Odd Loop Regions of XenA and XenB Enzymes Modulate Their Interaction with Nitro-explosives Compounds and Provide Structural Support for Their Regioselectivity.

The nitro-explosive compounds 2,4,6-trinitrotoluene, 2,4,6-trinitrophenol, and 1,2,3-trinitroglycerol are persistent environmental contaminants. The presence of different functional groups in these molecules represents a great challenge to enzymatic catalysis. The chemical variety of these three substrates is such that they do not bind and interact with catalytic residues within an enzyme with the same affinity. In this context, two Xenobiotic Reductase enzymes produced by the bacteria Pseudomonas putida can catalyze the reduction of these compounds with different affinities and regioselectivity. The structural bases that support this substrate promiscuity and catalytic preferences are unknown. Therefore, through molecular dynamics simulations and free energy calculations, we explored the structural properties driving the specific interactions of these enzymes with their substrates and cofactor. Models of Xenobiotic Reductase A and B enzymes in complex with 2,4,6-trinitrotoluene, 2,4,6-trinitrophenol, or 1,2,3-trinitroglycerol were built, and the ligand enzyme interaction was simulated by molecular dynamics. The structural analysis of the molecular dynamics simulations shows that loops 3, 5, 7, 9, 11, and 13 of Xenobiotic Reductase B, and loops 4, 5, 7, 11, 13, and 15 Xenobiotic Reductase A, are in contact with the ligands during the first stages of the molecular recognition. These loops are the most flexible regions for both enzymes; however, Xenobiotic Reductase B presents a greater range of movement and a higher number of residues interacting with the ligands. Finally, the distance between the cofactor and the different reactive groups in the substrate reflects the regioselectivity of the enzymes, and the free energy calculations are consistent with the substrate specificity of both enzymes studied. The simulation shows a stable interaction between the aromatic ring of the substrates and Xenobiotic Reductase B. In contrast, a less stable interaction with the different nitro groups of the aromatic ligands was observed. In the case of 1,2,3-trinitroglycerol, Xenobiotic Reductase B interacts more closely with the nitro groups of carbon 1 or 3, while Xenobiotic Reductase A is more selective by nitro groups of carbon 2. The obtained results suggest that the flexibility of the loops in Xenobiotic Reductase B and the presence of polar and aromatic residues present in loops 5 and 7 are fundamental to determine the affinity of the enzyme with the different substrates, and they also contribute to the proper orientation of the ligands that directs the catalytic reaction.

Rapid separation of post-blast explosive residues on glass electrophoresis microchips.

This study describes the development of an analytical methodology based on the use of microchip electrophoresis (ME) devices integrated with capacitively coupled contactless conductivity detection (C4 D) for the separation and detection of inorganic anions in post-blast explosive residues. The best separation condition was achieved using a running buffer composed of 35 mmol/L lactic acid, 10 mmol/L histidine and 0.070 mmol/L cetyl(trimethyl ammonium) bromide. For C4 D measurements, the highest sensitivity was obtained applying a 700 kHz sinusoidal wave with excitation voltage of 20 Vpp . The separation of Cl- , NO3- , NO2- , SO42- , ClO4- and ClO3- was performed within ca. 150 s with baseline resolution and efficiencies between 4.4 × 104 and 1.7 × 105 plates/m. The found limits of detection ranged between 2.5 and 9.5 µmol/L. Last, real samples of post-blast explosive residues were analyzed on the ME-C4 D devices obtaining successfully the determination of Cl- , NO3- and SO42- . The achieved concentration values varied between 12.8-72.5 mg/L for Cl- , 1.7-293.1 mg/L for NO3- and 1.3-201.3 mg/L for SO42- . The data obtained using ME-C4 D devices were in good agreement with the concentrations found by ion chromatography. The approach reported herein has provided short analysis time, instrumental simplicity, good analytical performance and low cost. Furthermore, the ME-C4 D devices emerge as a powerful and portable analytical platform for on-site analysis demonstrating to be a promising tool for the crime scene investigation.

Fate and transport of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and its degradation products in sedimentary and volcanic rocks, Los Alamos, New Mexico.

High-explosive compounds including hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) were used extensively in weapons research and testing at Los Alamos National Laboratory (LANL). Liquid effluents containing RDX were released to an outfall pond that flowed to Cañon de Valle at LANL's Technical Area 16 (TA-16), resulting in the contamination of the alluvial, intermediate and regional groundwater bodies. Monitoring of groundwater within Cañon de Valle has shown persistent RDX in the intermediate perched zone located between 225 and 311 m below ground surface. Monitoring data also show detectable levels of RDX putative anaerobic degradation products. Batch and column experiments were conducted to determine the extent of adsorption-desorption and transport of RDX and its degradation products (MNX, DNX, and TNX) in major rock types that are within the RDX plume. All experiments were performed in the dark using water obtained from a well located at the center of the plume, which is fairly oxic and has a neutral pH of 7.5. Retardation factors and partitioning coefficient (Kd) values for RDX were calculated from batch experiments. Additionally, retardation factors and Kd values for RDX and its degradation products were calibrated from column experiments using a one-dimensional transport model with equilibrium sorption (linear isotherm). Results from the column and batch experiments showed little to no sorption of RDX to the aquifer materials tested, with retardation factors ranging from 1.0 to 1.8 and Kd values varying from 0 to 0.70 L/kg. Results also showed no measurable differences between the transport properties of RDX and its degradation products.

Flow injection analysis of picric acid explosive using a copper electrode as electrochemical detector.

A simple and fast electrochemical method for quantitative analysis of picric acid explosive (nitro-explosive) based on its electrochemical reduction at copper surfaces is reported. To achieve a higher sample throughput, the electrochemical sensor was adapted in a flow injection system. Under optimal experimental conditions, the peak current response increases linearly with picric acid concentration over the range of 20-300 µmol L(-1). The repeatability of the electrode response in the flow injection analysis (FIA) configuration was evaluated as 3% (n=10), and the detection limit of the method was estimated to be 6.0 µmol L(-1) (S/N=3). The sample throughput under optimised conditions was estimated to be 550 samples h(-1). Peroxide explosives like triacetone triperoxide (TATP) and hexamethylene triperoxide diamine (HMTD) were tested as potential interfering substances for the proposed method, and no significant interference by these explosives was noticed. The proposed method has interesting analytical parameters, environmental applications, and low cost compared with other electroanalytical methods that have been reported for the quantification of picric acid. Additionally, the possibility to develop an in situ device for the detection of picric acid using a disposable sensor was evaluated.

Combined zero-valent iron and fenton processes for the treatment of Brazilian TNT industry wastewater.

The environmental impact caused by the production of explosives made from nitroaromatic compounds such as 2,4,6-trinitrotoluene (TNT) is currently a major concern, mainly due to their toxic nature, a fact that makes these compounds highly harmful. This work evaluated a continual system treatment reactor (CSTR) consisting of column zero-valent iron and a system to promote a fenton reaction in order to create possible definitive routines for treating effluents originating from the TNT production process. The spectrophotometric results demonstrated that this combination of processes was highly efficient in promoting the removal of all the absorbed species at 290 nm and the visible region of the specter. The results also revealed that the combination of treatments was significantly efficient in terms of correcting the effluent's main parameters of relevance, mainly COD (95.5% reduction) and TNT concentration, whose total was converted into nitrous and phenolic compounds and, additionally, the acute toxicity was also significantly reduced (95%). These results indicate that the strategy can serve as an efficient option for effluent treatment, for release into the receiving body, or eventually for use as industrial reuse water.

Estimating the TNT equivalence of a 15-ton single base powder explosion through damaged building profiles analyses.

Back in 1964 President Vargas Works was the only place in the country which processed single base powder for the Brazilian Armed Forces. Then its industrial activity was quite strong and around 4:45 a.m. of 23rd September an intense decomposition of nearly 15 ton of that material took place in one of the production lines workshops. The consequences of this explosion were the destruction and extensive damage to the workshops around its epicenter. At that time pictures of all affected buildings were taken and their damages fully described. This led to the present work which consists in the evaluation of the TNT equivalent charge of the explosion using the concept of damage category developed by UK engineers based on the WWII damaging bombing data.