Methyl Triclosan Research Articles

Electrochemical reduction of 2,4-dichloro-1-(4-chloro-2-methoxyphenoxy)benzene (methyl triclosan) at glassy carbon cathodes in dimethylformamide

Methyl triclosan [2,4-dichloro-1-(4-chloro-2-methoxyphenoxy)benzene, 1], which is now considered an environmental pollutant, has been shown to be formed via microbial methylation of triclosan [5-chloro-2-(2,4-dichlorophenoxy)phenol, 2]. In the present work, cyclic voltammetry and controlled-potential (bulk) electrolysis have been employed to investigate the direct reduction of 1 at glassy carbon electrodes in dimethylformamide (DMF) containing 0.050M tetramethylammonium tetrafluoroborate (TMABF4). Cyclic voltammograms for the reduction of 1 at a glassy carbon electrode exhibit three irreversible cathodic peaks with peak potentials of −1.72, −2.07, and −2.35V (versus a cadmium-saturated mercury amalgam reference electrode). These three peaks correspond, respectively, to (a) scission of the ortho C–Cl bond, (b) cleavage of the two para C–Cl bonds, and (c) rupture of the C–O bond. Controlled-potential (bulk) electrolyses of 1 at reticulated vitreous carbon electrodes result in the formation of mixtures of 4-chloro-1-(4-chlorophenoxy)-2-methoxybenzene (3), 4-chloro-2-methoxy-1-phenoxybenzene (4a), 1-(4-chlorophenoxy)-2-methoxybenzene (4b), 1-methoxy-2-phenoxybenzene (5), anisole (6), and phenol.

Transformation products and human metabolites of triclocarban and triclosan in sewage sludge across the United States.

Removal of triclocarban (TCC) and triclosan (TCS) from wastewater is a function of adsorption, abiotic degradation, and microbial mineralization or transformation, reactions that are not currently controlled or optimized in the pollution control infrastructure of standard wastewater treatment. Here, we report on the levels of eight transformation products, human metabolites, and manufacturing byproducts of TCC and TCS in raw and treated sewage sludge. Two sample sets were studied: samples collected once from 14 wastewater treatment plants (WWTPs) representing nine states, and multiple samples collected from one WWTP monitored for 12 months. Time-course analysis of significant mass fluxes (α = 0.01) indicate that transformation of TCC (dechlorination) and TCS (methylation) occurred during sewage conveyance and treatment. Strong linear correlations were found between TCC and the human metabolite 2′-hydroxy-TCC (r = 0.84), and between the TCC-dechlorination products dichlorocarbanilide (DCC) and monochlorocarbanilide (r = 0.99). Mass ratios of DCC-to-TCC and of methyl-triclosan (MeTCS)-to-TCS, serving as indicators of transformation activity, revealed that transformation was widespread under different treatment regimes across the WWTPs sampled, though the degree of transformation varied significantly among study sites (α = 0.01). The analysis of sludge sampled before and after different unit operation steps (i.e., anaerobic digestion, sludge heat treatment, and sludge drying) yielded insights into the extent and location of TCC and TCS transformation. Results showed anaerobic digestion to be important for MeTCS transformation (37–74%), whereas its contribution to partial TCC dechlorination was limited (0.4–2.1%). This longitudinal and nationwide survey is the first to report the occurrence of transformation products, human metabolites, and manufacturing byproducts of TCC and TCS in sewage sludge.

Rotating-disk sorptive extraction: effect of the rotation mode of the extraction device on mass transfer efficiency

The extraction device used in rotating-disk sorptive extraction consists of a Teflon disk in which a sorptive phase is fixed on one of its surfaces. Depending on the configuration, the rotation axis of the disk device can be either perpendicular or parallel to its radius, giving rise to two different mass transfer patterns when rotating-disk sorptive extraction is applied in liquid samples. In the perpendicular case (configuration 1), which is the typical configuration, the disk contains an embedded miniature stir bar that allows the disk rotation to be driven using a common laboratory magnetic stirrer. In the parallel case (configuration 2), the disk is driven by a rotary rod connected to an electric stirrer. In this study, triclosan and its degradation product methyl triclosan were used as analyte models to demonstrate the significant effect of the rotation configuration of the disk on the efficiency of analyte mass transfer from water to a sorptive phase of polydimethylsiloxane. Under the same experimental conditions and at a rotation velocity of 1,250 rpm, extraction equilibrium was reached at 80 min when the disk was rotated in configuration 1 and at 30 min when the disk was rotated in configuration 2. The extraction equilibration time decreased to 14 min when the rotation velocity was increased to 2,000 rpm in configuration 2. Because the rotation pattern affects the mass transfer efficiency, each rotation configuration was characterized through the Reynolds number; Re values of 6,875 and 16,361 were achieved with configurations 1 and 2, respectively, at 1,250 rpm.

Fate of microconstituents in biosolids composted in an aerated silage bag

Although most composting studies report pathogen concentrations, little is known about the fate of Endocrine Disruptor Chemicals (EDCs) during composting. In this study, a positively aerated polyethylene bag composting system was filled with a mixture of woodchips and limed biosolids from a large Waste Water Treatment Plant (WWTP) to study the removal efficiency of two different groups of EDCs. Two antibacterial compounds, Triclocarban (TCC) and Triclosan (TCS), and a TCS byproduct, Methyltriclosan (MeTCS), as well as seven congeners of flame retardants known as PBDEs (Polybrominated Diphenyl Ethers) were studied during two phases of composting: 1) a thermophilic phase, in which positive mechanical aeration, pushing air into and through the materials matrix, was conducted for 2 months; and 2) a curing and stabilization phase in which no mechanical aeration was provided and the bag was opened to ambient passive aeration to simulate storage conditions for seven months. Our results showed that while TCC concentrations remained constant, TCS degradation took place during both phases. The degradation of TCS was corroborated by the formation of MeTCS in both phases. The TCS concentrations decreased from 18409 ± 1,877 to 11955 ± 288 ng g−1 dry wt. during the thermophilic phase and declined from 11,955 ± 288 to 7,244 ± 909. ng g−1 dry wt. by the end of the curing phase. Thus, slightly greater TCS transformation occurred during the second than during the first (35.1 vs. 39.4%). MeTCS concentrations increased from 189.3 ± 8.6 to 364.6 ± 72.5 ng g−1 dry wt. during the first phase and reached 589.0 ± 94.9 ng g−1 dry wt. at the end of the second phase. PBDEs concentrations were below quantification limits for all but two of the congeners analyzed (BDE-47 and BDE-99). PBDE concentrations were measured at the end of the first phase only and were comparable to initial concentrations.

Experimental and predicted acute toxicity of antibacterial compounds and their mixtures using the luminescent bacterium Vibrio fischeri

This article investigates the bioluminescence inhibition effects of the antimicrobials triclocarban, triclosan and its metabolite methyl triclosan, using the marine bacterium Vibrio fischeri as the test organism (Microtox©). The concentration response analysis was performed for the three individual substances and for a mixture in which the three compounds were mixed in a ratio of the IC50 of the individual components (equitoxic ratio). Toxicity values (the median inhibitory concentration value, in mgL−1) in the decreasing order of sensitivity were triclosan (0.73)>triclocarban (0.91)>methyl-triclosan (1.76). The comparison of the experimental data with those obtained by using Quantitative Structure–Activity Relationship (QSAR) equations indicated that triclosan and triclocarban act as polar narcotic compounds towards V. fischeri, whereas methyl-triclosan acts as a narcotic (baseline toxicity). The toxicity of the mixture was measured experimentally and predicted by two models (CA: concentration addition; IA: independent action). The results showed that the observed mixture toxicity (IC50=0.23mgL−1) had no significant differences from those predicted by both CA and IA models.

Triclocarban, triclosan and its transformation product methyl triclosan in native earthworm species four years after a commercial-scale biosolids application

Triclocarban (TCC), triclosan (TCS) and methyl triclosan (Me-TCS) were detected in soil and the native population of earthworms of an agricultural field in Ottawa, Canada, about four years after a commercial-scale application of biosolids. In soil that received biosolids, TCC and TCS were detected at median concentrations of 13.0 and 1.5ng/g soil (d.w.), respectively, while Me-TCS, the transformation product of triclosan, was detected at a six-fold higher median concentration than its precursor. In earthworms collected at the biosolids-amended field-plot about four years post application, Me-TCS was also detected at higher concentrations (26 to 114ng/g tissue d.w.) than TCS (16–51ng/g) and TCC (4–53ng/g). These data provide evidence that not only parent compounds but also their transformation products need to be considered in faunal bioaccumulation studies. Moreover, the preliminary results for pooled earthworm samples from different ecological groups suggest that the degree of bioaccumulation of biosolids-associated contaminants may depend on the habitat and feeding behavior of the organisms.

Determining the distribution of triclosan and methyl triclosan in estuarine settings

We have developed a method for the analysis of two sewage-derived contaminants: triclosan (TCS), an antibacterial agent, and methyl triclosan (MTCS), a TCS metabolite. For solid samples (4g), extraction and cleanup were integrated into the same step using pressurized liquid extraction (PLE) with in-cell-clean-up (1g of florisil). The extraction was performed using dichloromethane at 100°C, 1500 psi and 3 static extraction cycles of 5min each. For water samples (100mL), stir bar sorptive extraction–liquid desorption (SBSE–LD) was used. Bars were stirred for 10h and analytes were later desorbed using acetonitrile. Finally, MTCS and a silylated derivative of TCS were determined by gas chromatography–mass spectrometry (GC–MS). Recovery experiments in water and sediments were performed and the results ranged from 67% to 78%. Limits of detection (LODs) were 5ngL−1 for TCS and 1ngL−1 for MTCS, in water samples, and 0.1ngg−1 for TCS and MTCS in solid samples. The method was applied then to determine the levels of these compounds in the estuary of Guadalete River (SW Spain). TCS and MTCS concentrations up to 9.6ngg−1 in sediments and 310ngL−1 in water were measured. Their distribution was strongly influenced by the presence of wastewater sources, treated and untreated, along the sampling area, where maximum concentrations were detected. Highest values were reached in the water column during low tides as the water volume in the estuary becomes lower.

Bioconcentration of triclosan and methyl-triclosan in marine mussels (Mytilus galloprovincialis) under laboratory conditions and in metropolitan waters of Gulf St Vincent, South Australia

The anti-microbial agent triclosan (TCS), and its derivative methyl-triclosan (Me-TCS), are discharged with treated effluents from wastewater treatment plants to receiving environments. We investigated the bioconcentration of TCS and Me-TCS in mussels (Mytilus galloprovincialis) exposed to TCS (100ngL−1) for 30days in seawater aquaria (19±2°C) with fresh phytoplankton as a food source. Bioconcentration increased with time reaching a steady-state around 24–30days. The bioconcentration factor (log BCF) for TCS were 2.81Lkg−1 (dry weight) and 4.13Lkg−1, when lipid normalised concentrations were used. Mussels were also deployed in cages at four marine locations receiving effluents from WWTPs. The mean (±SD) TCS and Me-TCS concentrations for mussels from these sites were 9.87 (±1.34) and 6.99 (±2.44)μgkg−1. The study showed that mussels can be a useful tool for monitoring pollution of TCS and Me-TCS in marine and estuarine environments.

Methyl-triclosan binding to human serum albumin: Multi-spectroscopic study and visualized molecular simulation

Methyl-triclosan (MTCS), a transformation product and metabolite of triclosan, has been widely spread in environment through the daily use of triclosan which is a commonly used anti-bacterial and anti-fungal substance in consumer products. Once entering human body, MTCS could affect the conformation of human serum albumin (HSA) by forming MTCS–HSA complex and alter function of protein and endocrine in human body. To evaluate the potential toxicity of MTCS, the binding mechanism of HSA with MTCS was investigated by UV–vis absorption, circular dichroism and Fourier transform infrared spectroscopy. Binding constants, thermodynamic parameters, the binding forces and the specific binding site were studied in detail. Binding constant at room tempreture (T=298K) is 6.32×103Lmol−1; ΔH0, ΔS0 and ΔG0 were 22.48kJmol−1, 148.16Jmol−1K−1 and −21.68kJmol−1, respectively. The results showed that the interactions between MTCS and HSA are mainly hydrophobic forces. The effects of MTCS on HSA conformation were also discussed. The binding distance (r=1.2nm) for MTCS–HSA system was calculated by the efficiency of fluorescence resonance energy transfer. The visualized binding details were also exhibited by molecular modeling method and the results could agree well with that from the experimental study.

Rotating disk sorptive extraction of triclosan and methyl-triclosan from water samples

A method of sample preparation based on use of rotating disk sorptive extraction (RDSE) has been developed for determination of triclosan (TCS) and methyl-triclosan (MTCS) in water samples. The sorptive and desorptive behavior of the analytes was studied by use of a rotating disk coated with polydimethylsiloxane (PDMS) on one of its surfaces. Chemical and extraction behavior were studied to establish the best conditions for extraction. The optimum conditions for both analytes were: sample volume 25 mL, pH 4.5, NaCl concentration 6% (w/v), disk rotational velocity 1,250 rpm, and extraction time 80 min. A desorption time of 30 min was used with 5 mL methanol. The detection limits for TCS and MTCS were 46 and 34 ng L(-1), respectively. Recovery was evaluated at two concentrations, 160 and 800 ng L(-1), and the values obtained were between 80 and 100%. The method was applied to analysis of influent water at two treatment plants in Santiago, Chile.

Determination of Triclocarban, Triclosan and Methyl-Triclosan in Environmental Water by Silicon Dioxide/Polystyrene Composite Microspheres Solid-Phase Extraction Combined with HPLC-ESI-MS

This paper developed a sensitive and efficient analytical method for triclocarban (TCC), triclosan (TCS) and Methyl-triclosan (MTCS) determination in environmental water, which involves enrichment by using silicon dioxide/polystyrene composite microspheres solid-phase extraction and detection with HPLC-ESI-MS. The influence of several operational parameters, including the eluant and its volume, the flow rate and acidity of water sample were investigated and optimized. Under the optimum conditions, the limits of detection were 1.0 ng/L, 2.5 and 4.5 ng/L for TCC, TCS, and MTCS, respectively. The linearity of the method was observed in the range of 5-2000 ng/L, with correlation coefficients (r2) >.99. The spiked recoveries of TCC, TCS and MTCS in water sampleswereachieved in the range of 89.5% -96.8% with RSD below 5.7%. The proposed method has been successfully applied to analyze real water samples and satisfactory results were achieved.

Metabolization of the Bacteriostatic Agent Triclosan in Edible Plants and its Consequences for Plant Uptake Assessment

Persistent environmental contaminants may enter agricultural fields via the application of sewage sludge, by irrigation with treated municipal wastewater or by manuring. It has been shown that such contaminants can be incorporated into crop plants. The metabolism of the bacteriostatic agents triclocarban, triclosan, and its transformation product methyl triclosan was investigated after their uptake into carrot cell cultures. A fast metabolization of triclosan was observed and eight so far unknown phase II metabolites, conjugates with saccharides, disaccharides, malonic acid, and sulfate, were identified by liquid chromatography-mass spectrometry. Triclocarban and methyl triclosan lack a phenolic group and remained unaltered in the cell cultures. Phase I metabolization was not observed for any of the compounds. All eight triclosan conjugates identified in the cell cultures were also detected in extracts of intact carrot plants cultivated on triclosan contaminated soils. Their total amount in the plants was assessed to exceed the amount of the triclosan itself by a factor of 5. This study shows that a disregard of conjugates in studies on plant uptake of environmental contaminants may severely underestimates the extent of uptake into plants and, eventually, the potential human exposure to contaminants via food of plant origin.

Slow O-demethylation of methyl triclosan to triclosan, which is rapidly glucuronidated and sulfonated in channel catfish liver and intestine

The antibacterial personal care product triclosan is discharged in municipal waste, and converted in part by bacteria in sewage sludge and soil to its more lipid-soluble methyl ether, methyl triclosan. Triclosan and methyl triclosan have been detected in water, sediment, fish and invertebrates near sewage treatment facilities. Understanding the biotransformation of methyl triclosan and triclosan in a model food fish, the channel catfish, will be of value in assessing the likelihood that these compounds will bioaccumulate in exposed fish, and therefore potentially pass up the food chain. We hypothesize that cytochrome P450 will catalyze the O-demethylation of methyl triclosan to yield triclosan, which is likely to undergo glucuronidation or sulfonation of the phenolic hydroxyl group. Conversion of methyl triclosan to triclosan was measured by LC/MS/MS following aerobic incubation of varying concentrations of methyl triclosan with NADPH and hepatic and intestinal microsomes from untreated, 3-methylcholanthrene-treated (10mg/kg, i.p.) or PCB-126-treated (0.1mg/kg, i.p.) channel catfish (n=4 per treatment group). The Km values for methyl triclosan were similar for untreated, 3-methylcholanthrene-treated and PCB-126-treated catfish liver microsomes, ranging from 80 to 250μM. Vmax values for O-demethylation ranged from 30 to 150pmol/min/mg protein, with no significant differences between controls, PCB-126-treated or 3-methylcholanthrene-treated fish, suggesting that methyl triclosan O-demethylation was not a CYP1-catalyzed reaction. Methyl triclosan O-demethylation activities in intestinal microsomes were similar to or lower than those found with liver microsomes. The calculated rate of O-demethylation of methyl triclosan in catfish liver at 1μM, a concentration reported in exposed fish, and 21°C, an early summer water temperature, is 0.10pmol/min/mg protein. This slow rate of metabolism suggests that upon continued exposure, methyl triclosan may bioaccumulate in the channel catfish. Triclosan itself, however, was readily glucuronidated by hepatic and intestinal microsomes and sulfonated by hepatic and intestinal cytosol. Triclosan glucuronidation followed Michaelis-Menten kinetics when rates were measured across a concentration range of 5–1000μM, whereas triclosan sulfonation exhibited substrate inhibition at concentrations above 10–20μM in both intestinal and hepatic cytosol. Based on the enzyme kinetic constants measured in hepatic and intestinal fractions at 21°C, triclosan at 1μM could be glucuronidated at rates of 23 and 3.2pmol/min/mg protein respectively in liver and intestine, and sulfonated at rates of 277 (liver) and 938 (intestine) pmol/min/mg protein. These rates are much higher than the rates of demethylation of methyl triclosan, and suggest that triclosan would be rapidly cleared and unlikely to bioaccumulate in catfish tissues

Qualitative Analysis of Some Emerging Halogenous Pollutions in Fish Sample by Comprehensive Two-Dimensional Gas Chromatography/Time-of-Flight Mass Spectrometry

Abstract Numerous halogenated organic compounds have been identified as pollutants of concern, however, the release to the environment of many is still unrecognized. In this study, the comprehensive two dimensional gas chromatography/time of flight mass spectrometry (GC×GC-TOFMS) was applied to screen no-target halogenous pollutions in the fish sample. After extrated with hexane/actone (1:1, V/V ), cleaned-up with Gel Permeation Chromatography (GPC) and silica gel packed column, the residues were re-dissolved in the hexane and analyzed by GC×GC-TOFMS coupled with DB-5MS column and HT-8 column. In total, 72 compounds were identified as organohalogens, which including 33 polychlorinated biphenyls, 9 organochlorine pesticides, 4 polybrominated diphenyl ethers, 4 metabolites of DDT, 2 chlorinated anisole, 2 chlorostyrene, 1 thioanisole, and methyl triclosan. The rest 16 compounds were tentatively identified as halogenous pollutions, however, these compounds could not be exactly identified due to the lack of authentic standard reference mass spectrum in the library. Using external standard method, 10 dominating PCBs and 1 PBDE were quantified in 5 fish samples with S 10 PCBs and BDE-47 concentration ranged from 52-332 ng g −1 lipid and 4.8-17 ng g −1 lipid, respectively.

Occurrence and toxicity of antimicrobial triclosan and by-products in the environment

A review was undertaken on the occurrence, toxicity, and degradation of triclosan (TCS; 5-chloro-2,4-dichlorophenoxy)phenol) in the environment. TCS is a synthetic, broad-spectrum antibacterial agent incorporated in a wide variety of household and personal care products such as hand soap, toothpaste, and deodorants but also in textile fibers used in a range of other consumer products (e.g., toys, undergarments and cutting boards among other things). Because of its partial elimination in sewage treatment plants, most reports describe TCS as one of the most commonly encountered substances in solid and water environmental compartments. It has been detected in a microgram per liter or microgram per kilogram level in sewage treatment plants (influents, effluents, and sludges), natural waters (rivers, lakes, and estuarine waters), and sediments as well as in drinking water. Moreover, due to its high hydrophobicity, TCS can accumulate in fatty tissues and has been found in fish and human samples (urine, breast milk, and serum). TCS is known to be biodegradable, photo-unstable, and reactive towards chlorine and ozone. As a consequence, it can be transformed into potentially more toxic and persistent compounds, such as chlorinated phenols and biphenyl ethers after chlorination, methyl triclosan after biological methylation, and chlorinated dibenzodioxins after photooxidation. The toxicity of TCS toward aquatic organisms like fish, crustaceans, and algae has been demonstrated with EC50 values near TCS environmental concentrations. It has even been shown to produce cytotoxic, genotoxic, and endocrine disruptor effects. Furthermore, the excessive use of TCS is suspected to increase the risk of emergence of TCS-resistant bacteria and the selection of resistant strains.

Reply to 2nd Comment on “Effects of Triclocarban, Triclosan, And Methyl Triclosan on Thyroid Hormone Action and Stress in Frog and Mammalian Culture Systems”

Reply to 2nd Comment on “Effects of Triclocarban, Triclosan, And Methyl Triclosan on Thyroid Hormone Action and Stress in Frog and Mammalian Culture Systems”

Comment on “Effects of Triclocarban, Triclosan, and Methyl Triclosan on Thyroid Hormone Action and Stress in Frog and Mammalian Culture Systems”

Comment on “Effects of Triclocarban, Triclosan, and Methyl Triclosan on Thyroid Hormone Action and Stress in Frog and Mammalian Culture Systems”

Fate of Triclosan and Methyltriclosan in soil from biosolids application

This study investigates the persistence of Triclosan (TCS), and its degradation product, Methyltriclosan (MeTCS), after land application of biosolids to an experimental agricultural plot under both till and no till. Surface soil samples ( n = 40) were collected several times over a three years period and sieved to remove biosolids. Concentration of TCS in the soil gradually increased with maximum levels of 63.7 ± 14.1 ng g −1 dry wt., far below the predicted maximum concentration of 307.5 ng g −1 dry wt. TCS disappearance corresponded with MeTCS appearance, suggesting in situ formation. Our results suggest that soil incorporation and degradation processes are taking place simultaneously and that TCS background levels are achieved within two years. TCS half-life ( t 0.5) was determined as 104 d and MeTCS t 0.5, which was more persistent than TCS, was estimated at 443 d.

Biodegradation of triclosan in biosolids‐amended soils

Land application of biosolids can constitute an important source of triclosan (TCS) input to soils, with uncertain effects. Several studies have investigated the degradation potential of TCS in biosolids-amended soils, but the results vary widely. We conducted a laboratory degradation study by mixing biosolids spiked with [¹⁴C]-TCS (final concentration = 40 mg/kg) with Immokalee fine sand and Ashkum silty clay loam soils at an agronomic application rate (22 Mg/ha). Biosolids-amended soils were aerobically incubated in biotic and inhibited conditions for 18 weeks. Subsamples removed at 0, 2, 4, 6, 9, 12, 15, and 18 weeks were sequentially extracted with an operationally defined extraction scheme to determine labile and nonlabile TCS fractions. Over the 18-week incubation, the proportion of [¹⁴C] in the nonlabile fraction increased and the labile fraction decreased, suggesting decreasing availability to biota. Partitioning of TCS into labile and nonlabile fractions depended on soil characteristics. Less than 0.5% of [¹⁴C]-TCS was mineralized to carbon dioxide (¹⁴CO₂) in both soils and all treatments. A degradation metabolite, methyl triclosan (Me-TCS), was identified in both soils only in the biotic treatment, and increased in concentration over time. Even under biotic conditions, biosolids-borne TCS is persistent, with a primary degradation (TCS to Me-TCS) half-life of 78 d in the silty clay loam and 421 d in the fine sand. A half-life of approximately 100 d would be a conservative first approximation of TCS half-life in biosolids-amended soils for risk estimation.

Comment on “Effects of Triclocarban, Triclosan, And Methyl Triclosan on Thyroid Hormone Action and Stress in Frog and Mammalian Culture Systems”

Comment on “Effects of Triclocarban, Triclosan, And Methyl Triclosan on Thyroid Hormone Action and Stress in Frog and Mammalian Culture Systems”