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The research introduces a novel method for creating drug-loaded hydrogel beads that target anti-aging, anti-oxidative, and anti-inflammatory effects, addressing the interconnected processes underlying various pathological conditions. The study focuses on the development of hydrogel beads containing anti-aging compounds, antioxidants, and anti-inflammatory drugs to effectively mitigate various processes. The synthesis, characterization and in vitro evaluations, and potential applications of these multifunctional hydrogel beads are discussed. A polymeric alginate-orange peel extract (1:1) hydrogel was synthesized for encapsulating fish oil. Beads prepared with variable fish oil concentrations (0.1, 0.3, and 0.5 ml) were characterized, showing no significant decrease in size i.e., 0.5 mm and a reduction in pore size from 23 to 12 µm. Encapsulation efficiency reached up to 98% within 2 min, with controlled release achieved upto 45 to 120 min with increasing oil concentration, indicating potential for sustained delivery. Fourier-transform infrared spectroscopy confirmed successful encapsulation by revealing peak shifting, interaction between constituents. In vitro degradation studies showed the hydrogel's biodegradability improved from 30 to 120 min, alongside anti-inflammatory, anti-oxidative, anti-collagenase and anti-elastase activities, cell proliferation rate enhanced after entrapping fish oil. In conclusion, the synthesized hydrogel beads are a promising drug delivery vehicle because they provide stable and effective oil encapsulation with controlled release for notable anti-aging and regenerative potential. Targeted delivery for inflammatory and oxidative stress-related illnesses is one set of potential uses. Further research may optimize this system for broader applications in drug delivery and tissue engineering.

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There is great interest in developing effective therapies for the treatment of skin wounds accompanied by deep tissue losses and severe infections. We have attempted to prepare biohybrids formed of agglomerates of mesenchymal stem cells (MSCs) with gelatin hydrogel beads (GEL beads) delivering bacteriophages (phages) as antibacterial agents and/or basic fibroblast growth factor (bFGF) for faster and better healing, providing combined therapies for these types of skin wounds. The gelatin beads were produced through a two-step process using basic and/or acidic gelatins with different isoelectric points. Escherichia coli (E. coli) and its specific T4 phages were propagated. Phages and/or bFGF were loaded within the GELs and their release rates and modes were obtained. The phage release from the basic GEL beads was quite fast; in contrast, the bFGF release from the acidic GEL beads was sustained, as anticipated. MSCs were isolated from mouse adipose tissues and 2D-cultured. Agglomerates of these MSCs with GEL beads were formed and maturated in 3D cultures, and their time-dependent changes were followed. In these 3D culture experiments, it was observed that the agglomerates with GEL beads were very healthy and the MSCs formed tissue-like structures in 7 days, while the MSC agglomerates were not healthy and shrunk considerably as a result of cell death.

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Carotenoids, the natural pigments that confer the bright orange color of sea buckthorn berries, are also associated with several health benefits, such as antioxidant activity and skin and eye protection. Due to their lipophilic nature and localization, carotenoids are largely retained in the sea buckthorn pomace (SBP) resulting from juice production. Carotenoids from SBP (70.03 mg/100 g DW), extracted and characterized by HPLC-PDA, contained zeaxanthin (free and esterified) and beta-carotene as major compounds. The SBP carotenoids-enriched sunflower oil was further encapsulated in Ca-alginate hydrogel beads (98.4% encapsulation efficiency) using ionotropic gelation. The hydrogel beads were characterized by confocal laser scanning microscopy and scanning electron microscopy. Fairly good stability (>64%) of the encapsulated carotenoids in the alginate hydrogel beads during storage (30 days, 4 °C and 25 °C) was found, with zeaxanthin esters being the most stable compounds, for all the experimental conditions. The bioaccessibility of the total carotenoids (INFOGEST protocol) was 42.1 ± 4.6% from hydrated, and, respectively, 40.8 ± 4% from dehydrated SBP alginate hydrogel beads. The addition of yogurt to the dehydrated hydrogel beads had a positive effect on the bioaccessibility of free and esterified zeaxanthin, but not on that of the carotenes. In conclusion, SBP is a valuable source of carotenoids which can be protected by encapsulation in alginate hydrogel beads, thus still retaining a good bioaccessibility.

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Red Monascus Pigment (RMP), a natural pigment, has attracted significant attention due to its suitability for food use and potential health benefits. However, preserving its stability and exploring value-added development opportunities remain crucial challenges. This study outlined the utilization of RMP, by successfully preparing hydrogel beads encapsulating RMP crude extract (RMPCE) through Ca2+-mediated chitosan (CS)/sodium alginate (SA) encapsulation (CO-RMPHB). A systematic investigation into the fabrication and stability parameters, including preparation conditions, temperature, monochromatic light and storage time, was undertaken. Through optimization (SA: 2.50 wt%; CaCl2: 6.00 wt%; CS: 0.50 wt%), maximum encapsulation efficiency of 73.54 ± 2.16 % was achieved. The maximum swelling degree of blank hydrogel beads (BHB) in simulated gastric solution (pH = 1.2, 1.50 ± 0.97 %) was significantly lower than in simulated intestinal solution (pH = 7.0, 28.05 ± 1.43 %), confirming their sensitivity to pH changes. Additionally, the CO-RMPHB (66.08 %, 1000 µL) exhibited superior DPPH radical scavenging capability compared to individual RMPCE or BHB. Furthermore, analysis of the release kinetics based on zero-order, first-order, Higuchi, and Ritger-Peppas models revealed that RMPCE release from CO-RMPHB under in vitro digestion models followed non-Fickian diffusion. This discovery effectively addresses the challenges of the stability and controlled release of RMP, expanding its applications in the food and pharmaceutical industries.

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A new guar gum hydrogel beads were fabricated by dropping method from an aqueous solution of guar gum (GG) using ammonium persulphate and polyethylene glycol as initiator and crosslinker respectively, for the adsorption of chlorpyrifos (CP) from water. The semi-crystalline nature of the synthesized beads was confirmed by FESEM analysis. The TGA studies implied that the beads were thermally stable up to 600 °C. The maximum swelling ratio of 1400 gg-1 was attained at pH 9.2 and 80 min. The evidence of a strong absorption band was found in FTIR spectrum at 584 cm-1 due to -P=S of the adsorbed pesticide CP. The maximum adsorption of CP was found to be 220.97 mgg-1. The adsorption followed pseudo second-order kinetics and Langmuir adsorption isotherm with regression coefficients 0.9998 and 0.9938 which followed the chemisorption process. It is due to the hydrolysis of CP at pH 9.2 to yield 3,5,6-trichloropyridinol which in turn reacts with the carboxylic group present in GG giving -N-C=O linkage. A -ΔG indicates that the process is spontaneous and involves chemisorption which is thermodynamically and kinetically favorable and a -ΔH value (-10.37 kJ/mol) suggests that the adsorption is exothermic.

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A novel core-shell structured alginate-based hydrogel bead modified by co-gelatinizing with starch and protocatechuic acid (PA), was designed to modulate physical properties of beads, release behavior and antioxidant stability of encapsulated bioactives. Core was fabricated by ionotropic gelation, and its formulation (ratio of sodium alginate/starch) was determined by particle size/starch distribution, texture and bioactive encapsulation capacity of core. Then, coating core with shell-forming solution co-gelatinized with different doses of PA, and subsequently cross-linked with Ca2+ to obtain core-shell structured beads. Surface microstructure, mechanical characteristics, and swelling ratio of beads were affected by concentrations of PA. Besides, core-shell structure containing PA could enhance delivery and sustained release of encapsulated phenolic bioactives during in vitro digestion, and improve their antioxidant potential stability. Furthermore, interaction between PA and polysaccharide components was elucidated by FTIR and TGA. The present information was beneficial for the advancement of functional food materials and bioactive delivery systems.

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Perilla frutescens L. essential oil (PLEO) has antibacterial and antioxidant properties, which can effectively maintain the quality of fruits and extend their shelf life. In this study, sodium alginate and chitosan were used as wall materials, and PLEO microcapsule powder was used as the core materials to prepare PLEO hydrogel beads. The best results were obtained by using 2%w/v sodium alginate and 1.5%w/v chitosan as wall materials, with a core-to-wall ratio of 2:1 and homogenized for 15 min producing PLEO hydrogel beads with encapsulation efficiency of 82.61 %. For strawberries preservation, PLEO hydrogel beads preservation group had a better effect after 5 d of storage, showing a lower decay rate (15.71 %), better maintaining the hardness of 1.75 kg/cm2, and a weight loss of 3.29 %. Furthermore, organic acids and total phenols were retained more in this group, the number of microorganisms was significantly reduced, and sensory qualities were improved, especially taste and color. This study provides important insights into the application of natural preservatives in the food industry and promotes sustainable practices in food preservation.

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In this study, vitamins C and E were simultaneously encapsulated in water-in-oil-in-water (W/O/W) emulsion-filled sodium alginate (SA) hydrogel beads, as well as the effects of SA concentrations (0.5%, 1.0%, 1.5%, and 2.0%) on the structures and lipolysis the of hydrogel beads were investigated. With increasing SA concentration, the beads showed larger sizes, denser structures and better textures. The droplets tightly penetrated the gel network at high SA concentrations. Digestion behavior revealed the disintegrated intramolecular structure at low SA concentrations. The beads with 0.5% SA were fragmented, losing the initial shape during digestion in the intestinal fluid. Additionally, lipid phases were released as W/O/W and O/W emulsion droplets after digestion. However, the high SA concentration-containing beads exhibited a well-preserved morphological structure after digestion, and the release profiles of lipid phase were mainly O/W emulsion droplets. Furthermore, vitamins C and E encapsulated in the beads exhibited high bioaccessibility (vitamin C: 90.20% and vitamin E: 95.19%).

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Cellulose-based adsorbents have been extensively developed in heavy metal capture and wastewater treatment. However, most of the reported powder adsorbents suffer from the difficulties in recycling due to their small sizes and limitations in detecting the targets for the lack of sensitive sensor moieties in the structure. Accordingly, carbon dots (CDs) were proposed to be encapsulated in cellulosic hydrogel beads to realize the simultaneous detection and adsorption of Hg (II) in water due to their excellent fluorescence sensing performance. Besides, the molding of cellulose was beneficial to its recycling and further reduced the potential environmental risk generated by secondary pollution caused by adsorbent decomposition. In addition, the detection limit of the hydrogel beads towards Hg (II) reached as low as 8.8 × 10-8 M, which was below the mercury effluent standard declared by WHO, exhibiting excellent practicability in Hg (II) detection and water treatment. The maximum adsorption capacity of CB-50 % for Hg (II) was 290.70 mg/g. Moreover, the adsorbent materials also had preeminent stability that the hydrogel beads could maintain sensitive and selective sensing performance towards Hg (II) after 2 months of storage. Additionally, only 3.3% of the CDs leaked out after 2 weeks of immersion in water, ensuring the accuracy of Hg (II) evaluation. Notably, the adsorbent retained over 80% of its original adsorption capacity after five consecutive regeneration cycles, underscoring its robustness and potential for sustainable environmental applications.

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Antibody drugs play a vital role in diagnostics and therapy. However, producing antibodies from mammalian cells is challenging owing to cellular heterogeneity, which can be addressed by applying droplet-based microfluidic platforms for high-throughput screening (HTS). Here, we designed an integrated system based on disulfide-bonded redox-responsive hydrogel beads (redox-HBs), which were prepared through enzymatic hydrogelation, to compartmentalize, screen, select, retrieve, and recover selected Chinese hamster ovary (CHO) cells secreting high levels of antibodies. Moreover, redox-HBs were functionalized with protein G as an antibody-binding module to capture antibodies secreted from encapsulated cells. As proof-of-concept, cells co-producing immunoglobulin G (IgG) as the antibody and green fluorescent protein (GFP) as the reporter molecule, denoted as CHO(IgG/GFP), were encapsulated into functionalized redox-HBs. Additionally, antibody-secreting cells were labeled with protein L-conjugated horseradish peroxidase using a tyramide amplification system, enabling fluorescence staining of the antibody captured inside the beads. Redox-HBs were then applied to fluorescence-activated droplet sorting, and selected redox-HBs were degraded by reducing the disulfide bonds to recover the target cells. The results indicated the potential of the developed HTS platform for selecting a single cell viable for biopharmaceutical production.

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Novel hydrogel beads based on nanocomposite with outstanding antibacterial and swelling capabilities have been successfully produced as an efficient drug carrier for potential drug delivery systems in wound healing applications. The beads were characterized by Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and EDX-Mapping analysis. Then, using tetracycline hydrochloride (TCH) as a model drug system, they were studied in vitro for their potential efficiency as pH and temperature dependent sustained drug delivery carriers. Moreover, they were assessed in terms of porosity, swelling degree, encapsulation efficiency, and in vitro release kinetics. Beads released drugs at their highest levels under alkaline circumstances (pH = 8) and at a temperature of 39 °C, with a cumulative TCH release of 96.2 % at 36 h and in accordance with the Weibull kinetics model (R2 = 0.98). Additionally, the disc diffusion experiment demonstrated the strong antibacterial activity of the synthesized beads and offered a feasible and cost-effective wound dressing material for treating infected wounds.

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In this study, hydrogel beads [SPI/HP-Fe (II)] were prepared by cross-linking soybean isolate protein (SPI) and hawthorn pectin (HP) with ferrous ions as a backbone, and the effects of ultrasound and Fe2+ concentration on the mechanical properties and the degree of cross-linking of internal molecules were investigated. The results of textural properties and water-holding capacity showed that moderate ultrasonic power and Fe2+ concentration significantly improved the stability and water-holding capacity of the hydrogel beads and enhanced the intermolecular interactions in the system. Scanning electron microscopy (SEM) confirmed that the hydrogel beads with 60% ultrasonic power and 8% Fe2+ concentration had a denser network. X-ray photoelectron spectroscopy (XPS) and atomic absorption experiments demonstrated that ferrous ions were successfully loaded into the hydrogel beads with an encapsulation efficiency of 82.5%. In addition, in vitro, simulated digestion experiments were performed to understand how the encapsulated Fe2+ is released from the hydrogel beads, absorbed, and utilized in the gastrointestinal environment. The success of the experiments demonstrated that the hydrogel beads were able to withstand harsh environments, ensuring the bioactivity of Fe2+ and improving its bioavailability. In conclusion, a novel and efficient ferrous ion delivery system was developed using SPI and HP, demonstrating the potential application of SPI/HP-Fe (II) hydrogel beads as an iron supplement to overcome the inefficiency of intake of conventional iron supplements.

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The objective of the present study was to develop and evaluate NLC-chitosan hydrogel beads for topical administration. The feasibility of the preparation technology was verified by investigating various formulation factors and the impact of chitosan hydrogel beads on the NLC. The encapsulation efficiency of NLC-chitosan hydrogel beads was above 95% in optimized process conditions. The physical characterization of the NLC-chitosan hydrogel beads showed that the NLC was distributed within the network of the chitosan hydrogel beads. Furthermore, the incorporation of NLC into the chitosan hydrogel beads was related to the electrostatic interaction between the surface of the NLC and chitosan, which influenced the lipid ordering degree of the NLC and contributed to the stability. The stability studies showed that the retention rate of quercetin in the NLC-chitosan hydrogel beads was 88.63 ± 2.57% after 10 months of storage under natural daylight. An in vitro permeation study showed that NLC-chitosan hydrogel beads exhibited superior ability in enhancing skin permeation by hydrophobic active ingredients compared to the NLC and significantly increased skin accumulation. These studies demonstrated that the use of NLC-chitosan hydrogel beads might be a promising strategy for the delivery of hydrophobic active ingredients in topical administration.

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Polycyclic aromatic hydrocarbons (PAHs) are prevalent environmental contaminants that are harmful to ecological and human health. Bioremediation is a promising technique for remediating PAHs in the environment, however bioremediation often results in the accumulation of toxic PAH metabolites. The objectives of this research were to demonstrate the cometabolic treatment of a mixture of PAHs by a pure bacterial culture, Rhodococcus rhodochrous ATCC 21198, and investigate PAH metabolites and toxicity. Additionally, the surfactant Tween ® 80 and cell immobilization techniques were used to enhance bioremediation. Total PAH removal ranged from 70-95% for fluorene, 44-89% for phenanthrene, 86-97% for anthracene, and 6.5-78% for pyrene. Maximum removal was achieved with immobilized cells in the presence of Tween ® 80. Investigation of PAH metabolites produced by 21198 revealed a complex mixture of hydroxylated compounds, quinones, and ring-fission products. Toxicity appeared to increase after bioremediation, manifesting as mortality and developmental effects in embryonic zebrafish. 21198's ability to rapidly transform PAHs of a variety of molecular structures and sizes suggests that 21198 can be a valuable microorganism for catalyzing PAH remediation. However, implementing further treatment processes to address toxic PAH metabolites should be pursued to help lower post-remediation toxicity in future studies.

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Currently, the presence of drugs used in the COVID-19 pandemic in water bodies is worrisome due to their high toxicity, which necessitates their critical removal by developing highly efficient adsorbents. Hence, in this study, alginate hydrogel beads of magnetic graphene oxide@MIL-88 metal-organic framework (GO@Fe3O4@MIL-88@Alg) were prepared for the first time and then utilized as a new absorption system for the removal of COVID-19 drugs such as doxycycline (DOX), hydroxychloroquine (HCQ), naproxen (NAP), and dipyrone (DIP) from aqueous solutions by batch adsorption manner. The effects of different experimental factors, such as adsorbent dosage, contact time, pH, drug concentration, temperature, ionic strength, presence of an external magnetic field (EMF), and magnet distance from the adsorption flask were optimized for the removal of COVID-19 drugs. The adsorption equilibrium isotherm proved that the adsorption process of DOX, HCQ, NAP, and DIP drugs on GO@Fe3O4@MIL-88@Alg hydrogel beads conformed to the Langmuir model and followed the pseudo-second-order adsorption kinetics. The maximum adsorption capacities of DOX, HCQ, NAP, and DIP drugs obtained for GO@Fe3O4@MIL-88@Alg hydrogel beads with the Langmuir model were 131.57, 79.92, 55.55, and 49.26 mg/g at 298 K, respectively. The thermodynamic study suggested a spontaneous endothermic adsorption process. Also, the conclusion from this study confirmed the validity of GO@Fe3O4@MIL-88@Alg hydrogel beads for excellent removal of COVID-19 drugs from water samples. It was also found that the GO@Fe3O4@MIL-88@Alg hydrogel beads could be reused with satisfactory removal efficiency in six cycles. Based on the study, the GO@Fe3O4@MIL-88@Alg hydrogel beads could be considered a sustainable, simple, economical, environmentally friendly absorption system for the removal of pharmaceutical contaminants from water.

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Acid-sensitive CdTe quantum dots-loaded alginate hydrogel (CdTe QDs-AH) beads were designed for the visual detection of SO2 residues. As proof of concept, two types of CdTe QDs were selected as model probes and embedded in AH beads. The entire test was performed within 25 min in a modified double-layer test tube with one bead fixed above the sample solution. Adding citric acid and heating at 70 ℃ for 20 min transformed the sulfites in the solution into SO2 gas, which then quenched the fluorescence of the CdTe QDs-AH beads. Using this assay, qualitative, naked-eye detection of SO2 residues was achieved in the concentration range of 25-300 ppm, as well as precise quantification was possible based on the difference in the average fluorescence brightness of the beads before and after the reaction. Five food types were successfully analysed using this method, which is simpler and more economical than existing methods, and does not require complex pretreatment.

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BACKGROUND: Fallen young rambutan fruit is an underrated agricultural waste which may contain several bioactive compounds. In this study, fallen young rambutan fruit was assessed regarding its phenolic contents and antioxidant activities. In order to expand its utilization, rambutan extract-loaded hydrogel beads were developed by a basic spherification technique using sodium alginate. The effect of ratios of polymer and extract and different calcium sources were evaluated. The recovery of bioactive compounds from the hydrogel beads was determined using in vitro gastrointestinal digestion models. RESULTS: Use of 50% (v/v) ethanol yielded rambutan extract with good chemical properties. The production of hydrogel beads using a ratio of 1:3 with calcium lactate provided the highest production yield of 122.94%. The hydrogel beads developed using the ratio of 1:3 with a combination of calcium lactate and calcium chloride showed high recovery of phenolic compounds and antioxidant activity after simulated intestinal digestion, which were greater compared to unencapsulated extract. CONCLUSION: The findings demonstrate that the ratio of wall material to rambutan extract and the calcium source influence the physical properties, chemical properties and in vitro gastrointestinal digestion stability of alginate beads. The obtained hydrogel beads may have potential for application in the food or pharmaceutical industries. © 2024 Society of Chemical Industry.

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BACKGROUND: Hyaluronic acid liquid-core hydrogel beads (HA-LHB) is a good way for oral intake of HA. However, HA may affect the reaction-diffusion of sodium alginate (SA) and Ca2+ leading to poor mechanical properties, since HA is a polyanionic electrolyte having electrostatic effect and a certain spatial site-blocking effect. RESULTS: The mechanical properties of HA-LHB were modified from bathing solution, core solution and secondary calcium bath time. The mechanical properties varied with the SA structure and concentration in bathing solution, where SA with high G (guluronic acid) segment compounded with SA with high M (mannuronic acid) segment at a mass ratio of 7:3 with a 11 g kg-1 concentration showed the best mechanical properties. The secondary calcium bath can greatly improve the mechanical properties due to the tight network formed by bidirectional crosslinking, and 15 min reaction reached the plateau if Ca2+ is sufficient. And the mechanical properties were positively correlated with calcium lactate concentration only at <70 g kg-1 in core solution, but the diffusion of Ca2+ was hindered by the tight gel network at higher concentrations. Moreover, the mechanical properties can be maintained during heat treatment, due to the rearrangement of alginate network structure. CONCLUSION: Our results suggested that the problem of poor mechanical properties of LHB in the presence of high HA concentration can be avoided by process control, which may broaden the development of HA and popping boba market. © 2024 Society of Chemical Industry.

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This study details the synthesis and characterization of surfactant-modified sodium alginate hydrogel beads crosslinked with Ba2+ ions through ionotropic gelation. Cationic surfactants such as, dodecyltrimethylammonium bromide (DTAB), didodecyldimethylammonium bromide (DDAB), and butanediyl-α,ω-bis-(dimethyldodecylammonium bromide) (GEM), were employed in the modification process. The surfactant-modified ALG-DTAB, ALG-DDAB, and ALG-GEM beads were investigated for the removal of cationic dye Malachite Green (MG) to elucidate the impact of hydrophobicity of amphiphiles on the adsorption process. The characterizations were carried out using Rheometry, Field Emission Scanning Electron Microscopy (FESEM), Infrared Spectroscopy (IR), and Energy Dispersive X-ray Spectroscopy (EDX). Under optimized conditions, ALG-GEM and ALG-DDAB demonstrated highest maximum adsorption capacity (Qmax > 700 mgg-1). The adsorption data fitted well to pseudo-second order kinetic and Langmuir adsorption models, suggesting the involvement of chemisorption phenomena with notable contributions from pore diffusion. The effects of pH, initial dye concentration, adsorbent dose, temperature, and competing ions on the removal of MG were investigated. Interestingly, ALG-GEM beads exhibited an increase in adsorption capacity with rising pH and a subsequent decrease with increasing temperature, showcasing optimal adsorption at pH 7.0 and 25 °C. The study proposes that ALG beads modified with cationic surfactants with higher hydrophobicity could offer a promising avenue in wastewater treatment processes.

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The current study effectively designed novel cross-linked tosyl-carrageenan/alginate (Ts-Car/Alg) beads to remove Pb2+ ions from their aqueous solutions. To confirm the structure of the produced matrix, characterization methods such as XRD, SEM, FTIR, and EDX were used. Batch experiments were employed in order to further evaluate the adsorption efficiency of Pb2+ ions. Additionally, various variables, including contact time, solution pH, adsorbent dosage, and initial concentration of Pb2+ ions were investigated using atomic absorption. The results of this study showed that the adsorption equilibrium increased as Pb2+ ions concentration increased at pH = 5.3 after a contact time of 120 min, with 0.3 g of Ts-Car/Alg that having the best adsorption capacity at 74 mg/g. The adsorption progression was further examined using the kinetic and isothermal models. With a correlation coefficient of 0.975, the Freundlich model was thought to better fit Pb2+ ions adsorption from the isotherm investigation. Also, the adsorption kinetics were investigated using a pseudo-second-order model with 1/n ratio of 0.683. This Ts-Car/Alg adsorbent is regarded as an effective candidate to be used for water treatment because the reusability process of produced beads was successfully completed twice, and the adsorbent maintained its ability to remove Pb2+ ions. The prepared Ts-Car/Alg beads are therefore excellent candidates to be used as potent Pb2+ ions adsorbents from their aqueous solutions. The Ts-Car/Alg beads' regeneration and reusability investigation for the removal of heavy metal ions was completed in at least two successful cycles.