Concentrations Of Sarin Research Articles

UiO-66-NH2 initiated cascade reaction: Constructing a ratiometric fluorescence sensor for ultrasensitive detection of nerve agent simulant

BackgroundHighly toxic organophosphorus nerve agents often exist in the form of gas in the environment and can damage human neuroregulatory system by inhibiting the activity of acetylcholinesterase (AChE). However, fluorescent probes based on small organic molecules bring a secondary burden to environment, and their sensitivity and specificity for sarin simulant diethyl chlorophosphate (DCP) detection are unsatisfactory. Nanozyme cascade systems with signal amplification can be used for highly sensitive identification of analytes, but are rarely used in ratiometric analysis of DCP. Combination of enzyme cascades and ratiometric fluorescence ensures the accuracy and sensitivity of the output signal. ResultsWe prepared a self-assembled nanohybrid (Ag–AuNCs@UiO-66-NH2) by metal-organic framework material and gold nanoclusters. On the one hand, UiO-66-NH2 with enzyme-like activity was used to hydrolyze DCP into diethyl phosphate (DEP) and chloridion (Cl−). Cl− hindered aggregation-induced enhanced emission (AIEE) of AuNCs by binding with Ag+ and decreased the fluorescence of AuNCs. On the other hand, ligand metal charge transfer effect (LMCT) of UiO-66-NH2 was blocked by DCP to enhance the fluorescence of UiO-66-NH2. Combining ratiometric analysis and nanozyme cascade reaction, an ultra-sensitive fluorescence sensor for detecting DCP was constructed, and ensured the accuracy of experimental results. In addition, Ag–AuNCs@UiO-66-NH2 was embedded into the agarose hydrogel substrate, the resulting agarose hydrogel film allowed quantitative assessment of DCP vapor and high sensitivity was demonstrated (detection limit as low as 1.02 ppb). SignificanceA strategy combining enzyme cascade with ratiometric fluorescence was proposed, which improved the accuracy and sensitivity of the analysis results. The soft-solid platform based on agarose hydrogel film was constructed to realize the quantitative monitoring of sarin simulant gas. The LOD value obtained in this work is much lower than the immediately life-threatening or health threatening concentration of sarin.

Improving Quantification of tabun, sarin, soman, cyclosarin, and sulfur mustard by focusing agents: A field portable gas chromatography-mass spectrometry study

Commercial gas chromatograph-mass spectrometers, one of which being Inficon's HAPSITE® ER, have demonstrated chemical detection and identification of nerve agents (G-series) and blistering agents (mustard gas) in the field; however most analyses relies on self-contained or external calibration that inherently drifts over time. We describe an analytical approach that uses target-based thermal desorption standards, called focusing agents, to accurately calculate concentrations of chemical warfare agents that are analyzed by gas chromatograph-mass spectrometry. Here, we provide relative response factors of focusing agents (2-chloroethyl ethyl sulfide, diisopropyl fluorophosphate, diethyl methylphosphonate, diethyl malonate, methyl salicylate, and dichlorvos) that are used to quantify concentrations of tabun, sarin, soman, cyclosarin and sulfur mustard loaded on thermal desorption tubes (Tenax® TA). Aging effects of focusing agents are evaluated by monitoring deviations in quantification as thermal desorption tubes age in storage at room temperature and relative humidity. The addition of focusing agents improves the quantification of tabun, sarin, soman, cyclosarin and sulfur mustard that is analyzed within the same day as well as a 14-day period. Among the six focusing agents studied here, diisopropyl fluorophosphate has the best performance for nerve agents (G-series) and blistering agents (mustard gas) compared to other focusing agents in this work and is recommended for field use for quantification. The use of focusing agent in the field leads to more accurate and reliable quantification of Tabun (GA), Sarin (GB), Soman (GD), Cyclosarin (GF) and Sulfur Mustard (HD) than the traditional internal standard. Future improvements on the detection of chemical, biological, radiological, nuclear, and explosive materials (CBRNE) can be safely demonstrated with standards calibrated for harmful agents.

A New Portable Colorimetric Sensor Based on RGB Chromaticity for Quantitative Determination of Sarin in Water

Background: Sarin is a nerve agent which is lethal to people due to its high toxicity. According to its extreme toxicity, sarin, relatively lack of color, highly toxic, miscible in water, poses viable threats to potable water sources. Therefore, there is an urgent need for portable, rapid and yet reliable methods to monitor for adulteration of potable water sources by sarin on spot. Methods: A stock solution of 30 mg/L sarin was prepared daily by dissolving 300 μg of sarin in 10 mL isopropanol. A certain amount of sarin was added to the glass tube, and then o-dianisidine and hydrogen peroxide were added. The pH value of the solution was adjusted to 9.8. The solution was transferred to the test tube after 10 minutes. A test tube of 2 mL was placed between the light source and the RGB color sensor. The LED light source illuminates directly over the test tube while the RGB sensor obtained the generated spectral response. This RGB voltage output is connected to the ADC and microcontroller to convert these analog voltages to three digital data. This RGB digital data is linked to the microcomputer through the serial port that is interfaced with the user interface. The data thus obtained in the sensor can be processed to display the sarin concentration. Results: Under the optimum conditions as described above, the calibration curve of chromaticity value versus sarin concentration was linear in the range of 0.15 mg/L to 7.8 mg/L. According to the IUPAC definition, theoretical detection limits of this method were 0.147 mg/L and 0.140 mg/L for R and B values, respectively. The practical detection limit was 0.15 mg/L. The sensor was successfully applied to the determination of sarin in artificial water samples and the recoveries were between 86.0% to 95.9%. Conclusion: The results in the present work have demonstrated the feasibility to design a new portable colorimetric sensor based on the RGB chromaticity method for quantitative determination of sarin in water. The influences of chromogenic reagent, oxidant, reaction time, o-dianisidine concentration, hydrogen peroxide concentration, reaction temperature, pH on the chromaticity values were investigated. The results showed that the sensor possessed high selectivity, sensitivity and good repeatability. The method would be potentially applied to the analysis of other toxic compounds in environment, such as other chemical warfare agents.

Structural and kinetic evidence of aging after organophosphate inhibition of human Cathepsin A

Human Cathepsin A (CatA) is a lysosomal serine carboxypeptidase of the renin-angiotensin system (RAS) and is structurally similar to acetylcholinesterase (AChE). CatA can remove the C-terminal amino acids of endothelin I, angiotensin I, Substance P, oxytocin, and bradykinin, and can deamidate neurokinin A. Proteomic studies identified CatA and its homologue, SCPEP1, as potential targets of organophosphates (OP). CatA could be stably inhibited by low µM to high nM concentrations of racemic sarin (GB), soman (GD), cyclosarin (GF), VX, and VR within minutes to hours at pH 7. Cyclosarin was the most potent with a kinetically measured dissociation constant (KI) of 2 µM followed by VR (KI = 2.8 µM). Bimolecular rate constants for inhibition by cyclosarin and VR were 1.3 × 103 M−1sec−1 and 1.2 × 103 M−1sec−1, respectively, and were approximately 3-orders of magnitude lower than those of human AChE indicating slower reactivity. Notably, both AChE and CatA bound diisopropylfluorophosphate (DFP) comparably and had KIDFP = 13 µM and 11 µM, respectively. At low pH, greater than 85% of the enzyme spontaneously reactivated after OP inhibition, conditions under which OP-adducts of cholinesterases irreversibly age. At pH 6.5 CatA remained stably inhibited by GB and GF and <10% of the enzyme spontaneously reactivated after 200 h. A crystal structure of DFP-inhibited CatA was determined and contained an aged adduct. Similar to AChE, CatA appears to have a “backdoor” for product release. CatA has not been shown previously to age. These results may have implications for: OP-associated inflammation; cardiovascular effects; and the dysregulation of RAS enzymes by OP.

Determination of free G-type nerve agents in blood: in situ derivatization on a dried blood spot (DBS) paper followed by LC–MS/MS analysis

G-type nerve agents, including sarin, soman and cyclosarin are potent neurotoxic chemical weapons, sarin of which is the most used chemical warfare agent of our time. These compounds have limited stability, are highly reactive in the body and thus require the detection of stable metabolites, mainly their hydrolysis products or adducts to proteins. The aim of this study was to develop a new method for the determination of trace amounts of intact G-type nerve agents from small amounts of whole blood. The G-agents were derivatized on a dried blood spot (DBS) paper with 2-((dimethylamino)methyl)phenol. After drying, they were extracted from the paper and analyzed by liquid chromatography–tandem mass spectrometry (LC–MS/MS) or Orbitrap LC–MS/MS. Recoveries of ~ 34% were obtained in the range of 3–300 ng/mL with good linear response. A detection limit as low as 0.7 ng/mL in whole blood was demonstrated. Derivatized sarin was stable on the DBS paper for 19 days at room temperature before analysis. The method was applied for the short-time-course determination of free sarin in mouse blood after in vivo intranasal exposure to 3LD50 sarin. A fast-time-course decay (t1/2: ~ 2 min) was observed, with blood sarin concentrations in the range of 6−60 ng/mL in the post-exposure time range of 1−7 min. This method is simple and unique, enabling the monitoring of G-agents before their rapid conversion in the body. The technique can serve for the study of very fast decay mechanisms of drugs and toxic materials in the body.

Ocular surface histopathological insult following sarin and VX exposure and potential treatments in the rat model

Eye exposure to organophosphate (OP) chemical warfare irreversible acetylcholinesterase inhibitors, results in long-term miosis and impaired visual function. In contrast to the well-documented miotic and ciliary muscle spasm observed following chemical warfare, OP ocular exposure, little is known regarding the ocular surface histopathological insult. The aim of the present study was to determine the degree of the ocular surface insult following sarin or VX ocular exposure and to evaluate potential anti-cholinergic treatments in counteracting this insult.Rats that were whole body exposed to various sarin concentrations (0.049–43 μg/L; 5 min exposure), showed a dose-dependent miotic response and light reflex impairment.Following whole body sarin exposure, a dose dependent ocular surface histopathological insult was developed. A week following exposure to a low concentration of 0.05 μg/L, conjunctival pathology was observed, while corneal insult was noticed only following exposure to a concentration of 0.5 μg/L and above. Both tissues presented poorer outcomes when exposed to higher sarin concentrations. In contrast, eyes topically exposed to 1 μg sarin demonstrated no ocular insult a week following exposure. On the contrary, topical exposure to 1 μg VX resulted in a significant corneal insult. Anticholinergic treatments such as 0.1% atropine or 2% homatropine, given shortly following VX exposure, counteracted this insult.The results of this study show that not only do anti-cholinergic treatments counteract the miotic response, but also prevent the histopathological insult observed when given shortly following OP exposure.

Acetylcholinesterase-functionalized two-dimensional photonic crystal for the sensing of G-series nerve agents.

G-series nerve agents, such as sarin, tabun, and soman, would cause tremendous harm in military and terrorist attacks, so it is necessary to develop a simple method for the rapid and efficient detection of these hazardous substances. We have developed a tunable acetylcholinesterase (AChE)-functionalized two-dimensional photonic crystal (2D PhC) for the detection of a real nerve agent, sarin. In accordance with the 2D PhC previously prepared by our group, the AChE-functionalized 2D PhC was optimized by adjustment of the amount of monomer in the hydrogel, which not only increased the sensitivity of the 2D PhC, with the detection limit decreasing by two orders of magnitude, but also ensured the structural color spanned the whole visible region in the detection range. A linear relationship between the logarithm of the sarin concentration and the particle spacing of the AChE-functionalized 2D PhC was observed from 7.1 × 10-17 to 7.1 × 10-4 mol/L. The AChE-functionalized 2D PhC also responded to mimics of G-series nerve agents, including dimethyl methylphosphonate, diisopropyl methylphosphonate, and isodipropyl methylphosphonate, to various degrees. The proposed 2D-PhC hydrogel has potential for low-cost, trace-level, and on-site monitoring of other G-series nerve agents. Graphical abstract.

Search for Optimal Parameters for Gas Chromatographic Complex When Determining Sarin in Water Samples by Flame Photometric Detection Method

Sarin is considered to be an instrument for chemical terror by the extremist organizations due to the availability of technologies and components for its production. This fact became evident after the use of sarin by several members of the extremist organization «Aum Shinrikyo» for air infection of the capital`s subway in 1995 in Japan and by ISIL bands in Syria in 2013. Because of its physical and chemical properties, as well as its ability to be dissolved in water in any ratios, sarin can infect water sources for the long period of time. But the sensitivity of flame photometric detectors (FPD) is not always sufficient for the analysis of water samples for the presence of sarin, its precursors and products of destruction, especially in cases of necessity to detect the concentrations less than 0,03 ppm (3×10 -5 mg/ml). We have found out that higher sensitivity of the detector can be achieved in case of the increase of the gas carrier (helium) pressure in the injector up to 25 psi, and the temperature – to 250 °C, single-segment temperature rise in the thermostat with a speed of 10 °C/min from 40 °C (1 min) to 280 °C (5 min) at a hydrogen stream in PFD – 75 ml/min, air – 100 ml/min, helium – 60 ml/min. In comparison with the initial method, the offered one is more sensitive in case of sarin concentration in water in the range from 6×10 -6 to 1×10 -4 mg/ml to 1,7 times. The approbation of the offered optimal parameters was carried out during the gas chromatographic detection of sarin in water samples during the OPCW Proficiency Tests, during the analysis of arbitration samples, during the scientific research and the obtaining of national accreditation of the Laboratory of Chemical Analytical Control of the 27 Scientific Centre of the Minstry of Defence of the Russian Federation. These parameters can be used successfully during the studies of any organophosphorous chemicals by GC analysis with flame photometric detection

Maximum acceptable concentration levels of the chemical agent sarin in Canadian and Russian soil types

The lowest-observed-adverse-effect and no-observed-adverse-effect concentrations of sarin were determined in soils typical to Canada and Russia. Maximum acceptable concentrations were established to be 3.0 × 10−4 mg kg−1 for the standard reference soil, 2.0 × 10−3 mg kg−1 for prairie soil, and 1.0 × 10−2 mg kg−1 for forest soil.

Younger rats are more susceptible to the lethal effects of sarin than adult rats: 24 h LC50 for whole-body (10 and 60 min) exposures

Chemical warfare nerve agents (CWNA) inhibit acetylcholinesterase and are among the most lethal chemicals known to man. Children are predicted to be vulnerable to CWNA exposure because of their smaller body masses, higher ventilation rates and immature central nervous systems. While a handful of studies on the effects of CWNA in younger animals have been published, exposure routes relevant to battlefield or terrorist situations (i.e. inhalation for sarin) were not used. Thus, we estimated the 24 h LC50 for whole-body (10 and 60 min) exposure to sarin using a stagewise, adaptive dose design. Specifically, male and female Sprague-Dawley rats were exposed to a range of sarin concentrations (6.2–44.0 or 1.6–12.5 mg/m³) for either 10 or 60 min, respectively, at six different times during their development (postnatal day [PND] 7, 14, 21, 28, 42 and 70). For male and female rats, the lowest LC50 values were observed for PND 14 and the highest LC50 values for PND 28. Sex differences were observed only for PND 42 for the 10 min exposures and PND 21 and 70 for the 60 min exposures. Thus, younger rats (PND 14) were more susceptible than older rats (PND 70) to the lethal effects of whole-body exposure to sarin, while adolescent (PND 28) rats were the least susceptible and sex differences were minimal. These results underscore the importance of controlling for the age of the animal in research on the toxic effects associated with CWNA exposure.

Selective Surface Acoustic Wave-Based Organophosphorus Sensor Employing a Host-Guest Self-Assembly Monolayer of β-Cyclodextrin Derivative.

Self-assembly and molecular imprinting technologies are very attractive technologies for the development of artificial recognition systems and provide chemical recognition based on need and not happenstance. In this paper, we employed a β-cyclodextrin derivative surface acoustic wave (SAW) chemical sensor for detecting the chemical warfare agents (CWAs) sarin (O-Isoprophyl methylphosphonofluoridate, GB). Using sarin acid (isoprophyl hydrogen methylphosphonate) as an imprinting template, mono[6-deoxy-6-[(mercaptodecamethylene)thio]]-β-cyclodextrin was prepared by self-assembled method on one of the SAW oscillators. After templates’ removal, a sensitive and selective molecular imprinting (MIP) monolayer for GB was prepared. Electrochemical impedance spectroscopy and atomic force microscope (AFM) were used to characterize this film. Comparing the detection results to GB by MIP film and non-MIP film, the molecularly imprinting effect was also proved. The resulting SAW sensor could detect sarin as low as 0.10 mg/m3 at room temperature and the frequency shift was about 300 Hz. The response frequency increased linearly with increasing sarin concentration in the range of 0.7 mg/m3~3.0 mg/m3. When sarin was detected under different temperatures, the SAW sensor exhibited outstanding sensitivity and reliability.

Synthesis and evaluation of a new phthalocyanine as sensor material for sarin detection

A new phthalocyanine material, 2,9,16,23-tetrakis [4-(hexafluoroisopropanol)-anilinomethyl]phthalocyaninato zinc (ZnPcF24N4O4) was designed and synthesized. The chemoselective material was coated on a 300MHz surface acoustic wave (SAW) device using cyclic voltammetry for the detection of sarin (GB) vapor. It was found that the observed frequency shifts were linear to the concentrations of sarin in the range of 0.6–43.4mg/m3 with a high sensitivity of 205.8Hz/mg/m3. The influences of other interfering vapors on the sensor response were also examined. The results showed that the phthalocyanine-based SAW sensor exhibited a fairly fast response, excellent reversibility, linearly sensitivity, and a high selectivity to vapor of the organophosphorus nerve agent GB.

Interactions of Sarin with Polyelectrolyte Membranes: A Molecular Dynamics Simulation Study

Nanostructured polyelectrolyte membranes (PEMs), which are widely used as permselective diffusion barriers in fuel cell technologies and electrochemical processing, are considered as protective membranes suitable for blocking warfare toxins, including water-soluble nerve agents such as sarin. In this article, we examine the mechanisms of sorption and diffusion of sarin in hydrated PEMs by means of atomistic molecular dynamics simulations. Three PEMs are considered: Nafion, sulfonated polystyrene (sPS) that forms the hydrophilic subphase of segregated sPS-polyolefin block copolymers, and random sPS-polyethylene copolymer. We found that sarin concentrates at the interface between the hydrophilic and hydrophobic subphases of hydrated Nafion acting as a surfactant. In hydrated sPS, where the scale of water-polymer segregation is much smaller (1-2 nm), sarin also interacts favorably with hydrophobic and hydrophilic components. Water diffusion slows as the sarin content increases despite the overall increase in solvent content, which suggests that sarin and water have somewhat different pathways through the segregated membrane. Upon replacement of counterions of monovalent potassium with those of divalent calcium, sarin diffusion slows but remains substantial in all ionomers considered, especially at high sarin concentrations. The behavior of sarin is similar to that of its common simulant, dimethyl methylphosphonate.

A comparative analysis of mathematical models for relating indoor and outdoor toxic gas concentrations in accidental releases

This paper surveys various models for estimating indoor concentration as a function of outdoor concentration in the event of an accidental release or chemical attack involving toxic substances. It is essential to know indoor concentration in order to estimate the effectiveness of shelter in place as a protective measure. Several models—deposition, one-sink, sink-diffusion and two-sink—were considered. These models can be derived from a general model by making different assumptions. The models showed significant variations in terms of the adsorption/desorption considerations. Since indoor materials act as reservoirs, adsorption may lead to a significant decrease in indoor concentration, but subsequent desorption may also take place. The models require the knowledge of a set of parameters that are specific to each compound and material, which are currently scarce in the literature. As a result, the more complex and complete the model, the more limited its applicability. Outdoor concentrations obtained from a Gaussian model and originating from three source types (continuous, temporary and instantaneous) were used as inputs in the reviewed models. Indoor concentrations of chlorine and sarin from a temporary source were estimated in order to compare the predictions of the models.

Advantages of the WRAIR whole blood cholinesterase assay: Comparative analysis to the micro-Ellman, Test-mate ChE™, and Michel (ΔpH) assays

Red blood cell AChE (RBC-AChE) and plasma BChE can be used as sensitive biomarkers to detect exposure to OP nerve agents, pesticides, and cholinergic drugs. In a comparative study, RBC-AChE and serum BChE activities in whole blood was obtained from forty seven healthy male and female human volunteers, and then exposed separately ex vivo to three OP nerve agents (soman (GD), sarin (GB) and VX) to generate a wide range of inhibition of AChE and BChE activity (up to 90% of control). These samples were measured using four different ChE assays: (i) colorimetric microEllman (using DTNB at 412 nm), (ii) Test-mate ChE field kit (also based on the Ellman assay), (iii) Michel (delta pH), and (iv) the Walter Reed Army Institute of Research Whole Blood (WRAIR WB) cholinesterase assay. The WRAIR assay is a modified Ellman method using DTP at 324 nm (which minimizes hemoglobin interference and improves sensitivity), and determines AChE and BChE in a small whole blood sample simultaneously. Scatter plots of RBC-AChE activities were determined using the WRAIR ChE assay versus the micro-Ellman, Test-mateTM and Michel after exposure to varying concentrations of soman, sarin and VX. Regression analyses yielded mostly linear relationships with high correlations (r2 = 0.83–0.93) for RBC-AChE values in the WRAIR assay compared to the alternate methods. For the plasma BChE measurements, individual human values were significantly more variable (as expected), resulting in lower correlations using WRAIR ChE versus the alternate assays (r2 values 0.5 – 0.6). To circumvent the limitations of simple correlation analysis, Bland and Altman analysis for comparing two independent measurement techniques was performed. For example, a Bland and Altman plot of the ratio of the WRAIR whole blood AChE and Michel AChE (plotted on the y-axis) vs. the average of the two methods (x-axis) shows that the majority of the individual AChE values are within ± 1.96 S.D. of the mean difference, indicating that the two methods may be used interchangeably with a high degree of confidence. The WRAIR ChE assay can be thus be used as a reliable inter-conversion assay when comparing results from laboratory-based (Michel) and field-based (Test-mateTM ChE kit), which use different methodology and report in different units of AChE activity.

Determination of miosis threshold from whole-body vapor exposure to sarin in African green monkeys

We determined the threshold concentration of sarin vapor exposure producing miosis in African green monkeys ( Chlorocebus aethiops). Monkeys ( n = 8) were exposed to a single concentration of sarin (0.069–0.701 mg/m 3) for 10 min. Changes in pupil size were measured from photographs taken before and after the exposure. Sarin EC 50 values for miosis were determined to be 0.166 mg/m 3 when miosis was defined as a 50% reduction in pupil area and 0.469 mg/m 3 when miosis was defined as a 50% reduction in pupil diameter. Monkeys were also evaluated for behavioral changes from sarin exposure using a serial probe recognition test and performance remained essentially unchanged for all monkeys. None of the concentrations of sarin produced specific clinical signs of toxicity other than miosis. Sarin was regenerated from blood sampled following exposure in a concentration-dependent fashion. Consistent with a predominant inhibition of acetylcholinesterase (AChE), more sarin was consistently found in RBC fractions than in plasma fractions. Further, elimination of regenerated sarin from RBC fractions was slower than from plasma fractions. Blood samples following exposure also showed concentration-dependent inhibition of AChE activity and, to a lesser extent, butyrylcholinesterase activity. At the largest exposure concentration, AChE inhibition was substantial, reducing activity to approximately 40% of baseline. The results characterize sarin exposure concentrations that produce miosis in a large primate species in the absence of other overt signs of toxicity. Further, these results extend previous studies indicating that miosis is a valid early indicator for the detection of sarin vapor exposure.

Changes of Cholinesterase Activities in the Rat Blood and Brain After Sarin Intoxication Pretreated with Butyrylcholinesterase

After sarin inhalation exposure of rats pretreated with equine serum butyrylcholinesterase (EqBuChE), cholinesterase activities of the whole blood, acetylcholinesterase (AChE) in erythrocytes, pontomedullar area, frontal cortex, and striatum of the brain, and plasma butyrylcholinesterase (BuChE) were determined. Using different doses of EqBuChE as a pretreatment (intraperitoneal injection), dose-dependent increases in plasma BuChE activity and no changes in the erythrocyte and brain AChE activities were demonstrated. Decreases in plasma BuChE activity and red blood cells (RBC) and brain AChE activities were observed in control rats after sarin inhalation exposure without EqBuChE pretreatment. In rats pretreated with EqBuChE, this inhibition was lower compared with control animals not only in the blood but also in the brain structures studied. These results demonstrate protective effects of EqBuChE pretreatment in rats intoxicated with sublethal concentrations of sarin by inhalation.

Evaluation of cognitive and biochemical effects of low-level exposure to sarin in rhesus and African green monkeys

We investigated the potential of low-level exposures to the chemical warfare nerve agent, sarin, to produce adverse effects. Rhesus ( Macaca mulatta) and African green monkeys ( Chlorocebus acthiops) were trained on a serial probe recognition (SPR) task before IM administration of a low-level concentration (5.87 μg/kg or 2.93 μg/kg) of sarin. Blood was sampled before agent administration and at various times following administration. Sarin administration did not disrupt performance on the SPR task in either species. Major dependent measures characterizing performance (accuracy, number of completed trials per session, average choice response time) were largely unaffected on the day sarin was administered as well as on subsequent testing sessions occurring over several weeks following administration. Analyses of red blood cell (RBC) and plasma samples revealed that sarin administration produced a substantial degree of inhibition of circulating acetylcholinesterase (AChE) in RBC fractions and butyrylcholinesterase (BChE) in plasma fractions, which only slowly recovered. In this regard, AChE activity was inhibited to a greater extent than BChE activity. Blood samples were also evaluated for regenerated sarin, which was found in RBC and plasma fractions in both species and showed orderly elimination functions. More sarin was regenerated from RBC fractions than from plasma fractions. Elimination of regenerated sarin was much slower in RBC than plasma and exceeded the expected time of AChE aging, suggesting the presence of additional sarin binding sites. In general, effects were similar in both species. Taken together, our results show that while the concentrations of sarin administered were clearly biochemically active, they were below those that are required to produce a disruption of behavioral performance.

Detection of sarin in plasma of rats after inhalation intoxication

Presently used methods for detection and diagnosis of the severity of intoxication with organophosphorus (OP) compounds are mostly those that quantify inhibition of blood cholinesterases. It was found that when plasma inhibited with OP compounds is incubated in the presence of a high concentration of fluoride ions, the organophosphate is released from the enzyme thus yielding a phosphofluoridate, which can be analyzed by gas chromatography and NP detection. In our study, the concentration of sarin released after fluoride ions were added to the plasma of sarin-poisoned rats was determined. Sarin amounts in plasma measured after refluoridation and plasma butyrylcholinesterase activity in ten rats, that were exposed to sarin vapors at concentration of 1.25 μg/L (E1 group) and 2.5 μg/L (E2 group) respectively, for 60 min. In the E2 group the concentration of sarin in plasma was nearly 2-fold higher than in the E1 group. These results correspond well with the concentrations of sarin vapors to which the animals were exposed. Both experimental groups of animals showed significant decreases in butyrylcholinesterase activity by more than 30%–36.4% (E1 group) and 47.0% (E2 group). The method of fluoride-induced reactivation provides a very good marker for monitoring sarin intoxication in laboratory animals determined previously mostly by ChE determination which does not allow any information on sarin amounts in plasma.

Changes of acetylcholinesterase activity after long-term exposure to sarin vapors in rats

In our study, rats were exposed to sarin vapors for 240 min at four different concentrations (0.30, 0.43, 0.58 and 0.82 microg/L) in a whole-body inhalation chamber. The acetylcholinesterase activity (AChE, EC 3.1.1.7) was measured in the whole blood, frontal cortex (FC), pontomedullar area (PM) and basal ganglia (BG). Convulsions and hypersalivation were observed in only one animal of the group exposed to the highest sarin concentration. The decrease in blood AChE activity was significant in all animals exposed to sarin vapors. The highest inhibition of AChE activity (61%) was determined in animals exposed to sarin vapors at a concentration of 0.82 microg/L. In the PM, AChE activity was decreased in all experimental groups, significantly only in the group exposed to sarin vapors at a concentration of 0.58 microg/L. Our results show that in long-term exposure to low concentrations of sarin, the significant decrease in AChE activity in the blood is followed by significant changes of AChE activity in the PM only. This part of the brain seems to be more sensitive than the FC or BG.