mine Drainage Galleries via Ion Exchange: A Case Study from Freiberg Mining Area, Germany

The discharge of metal-loaded mining-influenced waters can significantly pollute downstream water bodies for many kilometers. Addressing this issue at the earliest discharge point is crucial to prevent further contamination of the natural environment. Additionally, recovering metals from these discharges and other sources of contamination can reduce the environmental impacts of mining and support the circular economy by providing secondary raw materials. This study focused on optimizing zinc recovery from mining-influenced water in the Freiberg mining region in Germany, where significant loads of zinc are released into the Elbe River. By employing pretreatment techniques, conducting 100 mL scale ion-exchange column experiments, and refining the regeneration process, we aimed to identify optimal conditions for efficient zinc removal and recovery. Initial tests showed that aminophosphonic functionalized TP 260 resin had a high affinity for aluminum, occupying 93% of the resin’s capacity, while zinc capacity was limited to 0.2 eq/L. To improve zinc recovery, selective precipitation of aluminum at pH 6.0 was introduced as a pretreatment step. This significantly increased the zinc loading capacity of the resin to 1 eq/L. Under optimal conditions, a concentrated zinc solution of 18.5 g/L was obtained with 100% recovery. Sulfuric acid proved more effective than hydrochloric acid in eluting zinc from the resin. Further analysis using SEM-EDX revealed residual acid on the resin, indicating a need for additional study on long-term resin performance and capacity variation. The research also highlighted the environmental impact of the Freiberg mining area, where three drainage galleries currently contribute nearly 85 tons of zinc annually to the Elbe River. This study underscores the feasibility of efficient zinc recovery from these point sources of pollution using advanced ion-exchange processes, contributing to circular economy efforts and environmental conservation.

Ultrafast and Highly Selective Sequestration of Radioactive Barium Ions by a Layered Thiostannate.

As a simulant of hazardous 226Ra2+, the simultaneously selective and rapid elimination of radioactive 133Ba2+ ions from geothermal water is necessary but still challenging. In this paper, we demonstrated the usability of a layered thiostannate with facile synthesis and inexpensive cost, namely, K2xSn4-xS8-x (KTS-3, x = 0.65-1), for the remediation of radioactive 133Ba2+ in multiple conditions, including sorption isotherm, kinetics, and the influences of competitive inorganic/organic ions, pH values, and dosages. KTS-3 has a strong barium uptake ability (171.3 mg/g) and an ultrafast adsorption kinetics (about 2 min). Impressively, it can achieve a high preference for barium regardless of the excessive interference ions (Na+, K+, Mg2+, Ca2+, and humic acid) and acidic/alkaline environments, with the largest distribution coefficient Kd value reaching 6.89 × 105 mL/g. Also, the Ba2+-laden products can be easily eluted by a concentrated KCl solution, and its adsorption performances for barium resist well even after five consecutive cycles. In addition, owing to the regular appearance and excellent mechanical strength, the prepared KTS-3/PAN (PAN = polyacrylonitrile) granule displays a good removal efficiency in the flowing ion-exchange column. These advantages mentioned above render it very promising for the effective and efficient cleanup of radioactive 133Ba2+-contaminated wastewater.

Ion exchange columns. A promising technology for nitrogen and phosphorus recovery in the main line of a wastewater treatment plant

Anaerobic membrane bioreactor (AnMBR) technology has great advantages for treating urban wastewaters, but, when irrigation cannot be applied and the effluent is discharged in a sensitive zone, a post-treatment of this effluent is needed for nitrogen and phosphorus removal. Under this scenario, ion exchange processes represent one of the most promising technologies for treating this effluent. Ion exchange technology allows to meet discharge limits and to recover these nutrients in a highly concentrated stream. In this work, the technical feasibility of using a commercial resin for phosphorus recovery and a natural zeolite (clinoptilolite) for nitrogen recovery was evaluated. Purolite FerrIX A33E resin removed phosphate from the AnMBR permeate within 500 Bed Volumes (BVs) with a maximum adsorption capacity (qmax) of 2,1 mg P-PO4/g resin. Regeneration of the resin (2% NaOH 2% NaCl) recovered over 95% of the phosphorous retained, achieving a concentration of 316,7 mg P-PO4/L in the regeneration solution. In the absence of a long-term study, the resin showed a stable adsorption capacity during 16 cycles of saturation-regeneration. Clinoptilolite removed nitrogen within 139 BVs obtaining a qmax of 3,68 mg N-NH4/g zeolite. 97 % of the retained N-NH4 was recovered in the regeneration stage (0,8% NaOH) with an average concentration of 577 mg N-NH4/L. Continuous exposure of the zeolite to alkaline solutions led to reduction of 50% of the adsorption capacity after 17 cycles.

Recovering Nitrogen from Anaerobic Membrane Bioreactor Permeate Using a Natural Zeolite Ion Exchange Column

In the framework of a circular economy, wastewater treatment should be oriented toward processes that allow the recovery of the resources present in the wastewater while ensuring good effluent quality. Nitrogen recovery is usually carried out in streams concentrated in this nutrient because these high concentrations facilitate nitrogen valorization. On the other hand, the mainstream of a wastewater treatment plant (WWTP) has a high potential for nitrogen recovery, but it is not usually considered because it is hard to manage due to its low nitrogen concentration. To solve this problem and facilitate the recovery of nitrogen in the mainstream, this work proposes ion exchange with zeolites as a stage of ammonium concentration, to provide a nitrogen-concentrated stream that could be valorized by another technology, while obtaining a nitrogen-free effluent. The working stream, the permeate of an AnMBR process in the mainstream, has suitable characteristics to be treated in an ion exchange column (free of suspended solids and with very low organic matter content). To this end, the effect of the working flow rate (17.5 to 4.4 BV/h) and the ammonium concentration (54 to 17 mg NH4-N/L) on the adsorption capacity of the zeolite in the loading phase was evaluated. The adsorption curves were fitted to three mathematical models: Thomas, Bohart–Adams, and Yoon–Nelson. The effect of the regeneration flow rate (from 8.7 to 2.2 BV/h) and the regenerant concentration (NaOH at 0.2, 0.1, and 0.05 M) on regeneration capacity and efficiency were also studied. A novel control strategy based on effluent conductivity was used in both phases to control the duration of the adsorption and regeneration phases.

Anti-Melanogenic Effects of a Polysaccharide Isolated from Undaria pinnatifida Sporophyll Extracts.

Undaria pinnatifida is a temperate brown alga known to exert free radical-scavenging and anti-inflammatory effects. In this study, we investigated the skin-whitening effects of U. pinnatifida sporophyll extracts (UPEs) in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 melanoma cells. The crude polysaccharide fraction (UPF) was obtained via ethanol precipitation. Four polysaccharide fractions (UPF1-4) were isolated and purified using ion-exchange column chromatography, and their anti-melanogenic activity was evaluated. UPF3 exhibited the highest anti-melanogenic activity, showing the highest sulfate (39.79%), fucose (143 μg/mg), and galactose (208 μg/mg) contents. UPF3 significantly inhibited intracellular tyrosinase activity in B16F10 cells. We also evaluated the melanogenic signaling pathway to determine the mechanism of action of UPF3 in melanongenesis. UPF3 reduced the expression of tyrosinase-related protein-1 (TRP-1), tyrosinase-related protein-2 (TRP-2), and tyrosinase, which play important roles in melanin production. Therefore, UPF3 has high potential for use in skin-whitening functional pharmaceuticals and cosmetics.

Central composite experimental design for Indigo Carmine dye removal from solutions by applying electro-coagulation and electro-oxidation processes

Textile and food industries produce toxic or harmfull dye-containing wastewaters that should be treated for safely discharge. The electro-coagulation (EC) and the electro-oxidation (EO) methods for removal of dyes from wastewaters are being investigated nowadays by researchers. In this study, the Indigo Carmine, a textile and food coloring agent, removal was investigated by applying the electro-coagulation and the electro-oxidation methods. For this purpose, aluminum electrodes for EC and graphite plates for EO were used. The central composite experimental design was applied as the optimization method for the EC and the EO processes. The optimization parameters were selected as time (10-30 minutes), current density (0.4-2 Ampere/500 mL) and concentration (50-250 mg/L), natural pH (5.78-6.90) and room temperature (20-25 °C). The EO process was determined to be effective than the EC process. Statistically important parameters were concentration and time-current density interaction for the EC, but all the parameters were statistically unimportant for the EO. Dye removal percentages by the EC were calculated between 82.75% and 98.38%, and dye removal percentages by the EO were calculated between 46.88% and 100% for the determined experimental matrix. Electrical consumptions were almost equal for the EO and the EO prosesses. A column ion exchange process (Selion SBA 2000 resin) was applied to the dye residue after the EO treatment. From the oxidation reduction measurements, the treated solutions were determined as dischargeable.

Direct filtration of low-turbidity water by fixed-bed ion exchange treatment

In drinking water treatment, low-turbidity water (e.g., < 5 NTU) is often purified by direct filtration comprising coagulation, flocculation, and rapid sand filtration. This study assessed the potential of replacing the rapid sand filter with a fixed-bed ion exchange column. Two types of anion exchange resins, polyacrylic (Purolite A860) and polystyrene (Purolite A502PS), were used in fixed-bed columns without preceding coagulation and flocculation. After the first use of the new resins (14-h operation), the A860 column showed more significant head loss buildup than the A502PS column (151 and 101 cm, respectively). This can be attributed to the effective size of A860 resins (0.38 mm), which was smaller than that of A502PS resins (0.52 mm). A860 resins were superior to A502PS resins in filtrate quality. The A860 resins achieved filtrate turbidity (0.16–0.26 NTU) lower than the A502PS resins (0.19–0.35 NTU). Similarly, dissolved organic carbon removal by A860 resins (25–37 %) was higher than that by A502PS resin (15–25 %) due to the low hydrophobicity of the feed water. Air scouring and water backwashing were ineffective in regenerating A860 resins. In contrast, cleaning with a NaCl solution (pH = 12) partially recovered the capacity of A860 resins for turbidity removal.

Isolation, Characterization, and Functional Properties of Antioxidant Peptides from Mulberry Leaf Enzymatic Hydrolysates.

Recent evidence suggests that mulberry leaves have good antioxidant activity. However, what the antioxidant ingredient is and how the ingredient works are still not well understood. In this study, we enzymatically hydrolyze mulberry leaf proteins (MLPs) using neutral protease and find that the mulberry leaf protein hydrolysates (MLPHs) have stronger antioxidant activity compared to MLPs. We separate the core antioxidant components in MLPHs by ion-exchange columns and molecular sieves and identify 798 antioxidant peptides by LC-MS/MS. Through bioinformatics analysis and biochemical assays, we screen two previously unreported peptides, P6 and P7, with excellent antioxidant activities. P6 and P7 not only significantly reduce ROS in cells but also improve the activities of the antioxidant enzymes SOD and CAT. In addition, both peptides are found to exert protective effects against H2O2-induced chromatin damage and cell apoptosis. Collectively, these results provide support for the application of mulberry leaf peptides as antioxidants in the medical, food and livestock industries.

Toxic Peptides from the Mexican Scorpion Centruroides villegasi: Chemical Structure and Evaluation of Recognition by Human Single-Chain Antibodies.

Alternative recombinant sources of antivenoms have been successfully generated. The application of such strategies requires the characterization of the venoms for the development of specific neutralizing molecules against the toxic components. Five toxic peptides to mammals from the Mexican scorpion Centruroides villegasi were isolated by chromatographic procedures by means of gel filtration on Sephadex G-50, followed by ion-exchange columns on carboxy-methyl-cellulose (CMC) resins and finally purified by high-performance chromatography (HPLC) columns. Their primary structures were determined by Edman degradation. They contain 66 amino acids and are maintained well packed by four disulfide bridges, with molecular mass from 7511.3 to 7750.1 Da. They are all relatively toxic and deadly to mice and show high sequence identity with known peptides that are specific modifiers of the gating mechanisms of Na+ ion channels of type beta-toxin (β-ScTx). They were named Cv1 to Cv5 and used to test their recognition by single-chain variable fragments (scFv) of antibodies, using surface plasmon resonance. Three different scFvs generated in our laboratory (10FG2, HV, LR) were tested for recognizing the various new peptides described here, paving the way for the development of a novel type of scorpion antivenom.

Fast and reliable method for analysis of derivatized plasma amino acids by liquid chromatography-single quadrupole-mass spectrometry

The pool of free, genetically encoded AA in plasma plays an essential role not only as substrate for every protein synthesized in the body but also as signaling molecules that regulate a wide range of physiological processes. Here we present a method for the analysis of 19 of the 20 encoded AA (except Cys) in dairy cow plasma. Isolated plasma or standards for the 19 AA were gravimetrically mixed with an internal standard mix consisting of 13C isotopes of each AA. Plasma proteins were precipitated on acetonitrile and supernatants transferred to glass vials. For pre-column derivatization, plasma supernatants were buffered with sodium borate (pH 9.5 −10), and AA were derivatized with 9-fluorenylmethoxycarbonyl (Fmoc) chloride. Analytes were isolated by solid-phase extraction using a strong-anionic ion exchange column and dry eluates were reconstituted in mobile phase consisting of 70% water solution of ammonium formate and 30% acetonitrile. Amino acid derivatives were separated by reverse phase liquid chromatography over 17.5 min with a C18 column in which acetonitrile increased to 80% over the first 11 min of the method, before returning to initial levels. Electrospray ionization on negative mode was used for most AA, except Arg and Pro, for which positive mode yielded superior results. Single or double (only Lys) derivatives were measured by single quadrupole mass spectrometry. We hypothesized that pre-column Fmoc derivatization would yield optimal resolution for quantitative analysis of the 19 targeted AA and their respective 13C internal standards, with limits of quantitation beyond physiological ranges. All the 19 AA were detected with minimal background noise. An 11-point standard curve was developed for each AA. Limits of quantitation were beyond concentrations observed in plasma samples of lactating dairy cows, except for Gly that upper curve points had to be removed to maintain linearity, limiting quantitation to the upper range of physiological concentration. After removing the 4 highest concentrations from the Gly standard curve, coefficients of determination were greater than 0.999 for all the AA. Recovery of spiked AA from plasma samples ranged from 89.9% for Phe to 100.3% for Trp. Instrument repeatability averaged 0.91 and ranged from 0.33 for Val to 2.29 for Arg. Meanwhile, sample preparation method repeatability averaged 2.02 and ranged from 1.14 for Tyr to 3.34 for Arg. While robust methods have been developed, they depend on either availability of sophisticated instruments, mostly limited to core facilities (i.e., tandem MS methods), long and expensive chromatography without specific internal standards for each AA (i.e., HPLC-UV and HPLC-FLD), or unstable derivatization (GC-MS). Here we describe a method with high throughput, stable derivatization, high precision and recovery, and potentially more affordable than most existing methods. This method could help dairy nutritionists to consider plasma AA information for diet formulation strategies, potentially reducing feeding costs and N emissions.

Continuous multicolumn ion exchange process for spent lithium-ion battery leachate: Recovery and purification of a Li+Ni+Co mixture

A continuously operated ion exchange process scheme for the recovery and purification of valuable metals from acid leachates of spent Lithium-ion battery cathodes was developed. The aim is to provide a versatile and industrially feasible alternative for liquid–liquid extraction and precipitation for recycling of spent Li-ion batteries. A chelating resin with aminomethylphosphonic acid functional group (Lewatit® MDS TP 260) was used in a laboratory-scale multicolumn configuration that combined counter-current and cross-current operations. The process was operated at 60 °C with a synthetic leachate with pH 1.8 as the feed. A two-step desorption procedure (1.0 M sulfuric acid (H2SO4) followed by 0.4 M potassium oxalate (K2C2O4)) was found to prevent accumulation of strongly bound metals in the resin. 1 M H2SO4 was found sufficient for regeneration. Pure raffinate containing Li, Ni, and Co was successfully separated from impurity metals (Mn, Cu, Al, and Fe). In steady state operation, no loss of ion exchange capacity was observed. The process was optimized experimentally with focus on the effect of fresh feed and eluent flow rates to increase the productivity and improve the recovery rates of the target metals while maintaining a raffinate purity of 100 %. Increasing the amount of leachate processed per switch was found to enhance the recovery of Co, Li and Ni due to the displacement effect. Very high recovery yields (96.97 % for Co, 99.15 % for Ni, and 100 % for Li) were achieved. The laboratory scale unit could process 1.64L(L·h)−1 of leachate to yield 34.04 g(L·h)−1 of Li+Ni+Co mixture in the raffinate. Moreover, Cu+Mn and Al+Fe rich extract streams with 84.74 % and 97.46 % purities were obtained as by-products. With respect to the recovery rates, the continuous ion exchange process is superior to batchwise operation of ion exchange columns, conventional solvent extraction and not to mention precipitation processes making it a viable alternative in hydrometallurgical LIB recycling.

Purification and identification of novel osteogenic peptides from sea cucumber intestine hydrolysates and their pro-osteogenesis effects on MC3T3-E1 cells

The present study aimed to purify and identify osteogenic peptides from sea cucumber intestine hydrolysates (Mw＜3 kDa) and further evaluate their pro-osteogenesis properties on MC3T3-E1 cells. Two novel osteogenic peptides, RPQYPQYPS and LSFSPY, were purified and identified from the alcalase hydrolysate of sea cucumber intestine by gel filtration chromatography, ion exchange column chromatography, RP-HPLC, which were further sequenced by LC-MS/MS and synthesized with solid-phase synthesis method. Peptides RPQYPQYPS and LSFSPY significantly increased the proliferation of MC3T3-E1 cells by 38.7% and 32.8%. Alizarin red staining also confirmed that peptides RPQYPQYPS and LSFSPY significantly promoted MC3T3-E1 cell mineralization in a dose-dependent manner. Further, molecular docking results showed that RPQYPQYPS and LSFSPY had a higher affinity with integrin α5β1. Western blot results confirmed that RPQYPQYPS and LSFSPY significantly upregulated the expression of integrin downstream signals PI3K, ERK and Wnt10b. These findings provided evidence for the osteogenic peptides from sea cucumber intestines as a functional food ingredient.

Kenaf Seed Cysteine Protease (KSCP) Inhibits the Intrinsic Pathway of the Blood Coagulation Cascade and Platelet Aggregation.

Thrombosis is the key event that obstructs the flow of blood throughout the circulatory system, leading to stroke, myocardial infarction and severe cardiovascular complications. Currently, available antithrombotic drugs trigger several life-threatening side effects. Antithrombotic agents from natural sources devoid of adverse effects are grabbing high attention. In our previous study, we reported the antioxidant, anticoagulant and antiplatelet properties of kenaf seed protein extract. Therefore, in the current study, purification and characterization of cysteine protease from kenaf seed protein extract responsible for potential antithrombotic activity was undertaken. Purification of KSCP (Kenaf Seed Cysteine Protease) was carried out using gel permeation and ion exchange column chromatography. The purity of the enzyme was evaluated by SDS PAGE (Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis). RP-HPLC (Reverse Phase High-Performance Liquid Chromatography), MALDI-TOF (Matrix-Assisted Laser Desorption Ionization Time-Of-Flight) and CD (Circular Dichroism techniques) were employed for its characterization. Proteolytic, fibrinolytic and kinetic study was done using spectroscopy. Plasma recalcification time, Prothrombin Time (PT), Thrombin clotting time (TCT), Activated Partial Thromboplastin Time (APTT), bleeding time and platelet aggregation studies were carried out for antithrombotic activity of KSCP. A single sharp band of KSCP was observed under both reduced and non-reduced conditions, having a molecular mass of 24.1667kDa. KSCP was found to contain 30.3% helix turns and 69.7% random coils without a beta-pleated sheet. KSCP digested casein and fibrin, and its activity was inhibited by iodoacetic acid (IAA). KSCP was optimally active at pH 6.0 at the temperature of 40°C. KSCP exhibited anticoagulant properties by interfering in the intrinsic pathway of the blood coagulation cascade. Furthermore, KSCP dissolved both whole blood and plasma clots and platelet aggregation. KSCP purified from kenaf seed extract showed antithrombotic potential. Hence, it could be a better candidate for the management of thrombotic complications.

Precise measurement of Fe isotopes in marine and biological samples by pseudo-high-resolution multi-collector inductively coupled plasma mass spectrometry (MC-ICPMS).

This paper introduces an enhanced technique for analyzing iron isotopes in complex marine and biological samples. A dedicated iron purification method for biological marine matrices, utilizing three ion exchange columns, is validated. The MC-ICPMS in pseudo-high-resolution mode determines precise iron isotopic ratios, with sensitivity improved through the DSN-100 desolvating nebulizer system and Apex-IR. Only 2µg of iron on DSN versus 1µg on Apex is needed for six replicates (30-60 times improvement) while 10 to 20µg is required for a single measurement on awet system considering the resolution power (Rp) is maintained at 11,000-13,000. The Ni-doping method with a Fe/Ni ratio of 1 yields more accurate isotopic ratios than standard-sample bracketing alone. Measurement reproducibility of triplicate samples from marine biological experiments on MC-ICPMS is ± 0.03‰ (2SD) for δ56Fe and ± 0.07‰ for δ57Fe (2SD). This study introduces a novel iron purification process specifically designed for marine and biological samples, enhancing sensitivity and enabling more reliable measurements with smaller sample sizes and reduced uncertainties. It proposes iron isotopic compositions for biological reference materials, offering a valuable reference dataset in diverse scientific disciplines.

“Ion-imprinting” strategy towards metal sulfide scavenger enables the highly selective capture of radiocesium

Highly selective capture of radiocesium is an urgent need for environmental radioactive contamination remediation and spent fuel disposal. Herein, a strategy is proposed for construction of “inorganic ion-imprinted adsorbents” with ion recognition-separation capabilities, and a metal sulfide Cs2.33Ga2.33Sn1.67S8·H2O (FJSM-CGTS) with “imprinting effect” on Cs+ is prepared. We show that the K+ activation product of FJSM-CGTS, Cs0.51K1.82Ga2.33Sn1.67S8·H2O (FJMS-KCGTS), can reach adsorption equilibrium for Cs+ within 5 min, with a maximum adsorption capacity of 246.65 mg·g−1. FJMS-KCGTS overcomes the hindrance of Cs+ adsorption by competing ions and realizes highly selective capture of Cs+ in complex environments. It shows successful cleanup for actual 137Cs-liquid-wastes generated during industrial production with removal rates of over 99%. Ion-exchange column filled with FJMS-KCGTS can efficiently treat 540 mL Cs+-containing solutions (31.995 mg·L−1) and generates only 0.12 mL of solid waste, which enables waste solution volume reduction. Single-crystal structural analysis and density functional theory calculations are used to visualize the “ion-imprinting” process and confirm that the “imprinting effect” originates from the spatially confined effect of the framework. This work clearly reveals radiocesium capture mechanism and structure-function relationships that could inspire the development of efficient inorganic adsorbents for selective recognition and separation of key radionuclides.

Purification and Biochemical Characterization of Polyphenol Oxidase Extracted from Wheat Bran (Wan grano).

Currently, little is known about the characteristics of polyphenol oxidase from wheat bran, which is closely linked to the browning of wheat product. The wheat PPO was purified by ammonium sulfate precipitation, DEAE-Sepharose ion-exchange column, and Superdex G-75 chromatography column. Purified wheat PPO activity was 11.05-fold higher, its specific activity was 1365.12 U/mg, and its yield was 8.46%. SDS-PAGE showed that the molecular weight of wheat PPO was approximately 21 kDa. Its optimal pH and temperature were 6.5 and 35 °C for catechol as substrate, respectively. Twelve phenolic substrates from wheat and green tea were used for analyzing the substrate specificity. Wheat PPO showed the highest affinity to catechol due to its maximum Vmax (517.55 U·mL-1·min-1) and low Km (6.36 mM) values. Docking analysis revealed strong affinities between catechol, gallic acid, EGCG, and EC with binding energies of -5.28 kcal/mol, -4.65 kcal/mol, -4.21 kcal/mol, and -5.62 kcal/mol, respectively, for PPO. Sodium sulfite, ascorbic acid, and sodium bisulfite dramatically inhibited wheat PPO activity. Cu2+ and Ca2+ at 10 mM were considered potent activators and inhibitors for wheat PPO, respectively. This report provides a theoretical basis for controlling the enzymatic browning of wheat products fortified with green tea.

Electrostatic Repulsion Hydrophilic Interaction Liquid Chromatography (ERLIC) for the Quantitative Analysis of Polyamines

Highly polar low molecular weight organic molecules are still very challenging to analyze by liquid chromatography. Yet, with the steadily increasing application of metabolomics and similar approaches in chemical analysis, separating polar compounds might be even more important. However, almost all established liquid chromatography techniques (i.e., normal and reversed phase, hydrophilic interaction liquid chromatography (HILIC), ion chromatography) struggle with either carry-over, low sensitivity, or a lack of retention. For improving these shortcomings, electrostatic repulsion hydrophilic interaction chromatography (ERLIC) might be an alternative. By combining a HILIC mobile phase, that is highly organic with a low water content, and an ion exchange column, a distinct layer system develops. When the analyte's charge is of the same direction as the stationary phase, retention and elution are determined by two antagonistic forces: electrostatic repulsion and hydrophilicity. One prominent group of challenging polar analytes are the polyamines cadaverine, putrescine, spermidine, and spermine. Carrying charges from +2 to +4 at physiological pH, these compounds are essential cell constituents and found in all living organisms. However, they are still notoriously challenging to analyze via the established liquid chromatography methods. In the present work, an ERLIC tandem mass spectrometry method has been exemplarily developed, optimized, and validated for the quantitative determination of cadaverine, putrescine, spermidine, and spermine. This method enables symmetrical peak shapes and good separation of analytes with different charges while simultaneously selectively detecting the co-eluting diamines by MS/MS. Furthermore, high linearity (R > 0.998) and sensitivity (LODs ≤ 2 ng/mL) have been proven. Thus, ERLIC may be interesting for both targeted and untargeted analysis approaches of highly charged low molecular weight organic molecules.

Characterization of Sodium Channel Peptides Obtained from the Venom of the Scorpion Centruroides bonito.

Five peptides were isolated from the venom of the Mexican scorpion Centruroides bonito by chromatographic procedures (molecular weight sieving, ion exchange columns, and HPLC) and were denoted Cbo1 to Cbo5. The first four peptides contain 66 amino acid residues and the last one contains 65 amino acids, stabilized by four disulfide bonds, with a molecular weight spanning from about 7.5 to 7.8 kDa. Four of them are toxic to mice, and their function on human Na+ channels expressed in HEK and CHO cells was verified. One of them (Cbo5) did not show any physiological effects. The ones toxic to mice showed that they are modifiers of the gating mechanism of the channels and belong to the beta type scorpion toxin (β-ScTx), affecting mainly the Nav1.6 channels. A phylogenetic tree analysis of their sequences confirmed the high degree of amino acid similarities with other known bona fide β-ScTx. The envenomation caused by this venom in mice is treated by using commercially horse antivenom available in Mexico. The potential neutralization of the toxic components was evaluated by means of surface plasmon resonance using four antibody fragments (10FG2, HV, LR, and 11F) which have been developed by our group. These antitoxins are antibody fragments of single-chain antibody type, expressed in E. coli and capable of recognizing Cbo1 to Cbo4 toxins to various degrees.

Purification, kinetic characterization of thermostable multicopper oxidase from the oyster mushroom and its versatility for greener agro-pulp bio bleaching in the paper industry.

Production of a thermostable laccase from Pleurotus florida was reported for the first time, both in submerged and solid-state fermentation using agro-industrial residues. This enzyme was purified using ammonium sulphate precipitation (60-90%), Sephadex G-100 and DEAE column ion exchange chromatography, respectively. The laccase was purified to 21.49 fold with an apparent molecular weight of 66 kDa and had an optimal pH of 5 with temperature stability at 60°C. Metal ions such as Cu2+ (91.26 µmole/mL/min), Mg2+ (68.15 µmole/mL/min), and Fe2+ (1.73 µmole/mL/min) enhanced the laccase activity, but Fe2+ (1.73µmole/mL/min) inhibited the enzyme activity. The purified laccase had Km and Vmax of 16.68 mM and 26.73 µmole/mL/min for guaiacol as a substrate. The isolated enzyme was characterized by FT-IR which revealed bands at 3655.0 cm-1, 2894.7 cm-1, and 1151.7 cm-1 corresponding to primary amines, C-H stretch, and amide -III, respectively. The enzymatic bio bleaching of paddy straw pulp was found to be most effective which resulted in a lowering of kappa number and yellowness by 19.47% & 17.84% whereas an increase in brightness and whiteness by 41.92%. & -19.61%. Thus, this might be stated that the crude laccase from P. florida can be exploited to reduce the toxic waste load for managing environmental pollution and helps in enhancing the yield and quality of the paper.

Extraction, purification, and investigation of the antibacterial potential of lysozyme isolated from the latex of Calotropis procera

Calotropis procera produces a latex used in traditional medicine because of its components which are found to be biologically active. The latex of C. procera has many benefits such as pain reduction, antimicrobial attitude and other precious attributes. Latex lysozyme (LL) is a particular protein contained in C. procera plant having a promising antibacterial property. The process applied for extraction implicated ammonium sulfate precipitation, water extraction, ion exchange and gel filtration column chromatography evidencing 14.4 kDa for LL as a molecular mass. LL after purification showed 397.18 U mg−1 as specific activity (10.30 purification fold), and with 37.9% as a yield. In addition, the chemical and physical analyses showed that LL had its best realization when the pH was 5.5 at 50 °C. Moreover when metal ions such as Ca2+ and Mg2+ were incorporated the relative activity of the enzyme resulted ampliflied. When a purification was conducted with CM-Cellulose and Sephadex G-100 chromatography, LL revealed an antibacterial property versus both Escherichia coli (MICs 14 µg ml−1) and Bacillus cereus (MICs 13 µg ml−1). HR-TEM analysis displayed an antimicrobial potential of LL after its purification; that analysis showed the deterioration of the cell wall (external membrane of the cell) of both E. coli and B. cereus. Therefore, the LL obtained from C. procera maintains ability as an antibacterial enzyme applicable in various domains such as biotechnological and pharmaceutical industries.Graphical abstract