Quantification of sarin and cyclosarin metabolites isopropyl methylphosphonic acid and cyclohexyl methylphosphonic acid in minipig plasma using isotope-dilution and liquid chromatography- time-of-flight mass spectrometry.

An analysis method for determining isopropyl methylphosphonic acid (IMPA) and cyclohexyl methylphosphonic acid (CMPA), the metabolic hydrolysis products of toxic organophosphorus nerve agents isopropyl methylphosphonofluoridate (sarin, GB) and cyclohexyl methylphosphonofluoridate (cyclosarin, GF), respectively, has been developed and validated using high-performance liquid chromatography-mass spectrometry with negative ion electrospray ionization with time-of-flight detection (LC-ESI-MS-TOF). The linear range of quantitation was 5 to 125 ng/mL in plasma with a method detection limit of 2 ng/mL for each compound. This method was developed to determine the amount of metabolic hydrolysis that was formed during and after nerve agent exposure in minipigs to account for a major pathway of GB and GF elimination that had not been previously characterized in the bloodstream, particularly during low-level whole-body inhalation experiments. Metabolic hydrolysis accounted for 70% to 90% of the recoverable agent in the bloodstream during exposure, when compared to both unbound and cholinesterase bound agent recovered by fluoride ion reactivation analysis for the same samples. The estimated half-life of IMPA and CMPA in plasma was determined to be 44 and 61 min, respectively. The method utilizes the mass selectivity of LC-ESI-MS-TOF using a bench-top instrument to achieve a detection limit that is consistent with reported LC-MS-MS methods analyzing blood samples.

Quantitation of fluoride ion released sarin in red blood cell samples by gas chromatography-chemical ionization mass spectrometry using isotope dilution and large-volume injection.

A new method for measuring fluoride ion released isopropyl methylphosphonofluoridate (sarin, GB) in the red blood cell fraction was developed that utilizes an autoinjector, a large-volume injector port (LVI), positive ion ammonia chemical ionization detection in the SIM mode, and a deuterated stable isotope internal standard. This method was applied to red blood cell (RBC) and plasma ethyl acetate extracts from spiked human and animal whole blood samples and from whole blood of minipigs, guinea pigs, and rats exposed by whole-body sarin inhalation. Evidence of nerve agent exposure was detected in plasma and red blood cells at low levels of exposure. The linear method range of quantitation was 10-1000 pg on-column with a detection limit of approximately 2-pg on-column. In the course of method development, several conditions were optimized for the LVI, including type of injector insert, injection volume, initial temperature, pressure, and flow rate. RBC fractions had advantages over the plasma with respect to assessing nerve agent exposure using the fluoride ion method especially in samples with low serum butyrylcholinesterase activity.

The toxicity of brass dust to the microalgae Ankistrodesmus falcatus and Selenastrum capricornutum.

The toxicity of brass dust was examined by conducting 96 h growth inhibition tests. Two species of algae were used, Ankistrodesmus falcatus (EC50 = 0.316 mg brass/l) and Selenastrum capricornutum (EC50 = 0.056 mg brass/l). Brass dissociates into two components, Cu (68.5%) and Zn (27.5%). Enhanced algal growth was exhibited at concentrations of 0.01 and 0.001 mg brass/l. Available literature on the toxicity of copper to S. capricornutum (EC50 = 0.047 mg Cu/l), indicate that the toxicity of brass dust is due to the ionized copper. Reported toxicities of zinc are orders of magnitude lower than copper. The ionization of the brass is dependent on pH and hardness. The literature cites cases in which copper toxicity varies with pH, clay content and dissolved organics. At present little is known of the fate and distribution of brass dust upon the release into the environment. However, the presence of heavy metals has consistently been shown to impact aquatic systems.

Acute toxicity of brass particles to Daphnia magna.

The aquatic toxicity of brass particles was examined. Acute, 48 hour bioassays were performed using the water flea, Daphnia magna. Tests were conducted with uniform suspensions of uncoated brass particles, brass particles coated with a Teflon solution, silica particles, and titanium dioxide particles. The Teflon coating solution and the supernatant of the brass suspension (after settling of the brass) also were tested. All tests were conducted according to guidelines set forth by the US Environmental Protection Agency and the Organization for Economic Cooperation and Development. Mean EC50 determinations of 20.0 micrograms l-1 and 23.6 micrograms l-1 were calculated for uncoated brass particles and coated brass particles, respectively. The silica, titanium dioxide, and Teflon each had an EC50 greater than 1 g l-1. Chemical fate studies demonstrated that the brass dissociated to its ionic components of copper and zinc quickly at pH 2.0. At pH 5.0 and 6.5, the dissociation occurred too slowly to account for the observed toxicity. The data suggested that the toxicity is due to filtration by the daphnids and subsequent ingestion. EC50 determinations for the brass particles are nearly identical with published EC50 values for copper salts.

Chemical and toxicological evaluation of pyrotechnically disseminated terephthalic acid smoke.

The terephthalic acid (TPA) smoke obscurants (M-83 grenade and M-8 smoke pot) were developed by the U.S. Army for training purposes to replace the more toxic hexachloroethane (HC) smoke. Inhalation toxicity testing and chemical characterization of pyrotechnically generated TPA was conducted to assess the health hazard potential of TPA and its combustion products. Fisher 344 rats were subjected to acute and repeated exposures to TPA smoke generated from the M-83 grenade. Acute exposure levels ranged from 150-1,900 mg/m3 for 30 minutes and repeated dose exposures ranged from 128-1,965 mg/m3 for 30 min/day for 5 days. Exposed and control rats were evaluated for toxic signs, and histopathologic changes. During exposure, the rats exhibited slight to moderate lacrimation, rhinorrhea, lethargy and dyspnea, which reversed within 1-hr post-exposure. No deaths occurred, even at the highest smoke concentrations. Histopathological changes were confined to exposure related nasal necrosis and inflammation in both the acute and repeated dose exposures at levels above 900 mg/m3. Chemical characterization of the M-83 grenade and the M-8 smoke pot showed that formaldehyde, benzene and carbon monoxide were the major organic vapor by-products formed. These by-products were above their respective ACGIH threshold limit values at various concentrations, but should not pose a hazard if the smoke is deployed in an open area. Overall, TPA is a safer training smoke to replace the HC smoke.