EtBr Removal Research Articles

Chromatin extracted from common carp testis as an economical and easily available adsorbent for ethidium bromide decontamination

The waste of ethidium bromide (EtBr) used in the laboratory will bring a great burden to the environment, which need to be solved urgently. In the present paper, an efficient and inexpensive method for EtBr removal using chromatin extracted from common carp testis was investigated. The observation of fluorescence microscopy showed that chromatin had similar property to DNA for selective adsorption of EtBr. The results of batch adsorption showed that the removal efficiency of EtBr by chromatin exceeded 99% at pH 7.4 and 30 °C for 3 min with the EtBr concentration of 2 mg L−1 and the chromatin dosage of 0.5 g L−1, and the maximum adsorption amount of chromatin was 45.73 mg g−1. Further, the analysis of kinetic and isotherm suggested that the adsorption followed Pseudo-second-order kinetics and Langmuir isotherm model, and the calculated maximum theoretical adsorption amount of chromatin to EtBr was 48.08 mg g−1. According to thermodynamic analysis, chromatin adsorption of EtBr was a spontaneous process dominated by hydrogen bonding and van der Waals forces. This work will not only offer an adsorbent for EtBr decontamination, also provide a possibility for EtBr analogs removal.

High capacity ethidium bromide removal by montmorillonites

Ethidium bromide (EtBr) is commonly used as a reagent to investigate DNA and RNA bonding in biochemistry. However, it is mutagenic and toxic; thus, its removal from the waste solution is of the top priority in lab safety practice. Although many products with high EtBr removal capacities are available on the market, developing new products with low material costs and high removal capacities is still an urgent priority. As the EtBr is in a cationic form Et+ balanced by counterion Br in aqueous solution, materials with high cation exchange capacity and large specific surface area may have great potential for efficient EtBr removal, Thus, several montmorillonites (MMTs) were evaluated for their EtBr removal capacity and methods of regeneration in this study. Results showed that both external and internal surfaces of MMTs were effective sorption sites for EtBr with a capacity up to 600mg/g. And the waste-laden materials could be regenerated or safely disposed after incineration at 500 °C for 2 h. As such, further tests on optimization and manufacturing kits or devices for practical EtBr removal in routine lab practice is of engineering priority, should MMTs be further explored as an effective material for EtBr removal.

Mechanistic insights into ethidium bromide removal by palygorskite from contaminated water

Ethidium bromide (EtBr)-containing wastewater can be hazardous to biodiversity when released into the soil and water bodies without treatment. EtBr can mutate living microbial cells and pose toxicity to even higher organisms. This work investigated the removal of EtBr from aqueous solutions by a naturally occurring palygorskite (PFl-1) clay mineral via systematic batch adsorption experiments under different physicochemical conditions. EtBr existed in an undissociated form at pH ~7, and was adsorbed on PFl-1 obeying the Freundlich isotherm model. The maximum EtBr adsorption capacity was 285 mmol/kg. The best fitted kinetic model for EtBr adsorption was the pseudo-second order model. The amounts of exchangeable cations desorbed from PFl-1 during EtBr adsorption was linearly correlated to the amounts of EtBr adsorbed, with a slope of 0.97, implying that a cation exchange-based adsorption mechanism was dominating. Additionally, dimerization of EtBr molecules via bromide release assisted an increased EtBr removal by PFl-1 at high adsorbate concentrations. Detailed x-ray diffraction, Fourier transform infrared, scanning electron imaging and energy dispersive x-ray analyses confirmed that EtBr adsorption occurred dominantly on the surface of palygorskite which mineralogically constituted 80% of the bulk PFl-1 adsorbent. A small portion of EtBr was also adsorbed by PFl-1 through intercalation onto the smectite impurity (10%) in PFl-1. This study suggested that PFl-1 could be an excellent natural material for removing EtBr from pharmaceutical and laboratory wastewater.

Mechanisms and pathways of ethidium bromide Fenton-like degradation by reusable magnetic nanocatalysts

Ethidium bromide (3,8-diamino-6-phenyl-5-ethylphenanthridinium bromide, EtBr) is a carcinogenic compound widely used for staining nucleic acids that is difficult to treat. In this study, magnetic nanocatalysts (MNCs) were synthesized for the heterogeneous Fenton-like degradation of EtBr. The initial pH, MNC content, and H2O2 concentration were the key factors affecting the EtBr degradation performance and dynamics. An EtBr removal efficiency of 98.97% was achieved within 4 h under optimal conditions (initial pH, 3.0; MNC content, 1 g/L; H2O2 concentration, 50 mM), and the degradation followed the ring-open pathway with (2E,4Z,8E)-3-amino-N-ethyl-7,9-dihydroxynona-2,4,8-trienamide as an intermediate, as determined by liquid chromatography and mass spectrometry (LC/MS). Unexpected and satisfactory Fenton-like oxidation of EtBr occurred under basic conditions, which was explained by a novel denitration pathway with 2-[nitro(phenyl)methyl]-(1,1′-biphenyl)-4,4′-diamine as an intermediate. The MNCs retained 62.17% of their degradation efficiency after five consecutive reaction and harvest cycles. Our work elucidated the mechanisms and pathways of EtBr removal in a Fenton-like reaction using MNCs, and comprehensively discussed the optimal reaction conditions and its potential for re-use.

Phycoremediation potential of microalgae species for ethidium bromide removal from aqueous media

Ethidium Bromide (EtBr) is an organic compound used in molecular biology investigations. EtBr ability of intercalating in the DNA molecule makes it a toxic substance. The objective was to evaluate the phycoremediation potentials of Chlorella vulgaris, Desmodesmus subspicatus and Raphidocelis subcapitata tested separately and in a mixture (Mix) for EtBr removal from the aqueous medium. Experiments were conducted using an initial algae biomass of 106 cell/mL, exposed to 500 µg/L of EtBr. The removal efficiency (µg EtBr L−1) after 3 h in each treatment were: Mix (72.8 µg.L−1) >D. subspicatus (48.4 µg.L−1) >R. subcapitata (24.6 µg.L−1) >C. vulgaris (19.9 µg.L−1). However, when EtBr mass reduction per microalgae density is considered (ng.algae−1), the efficiency ranking changes to: D. subspicatus (1.9 × 10−5 ng.algae−1) >C. vulgaris (1.4 × 10−5 ng.algae−1) >Mix (9.8 × 10−6 ng.algae−1) >R. subcapitata (2.8 × 10−6 ng.algae−1). The results suggest that initial algal population density is a determinant factor for efficient EtBr removal by microalgae species in short term treatments. In order to obtain 100% of EtBr removal, it should be necessary 1010, 1010 and 1011 algae.mL−1 of C. vulgaris, D. subspicatus and R. subcapitata, respectively. The results strongly suggest phycoremediation can be explored as an alternative method for EtBr removal.

Enhanced removal of ethidium bromide (EtBr) from aqueous solution using rectorite

Ethidium bromide (EtBr) is an intercalating agent commonly used as nucleic acid fluorescent tag in various techniques of life science field. It is considered as a serious biohazard due to its mutagenicity and carcinogenicity. As such, developing high efficiency and low cost materials as cleanup kits is in urgent need although many methods have already been developed. In this study we take use of the affinity of organic cations for clay minerals of high cation exchange capacity (CEC) and large specific surface area (SSA) and tested the removal of EtBr using rectorite, a type of clay mineral made of 1:1 regularly mixed layers of illite and montmorillonite. Our results showed that the uptake of Et+ on rectorite could be as high as 400 mmol/kg and the removal of Et+ was extremely fast. Desorption of inorganic cation Ca2+ and sorption of counterion Br− revealed that cation exchange was the dominating mechanism of Et+ removal using rectorite. Thermal analyses revealed that the EtBr could be thermally destructed inside the interlayer of rectorite and the material could be thermally regenerated. Thus, clay minerals could have a great potential to be fabricated into cleanup kits for the removal of EtBr in case of spill.

Efficient removal of ethidium bromide from aqueous solution by using DNA-loaded Fe3O4 nanoparticles.

Ethidium bromide (EtBr) is widely used as DNA-staining dyes for the detection of nucleic acids in laboratories and known to be powerful mutagens and carcinogens. In the present paper, the removal of EtBr from aqueous solutions in a batch system using DNA-loaded Fe3O4 nanoparticles as a simple and efficient method was investigated. DNA was covalently loaded on the surface of Fe3O4 magnetic nanoparticles, which was confirmed by FT-IR analysis and zeta potential measurements. The morphology and crystal structure were characterized by SEM, TEM, and XRD. The influence factors on the removal efficiency such as initial EtBr concentration, contact time, adsorbent dose, pH, and temperature were also studied. The removal process of EtBr can be completed quickly within 1min. The removal efficiency was more than 99% while the EtBr concentration was routinely used (0.5mgL-1) in biology laboratories and the dosages of nanoparticles were 1gL-1. For the different EtBr concentrations from 0.5 to 10mgL-1 in aqueous solution, the goal of optimized removal was achieved by adjusting the dosage of DNA-loaded Fe3O4 nanoparticles. The optimum pH was around 7 and the operational temperature from 4 to 35°C was appropriate. Kinetic studies confirmed that the adsorption followed second-order reaction kinetics. Thermodynamic data revealed that the process was spontaneous and exothermic. The adsorption of EtBr on DNA-loaded Fe3O4 nanoparticles fitted well with the Freundlich isotherm model. These results indicated that DNA-loaded Fe3O4 nanoparticles are a promising adsorbent for highly efficient removal of EtBr from aqueous solution in practice.

Adsorption of ethidium bromide (EtBr) from aqueous solutions by natural pumice and aluminium-coated pumice

In the present paper, the removal of ethidium bromide (EtBr) from aqueous solutions in a batch system using natural (NP) and aluminium-coated pumice (ACP) as alternative low-cost adsorbents was investigated. The maximum adsorption capacity, qm (mg/g) was 58.82 and 76.92mg/g for NP and ACP, respectively, operating at an initial pH of 8, an adsorbent dose of 8g/L, a contact time of 210min and an initial EtBr concentration of 30mg/L. The equilibrium data of both adsorbents fitted the Freundlich isotherm model, indicating the heterogeneity of the adsorbent surface. In addition, the adsorption rate of both adsorbents was well described by the pseudo-second-order kinetics model. This indicated chemisorption was the rate-controlling step of the adsorption process which occurred by ion exchange. Within the performed study, a three-layer artificial neural network (ANN) model was also developed to predict the efficiency of EtBr removal. Computational results clearly demonstrated that the ANN model was able to predict the combined effect of initial pH, adsorbent dose, contact time and initial EtBr concentration on the adsorption efficiency with a very high determination coefficient (R2=0.998) and a low relative error (RE=0.037).

A modeling study by artificial neural network on ethidium bromide adsorption optimization using natural pumice and iron-coated pumice

In this study, the potential of natural pumice (NP) and iron-coated pumice stone (Fe-CP) as novel low-cost adsorbents to remove ethidium bromide (EtBr) from aqueous solutions was investigated. The operational parameters affecting removal efficiency and adsorption capacity such as adsorbent dose, initial EtBr concentration, pH, and contact time were studied in order to maximize EtBr removal. The maximum amount of adsorbed EtBr (qm) using NP and Fe-CP was 40.25 and 45.08 mg g‒1, respectively. It was found that EtBr adsorption followed the Freundlich isotherm model and fitted the pseudo-second-order kinetics equation for both adsorbents. In addition, the experimental system could be easily modeled by artificial neural network calculations.

Removal of ethidium bromide by carbon nanotube in aqueous solution: isotherms, equilibrium mechanism studies, and its comparison with nanoscale of zero valent iron as adsorbent

The adsorption of ethidium bromide (EtBr) by single-walled carbon nanotubes (SWCNTs) and nanoscale of zero valent iron (NZVI) were investigated to assess its possible use as adsorbents. The effect of various factors, namely initial adsorbate concentration, adsorbent dosage, and contact time, were studied to identify adsorption capacity of SWCNTs and NZVI surfaces. The experiment demonstrated the maximum EtBr which was obtained at 5 min to attain equilibrium for SWCNTs and NZVI surfaces. Adsorption data were modeled with the Langmuir, Freundlichand, Temkin isotherms. Langmuir adsorption model was used for the mathematical description of the adsorption equilibrium, and the equilibrium data fitted very well with this model for both surfaces as adsorbents. The study showed that SWCNTs and NZVI surfaces could be used as new and efficient adsorbent materials for the removal of EtBr from aqueous solution. Also, the result showed that the SWCNTs were more effective than NZVI in the removal of EtBr from aqueous solution.

Direct observation of the reversible unwinding of a single DNA molecule caused by the intercalation of ethidium bromide

Ethidium bromide (EtBr) is the conventional intercalator for visualizing DNA. Previous studies suggested that EtBr lengthens and unwinds double-stranded DNA (dsDNA). However, no one has observed the unwinding of a single dsDNA molecule during intercalation. We developed a simple method to observe the twisting motions of a single dsDNA molecule under an optical microscope. A short dsDNA was attached to a glass surface of a flow chamber at one end and to a doublet bead as a rotation marker at the other end. After the addition and removal of EtBr, the bead revolved in opposite directions that corresponded to the unwinding and rewinding of a dsDNA, respectively. The amount of intercalating EtBr was estimated from the revolutions of the bead. EtBr occupied 57% of base pairs on a single dsDNA at 1 mM of EtBr, indicating that EtBr molecules could bind at contiguous sites to each other. The isotherm of intercalation showed that negative cooperativity existed between adjoining EtBr molecules. The association constant of EtBr and dsDNA (1.9 (±0.1) × 105 M−1) was consistent with that of previous results. Our system is useful to investigate the twisting of a single dsDNA interacting with various chemicals and biomolecules.