Amyloid-Templated Palladium Nanoparticles for Water Purification by Electroreduction.

Electrocatalysis offers great promise for water purification but is limited by low active area and high uncontrollability of electrocatalysts. To overcome these constraints, we propose hybrid bulk electrodes by synthesizing and binding a Pd nanocatalyst (nano-Pd) to the electrodes via amyloid fibrils (AFs). The AFs template is effective for controlling the nucleation, growth, and assembly of nano-Pd on the electrode. In addition, the three-dimensional hierarchically porous nanostructure of AFs is beneficial for loading high-density nano-Pd with a large active area. The novel hybrid cathodes exhibit superior electroreduction performance for the detoxification of hexavalent chromium (Cr6+ ), 4-chlorophenol, and trichloroacetic acid in wastewater and drinking water. This study provides a proof-of-concept design of an AFs-templated nano-Pd-based hybrid electrode, which constitutes a paradigm shift in electrocatalytic water purification, and broadens the horizon of its potential engineered applications.

Fabrication of carbonyl-functional hypercrosslinked polymers as solid-phase extraction sorbent for enrichment of chlorophenols from water, honey and beverage samples.

Three carbonyl-functional novel hypercrosslinked polymers (HCP-TPS, HCP-TPA, and HCP-TPP) were successfully fabricated through an one-step Friedel-Crafts acylation reaction by copolymerizing paraphthaloyl chloride with triphenylsilane, triphenylamine, and triphenylphosphine, respectively. The resultant HCPs contained plenty of carbonyl-functional groups. Among the series of such HCPs, HCP-TPS displayed the best adsorption capability to chlorophenols (CPs), and thus it was employed as solid-phase extraction (SPE) adsorbent for enrichment of chlorophenols from water, honey, and white peach beverage prior to determination by high-performance liquid chromatography. Under the optimal conditions, the detection limits of the method (S/N = 3) were 0.15-0.3 ng mL-1 for tap water and leak water, 2.5-6.0 ng g-1 for honey sample and 0.4-0.6 ng mL-1 for white peach beverage sample. The recoveries of CPs in the spiked water, honey samples, and white peach beverage were in the range of 89.0-108.4%, 81.4-118.2%, and 85.0-113.5%, respectively. This work provides a new strategy for constructing functionalized HCPs as efficient SPE adsorbents. In this work, three novel hypercrosslinked polymers (HCPs) were synthesized by the Friedel-Crafts alkylation reaction (paraphthaloyl chloride as the alkylating agent, triphenylsilane, triphenylamine, and triphenylphosphine as the aromatic units). Then, HCP-TPS was applied to soild-phase extraction sorbent for enrichment CPs from water, honey, and white peach beverage samples.

Synthesis, characterization, and application of chemically interconnected carbon nanotube monolithic sorbents by photopolymerization in polypropylene caps.

A facile and convenient approach for the preparation of interconnected multiwalled carbon nanotube (MWCNT) monolithic sorbents in recycled plastic caps has been developed. The method, which was based on the photopolymerization of the individual MWCNTs via the formation of a W/O medium internal phase emulsion (40/60 w/w%), provides control over the size of pores, rigidity, and the mechanical stability of the final solid. Pluronic L121 was used as a surfactant containing the water phase inside it and, consequently, the organic and non-polar phase, in which the MWCNTs and the cross-linker were trapped, remained on the outside of the droplets. Optical microscopy and scanning electron microscopy (SEM) were employed to characterize the morphology of both the emulsions and the final solids, respectively. In addition, nitrogen intrusion porosimetry was performed in order to study how the specific surface area of the final monolithic solid changed (from 19.6 to 372.2 m2 g-1) with the variables involved in the polymerization step. To exemplify the great sorbent potential of the synthesized material, a colorimetric assay based on the retention of methylene blue within the interconnected MWCNT monolithic structure was carried out. Finally, following the positive results, the carbon nanotube-monolithic stirred caps were applied for the determination of chlorophenols in a biological matrix such as human urine, obtaining excellent recovery values (91-98%) and good precision (5.4-9.1%) under optimized extraction conditions. Graphical abstract.

Electronic Tongue Coupled to an Electrochemical Flow Reactor for Emerging Organic Contaminants Real Time Monitoring.

Triclosan, which is a bacteriostatic used in household items, has raised health concerns, because it might lead to antimicrobial resistance and endocrine disorders in organisms. The detection, identification, and monitoring of triclosan and its by-products (methyl triclosan, 2,4-Dichlorophenol and 2,4,6-Trichlorophenol) are a growing need in order to update current water treatments and enable the continuous supervision of the contamination plume. This work presents a customized electronic tongue prototype coupled to an electrochemical flow reactor, which aims to access the monitoring of triclosan and its derivative by-products in a real secondary effluent. An electronic tongue device, based on impedance measurements and polyethylenimine/poly(sodium 4-styrenesulfonate) layer-by-layer and TiO2, ZnO and TiO2/ZnO sputtering thin films, was developed and tested to track analyte degradation and allow for analyte detection and semi-quantification. A degradation pathway trend was observable by means of principal component analysis, being the sample separation, according to sampling time, explained by 77% the total variance in the first two components. A semi-quantitative electronic tongue was attained for triclosan and methyl-triclosan. For 2,4-Dichlorophenol and 2,4,6-Trichlorophenol, the best results were achieved with only a single sensor. Finally, working as multi-analyte quantification devices, the electronic tongues could provide information regarding the degradation kinetic and concentrations ranges in a dynamic removal treatment.

Enzyme-Nanowire Mesocrystal Hybrid Materials with an Extremely High Biocatalytic Activity.

The laccase-Cu2O-nanowire mesocrystal hybrid materials were developed with a superior catalytic activity inspired by natural biocatalysis processes in living cells that highly resemble the metal ions activation and the well-organized spatial structure of the natural rough endoplasmic reticulum. The enzyme and nanobiocatalyst activities of the obtained hybrid material exhibited an approximate 10-fold and 2.2-fold increase than the free enzyme, surpassing the currently available nanobiocatalysts. The comprehensive catalytic performance of the hybrid materials has been further demonstrated using a prototype continuous-flow reactor for the bioremediation of 2,4-dichlorophenol-contaminated water, which showed a high degradation efficiency and remarkable reusability. These new highly efficient nanobiocatalysts are expected to be used for diverse applications in biotechnology, biosensing, and environmental remediation.

Laccase removal of 2-chlorophenol and sulfamethoxazole in municipal wastewater.

Laccases were studied for their ability to remove two compounds, 2-chlorophenol and sulfamethoxazole, in batch studies, both in buffered solutions and in wastewater samples from different points in a municipal water resource recovery facility. Two enzymes with and without a mediator (acetosyringone) were investigated: a commercial product derived from Myceliphthora thermophile and a laboratory-generated enzyme mix derived from Tramates versicolor. The chlorophenol was removed rapidly by the commercial enzyme in the presence of acetosyringone, but the primary products were coupling complexes of the reactants. Excellent removal was achieved without acetosyringone by the natural enzyme mix. Sulfamethoxazole was poorly removed in all laboratory-generated chemically buffered solutions, but was very well removed, without the addition of mediators, in secondary effluent suspensions from a municipal water resource recovery facility. Mechanistic studies are still required, but the results suggest that treatment via direct addition of enzymes is feasible to remove recalcitrant compounds in municipal wastewater.

Poly (Octadecyl Methacrylate-Co-Trimethylolpropane Trimethacrylate) Monolithic Column for Hydrophobic in-Tube Solid-Phase Microextraction of Chlorophenoxy Acid Herbicides.

Chlorophenoxy acid herbicides (CAHs), which are widely used on cereal crops, have become an important pollution source in grains. In this work, a highly hydrophobic poly (octadecyl methacrylate-co-trimethylolpropane trimethacrylate) [poly (OMA-co-TRIM)] monolithic column has been specially prepared for hydrophobic in-tube solid-phase microextraction (SPME) of CAHs in rice grains. Due to the hydrophobicity of CAHs in acid conditions, trace CAHs could be efficiently extracted by the prepared monolith with strong hydrophobic interaction. Several factors for online hydrophobic in-tube SPME, including the length of the monolithic column, ACN and trifluoroacetic acid percentage in the sampling solution, elution volume, and elution flow rate, were investigated with respect to the extraction efficiencies of CAHs. Under the optimized conditions, the limits of detection of the four CAHs fell in the range of 0.9-2.1 µg/kg. The calibration curves provided a wide linear range of 5-600 µg/kg and showed good linearity. The recoveries of this method ranged from 87.3% to 111.6%, with relative standard deviations less than 7.3%. Using this novel, highly hydrophobic poly (OMA-co-TRIM) monolith as sorbent, a simple and sensitive online in-tube SPME-HPLC method was proposed for analysis of CAHs residue in practical samples of rice grains.

Dispersive liquid-liquid microextraction of five chlorophenols in water samples followed by determination using capillary electrophoresis.

Dispersive liquid-liquid microextraction (DLLME) coupled with CE was developed for simultaneous determination of five types of chlorophenols (CPs), namely 2-chlorophenol (2-CP), 4-chlorophenol (4-CP), 2,4-dichlorophenol (2,4-DCP), 2,6-dichlorophenol (2,6-DCP), and 2,4,6-trichlorophenol (2,4,6-TCP) in water samples. Several parameters affecting DLLME and CE conditions were systematically investigated. Under the optimized DLLME-CE conditions, the five CPs were separated completely within 7.5 min and good enrichment factors were obtained of 40, 193, 102, 15, and 107 for 4-CP, 2,4,6-TCP, 2,4-DCP, 2-CP, and 2,6-DCP, respectively. Good linearity was attained in the range of 1-200 µg/L for 2,4,6-TCP, 2,4-DCP, 2-300 µg/L for 4-CP and 2-CP, and 1-300 µg/L for 2,6-DCP, with correlation coefficients (r) over 0.99. The LOD (S/N = 3) and the LOQ (S/N = 10) were 0.31-0.75 µg/L and 1.01-2.43 µg/L, respectively. Recoveries ranging from 60.85 to 112.36% were obtained with tap, lake, and river water spiked at three concentration levels and the RSDs (for n = 3) were 1.31-11.38%. With the characteristics of simplicity, cost-saving, and environmental friendliness, the developed DLLME-CE method proved to be potentially applicable for the rapid, sensitive, and simultaneous determination of trace CPs in complicated water samples.

Removal of dichlorophenol by Chlorella pyrenoidosa through self-regulating mechanism in air-tight test environment.

Microalgae are surprisingly efficient to remove pollutants in a hermetically closed environment, though its growth is inhibited in the absence of pollutants. The final pH, algal density, Chl-a content, and the removal efficiency of 2,4-dichlorophenol (2,4-DCP) by Chlorellar pyrenoidosa in a closed system were observed under different initial pH, lighting regimes, and various carbon sources. The optimal condition for 2,4-DCP removal was obtained, and adopted to observe the evolution of above items by domesticated and origin strains. The results showed that both respiration and photosynthesis participated in the degradation of 2,4-DCP, and caused the changes of pH. The photosynthesis seemed to increase the solution pH, while the respiration and the biodegradation of 2,4-DCP to decrease the solution pH. The domesticated strain achieved nearly 100% removal when initial concentrations of 2,4-DCP lower than 200 µg L-1, due to providing a appropriate but narrow pH evolution range, mostly falling between 6.5 and 7.9. The research helps to understand the mechanism of biodegradation of chlorophenol compounds by green algae.

In-Vial Temperature Gradient Headspace Single Drop Microextraction Designed by Multiphysics Simulation.

Presented herein is a novel headspace single drop microextraction (HS-SDME) based on temperature gradient (TG) for an on-site preconcentration technique of volatile and semivolatile samples. First, an inner vial cap was designed as a cooling device for acceptor droplet in HS-SDME unit to achieve fast and efficient microextraction. Second, for the first time, an in-vial TG was generated between the donor phase in a sample vial at 80 °C and the acceptor droplet under the inner vial cap containing cooling liquid at -20 °C for a TG-HS-SDME. Third, a simple mathematic model and numerical simulations were developed by using heat transfer in fluids, Navier-Stokes and mass balance equations for conditional optimization, and dynamic illumination of the proposed extraction based on COMSOL Multiphysics. Five chlorophenols (CPs) were selected as model analytes to authenticate the proposed method. The comparisons revealed that the simulative results were in good agreement with the quantitative experiments, verifying the design of TG-HS-SDME via the numerical simulation. Under the optimum conditions, the extraction enrichments were improved from 302- to 388-fold within 2 min only, providing 3.5 to 4 times higher enrichment factors as compared to a typical HS-SDME. The simulation indicated that these improvements in the extraction kinetics could be attributed due to the applied temperature gap between the sample matrix and acceptor droplet within the small volume of headspace. Additionally, the experiments demonstrated a good linearity (0.03-100 µg/L, R2 > 0.9986), low limit of detection (7-10 ng/L), and fair repeatability (<5.9% RSD, n = 6). All of the simulative and experimental results indicated the robustness, precision, and usefulness of TG-HS-SDME for trace analyses of analytes in a wide variety of environmental, pharmaceutical, food safety, and forensic samples.

Magnetic porous carbon derived from a Zn/Co bimetallic metal-organic framework as an adsorbent for the extraction of chlorophenols from water and honey tea samples.

A novel magnetic porous carbon derived from a bimetallic metal-organic framework, Zn/Co-MPC, was prepared by introducing cobalt into ZIF-8. Magnetic porous carbon that possesses magnetic properties and a large specific surface area was firstly fabricated by the direct carbonization of Zn/Co-ZIF-8. The prepared magnetic porous carbon material was characterized by scanning electron microscopy, transmission electron microscopy, powder X-ray diffraction, N2 adsorption, and vibrating sample magnetometry. The prepared magnetic porous carbon was used as a magnetic solid-phase extraction adsorbent for the enrichment of chlorophenols from water and honey tea samples before high-performance liquid chromatography analysis. Several experimental parameters that could influence the extraction efficiency were investigated and optimized. Under the optimum conditions, good linearities (r > 0.9957) for all calibration curves were obtained with low limits of detection, which are in the range of 0.1-0.2 ng mL(-1) for all the analytes. The results showed that the prepared magnetic porous carbon had an excellent adsorption capability toward the target analytes.

New polyphenols from a deep sea Spiromastix sp. Fungus, and their antibacterial activities.

Eleven new polyphenols namely spiromastols A-K (1-11) were isolated from the fermentation broth of a deep sea-derived fungus Spiromastix sp. MCCC 3A00308. Their structures were determined by extensive NMR data and mass spectroscopic analysis in association with chemical conversion. The structures are classified as diphenyl ethers, diphenyl esters and isocoumarin derivatives, while the n-propyl group in the analogues is rarely found in natural products. Compounds 1-3 exhibited potent inhibitory effects against a panel of bacterial strains, including Xanthomanes vesicatoria, Pseudomonas lachrymans, Agrobacterium tumefaciens, Ralstonia solanacearum, Bacillus thuringensis, Staphylococcus aureus and Bacillus subtilis, with minimal inhibitory concentration (MIC) values ranging from 0.25 to 4 µg/mL. The structure-activity relationships are discussed, while the polychlorinated analogues 1-3 are assumed to be a promising structural model for further development as antibacterial agents.

Characterization and evaluation of the efficiency of SiO2/tetra-α-(2,4-di-tert-butylphenoxy)-phthalocyaninato zinc nanocomposite as photosensitizers for oxidation of 2,4,6-trichlorophenol.

A photosensitizer tetra-α-(2,4-di-tert-butylphenoxy)-phthalocyaninato zinc [ZnPc(OAr)4] was successfully encapsulated in SiO2 nanoparticle by the microemulsion method. The photosensitized composite nanoparticle was able to degrade 2,4,6-trichlorophenol (TCP) in aqueous solution. Under visible light irradiation, the nanoparticles efficiently generated reactive oxygen species; 95.4% of TCP was degraded after 270 min of reaction. Some aromatic compounds and aliphatic carboxylic acids were detected by mass spectrometry as the reaction intermediates. The results were different from those of previously reported photocatalytic reactions, in which valence band holes or hydroxyl radicals functioned as the main oxidants. The photosensitizing composite nanoparticle is potentially applicable to the oxidation of phenol.

Dynamics of a microbial community exposed to several concentrations of 2-chlorophenol in an anaerobic sequencing batch reactor.

The aim of this study was to contribute to the knowledge on the dynamic of the microbial community involved in anaerobic degradation of different concentrations of 2-chlorophenol (2CP, from 28 to 196 mg 2CP-C/L) and a mixture of 2CP and phenol (from 28 to 196 mg phenol-C/L) and its relationship with the respiratory process in two anaerobic sequencing batch reactors (ASBR). The dynamic of the microbial community was evaluated by denaturant gradient gel electrophoresis (DGGE) and ecological indices (S and J indices). The respiratory process was evaluated by means of substrate consumption efficiency, biogas yield, and specific consumption rates as response variables. The high consumption efficiency (90%) and the constant biogas yields obtained at concentrations up to 140 mg C/L may be related with the evenness of microbial populations (J index=0.97±0.2) present in both reactors. Pseudomonas genus was present in all concentrations tested, suggesting a possible relationship with the dehalogenation observed in both reactors. The decrease in specific consumption rate and biogas yield as well as the accumulation of phenol and volatile fatty acids observed in both reactors at 196 mg 2CP-C/L might be associated with the disappearance of the bands related to Caulobacter and Bacillus. At these conditions, the disappearance of fermentative or acetogenic bacteria resulted in reduction of substrates required to carry out methanogenesis, which eventually might cause the declination in methanogenic populations present in the reactors.

Effects of selected operational parameters on efficacy and selectivity of electromembrane extraction. Chlorophenols as model analytes.

Effects of organic solvent type, pH value, and composition of donor/acceptor solution on the efficacy of electromembrane extraction (EME) were examined. For the first time, a comprehensive quantitative study, based also on measurements of electric charge passed through the EME system, was carried out, which demonstrates that apart from the pH value, also the nature of counter-ions in donor and acceptor solution plays a significant role in the electrically induced transfer of charged analytes across supported liquid membranes (SLMs). The EME transfer of model analytes correlated well with electrophoretic mobilities of inorganic cations, which were added to acceptor solutions during their alkalization with alkali metal hydroxides, and were highest for counter-cations with highest mobilities. Up to a 53-fold improvement of extraction efficiency was achieved for EMEs using optimized composition of donor (alkalized with KOH to pH 7) and acceptor (10 mM CsOH, pH 12) solutions. Six chlorophenols (CPs) were selected as model analytes due to the wide range of pH values that are required for their ionization and due to their high environmental relevance; quantitative measurements were carried out by CE with UV detection. Extraction recoveries of the six CPs ranged between 14 and 25% for 5 min EMEs at 150 V and 750 rpm across SLMs impregnated with 1-ethyl-2-nitrobenzene. Calibration curves were strictly linear (r(2) ≥ 0.999) in 0.01-10 µg/mL range, repeatability values of peak areas were between 0.7 and 5.6% and LODs for standard solutions and environmental samples were better than 5 ng/mL.

Effect of copper oxide concentration on the formation and persistency of environmentally persistent free radicals (EPFRs) in particulates.

Environmentally persistent free radicals (EPFRs) are formed by the chemisorption of substituted aromatics on metal oxide surfaces in both combustion sources and superfund sites. The current study reports the dependency of EPFR yields and their persistency on metal loading in particles (0.25, 0.5, 0.75, 1, 2, and 5% CuO/silica). The EPFRs were generated through exposure of particles to three adsorbate vapors at 230 °C: phenol, 2-monochlorophenol (2-MCP), and dichlorobenzene (DCBz). Adsorption resulted in the formation of surface-bound phenoxyl- and semiquinoine-type radicals with characteristic EPR spectra displaying a g value ranging from ∼ 2.0037 to 2.006. The highest EPFR yield was observed for CuO concentrations between 1 and 3% in relation to MCP and phenol adsorption. However, radical density, which is expressed as the number of radicals per copper atom, was highest at 0.75-1% CuO loading. For 1,2-dichlorobenzene adsorption, radical concentration increased linearly with decreasing copper content. At the same time, a qualitative change in the radicals formed was observed--from semiquinone to chlorophenoxyl radicals. The two longest lifetimes, 25 and 23 h, were observed for phenoxyl-type radicals on 0.5% CuO and chlorophenoxyl-type radicals on 0.75% CuO, respectively.

Simultaneous derivatization and extraction of chlorophenols in water samples with up-and-down shaker-assisted dispersive liquid-liquid microextraction coupled with gas chromatography/mass spectrometric detection.

A new up-and-down shaker-assisted dispersive liquid-liquid microextraction (UDSA-DLLME) for extraction and derivatization of five chlorophenols (4-chlorophenol, 4-chloro-2-methylphenol, 2,4-dichlorophenol, 2,4,6-trichloro-phenol, and pentachlorophenol) has been developed. The method requires minimal solvent usage. The relatively polar, water-soluble, and low-toxicity solvent 1-heptanol (12 µL) was selected as the extraction solvent and acetic anhydride (50 µL) as the derivatization reagent. With the use of an up-and-down shaker, the emulsification of aqueous samples was formed homogeneously and quickly. The derivatization and extraction of chlorophenols were completed simultaneously in 1 min. The common requirement of disperser solvent in DLLME could be avoided. After optimization, the linear range covered over two orders of magnitude, and the coefficient of determination (r (2)) was greater than 0.9981. The detection limit was from 0.05 to 0.2 µg L(-1), and the relative standard deviation was from 4.6 to 10.8 %. Real samples of river water and lake water had relative recoveries from 90.3 to 117.3 %. Other emulsification methods such as vortex-assisted, ultrasound-assisted, and manual shaking-enhanced ultrasound-assisted methods were also compared with the proposed UDSA-DLLME. The results revealed that UDSA-DLLME performed with higher extraction efficiency and precision compared with the other methods.

pH-assisted homogeneous liquid-liquid microextraction using dialkylphosphoric acid as an extraction solvent for the determination of chlorophenols in water samples.

In this study, a new pH-assisted homogeneous liquid-liquid microextraction combined with HPLC with UV detection was developed for the determination of chlorophenols in water samples. In this approach, bis(2-ethylhexyl) phosphate was used for the first time as the low-density extraction solvent. In particular, 60 µL of bis(2-ethylhexyl) phosphate was injected into the sample solution (5 mL) and dissolved completely in the sample solution while the pH was increased to 9. Afterwards, the pH of the sample solution was lowered to 1, and a cloudy solution was formed. At this stage, hydrophobic interactions between the analytes and the long double hydrocarbon chains of extraction solvent were expected to be the main forces driving extraction. A series of parameters that influence extraction were investigated systematically. Under the optimized conditions, the LODs and LOQs for the chlorophenols were 1.4-2.7 and 4.7-9.1 ng/mL, respectively. RSDs based on five replicate extraction of 100 ng/mL of each chlorophenols were <4.7% for intraday and 7.4% for interday precision. This method has been also successfully applied to analyze real water samples at two different spiked concentrations, and satisfactory recoveries were achieved.

Molecular imprinted polymer of methacrylic acid functionalised ß-cyclodextrin for selective removal of 2,4-dichlorophenol.

This work describes methacrylic acid functionalized ß-cyclodextrin (MAA-ßCD) as a novel functional monomer in the preparation of molecular imprinted polymer (MIP MAA-ßCD) for the selective removal of 2,4-dichlorophenol (2,4-DCP). The polymer was characterized using Fourier Transform Infrared (FTIR) spectroscopy, Brunauer-Emmett-Teller (BET) and Field Emission Scanning Electron Microscopy (FESEM) techniques. The influence of parameters such as solution pH, contact time, temperature and initial concentrations towards removal of 2,4-DCP using MIP MAA-ßCD have been evaluated. The imprinted material shows fast kinetics and the optimum pH for removal of 2,4-DCP is pH 7. Compared with the corresponding non-imprinted polymer (NIP MAA-ßCD), the MIP MAA-ßCD exhibited higher adsorption capacity and outstanding selectivity towards 2,4-DCP. Freundlich isotherm best fitted the adsorption equilibrium data of MIP MAA-ßCD and the kinetics followed a pseudo-second-order model. The calculated thermodynamic parameters showed that adsorption of 2,4-DCP was spontaneous and exothermic under the examined conditions.

The use of used automobile tyres in a partitioning bioreactor for the biodegradation of xenobiotic mixtures.

Waste tyres were utilized as the sorption phase in a two-phase partitioning bioreactor (TPPB) for the biodegradation of a binary mixture of 2,4-dichlorophenol (DCP) and 4-nitrophenol (4NP). These compounds are extensively used in the chemical industry and are found in many industrial effluents. Although both compounds are toxic and are on the EPA list of priority pollutants, a higher inhibitory effect on microorganisms is exerted by DCP, and our experimental tests were focused on strategies to reduce its negative impact on microbial activity. Sorption/desorption tests for the DCP-4NP mixture were first performed to verify the related uptake/release rates by the tyres, which showed that the tyres had a higher capacity for DCP uptake and practically no affinity for 4NP. An acclimatized mixed culture was then utilized in a sequencing batch reactor (SBR) operated in conventional and two-phase mode. For the binary DCP-4NP mixture a significant reduction in DCP toxicity, and a concomitant enhancement in substrate removal efficiency (up to 83%for DCP and approximate 100% for 4NP) were clearly seen for the TPPB operated with 10% and 15% v/v tyres, for influent concentrations up to 180 mg/L, with practically negligible biodegradation in the conventional single phase reactor. The long-term utilization of tyres was confirmed at an influent loading of 180 mg/L with a test performed over 20 work cycles showing an improvement of the removal performance for both compounds.