Elucidating Factors Contributing to Dicamba Volatilization by Characterizing Chemical Speciation in Dried Dicamba-Amine Residues.

Dicamba is a semivolatile herbicide that has caused widespread unintentional damage to vegetation due to its volatilization from genetically engineered dicamba-tolerant crops. Strategies to reduce dicamba volatilization rely on the use of formulations containing amines, which deprotonate dicamba to generate a nonvolatile anion in aqueous solution. Dicamba volatilization in the field is also expected to occur after aqueous spray droplets dry to produce a residue; however, dicamba speciation in this phase is poorly understood. We applied Fourier transform infrared (FTIR) spectroscopy to evaluate dicamba protonation state in dried dicamba-amine residues. We first demonstrated that commercially relevant amines such as diglycolamine (DGA) and n,n-bis(3-aminopropyl)methylamine (BAPMA) fully deprotonated dicamba when applied at an equimolar molar ratio, while dimethylamine (DMA) allowed neutral dicamba to remain detectable, which corresponded to greater dicamba volatilization. Expanding the amines tested, we determined that dicamba speciation in the residues was unrelated to solution-phase amine pKa, but instead was affected by other amine characteristics (i.e., number of hydrogen bonding sites) that also correlated with greater dicamba volatilization. Finally, we characterized dicamba-amine residues containing an additional component (i.e., the herbicide S-metolachlor registered for use alongside dicamba) to investigate dicamba speciation in a more complex chemical environment encountered in field applications.

Ecotoxicity studies reveal that organic cations in dicamba-derived ionic liquids can pose a greater environmental risk than the herbicide itself.

The following research provides novel and relevant insights into potential environmental consequences of combination of various organic cations with commercial systemic herbicide (dicamba), in accordance with a 'herbicidal ionic liquids' (HILs) strategy. Toxicity assays of five dicamba-based HILs comprising different hydrophobic and hydrophilic cations, namely choline [CHOL][DIC], ethyl betainate [BETC2][DIC], decyl betainate [BETC10][DIC], hexadecyl betainate [BETC16][DIC] and didecyldimethylammonium [DDA][DIC]), have been tested towards bacteria (Pseudomonas putida, Escherichia coli, Bacillus subtilis), algae (Chlorella vulgaris), fresh and marine water crustaceans (Daphnia magna, Artemia franciscana). The structure of respective substituents in the cation emerged as a decisive determinant of toxicity in the case of tested species. In consequence, small ions of natural origin ([CHOL] and [BETC2]) demonstrated toxicity numerous orders of magnitude lower compared to fully synthetic [DDA]. These results emphasize the role of cations' hydrophobicity, as well as origin, in the observed acute toxic effect. Time-dependent toxicity assays also indicated that betaine-type cations comprising an ester bond can rapidly transform into less harmful substances, which can generally result in a reduction in toxicity by even several orders of magnitude. Nonetheless, these findings challenge the concept of ionic liquids with herbicidal activity and give apparent parallels to adjuvant-dependent toxicity issues recently noted in typical herbicidal formulations.

Evaluation of perinatal exposure of glyphosate and its mixture with 2,4-D and dicamba οn liver redox status in Wistar rats.

Wide-scale emergence of glyphosate-resistant weeds has led to an increase in the simultaneous application of herbicide mixtures exacerbated by the introduction of crops tolerant to glyphosate plus dicamba or glyphosate plus 2,4-D. This raises serious concerns regarding the environmental and health risks resulting from increased exposure to a mixture of herbicide active ingredients. We evaluated hepatotoxic effects following perinatal exposure to glyphosate alone or in combination with 2,4-D and dicamba from gestational day-6 until adulthood in Wistar rats. Animals were administered with glyphosate at the European Union (EU) acceptable daily intake (ADI; 0.5 mg/kg bw/day) and no-observed-adverse-effect level (NOAEL; 50 mg/kg bw/day). A mixture of glyphosate with 2,4-D (0.3 mg/kg bw/day) and dicamba (0.02 mg/kg bw/day) with each at their EU ADI was evaluated. Redox status was determined by measuring levels of reduced glutathione, decomposition rate of Η2Ο2, glutathione reductase, glutathione peroxidase, total antioxidant capacity, thiobarbituric reactive substances, and protein carbonyls. Gene expression analysis of Nr1d1, Nr1d2, Clec2g, Ier3, and Gadd45g associated with oxidative damage to DNA, was also performed. Analysis of liver samples showed that exposure to the mixture of the three herbicides induced a marked increase in the concentration of glutathione and malondialdehyde indicative of a disturbance in redox balance. Nevertheless, the effect of increased lipid peroxidation was not discernible following a 3-month recuperation period where animals were withdrawn from pesticide exposure post-weaning. Interestingly, toxic effects caused by prenatal exposure to the glyphosate NOAEL were present after the same 3-month recovery period. No statistically significant changes in the expression of genes linked with genotoxicity were observed. Our findings reinforce the importance of assessing the combined effects of chemical pollutants at doses that are asserted by regulatory agencies to be safe individually.

Amine Volatilization from Herbicide Salts: Implications for Herbicide Formulations and Atmospheric Chemistry.

Amines are frequently included in formulations of the herbicides glyphosate, 2,4-D, and dicamba to increase herbicide solubility and reduce herbicide volatilization by producing herbicide-amine salts. Amines, which typically have higher vapor pressures than the corresponding herbicides, could potentially volatilize from these salts and enter the atmosphere, where they may impact atmospheric chemistry, human health, and climate. Amine volatilization from herbicide-amine salts may additionally contribute to volatilization of dicamba and 2,4-D. In this study, we established that amines applied in herbicide-amine salt formulations undergo extensive volatilization. Both dimethylamine and isopropylamine volatilized when aqueous salt solutions were dried to a residue at ∼20 °C, while lower-vapor pressure amines like diglycolamine and n,n-bis-(3-aminopropyl)methylamine did not. However, all four amines volatilized from salt residues at 40-80 °C. Because amine loss typically exceeded herbicide loss, we proposed that neutral amines dominated volatilization and that higher temperatures altered their protonation state and vapor pressure. Due to an estimated 4.0 Gg N/yr applied as amines to major U.S. crops, amine emissions from herbicide-amine salts may be important on regional scales. Further characterization of worldwide herbicide-amine use would enable this contribution to be compared to the 285 Gg N/yr of methylamines emitted globally.

Design of a novel hapten and development of a sensitive monoclonal immunoassay for dicamba analysis in environmental water samples.

Weed resistance to glyphosate has been a driving force behind the increased use of alternative herbicides in agriculture. Recently, dicamba-tolerant recombinant plants were introduced to the market, which may result in residues of this agrochemical contaminating environmental waters. Given that restrictions on the use of dicamba have consequently been established by regulatory agencies, it is therefore also desirable to conduct extensive controls on dicamba residues. Immunoassays are currently the most powerful bioanalytical technology for the rapid monitoring of chemical residues and contaminants. In the present study, a novel hapten was designed maintaining unaltered all the antigenic moieties of the target molecule, and this was used to generate high-affinity monoclonal antibodies against dicamba for the first time. Additionally, a collection of haptens with different linker composition or linker tethering site was synthesized and conjugated to proteins. Using these novel immunoreagents, a direct competitive enzyme-linked immunosorbent assay with a limit of detection for dicamba of 0.24 ng/mL was developed and validated. Analysis of water samples from different origins afforded recovery values between 90 % and 120 %, and coefficients of variation below 20 % were obtained. These results indicate that the developed immunochemical assay is suitable for the rapid determination of dicamba residues in environmental water samples.

Physicochemical properties, droplet size and volatility of dicamba with herbicides and adjuvants on tank-mixture.

The adoption of dicamba-tolerant soybean varieties has increased the concern and demand for new drift and volatility reduction technologies. Potential spray nozzles and adjuvants should be studied to determine its effects on drift and volatility of dicamba tank-mixtures. The objective of this study was to evaluate physicochemical characteristics of spray solutions containing dicamba; to analyze droplet size effect with air induction nozzles; and to assess dicamba volatilization on soybean plants with a proposed methodology. Treatments included dicamba only and mixtures with herbicides and adjuvants. Dicamba mixed with lecithin + methyl soybean oil + ethoxylated alcohol adjuvant had the greatest efficacy potential among treatments considering tank-mixture pH, surface tension, contact angle and droplet size. The MUG11003 nozzle produced the coarsest droplet size and was better suited for drift management among nozzle types. The proposed volatilization methodology successfully indicated dicamba volatilization in exposed soybean plants and among the evaluated treatments, it showed greater volatilization for dicamba with glyphosate + lecithin + propionic acid adjuvant.

Roles of the Gentisate 1,2-Dioxygenases DsmD and GtdA in the Catabolism of the Herbicide Dicamba in Rhizorhabdus dicambivorans Ndbn-20.

3-Chlorogentisate is a key intermediate in the catabolism of the herbicide dicamba in R. dicambivorans Ndbn-20. In this study, we identified two gentisate 1,2-dioxygenases (GDOs), DsmD and GtdA, from Ndbn-20. The amino acid sequence similarity between DsmD and GtdA is 51%. Both of them are dimers and showed activities to gentisate and 3-chlorogentisate but not 3,6-dichlorogentisate (3,6-DCGA) or 6-chlorogentisate in vitro. The kcat/Km of DsmD for 3-chlorogentisate was 28.7 times higher than that of GtdA, whereas the kcat/Km of DsmD for gentisate was only one-fourth of that of GtdA. Transcription of dsmD was dramatically induced by 3-chlorogentisate but not gentisate, whereas gtdA was not induced. Disruption of dsmD resulted in a significant decline in the degradation rates of 3-chlorogentisate and dicamba but had no effect on the degradation of gentisate, whereas the result of disruption of gtdA was converse; the disruption of both dsmD and gtdA led to the inability to degrade 3-chlorogentisate and gentisate. This study revealed that 3-chlorogentisate but not gentisate or 3,6-DCGA is the ring-cleavage substrate in the dicamba degradation pathway in R. dicambivorans Ndbn-20; DsmD is specifically responsible for cleavage of 3-chlorogentisate, whereas GtdA is a general GDO involved in the catabolism of various natural aromatic compounds.

Dicamba-Based Herbicides: Herbicidal Ionic Liquids versus Commercial Forms.

Dicamba is a widely applied herbicide for crop protection and has potential for volatility. New formulations containing dicamba with greatly reduced volatility, introduced to the market in 2017, still caused foliar injury to crops and other plants in Arkansas and neighboring states in the United States. In response, we proposed the transformation of dicamba into protic as well as aprotic dicamba-based organic salts called herbicidal ionic liquids (HILs). All of the HILs were characterized by high stability, whereas the biological activity of the most effective products, evaluated during greenhouse studies, was found to be greater than that of currently used commercial analogues. Furthermore, the possibility of introducing an alkyl chain of a specific length allows one to obtain plant protection products with the desired physicochemical properties while maintaining herbicidal effectiveness. These studies are expected to aid in the design and development of new herbicidal formulations, which, depending on the weed species, could increase the efficacy of the applied active ingredient. Simultaneously, the volatility of the synthesized compounds, particularly those containing quaternary ammonium cations, was multiple times lower than that of the free acid of dicamba. This strategy minimizes the risk of off-site movement via volatilization, which may cause significant damage to neighboring broadleaf crops and pose a threat to existing ecosystems.

Hapten Synthesis, Antibody Development, and a Highly Sensitive Indirect Competitive Chemiluminescent Enzyme Immunoassay for Detection of Dicamba.

Although dicamba has long been one of the most widely used selective herbicides, some U.S. states have banned the sale and use of dicamba because of farmers complaints of drift and damage to nonresistant crops. To prevent illegal use of dicamba and allow monitoring of nonresistant crops, a rapid and sensitive method for detection of dicamba is critical. In this paper, three novel dicamba haptens with an aldehyde group were synthesized, conjugated to the carrier protein via a reductive-amination procedure and an indirect competitive chemiluminescent enzyme immunoassay (CLEIA) for dicamba was developed. The assay showed an IC50 of 0.874 ng/mL which was over 15 times lower than that of the conventional enzyme immunoassay. The immunoassay was used to quantify dicamba concentrations in field samples of soil and soybean obtained from fields sprayed with dicamba. The developed CLEIA showed an excellent correlation with LC-MS analysis in spike-and-recovery studies, as well as in real samples. The recovery of dicamba ranged from 86 to 108% in plant samples and from 105 to 107% in soil samples. Thus, this assay is a rapid and simple analytical tool for detecting and quantifying dicamba levels in environmental samples and potentially a great tool for on-site crop and field monitoring.

Synthesis of biomass tar-derived foams through spontaneous foaming for ultra-efficient herbicide removal from aqueous solution.

Pyrolysis is one of the most important approaches to convert waste biomass into renewable energy and biomaterials, and the tar is the inevitable by-product of this process. In this study, carbon foams were prepared innovatively with biomass tar as the precursor through spontaneous gas foaming approach and used for dicamba removal from aqueous solution. The results showed that prepared carbon foams had unique properties including rich microporous structure and high specific surface area (reaching 1667 m2/g). In addition, the prepared carbons had high thermal stability due to the high graphitic degree. The adsorption results indicated that pH showed a great effect on the adsorption of dicamba onto the prepared carbon foams. The carbon foam exhibited ultra-fast dicamba removal and ultra-high adsorption capacity of 891.74 mg/g at room temperature. The adsorption process was well described by pseudo-second-order kinetics and Langmuir isotherm models. The thermodynamic study indicated dicamba adsorption onto the prepared carbon foams was a spontaneous and exothermic process. In addition, the good reusability from recovery test demonstrated that the prepared carbon foams had promising potential for dicamba removal from aqueous solution.

Synergism of mixtures of dicamba and 2,4-dichlorophenoxyacetic acid herbicide formulations on the neotropical fish Cnesterodon decemmaculatus (Pisces, Poeciliidae).

Dicamba (DIC) and 2,4-dichlorophenoxyacetic acid (2,4-D) are two of the most applied auxinic herbicides worldwide, both individually and as part of a mixture. However, the toxicity and interactions achieved when applied as a mixture have not yet been characterised. The equitoxic and non-equitoxic acute toxicity exerted by binary mixtures of Banvel® (57.71% DIC) and DMA® (58.4% 2,4-D) on the Neotropical fish Cnesterodon decemmaculatus were evaluated. Results revealed mean values of 1.02 (range, 0.96-1.08) for the toxic unit (TU) that induced 50% mortality (TU50 96 h) to the fish exposed to binary equitoxic mixtures of the commercial formulations Banvel®-DMA®. These results suggest that the mixture is nearly concentration additive. Furthermore, results demonstrated the occurrence of synergistic interaction when non-equitoxic combinations of Banvel®-or DMA®-formulated herbicides were assayed. In this context and regardless of their concentrations, either Banvel®- or DMA®-induced toxicity were synergised by the presence of the counterpart within mixtures. The present study represents the first evidence of the lethality exerted by mixtures of two auxinic herbicides-namely, DIC and 2,4-D-reported to date for fish and other biotic matrices. When C. decemmaculatus is used as the target organism, a synergistic pattern is observed following exposure to a mixture of both herbicides.

Alkyl(C16, C18, C22)trimethylammonium-Based Herbicidal Ionic Liquids.

In the framework of this study a synthesis methodology and characterization of long alkyl herbicidal ionic liquids (HILs) based on four commonly used herbicides (2,4-D, MCPA, MCPP, and dicamba) are presented. New HILs were obtained with high efficiency (>95%) using an acid-base reaction between herbicidal acids and hexadecyltrimethylammonium, octadecyltrimethylammonium, and behenyltrimethylammonium hydroxides in alcoholic medium. Among all synthesized salts, only three compounds comprising the MCPP anion were liquids at room temperature. Subsequently, the influence of both the alkyl chain length and the anion structure on their physicochemical properties (thermal decomposition profiles, solubility in 10 representative solvents, surface activity, density, viscosity, and refractive index) was determined. All HILs exhibited high thermal stability as well as surface activity; however, their solubility notably depended on both the length of the carbon chain and the structure of the anion. The herbicidal efficacy of the obtained salts was tested in greenhouse and field experiments. Greenhouse testing performed on common lambsquarters (Chenopodium album L.) and flixweed (Descurainia sophia L.) as test plants indicated that HILs were characterized by similar or higher efficacy compared to commercial herbicides. The results of field trials confirmed the high activity of HILs, particularly those containing phenoxyacids as anions (MCPA, 2,4-D, and MCPP).

Influence of oligomeric herbicidal ionic liquids with MCPA and Dicamba anions on the community structure of autochthonic bacteria present in agricultural soil.

The aim of this study was to evaluate the impact of selected herbicidal ionic liquids (HILs), which exhibit high efficacy in terms of weed control and low toxicity, but may be persistent due to limited biodegradability, on the community structure of autochthonic bacteria present in agricultural soil. Four different oligomeric HILs (with two types of cations and different ratio of herbicidal anions) were synthesized and characterized by employing (1)H and (13)C NMR. The results of biodegradation assay indicated that none of the tested HILs could be classified as readily biodegradable (biodegradation rate ranged from 0 to 7%). The conducted field studies confirmed that the herbicidal efficacy of the HILs was higher compared to the reference herbicide mixture by 10 to 30%, depending on the dose and weed species. After termination of field studies, the soil treated with the tested HILs was subjected to next generation sequencing in order to investigate the potential changes in the bacterial community structure. Proteobacteria was the dominant phylum in all studied samples. Treatment with the studied HILs resulted in an increase of Actinobacteria compared to the reference herbicidal mixture. Differenced among the studied HILs were generally associated with a significantly higher abundance of Bacteroidetes in case of 1-HIL-Dicamba 1/3 and Firmicutes in case of 2-HIL-Dicamba 1/3.

Glyphosate and dicamba herbicide tank mixture effects on native plant and non-genetically engineered soybean seedlings.

Crops engineered to contain genes for tolerance to multiple herbicides may be treated with several herbicides to manage weeds resistant to each herbicide. Thus, nearby non-target plants may be subjected to increased exposure to several herbicides used in combination. Of particular concern are native plants, as well as adjacent crops which have not been genetically engineered for tolerance to herbicides. We evaluated responses of seven species of native plants grown in a greenhouse and treated less than field application rates of glyphosate and/or dicamba: Andropogon gerardii, Asclepias syriaca, Eutrochium purpureum, Oenothera biennis, Polyganum lapathifolium, Solidago canadensis and Tridens flavus, and non-herbicide resistant soybean (Glycine max, Oregon line M4). Herbicide concentrations were 0.03 or 0.1 × field application rates of 1122 g ha(-1) active ingredient (a.i) (831 g ha(-1) acid glyphosate) for glyphosate and 562 g ha(-1) a.i. for dicamba. In general, plant growth responses to combinations of glyphosate and dicamba were less than the sum of growth responses to the individual herbicides (i.e., antagonistic effect), primarily when one or both herbicides alone caused a large reduction in growth. E. purpureum, P. lapathifolium and S. canadensis were the most sensitive species to both herbicides, while A. gerardii was the most tolerant, with no response to either herbicide. The combinations of herbicides resulted in responses most similar to that from dicamba alone for G. max and from glyphosate alone for T. flavus. The results of this study indicated the need for more data such as effects on native plants in the field to assess risks to non-target plants from combinations of herbicides.

Modeling of degradation kinetic and toxicity evaluation of herbicides mixtures in water using the UV/H2O2 process.

The UV/H2O2 process was applied to the treatment of different mixtures of herbicides in water. Glyphosate, the herbicide most used in the world, was mixed with other hormonal herbicides with residual activity as 2,4-D and dicamba. The main goals of the study were to develop a kinetic model for interpreting the simultaneous oxidation of two mixtures (glyphosate plus 2,4-D and glyphosate plus dicamba). The model is based on a complete reaction mechanism, which comprises hydrogen peroxide photolysis and decomposition of both herbicides in each mixture studied. It takes into account the effect of non-uniform distribution of the local rate of absorbed photons. Good agreement of experimental data and the model is achieved in spite of differences in the reactivity between glyphosate and 2,4-D (or dicamba). Toxicity assays (employing Vibrio fischeri) were also performed, indicating that the toxicity of the mixture of glyphosate and 2,4-D was significantly reduced after the treatment.

On-line monitoring of the photocatalytic degradation of 2,4-D and dicamba using a solid-phase extraction-multisyringe flow injection system.

A fully automated on-line system for monitoring the photocatalytic degradation of herbicides was developed using multisyringe flow injection analysis (MSFIA) coupled to a solid phase extraction (SPE) unit with UV detection. The calibration curves were linear in the concentration range of 100-1000 µg L(-1) for 3,6-dichloro-2-methoxybenzoic acid (dicamba) and 500-3000 µg L(-1) for 2,4-dichlorophenoxyacetic acid (2,4-D), while the detection limits were 30 and 135 µg L(-1) for dicamba and 2,4-D, respectively. The monitoring of the photocatalytic degradation (TiO2 anatase/UV 254 nm) of these two herbicides was performed by MSFIA-SPE system using a small sample volume (2 mL) in a fully automated approach. The degradation was assessed in ultrapure and drinking water with initial concentrations of 1000 and 2000 µg L(-1) for dicamba and 2,4-D, respectively. Degradation percentages of approximately 85% were obtained for both herbicides in ultrapure water after 45 min of photocatalytic treatment. A similar degradation efficiency in drinking water was observed for 2,4-D, whereas dicamba exhibited a lower degradation percentage (75%), which could be attributed to the presence of inorganic species in this kind of water.

O-Demethylations catalyzed by Rieske nonheme iron monooxygenases involve the difficult oxidation of a saturated C-H bond.

Dicamba monooxygenase (DMO) catalyzes the O-demethylation of dicamba (3,6-dichloro-2-methoxybenzoate) to produce 3,6-dichlorosalicylate and formaldehyde. Recent crystallographic studies suggest that DMO catalyzes the challenging oxidation of a saturated C-H bond within the methyl group of dicamba to form a hemiacetal intermediate. Testing of this hypothesis was made possible by our development of two new independent techniques. As a novel method to allow use of (18)O2 to follow reaction products, bisulfite was used to trap newly formed (18)O-formaldehyde in the stable adduct, hydroxymethanesulfonate (HMS(-)), and thereby prevent the rapid exchange of (18)O in formaldehyde with (16)O in water. The second technique utilized unique properties of Pseudomonas putida formaldehyde dehydrogenase that allow rapid conversion of (18)O-formaldehyde into stable and easily detectable (18)O-formic acid. Experiments using these two new techniques provided compelling evidence for DMO-catalyzed oxidation of the methyl group of dicamba, thus validating a mechanism for DMO [and for vanillate monooxygenase, a related Rieske nonheme iron monooxygenase] that involves the difficult oxidation of a saturated C-H bond.

Dissipation of six acid herbicides in water and sediment of two Canadian prairie wetlands.

In the present study, an ephemeral (E) and a semipermanent (SP) wetland were divided into halves using a polyvinyl curtain and one-half of each wetland was treated with dicamba (3,6-dichloro-o-anisic acid), bromoxynil (3,5-dibromo-4-hydroxy-benzonitrile), MCPA [(4-chloro-2-methylphenoxy)acetic acid], 2,4-D [(2,4-dichlorophenoxy)acetic acid], mecoprop-P (R)-2-(4-chloro-o-tolyloxy)propionic acid], and dichlorprop [(RS)-2-(2,4-dichlorophenoxy)propionic acid] such that concentrations in the water simulated an overspraying event, thus representing a worst-case scenario for wetland contamination. Water and sediment samples were taken over the 77-d study period to monitor herbicide concentrations. The dissipation of all six herbicides could be described by first-order reaction kinetics. In water, the field half-life (DT50) values ranged from 2.3 d (bromoxynil) to 31 d (dichlorprop). All six herbicides were detected in sediment samples from both wetlands. Overall, the phenoxypropionic acids (mecoprop-P and dichlorprop) were more persistent than the phenoxyacetic acids (2,4-D and MCPA) in both sediment and water. Use of bromide ion as a conservative tracer indicated that infiltration through sediment was an important route of water loss in both wetlands, especially in wetland E. Because strong correlations were found between the mass of each herbicide and bromide ion mass in wetland SP (r(2) = 0.59-0.76) and wetland E (r(2) = 0.80-0.95), it is likely that herbicide dissipation was due, in part, to mass lost by way of infiltration through sediment.

Simultaneous determination of three acidic herbicide residues in food crops using HPLC and confirmation via LC-MS/MS.

2,4-D, dicamba and 4-CPA with auxin-like activity have been intensively used in agriculture, for the control of unwanted broadleaf weeds. An analytical method involving HPLC coupled with UVD was developed for the simultaneous analysis of these three analytes in Chinese cabbage, apple and pepper fruits (representative non-fatty samples) and brown rice and soybean (representative fatty samples) using liquid-liquid partitioning and column cleanup procedures. The residues were confirmed via tandem mass spectrometry (MS/MS) in ion electrospray ionization (ESI) mode. The standard curves were linear over the range of the tested concentrations (0.25-10 microg/mL), as shown by a marked linearity in excess of 0.9999 (r(2) ). The average recoveries (mean, n = 3) ranged from 94.30 to 102.63 in Chinese cabbage, from 94.76 to 108.47 in apple, from 97.52 to 102.27 in pepper, from 76.19 to 101.90 in brown rice, and from 74.60 to 107.39 in soybean. The relative standard deviations (RSDs) were <9% in all tested matrices. The limits of detection and quantitation were 0.006 and 0.02 mg/kg, respectively. Samples purchased from local markets were analyzed to evaluate the applicability of the methods developed herein. The concentration of the 2,4-D residue was measured at 0.102 mg/kg in the soybean sample; however, this level is exactly the same MRL set by the Korea Food and Drug Administration. This developed method deserves full and complete consideration, as it clearly displays the sensitivity, accuracy and precision required for residue analysis of 2,4-D, dicamba and 4-CPA in food crops.

Binding of dicamba to soluble and bound extracellular polymeric substances (EPS) from aerobic activated sludge: a fluorescence quenching study.

Binding of dicamba to soluble EPS (SEPS) and bound EPS (BEPS) from aerobic activated sludge was investigated using fluorescence spectroscopy. Two protein-like fluorescence peaks (peak A with Ex/Em=225 nm/342-344 nm and peak B with Ex/Em=275/340-344 nm) were identified in SEPS and BEPS. Humic-like fluorescence peak C (Ex/Em=270-275 nm/450-460 nm) was only found in BEPS. Fluorescence of the peaks A and B for SEPS and peak A for BEPS were markedly quenched by dicamba at all temperatures whereas fluorescence of peaks B and C for BEPS was quenched only at 298 K. A dynamic process dominated the fluorescence quenching of peak A of both SEPS and BEPS. Fluorescence quenching of peak B and C was governed a static process. The effective quenching constants (logK(a)) were 4.725-5.293 for protein-like fluorophores of SEPS and 4.23-5.190 for protein-like fluorophores of BEPS, respectively. LogK(a) for humic-like substances was 3.85. Generally, SEPS had greater binding capacity for dicamba than BEPS, and protein-like substances bound dicamba more strongly than humic-like substances. Binding of dicamba to SEPS and BEPS was spontaneous and exothermic. Electrostatic force and hydrophobic interaction forces play a crucial role in binding of dicamba to EPS.