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The use of recently described ionisation techniques for the rapid analysis of some common drugs and samples of biological origin.

Three ionisation techniques that require no sample preparation or extraction prior to mass analysis have been used for the rapid analysis of pharmaceutical tablets and ointments. These methods were (i) the novel direct analysis in real time (DART), (ii) desorption electrospray ionisation (DESI), and (iii) desorption atmospheric pressure chemical ionisation (DAPCI). The performance of the three techniques was investigated for a number of common drugs. Significant differences between these approaches were observed. For compounds of moderate to low polarity DAPCI produced more effective ionisation. Accurate DESI and DAPCI tandem mass spectra were obtained and these greatly enhance the selectivity and information content of the experiment. The detection from human skin of the active ingredients from ointments is reported together with the detection of ibuprofen metabolites in human urine.

Alkaloids↗

Liquid chromatography/electrospray ionization tandem mass spectrometry analysis of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX).

A quantitative liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method was developed for the analysis of the explosive, octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX). In negative ionization mode, HMX forms an acetate adduct ion [M + CH(3)COO](-), m/z 355, in the presence of a small amount of acetic acid in the mobile phase. The ESI collision-induced dissociation (CID) spectrum of m/z 355 was acquired and the transitions m/z 355 --> 147 and m/z 355 --> 174 were chosen for the determination of HMX in samples. Using this quantification technique, the method detection limit was 1.57 microg/L and good linearity was achieved in the range 5-500 microg/L. This method will help to unambiguously analyze environmentally relevant concentrations of HMX.

Azocines↗

Fragmentation study of hexanitrostilbene by ion trap multiple mass spectrometry and analysis by liquid chromatography/mass spectrometry.

The fragmentation pathways of three explosive compounds with similar structures, hexanitrostilbene (HNS), cyclotrimethylene trinitramine (RDX), and 2,4,6-trinitrotoluene (TNT), have been investigated by multiple mass spectrometry (MSn, n = 1, 2, 3) with electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) sources. The electron capture mechanism for these compounds in negative ion APCI and ESI mode differs from the usual negative ion mechanism, deprotonation or addition of other species. This was shown for HNS and TNT, which both gave a [M]- anion but not a [M-H]- ion in APCI, and the [M]- anion of HNS was observed in ESI. The quantitative analysis of HNS was performed by liquid chromatography (LC)/ESI-MS, and the results obtained by the internal standard (ISTD) method were compared with those from the external standard (ESTD) method, demonstrating that both quantitation approaches are useful, with good sensitivity, reproducibility and linearity, and ESTD is preferable in routine applications.

Chromatography, High Pressure Liquid↗

Vitreoretinal surgery of the posterior segment for explosive trauma in terrorist warfare.

BACKGROUND: To describe surgical management and establish anatomic and visual results of patients with explosive ocular trauma in terrorist attacks treated with extreme vitreoretinal surgery. METHODS: Retrospective study of clinical records (6-month follow-up) of patients with visual acuity (VA) of light perception or better with posterior segment injuries [vitreous hemorrhage, retinal detachment (RD), intra-ocular foreign bodies (IOFB), perforating trauma (PT)] from explosive weapons who underwent vitreoretinal surgery. We reviewed the demographic characteristics, type of weapon, time between injury and surgery, VA at arrival and 6 months after surgery, and type of trauma according to the International Trauma Classification. RESULTS: Fifty-seven out of 236 patients with ocular injuries from explosive weapons were included in the study; all of them were military men, average age 22 years (range 16-53 years). The average time between the blast and primary closing was 1 day, and 10 days between primary closing and vitreoretinal surgery. Open traumas by laceration accounted for 96% of cases and 4% were closed traumas; 76% of the eyes had IOFB, of which 18% involved PT; 5% had endophthalmitis. Contusion was the diagnosis for 100% of the closed traumas. Of the open traumas, 40% were localized at zone I, 44% at zone II, and 16% at zone III. Upon arrival, 98% of patients had VA 20/800-LP and 2% had >20/40. The patients with closed trauma had the injuries at zone III and presented VA 20/800-LP. All patients underwent posterior vitrectomy, scleral buckling, endotaponade and when required, lensectomy (82%), IOFB removal (72%), and/or retinectomy (25%). Postoperative VA improved in 43% of the patients, stabilized in 41% and evolved to NLP in 15% of the cases. Initial expressions of ocular trauma such as RD, PT and endophthalmitis suggest bad prognosis. CONCLUSIONS: We presented a series of patients with severe ocular trauma of the posterior segment from explosive weapons. These patients were treated according to our surgical protocol with extreme vitreoretinal surgery within the first 2 weeks after the blast; with our procedure we obtained stabilization or improvement of the VA for 84% of the cases.

Adolescent↗

An epidemic of acute haemorrhagic conjunctivitis caused by enterovirus 70 in Okinawa, Japan, in 1994.

BACKGROUND: Although enterovirus 70 (EV70) has been identified as the major aetiological agent of acute haemorrhagic conjunctivities (ACH),no EV70 strain has been isolated by cell culture method since 1988. Therefore, recent clinical and epidemiological characteristics of AHC caused by EV70 have not been clarified. METHODS: Clinical and serological studies were carried out on patients during the AHC epidemic in Okinawa, Japan, in 1994 in which 7509 cases were reported by national epidemiological surveillance. EV70 was confirmed as the causative agent by reverse-transcription polymerase chain reaction. RESULTS: The 11-15 years age group contained the highest number of cases (62% of the total). Conjunctival hyperaemia was present in all patients, and subconjunctival haemorrhage, superficial punctate keratitis and preauricular lymphadenopathy were present in 24.0%, 11.7% and 9.3% of AHC cases, respectively. No neurological complication was observed in this epidemic. Out of 31 paired serum samples, 10 pairs showed a fourfold rise in antibody level to EV70. None of the paired serum samples showed a fourfold rise in antibody level to Coxsackie A24 variant virus. CONCLUSION: These findings demonstrate that the clinical features of AHC observed in this study were milder than those reported previously, in contrast to the high transmission rate during an epidemic. Changes in clinical features of AHC, such as a low incidence of subconjunctival haemorrhage and disappearance of neurological complications, might be due to biological transformation of EV70. It should be noted that EV70 is still an important aetiological agent of explosive epidemics of AHC.

Adolescent↗

Blast injuries from Madrid terrorist bombing attacks on March 11, 2004.

Blast injuries after terrorist attacks are seen with increasing frequency worldwide. Thousands of victims were attended in the hospitals of Madrid, Spain, on March 11, 2004 after the bombing attacks against the commuter trains. Thirty-six patients were attended in our institution. Seventeen of them suffered from severe or life-threatening injuries, and 19 had mild injuries. The most common lesions were thoracic trauma and blast lung injury, acoustic trauma, and orbital and paranasal sinus fractures. Other findings were hepatic and splenic lacerations, and vertebral and limb fractures. Emergency radiology had an important role in the correct management of the victims. Prompt radiological diagnoses of these complex lesions are crucial to efficient treatment. Therefore, radiologists have to become familiar with the injury patterns and specific lesions caused by blast wave.

Adolescent↗

Growth rate and nutrient limitation affect the transport of Rhodococcus sp. strain DN22 through sand.

Rhodococcus strain DN22 grows on the nitramine explosive RDX as a sole nitrogen source, and is potentially useful for bioremediation of explosives-contaminated soil. In order for strain DN22 to be effectively applied in situ, inoculum cells must reach zones of RDX contamination via passive transport, a process that is difficult to predict at field-scale. We examined the effect of growth conditions on the transport of DN22 cells through sand columns, using chemostat-grown cultures. Strain DN22 formed smaller coccoid cells at low dilution rate (0.02 h(-1)) and larger rods at high dilution rate (0.1 h(-1)). Under all nutrient limitation conditions studied, smaller cells grown at low dilution rate were retained more strongly by sand columns than larger cells grown at high dilution rate. At a dilution rate of 0.05, cells from nitrate-limited cultures were retained more strongly than cells from RDX-limited or succinate-limited cultures. Breakthrough concentrations (C/C (0)) from sand columns ranged from 0.04 (nitrate-limited, D=0.02 h(-1)) to 0.98 (succinate-limited, D=0.1 h(-1)). The observed strong effect of culture conditions on transport of DN22 cells emphasizes the importance of physiology studies in guiding the development of bioremediation technologies.

Biodegradation, Environmental↗

Biodegradation kinetics of the nitramine explosive CL-20 in soil and microbial cultures.

The cyclic nitramine explosive CL-20 (C(6)H(6)N(12)O(12), 2,4,6,8,10,12-hexanitro-2,4,6,8,10,12 -hexaazaisowurtzitane) is a relatively new energetic compound which could be a persistent organic pollutant. To follow its biodegradation dynamics, CL-20 was added to soil alone or together with organic co-substrates and N-source and incubated under oxic and anoxic conditions. Without co-substrates, the CL-20 degradation was detectable only under anoxic conditions. The highest degradation rate was found under aerobic conditions and with the addition of co-substrates, succinate and pyruvate being more efficient than acetate, glucose, starch or yeast extract. When added to intact soil, CL-20 degradation was not affected by the N content, but in soil serially diluted with N-free succinate-mineral medium, the process became N-limited. About 40% of randomly selected bacterial colonies grown on succinate agar medium were able to decompose CL-20. Based on 16S rDNA gene sequence and cell morphology, they were affiliated to Pseudomonas, Rhodococcus, Ochrobactrum, Mycobacterium and Ralstonia. In the pure culture of Pseudomonas sp. MS-P grown on the succinate-mineral N(+) medium, the degradation kinetics were first order with the same apparent kinetic constant throughout growth and decline phases of the batch culture. The observed kinetics agreed with the model that supposes co-metabolic transformation of CL-20 uncoupled from cell growth, which can be carried out by several constitutive cellular enzymes with wide substrate specificity.

Aerobiosis↗

A morphological investigation of soot produced by the detonation of munitions.

The morphology of three different detonation soot samples along with other common soot materials such as carbon black, diesel soot and chimney soot was studied by elemental and proximate analysis, X-ray diffraction and electron microscopy. The goal of this study was to better define the morphology of the detonation soot in order to better assess the interactions of this type of soot with explosive residues. The detonation soot samples were obtained by the detonation of artillery 155mm projectiles filled with either pure TNT (2,4,6-trinitrotoluene) or composition B, a military explosive based on a mixture of TNT and RDX (trimethylentrinitramine). The carbon content of the soot samples varied considerably depending on the feedstock composition. Detonation soot contains less carbon and more nitrogen than the other carbonaceous samples studied, due to the molecular structure of the energetic materials detonated such as TNT and RDX. The ash concentration was higher for detonation soot samples due to the high metal content coming from the projectiles shell and to the soil contamination which occurred during the detonation. By X-ray diffraction, diamond and graphite were found to be the major crystalline carbon forms in the detonation soot. Two electron microscopy techniques were used in this study to visualise the primary particles and to try to explain the formation mechanism of detonation soot samples.

Carbon↗

Anaerobic biotransformation of explosives in aquifer slurries amended with ethanol and propylene glycol.

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), and 2,4,6-trinitrotoluene (TNT) are explosives that are frequently found as environmental contaminants on military installations. Hydrogen has been shown to support the anaerobic transformation of these explosives. We investigated ethanol and propylene glycol as electron donors for providing syntrophically produced H2 for stimulating the anaerobic biodegradation of explosives in contaminated soil. The study was conducted using anoxic microcosms constructed with slurries of the contaminated soil and groundwater. The addition of 5mM ethanol and propylene glycol enhanced the biodegradation of RDX and HMX relative to the control bottles. Ethanol was depleted within about 20 days, resulting in the transient formation of hydrogen, acetate, and methane. The hydrogen headspace concentration increased from 8 ppm to 1838 ppm before decreasing to background concentrations. Propylene glycol was completely degraded after 15 days, forming hydrogen, propionate, and acetate as end-products. The hydrogen headspace concentrations increased from 56 ppm to 628 ppm before decreasing to background concentrations. No methane formation was observed during the incubation period of 48 days. Our findings indicate the addition of ethanol and propylene to the aquifer slurries increased the hydrogen concentrations and enhanced the biotransformation of RDX and HMX in the explosive-contaminated soil.

Anaerobiosis↗

Degradation of explosives-related compounds using nickel catalysts.

We report the ability of nickel-based catalysts to degrade explosives compounds in aqueous solution. Several nickel catalysts completely degraded the explosives, although rates varied. Nearly all of the organic explosive compounds tested, including 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), were rapidly degraded to below detection limits by a powdered nickel on an alumina-silicate support (Aldrich nickel catalyst). Perchlorate degradation was minimal (<25%). Degradation of TNT by Aldrich nickel catalyst resulted in apparent first-order kinetics. Significant gaseous 14C was released and collected in an alkaline solution (most likely carbon dioxide) from [14C]RDX and [14C]HMX, indicating heterocyclic ring cleavage. Significant gaseous 14C was not produced from [14C]TNT, but spectrophotometric evidence indicated loss of aromaticity. Degradation occurred in low ionic strength solutions, groundwater, and from pH 3 to pH 9. Degradation of TNT, RDX, and HMX was maintained in flow-through columns of Aldrich nickel catalyst mixed with sand down to a hydraulic retention time of 4h. These data indicate that nickel-based catalysts may be an effective means for remediation of energetics-contaminated groundwater.

Azocines↗

On-line strategies for determining trace levels of nitroaromatic explosives and related compounds in water.

We report the development and tests of several systems for the simultaneous determination of 18 energetic compounds and related congeners in untreated water samples. In these systems a Restricted Access Material trap or liquid-chromatography precolumn (with a C(18) or porous graphitic carbon, PGC, stationary phase) followed by a PGC analytical column are used for sample clean-up, enrichment and separation of the trace level analytes, which are then analyzed by mass spectrometry (MS). The relative merits of two MS ionization interfaces (atmospheric pressure chemical ionization, APCI, and atmospheric pressure photoionization, APPI) were also compared for the MS identification and quantification of these analytes. APCI was found to be superior in cases where both alternatives are applicable. A major drawback when applying APPI is that no signal is obtained for the cyclic nitramines and nitrate esters. Using APCI, a wide spectrum of unstable compounds can be determined in a single analysis, and the feasibility of using large volume samples (up to 100 mL) in combination with the sensitivity of the MS detection system provide method detection limits ranging from 2.5 pg/mL (for 2,4-dinitrotoluene and 2,6-diamino-6-nitrotoluene) to 563 pg/mL (for pentaerythritol tetranitrate, PETN), with repeatability ranging from 2 to 7%. Other chemometric parameters such as robustness, selectivity, repeatability, and intermediate precision were also evaluated in the validation of the extraction methods for use in water analysis. Tests with untreated groundwater and drinking water samples, spiked with 20 ng of the analytes, yielded results similar to those obtained with high purity water samples.

Analytic Sample Preparation Methods↗

Effects of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) metabolites on cricket (Acheta domesticus) survival and reproductive success.

The effect of two major hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) metabolites, hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX) and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX), on cricket (Acheta domesticus) survival and reproduction was studied. RDX metabolites did not have adverse effects on cricket survival, growth, and egg production. However, MNX and TNX did affect egg hatching. MNX and TNX were more toxic in spiked-sand than in topical tests. TNX was more toxic to egg than MNX. Developmental stage and exposure time affected hatching. After 30 days exposure to MNX or TNX, the EC20, EC50, and EC95 were 47, 128, and 247 microg/g for TNX, and 65, 140, and 253 microg/g for MNX in topical tests. The ECs for 20, 50, and 95 were 21, 52, and 99 microg/g for MNX, and 12, 48, and 97 microg/g for TNX in sand. No gross abnormalities in cricket nypmhs were observed in all experiments indicating that neither TNX or MNX is teratogenic in this assay.

Animals↗

Phytotoxicity to and uptake of RDX by rice.

Phytoremediation is an emerging strategy to remediate soils contaminated with pollutants like explosives in which plants will uptake, degrade and/or accumulate pollutants. To implement this technology on a site contaminated with RDX, we chose rice, which is able to grow in lagoons, and we tested its ability to grow in soils with high levels of RDX and to decrease RDX concentrations in soil. Rice was grown for 40 days in soil contaminated with increasing [14C]RDX concentrations. Emergence and growth were not affected by RDX. Total chlorophyll content decreased with RDX concentrations of over 500 mg kg(-1). Amounts of chlorophyll were correlated with the appearance of necrosis in leaf extremities. After 40 days, rice translocated 89% of uptaken radioactivity to leaves with 90% in leaf extremities. Analyzes of leaf extracts showed that 95% of radioactivity was RDX in its parent form. Necrosis appears to be a phytotoxic symptom of RDX accumulation.

Biodegradation, Environmental↗

High uptake of 2,4,6-trinitrotoluene by vetiver grass--potential for phytoremediation?

2,4,6-Trinitrotoluene (TNT) is a potent mutagen, and a Group C human carcinogen that has been widely used to produce munitions and explosives. Vast areas that have been previously used as ranges, munition burning, and open detonation sites are heavily contaminated with TNT. Conventional remediation activities in such sites are expensive and damaging to the ecosystem. Phytoremediation offers a cost-effective, environment-friendly solution, utilizing plants to extract TNT from contaminated soil. We investigated the potential use of vetiver grass (Vetiveria zizanioides) to effectively remove TNT from contaminated solutions. Vetiver grass plants were grown in hydroponic systems containing 40 mg TNTL(-1) for 8d. Aqueous concentrations of TNT reached the method detection limit ( approximately 1 microg L(-1)) within the 8-d period, demonstrating high affinity of vetiver for TNT, without any visible toxic effects. Results from this preliminary hydroponic study are encouraging, but in need of verification using TNT-contaminated soils.

Biodegradation, Environmental↗

Successive explosions in Mumbai the economic center of India.

Terrorist activities in India are increasing day by day with sophistication in modus operandi. Mumbai the economic center of India was attacked by a series of bomb blasts at twelve different places within a span of an hour on 12th March 1993. The main explosive used was RDX [Krishnamurthy R, Malve MK, Shinde BM. J Indian Acad Forensic Sci 1996;35(1& 2):46-61.]. After about 10 years, terrorist activity of late has again erupted taking a toll on innocent lives, with the use of explosives causing death and destruction. On 2.12.02 a public bus at Ghatkopar was blown up by an improvised explosive device (IED) with two casualties. On 27.1.03 the public vegetable market at Vileparle was targeted causing heavy damage and panic among common people. On 13.3.03 a fully packed local train compartment at Mulund railway station was blown up by an improved explosive device and the casualties ranged up to 10. In most of the explosions the explosives used were RDX, NC-NG, etc. The blasts that occurred at the Zaveri bazaar and the gateway of India on 25/8/03 showed the presence of big craters at the blast site and on analysis the presence of RDX and petroleum oil.

Blast Injuries↗

Dissolution kinetics of high explosives particles in a saturated sandy soil.

Solid phase high explosive (HE) residues from munitions detonation may be a persistent source of soil and groundwater contamination at military training ranges. Saturated soil column tests were conducted to observe the dissolution behavior of individual components (RDX, HMX, and TNT) from two HE formulations (Comp B and C4). HE particles dissolved readily, with higher velocities yielding higher dissolution rates, higher mass transfer coefficients, and lower effluent concentrations. Effluent concentrations were below solubility limits for all components at superficial velocities of 10-50 cm day(-1). Under continuous flow at 50 cm day(-1), RDX dissolution rates from Comp B and C4 were 34.6 and 97.6 microg h(-1) cm(-2) (based on initial RDX surface area), respectively, significantly lower than previously reported dissolution rates. Cycling between flow and no-flow conditions had a small effect on the dissolution rates and effluent concentrations; however, TNT dissolution from Comp B was enhanced under intermittent-flow conditions. A model that includes advection, dispersion, and film transfer resistance was developed to estimate the steady-state effluent concentrations.

Algorithms↗

Comparison of biotic and abiotic treatment approaches for co-mingled perchlorate, nitrate, and nitramine explosives in groundwater.

Biological and abiotic approaches for treating co-mingled perchlorate, nitrate, and nitramine explosives in groundwater were compared in microcosm and column studies. In microcosms, microscale zero-valent iron (mZVI), nanoscale zero-valent iron (nZVI), and nickel catalyzed the reduction of RDX and HMX from initial concentrations of 9 and 1 mg/L, respectively, to below detection (0.02 mg/L), within 2 h. The mZVI and nZVI also degraded nitrate (3 mg/L) to below 0.4 mg/L, but none of the metal catalysts were observed to appreciably reduce perchlorate ( approximately 5 mg/L) in microcosms. Perchlorate losses were observed after approximately 2 months in columns of aquifer solids treated with mZVI, but this decline appears to be the result of biodegradation rather than abiotic reduction. An emulsified vegetable oil substrate was observed to effectively promote the biological reduction of nitrate, RDX and perchlorate in microcosms, and all four target contaminants in the flow-through columns. Nitrate and perchlorate were biodegraded most rapidly, followed by RDX and then HMX, although the rates of biological reduction for the nitramine explosives were appreciably slower than observed for mZVI or nickel. A model was developed to compare contaminant degradation mechanisms and rates between the biotic and abiotic treatments.

Aniline Compounds↗