RESUMEN
The verification of exposure to nerve agents is a serious challenge, especially in cases of soman (GD) poisoning. Protein adducts are reliable biomarkers, that provide forensic information and evidence during incidents of terrorism or sporadic poisoning. Mass spectrometry, coupled with a proteomics approach, was established for the forensic analysis of GD-based protein adducts. The fragmentation pathways of GD-based protein adducts were investigated for the first time using electrospray ionization tandem mass spectrometry. Three abundant natural loss product ions, [M+2H-54]2+ (loss of two carbon cations), [M+2H-72]2+ (loss of tert-butyl and methyl moieties), and [M+2H-84]2+ (loss of the pinacolyl moieties), were observed in each of the GD-labeled adducts, and the product ions were independent of protein structure and exposure route. A unique mechanism for the formation of product ions involving GD-protein adducts is proposed here. These findings support the development of a simple and precise forensic analysis technique to rapidly verify GD poisoning using these three GD-related product ions.
Asunto(s)
Biomarcadores/sangre , Sustancias para la Guerra Química/análisis , Sustancias para la Guerra Química/toxicidad , Medicina Legal/métodos , Proteínas/metabolismo , Soman/sangre , Soman/toxicidad , Animales , Modelos Animales de Enfermedad , Exposición a Riesgos Ambientales , Femenino , Humanos , Masculino , Proteómica , Conejos , Espectrometría de Masas en Tándem/métodosRESUMEN
A method was developed to detect and quantify organophosphate nerve agent (OPNA) metabolites in dried blood samples. Dried blood spots (DBS) and microsampling devices are alternatives to traditional blood draws, allowing for safe handling, extended stability, reduced shipping costs, and potential self-sampling. DBS and microsamplers were evaluated for precision, accuracy, sensitivity, matrix effects, and extraction recovery following collection of whole blood containing five OPNA metabolites. The metabolites of VX, Sarin (GB), Soman (GD), Cyclosarin (GF), and Russian VX (VR) were quantitated from 5.0 to 500â¯ngâ¯mL-1 with precision of ≤16% and accuracy between 93 and 108% for QC samples with controlled volumes. For unknown spot volumes, OPNA metabolite concentrations were normalized to total blood protein to improve interpretation of nerve agent exposures. This study provides data to support the use of DBS and microsamplers to collect critical exposure samples quickly, safely, and efficiently following large-scale chemical exposure events.
Asunto(s)
Pruebas con Sangre Seca , Agentes Nerviosos/análisis , Compuestos Organofosforados/sangre , Compuestos Organotiofosforados/sangre , Sarín/sangre , Soman/sangre , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Humanos , Agentes Nerviosos/metabolismo , Compuestos Organofosforados/metabolismo , Compuestos Organotiofosforados/metabolismo , Sarín/metabolismo , Soman/metabolismo , Espectrometría de Masas en TándemRESUMEN
A sensitive method for the purification and determination of two protein adducts, organophosphorus (OP)-BChE and OP-albumin adducts, in a single sample using a simultaneous sample preparation method was developed and validated using liquid chromatography-tandem mass spectrometry. First, we isolated O-ethyl S-2-diisopropylaminoethyl methyl phosphonothiolate (VX) and O-pinacolyl methylphosphonofluoridate (soman, GD)-BChE adducts using an immunomagnetic separation (IMS) method and the HiTrap™ Blue affinity column was subsequently used to isolate and purify VX and GD-albumin adducts from the plasma of rhesus monkeys exposed to nerve agents. Additionally, we examined the time-concentration profiles of two biomarkers, VX and GD-nonapeptides and VX and GD-tyrosines, derived from OP-BChE and OP-albumin adducts up to 8 weeks after exposure. Based on the results, we determined that VX and GD-tyrosine is more suitable than VX and GD-nonapeptide as a biomarker owing to its longevity. This integrated approach is expected to be applicable for the quantification of other OP-BChE and OP-albumin adducts in human plasma, thus serving as a potential generic assay for exposure to nerve agents.
Asunto(s)
Butirilcolinesterasa/sangre , Inhibidores de la Colinesterasa/toxicidad , Agentes Nerviosos/toxicidad , Compuestos Organotiofosforados/toxicidad , Albúmina Sérica/análisis , Soman/toxicidad , Tirosina/análogos & derivados , Animales , Biomarcadores Farmacológicos/sangre , Butirilcolinesterasa/química , Butirilcolinesterasa/aislamiento & purificación , Inhibidores de la Colinesterasa/administración & dosificación , Inhibidores de la Colinesterasa/sangre , Inhibidores de la Colinesterasa/química , Cromatografía de Afinidad , Cromatografía Líquida de Alta Presión , Separación Inmunomagnética , Inyecciones Intravenosas , Límite de Detección , Macaca mulatta , Masculino , Estructura Molecular , Agentes Nerviosos/análisis , Agentes Nerviosos/química , Agentes Nerviosos/aislamiento & purificación , Oligopéptidos/sangre , Oligopéptidos/química , Oligopéptidos/aislamiento & purificación , Compuestos Organotiofosforados/administración & dosificación , Compuestos Organotiofosforados/sangre , Compuestos Organotiofosforados/química , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/química , Fragmentos de Péptidos/aislamiento & purificación , Reproducibilidad de los Resultados , Albúmina Sérica/química , Albúmina Sérica/aislamiento & purificación , Soman/análogos & derivados , Soman/sangre , Soman/química , Espectrometría de Masa por Ionización de Electrospray , Espectrometría de Masas en Tándem , Toxicocinética , Tirosina/sangre , Tirosina/química , Tirosina/aislamiento & purificaciónRESUMEN
Soman is a highly toxic organophosphorus chemical warfare compound that binds rapidly and irreversibility to a variety of serine active enzymes, i.e., butyryl- and acetyl-cholinesterases and carboxylesterase. The in vivo toxicity of soman has been reported to vary significantly in different animal species, such as rats and guinea pigs or non-human primates. This species variation makes it difficult to identify appropriate animal models for therapeutic drug development under the US Food and Drug Administration (FDA) Animal Rule. Since species variation in soman toxicity has been correlated with species variation in serum carboxylesterase, we undertook to determine if serum from guinea pigs, rats and non-human primates bound different levels of soman in vitro in the presence of equimolar concentrations of soman. Our results demonstrated that the amount of soman bound in the serum of rats was 4 uM, but essentially null in guinea pigs or non-human primates. The results strongly correlate with the presence or absence of carboxylesterase in the serum of animals and the difference in the toxic dose of soman in various species. Our results support prior suggestions that guinea pigs and non-human primates may be better animal models for the development of antidotes under the FDA Animal Rule.
Asunto(s)
Armas Biológicas , Inhibidores de la Colinesterasa , Soman/sangre , Animales , Cobayas , Macaca mulatta , Masculino , Radiofármacos/metabolismo , Ratas , Ratas Sprague-Dawley , Especificidad de la EspecieRESUMEN
Organophosphorus nerve agent (OPNA) adducts formed with human butyrylcholinesterase (HuBuChE) can be used as biomarker of OPNA exposure. Indeed, intoxication by OPNAs can be confirmed by the LC/MS2 analysis of a specific HuBuChE nonapeptide on which OPNAs covalently bind. A fast, selective, and highly sensitive online method was developed to detect sarin and soman adducts in plasma, including immunoextraction by anti-HuBuChE antibodies, pepsin digestion on immobilized enzyme reactors (IMER), and microLC/MS2 analysis of the OPNA adducts. The potential of three different monoclonal antibodies, covalently grafted on sepharose, was compared for the extraction of HuBuChE. The online method developed with the most promising antibodies allowed the extraction of up to 100% of HuBuChE contained in plasma and the digestion of 45% of it in less than 40 min. Moreover, OPNA-HuBuChE adducts, aged OPNA adducts, and unadducted HuBuChE could be detected (with S/N > 2000), even in plasma spiked with a low concentration of OPNA (10 ng mL-1). Finally, the potential of this method was compared to approaches involving other affinity sorbents, already described for HuBuChE extraction. Graphical abstract Online coupling of immunoextraction, digestion, and microliquid chromatography-tandem mass spectrometry for the analysis of organophosphorous nerve agent adducts formed with human butyrylcholinesterase.
Asunto(s)
Butirilcolinesterasa/metabolismo , Sustancias para la Guerra Química/farmacocinética , Inhibidores de la Colinesterasa/sangre , Sarín/sangre , Soman/sangre , Butirilcolinesterasa/sangre , Sustancias para la Guerra Química/metabolismo , Inhibidores de la Colinesterasa/metabolismo , Cromatografía de Afinidad/métodos , Cromatografía Liquida/métodos , Humanos , Sarín/análogos & derivados , Sarín/metabolismo , Soman/análogos & derivados , Soman/metabolismo , Espectrometría de Masas en Tándem/métodosRESUMEN
This work describes a novel and sensitive non-isotope dilution method for simultaneous quantification of organophosphorus nerve agents (OPNAs) soman (GD) and VX adducts to butyrylcholinesterase (BChE), their aged methylphosphonic acid (MeP) adduct and unadducted BChE in plasma exposed to OPNA. OPNA-BChE adducts were isolated with an off-column procainamide-gel separation (PGS) from plasma, and then digested with pepsin into specific adducted FGES*AGAAS nonapeptide (NP) biomarkers. The resulting NPs were detected by UHPLC-MS/MS MRM. The off-column PGS method can capture over 90% of BChE, MeP-BChE, VX-BChE and GD-BChE from their respective plasma materials. One newly designed and easily synthesized phosphorylated BChE nonapeptide with one Gly-to-Ala mutation was successfully reported to serve as internal standard instead of traditional isotopically labeled BChE nonapeptide. The linear range of calibration curves were from 1.00-200ngmL-1 for VX-NP, 2.00-200ngmL-1 for GD-NP and MeP-NP (R2≥0.995), and 3.00-200ngmL-1 for BChE NP (R2≥0.990). The inter-day precision had relative standard deviation (%RSD) of <8.89%, and the accuracy ranged between 88.9-120%. The limit of detection was calculated to be 0.411, 0.750, 0.800 and 1.43ngmL-1 for VX-NP, GD-NP, MeP-NP and BChE NP, respectively. OPNA-exposed quality control plasma samples were characterized as part of method validation. Investigation of plasma samples unexposed to OPNA revealed no baseline values or interferences. Using the off-column PGS method combined with UHPLC-MS/MS, VX-NP and GD-NP adducts can be unambiguously detected with high confidence in 0.10ngmL-1 and 0.50ngmL-1 of exposed human plasma respectively, only requiring 0.1mL of plasma sample and taking about four hours without special sample preparation equipment. These improvements make it a simple, sensitive and robust PGS-UHPLC-MS/MS method, and this method will become an attractive alternative to immunomagnetic separation (IMS) method and a useful diagnostic tool for retrospective detection of OPNA exposure with high confidence. Furthermore, using the developed method, the adducted BChE levels from VX and GD-exposed (0.10-100ngmL-1) plasma samples were completely characterized, and the fact that VX being more active and specific to BChE than GD was re-confirmed.
Asunto(s)
Butirilcolinesterasa/sangre , Sustancias para la Guerra Química/farmacocinética , Inhibidores de la Colinesterasa/sangre , Compuestos Organofosforados/sangre , Compuestos Organotiofosforados/sangre , Soman/sangre , Espectrometría de Masas en Tándem/métodos , Butirilcolinesterasa/aislamiento & purificación , Inhibidores de la Colinesterasa/aislamiento & purificación , Cromatografía Líquida de Alta Presión/instrumentación , Cromatografía Líquida de Alta Presión/métodos , Diseño de Equipo , Geles/química , Humanos , Límite de Detección , Compuestos Organofosforados/aislamiento & purificación , Compuestos Organotiofosforados/aislamiento & purificación , Procainamida/química , Soman/aislamiento & purificación , Espectrometría de Masas en Tándem/instrumentaciónRESUMEN
This work describes a new specific, sensitive, and rapid stable isotope dilution method for the simultaneous detection of the organophosphorus nerve agents (OPNAs) tabun (GA), sarin (GB), soman (GD), cyclosarin (GF), VR, VX, and VM adducts to tyrosine (Tyr). Serum, plasma, and lysed whole blood samples (50 µL) were prepared by protein precipitation followed by digestion with Pronase. Specific Tyr adducts were isolated from the digest by a single solid phase extraction (SPE) step, and the analytes were separated by reversed-phase ultra high performance liquid chromatography (UHPLC) gradient elution in less than 2 min. Detection was performed on a triple quadrupole tandem mass spectrometer using time-triggered selected reaction monitoring (SRM) in positive electrospray ionization (ESI) mode. The calibration range was characterized from 0.100-50.0 ng/mL for GB- and VR-Tyr and 0.250-50.0 ng/mL for GA-, GD-, GF-, and VX/VM-Tyr (R(2) ≥ 0.995). Inter- and intra-assay precision had coefficients of variation of ≤17 and ≤10%, respectively, and the measured concentration accuracies of spiked samples were within 15% of the targeted value for multiple spiking levels. The limit of detection was calculated to be 0.097, 0.027, 0.018, 0.074, 0.023, and 0.083 ng/mL for GA-, GB-, GD-, GF-, VR-, and VX/VM-Tyr, respectively. A convenience set of 96 serum samples with no known nerve agent exposure was screened and revealed no baseline values or potential interferences. This method provides a simple and highly specific diagnostic tool that may extend the time postevent that a confirmation of nerve agent exposure can be made with confidence.
Asunto(s)
Análisis Químico de la Sangre/métodos , Sustancias para la Guerra Química/análisis , Cromatografía Líquida de Alta Presión , Espectrometría de Masa por Ionización de Electrospray , Análisis Químico de la Sangre/instrumentación , Humanos , Compuestos Organofosforados/sangre , Compuestos Organofosforados/química , Compuestos Organotiofosforados/sangre , Reproducibilidad de los Resultados , Sarín/sangre , Sarín/química , Soman/sangre , Soman/química , Factores de Tiempo , Tirosina/sangre , Tirosina/químicaRESUMEN
Although nerve agent use is prohibited, concerns remain for human exposure to nerve agents during decommissioning, research, and warfare. Exposure can be detected through the analysis of hydrolysis products in urine as well as blood. An analytical method to detect exposure to five nerve agents, including VX, VR (Russian VX), GB (sarin), GD (soman), and GF (cyclosarin), through the analysis of the hydrolysis products, which are the primary metabolites, in serum has been developed and characterized. This method uses solid-phase extraction coupled with high-performance liquid chromatography for separation and isotopic dilution tandem mass spectrometry for detection. An uncommon buffer of ammonium fluoride was used to enhance ionization and improve sensitivity when coupled with hydrophilic interaction liquid chromatography resulting in detection limits from 0.3 to 0.5 ng/mL. The assessment of two quality control samples demonstrated high accuracy (101-105%) and high precision (5-8%) for the detection of these five nerve agent hydrolysis products in serum.
Asunto(s)
Sustancias para la Guerra Química/análisis , Compuestos Organofosforados/sangre , Compuestos Organotiofosforados/sangre , Sarín/sangre , Soman/sangre , Compuestos de Amonio , Biotransformación , Tampones (Química) , Sustancias para la Guerra Química/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Fluoruros , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Técnicas In Vitro , Límite de Detección , Compuestos de Amonio Cuaternario , Extracción en Fase Sólida , Espectrometría de Masas en TándemRESUMEN
A sensitive method for the determination of the organophosphorus nerve agents sarin, soman and VX adducts with tyrosine residue of albumin in rat plasma has been developed and validated using liquid chromatography-isotope dilution tandem mass spectrometry (LC-IDMS/MS). O-(O-Alkyl methylphosphonyl) tyrosine adducts and their deuterated products that were used as the internal standards were synthesised to establish the quantitative isotope-dilution method. Protein purification and solid-phase extraction (SPE) were applied to improve the recovery efficiency, reduce interference and achieve high sensitivity. The method provided a detection limit of 0.01 ng/mL for sarin and soman adducts and 0.05 ng/mL for the VX adduct. The value of the intra-day relative standard deviation over the calibration range was less than 6.16% (n=6), and that of the inter-day was less than 12.7% (n=6). The recovery varied from 86% to 111%. This sensitive method was successfully applied to the analysis of adducts in rat plasma after nerve agent exposure, and the results demonstrated the dose-effect relationships.
Asunto(s)
Cromatografía Liquida/métodos , Compuestos Organotiofosforados/sangre , Sarín/sangre , Soman/sangre , Espectrometría de Masas en Tándem/métodos , Animales , Biomarcadores/sangre , Biomarcadores/química , Biomarcadores/metabolismo , Butirilcolinesterasa/metabolismo , Límite de Detección , Masculino , Compuestos Organotiofosforados/química , Compuestos Organotiofosforados/metabolismo , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Sarín/química , Sarín/metabolismo , Albúmina Sérica/metabolismo , Soman/química , Soman/metabolismo , Tirosina/metabolismoRESUMEN
The nerve agent O-pinacolyl methylphosphonofluoridate, also known as soman or by its military designation GD, is a highly toxic organophosphorous compound that exerts its effects through inhibition of the enzyme acetylcholinesterase (AChE). In the present study, a fluoride ion based regeneration assay was developed to quantify the level of soman present in the blood of rats following a low-level whole-body inhalation exposure. It was hypothesized that the amount of regenerated nerve agent in the blood would be dose dependent in rats subjected to a whole-body inhalation exposure to a low-level dose of soman vapor, and that the fluoride ion-based regeneration method would be more sensitive for the detection of a low-level exposure to soman vapor than the measurement of whole blood AChE activity. Regenerated soman was dose-dependently detected in both the red blood cells (RBCs) and plasma of exposed rats at all concentrations tested (0.033-0.280 mg/m(3) for a 240-min exposure). Significant inhibition of whole blood AChE activity did not occur below a concentration of 0.101 mg/m(3), and was only depressed by approximately 10-25% at concentrations ranging from 0.101 mg/m(3) to 0.280 mg/m(3). This study is the first to utilize a fluoride ion-based regeneration assay to demonstrate the dose-dependent increases in soman in the blood following whole-body inhalation exposure to low levels of vapor. Additionally, the results of the present study demonstrate that the fluoride ion based regeneration assay was approximately threefold more sensitive than the measurement of AChE activity in the blood for the detection of exposure to soman, and also that miosis is a more sensitive marker of soman exposure than inhibition of AChE activity.
Asunto(s)
Biomarcadores/sangre , Sustancias para la Guerra Química/metabolismo , Inhibidores de la Colinesterasa/sangre , Reactivadores Enzimáticos/farmacología , Fluoruro de Sodio/farmacología , Soman/sangre , Acetilcolinesterasa/sangre , Administración por Inhalación , Animales , Butirilcolinesterasa/sangre , Sustancias para la Guerra Química/toxicidad , Inhibidores de la Colinesterasa/toxicidad , Relación Dosis-Respuesta a Droga , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Femenino , Cromatografía de Gases y Espectrometría de Masas , Exposición por Inhalación , Masculino , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Soman/toxicidad , VolatilizaciónRESUMEN
A method for determining the chemical warfare agent soman (GD) in rat plasma has been validated and applied to low-level inhalation exposure studies currently being conducted. This method utilizes a fluoride ion-based regeneration assay with isotope dilution followed by large volume injection gas chromatography with ammonia chemical ionization mass spectrometric detection. Following sample preparation by solid phase extraction, chromatographic separation was achieved using a 14% cyanopropylphenyl/86% dimethyl polysiloxane capillary column with a total run time of 18.16 min. Soman and the deuterated isotope ((2)H(4)-soman) internal standard were detected using the selected ion monitoring mode and quantitated using the ammonia adduction ratio of m/z ions 200/204. A reproducible linear relationship was obtained for the quantitative concentration range of 10 pg on-column to 1000 pg on-column (r(2) = 0.9995) for standards in ethyl acetate with a detection limit of 5.65 pg on-column, and an average recovery of 93% in plasma. This sensitive method was successfully applied to the analysis of soman in rat plasma immediately post-exposure, resulting in the construction of dose-response plots.
Asunto(s)
Monitoreo del Ambiente/métodos , Cromatografía de Gases y Espectrometría de Masas/métodos , Soman/sangre , Administración por Inhalación , Animales , Calibración , Sustancias para la Guerra Química/análisis , Sustancias para la Guerra Química/farmacocinética , Inhibidores de la Colinesterasa/administración & dosificación , Inhibidores de la Colinesterasa/sangre , Inhibidores de la Colinesterasa/farmacocinética , Exposición a Riesgos Ambientales/análisis , Fluoruros/química , Masculino , Compuestos de Potasio/química , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los Resultados , Extracción en Fase Sólida , Soman/administración & dosificación , Soman/farmacocinéticaRESUMEN
Organophosphorus nerve agents (OPNAs) continue to pose a threat to military personnel and the general public because of their toxicity and their potential use as weapons of mass destruction. An effective method for the detection of human exposure to OPNAs involves the refluoridation of nerve agents adducted to the serum protein butyrylcholinesterase. The regenerated agents are then enriched by solid-phase extraction and quantified by isotope-dilution gas chromatography-mass spectrometry. We have previously reported improvements that resulted in a 10-fold increase in sensitivity. We have now made further changes to the method that include the addition of confirmation ions, the addition of soman (GD) to the assay, the expansion of the linear range, and the elimination of high-volume injection to decrease background noise and run time while improving sensitivity. This report includes the standard operating procedures for this method for tabun, sarin, soman, cyclohexylsarin, and VX and validation studies. The method's limits of detection ranged from 5.5 to 16.5 pg/mL for the G analogue of VX and GD, respectively. Characterization of quality control (QC) materials resulted in an average coefficient of variation of 15.1% for the five analytes in low QC pools and 11.7% in high QC pools.
Asunto(s)
Proteínas Sanguíneas/metabolismo , Inhibidores de la Colinesterasa/sangre , Monitoreo del Ambiente/métodos , Fluoruros/química , Compuestos Organofosforados/sangre , Compuestos de Potasio/química , Biomarcadores/sangre , Proteínas Sanguíneas/química , Calibración , Sustancias para la Guerra Química/análisis , Sustancias para la Guerra Química/química , Sustancias para la Guerra Química/metabolismo , Inhibidores de la Colinesterasa/química , Inhibidores de la Colinesterasa/metabolismo , Exposición a Riesgos Ambientales/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , Humanos , Organofosfatos/sangre , Organofosfatos/química , Organofosfatos/metabolismo , Compuestos Organofosforados/química , Compuestos Organofosforados/metabolismo , Compuestos Organotiofosforados/sangre , Compuestos Organotiofosforados/química , Compuestos Organotiofosforados/metabolismo , Reproducibilidad de los Resultados , Sarín/sangre , Sarín/química , Sarín/metabolismo , Extracción en Fase Sólida/métodos , Solventes/química , Soman/sangre , Soman/química , Soman/metabolismoRESUMEN
The organophosphorus nerve agent soman is an irreversible cholinesterase (ChE) inhibitor that can produce long-lasting seizures and brain damage in which the neurotransmitters acetylcholine and glutamate are involved. These same neurotransmitters play key-roles in the auditory function. It was then assumed that exploring the hearing function may provide markers of the central events triggered by soman intoxication. In the present study, distortion product otoacoustic emissions (DPOAEs), a non-invasive audiometric method, were used to monitor cochlear functionality in rats administered with a moderate dose of soman (45 microg/kg). DPOAEs were investigated either 4h or 24h post-challenge. In parallel, the effects of soman on whole blood and brain ChE activity and on brain histology were also studied. The first main result is that DPOAE intensities were significantly decreased 4h post-soman and returned to baseline at 24h. The amplitude changes were well related to the severity of symptoms, with the greatest change being recorded in the rats that survived long-lasting convulsions. The second main result is that baseline DPOAEs recorded 8 days before soman appear to predict the severity of symptoms produced by the intoxication. Indeed, the lowest baseline DPOAEs corresponded to the occurrence of long-lasting convulsions and brain damage and to the greatest inhibition in central ChE. These results thus suggest that DPOAEs represent a promising non-invasive tool to assess and predict the central consequences of nerve agent poisoning. Further investigations will be carried out to assess the potential applications and the limits of this non-invasive method.
Asunto(s)
Síndromes de Neurotoxicidad/etiología , Emisiones Otoacústicas Espontáneas/efectos de los fármacos , Soman/toxicidad , Estimulación Acústica , Animales , Audiometría/métodos , Audiometría de Respuesta Evocada/métodos , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/patología , Química Encefálica/efectos de los fármacos , Inhibidores de la Colinesterasa/administración & dosificación , Inhibidores de la Colinesterasa/sangre , Inhibidores de la Colinesterasa/toxicidad , Cóclea/efectos de los fármacos , Cóclea/fisiopatología , Inyecciones Subcutáneas , Masculino , Síndromes de Neurotoxicidad/patología , Síndromes de Neurotoxicidad/fisiopatología , Ratas , Ratas Wistar , Índice de Severidad de la Enfermedad , Soman/administración & dosificación , Soman/sangre , Análisis de Supervivencia , Factores de TiempoRESUMEN
A physiologically based pharmacokinetic (PB/PK) model has been developed in advanced computer simulation language (ACSL) to describe blood and tissue concentration-time profiles of the C(+/-)P(-) stereoisomers of soman after inhalation, subcutaneous and intravenous exposures at low (0.8-1.0 x LD(50)), medium (2-3 x LD(50)) and high (6 x LD(50)) levels of soman challenge in three species (rat, guinea pig, marmoset). Allometric formulae were used to compute the compartment volumes, blood flow rates, tidal volume and respiratory rate based upon total animal weight. Blood/tissue partition coefficients for soman, initial carboxylesterase and acetylcholinesterase levels and the rate constants for interactions between soman and these enzymes were species-dependent and were obtained from in vitro measurements reported in the literature. The model incorporated arterial and venous blood, lung, kidney, liver, richly perfused, poorly perfused and fat tissue compartments as well as subcutaneous and nasal exposure site compartments. First-order absorption from linearly filled soman deposits into metabolizing exposure site compartments was employed to model subcutaneous and inhalation exposures. The model was validated by comparing the predicted and observed values for C(+/-)P(-)-soman in arterial blood at various times following exposure and by regression analysis. Sensitivity analysis was used to determine the effects of perturbations in the model parameters on the time-course of arterial C(-)P(-)-soman concentrations for different exposure routes. In our evaluation of 28 datasets, predicted values were generally within 95% confidence limits of the observed values, and regression coefficients comparing predicted and observed data were greater than 0.85 for 95% of the intravenous and subcutaneous datasets and 25% of the inhalation datasets. We conclude that the model predicts the soman toxicokinetics for doses >or=1 x LD(50) for intravenous and subcutaneous exposures and inhalation exposures of 8 min or less sufficiently well to allow its use in the modeling of bioscavenger protection.
Asunto(s)
Sustancias para la Guerra Química/farmacocinética , Inhibidores de la Colinesterasa/farmacocinética , Modelos Biológicos , Soman/farmacocinética , Administración por Inhalación , Animales , Callithrix , Inhibidores de la Colinesterasa/sangre , Simulación por Computador , Cobayas , Inyecciones Intravenosas , Inyecciones Subcutáneas , Masculino , Ratas , Ratas Wistar , Soman/sangreRESUMEN
The authors summarized study results on chronic effects caused by products of leaching from bituminous-salt masses obtained through destruction of sarin, soman and RVX. State of experimental rats was evaluated with integral informative tests (physiologic, biochemical, hematologic and morphologic) presenting changes in objective health parameters and revealing every disorder in organs and systems functioning.
Asunto(s)
Sustancias para la Guerra Química/efectos adversos , Residuos Peligrosos/efectos adversos , Hidrocarburos/efectos adversos , Organotiofosfatos/efectos adversos , Sales (Química) , Sarín/efectos adversos , Soman/efectos adversos , Animales , Sustancias para la Guerra Química/análisis , Sustancias para la Guerra Química/metabolismo , Hidrocarburos/análisis , Hidrocarburos/sangre , Masculino , Organotiofosfatos/análisis , Organotiofosfatos/sangre , Ratas , Ratas Wistar , Sarín/análisis , Sarín/sangre , Soman/análisis , Soman/sangre , Agua/químicaRESUMEN
The fluoride reactivation process was evaluated for measuring the level of sarin or soman nerve agents reactivated from substrates in plasma and tissue from in vivo exposed guinea pigs (Cava porcellus), in blood from in vivo exposed rhesus monkeys (Macaca mulatta), and in spiked human plasma and purified human albumin. Guinea pig exposures ranged from 0.05 to 44 LD50, and reactivated nerve agent levels ranged from 1.0 ng/mL in plasma obtained from 0.05 LD50 sarin-exposed guinea pigs to an average of 147 ng/g in kidney tissue obtained from two 2.0 LD50 soman-exposed guinea pigs. Positive dose-response relationships were observed in all low-level, 0.05 to 0.4 LD50, exposure studies. An average value of 2.4 ng/mL for reactivated soman was determined in plasma obtained from two rhesus monkeys three days after a 2 LD50 exposure. Of the five types of guinea pig tissue studied, plasma, heart, liver, kidney and lung, the lung and kidney tissue yielded the highest amounts of reactivated agent. In similar tissue and with similar exposure procedures, reactivated soman levels were greater than reactivated sarin levels. Levels of reactivated agents decreased rapidly with time while the guinea pig was alive, but decreased much more slowly after death. This latter chemical stability should facilitate forensic retrospective identification. The high level of reactivated agents in guinea pig samples led to the hypothesis that the principal source of reactivated agent came from the agent-carboxylesterase adduct. However, there could be contributions from adducts of the cholinesterases, albumin and fibrous tissue, as well. Quantitative analysis was performed with a GC-MS system using selected ion monitoring of the 99 and 125 ions for sarin and the 99 and 126 ions for soman. Detection levels were as low as 0.5 ng/mL. The assay was precise and easy to perform, and has potential for exposure analysis from organophosphate nerve agents and pesticides in other animal species.
Asunto(s)
Sustancias para la Guerra Química/farmacocinética , Reactivadores de la Colinesterasa/farmacología , Fluoruros/farmacología , Sarín/farmacocinética , Soman/farmacocinética , Animales , Sustancias para la Guerra Química/toxicidad , Relación Dosis-Respuesta a Droga , Cobayas , Humanos , Riñón/efectos de los fármacos , Riñón/metabolismo , Dosificación Letal Mediana , Macaca mulatta , Sarín/sangre , Sarín/toxicidad , Soman/sangre , Soman/toxicidadRESUMEN
The effects of verapamil on the elimination of soman in rabbit blood and distribution in mouse brain and diaphragm by determining the concentration of P(-)soman using the chirasil capillary gas chromatographic analysis method were studied in order to study the effects of verapamil on the metabolic detoxification of soman. Verapamil (10 mg kg(-1), im, 30 min before soman administration) could significantly reduce the concentration of P(-)soman in rabbit blood at 15, 60, 90, 120, 180 and 240 s after soman injection (43.2 microg kg(-1), iv) as compared to soman-treated control animal respectively. Toxicokinetics parameters showed verapamil could increase clearance rate from 20.8+/-1.51 to 44.3+/-7.0 ml kg(-1)s(-1) and reduce AUC of P(-)soman from 2.08+/-0.151 to 0.996+/-0.172 mg s l(-1). For experiments in mice, verapamil could reduce the concentration P(-)soman in diaphragm from 74.7, 70.5, 88.7 to 41.1, 39.0, 49.3 ng g(-1) at the time of 30, 90, 120 s after intoxication of soman subcutaneously vs. soman control respectively, but it had no influence on the concentration of free P(-)soman in brain. Verapamil accelerated the elimination of P(-)soman in the rabbits blood and reduced the distribution of P(-)soman in the mouse diaphragm.
Asunto(s)
Inhibidores de la Colinesterasa/farmacocinética , Soman/farmacocinética , Vasodilatadores/farmacología , Verapamilo/farmacología , Animales , Área Bajo la Curva , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Bloqueadores de los Canales de Calcio/farmacología , Inhibidores de la Colinesterasa/toxicidad , Cromatografía de Gases , Diafragma/efectos de los fármacos , Diafragma/metabolismo , Semivida , Inactivación Metabólica , Ratones , Conejos , Soman/antagonistas & inhibidores , Soman/sangre , Soman/toxicidad , EstereoisomerismoRESUMEN
AIM: To investigate the effect of nimodipine on the elimination of soman in rabbit blood and distribution of [3H]soman in mice. METHODS: Chirasil capillary gas chromatographic analysis method with large volume injections was used to determine the concentration of C(+/-)P(-)soman in rabbit blood. [3H]soman trace method was used to study the effect of nimodipine on soman distribution in mice. RESULTS: Nimodipine (10 mg/kg, ip, 1 h pre-treated) could significantly reduce the concentration of C(+/-)P(-)soman in rabbit blood from (54+/-13) to (19+/-12) microg/L blood at 15 s after soman injection (43.2 microg/kg, iv). Nimodipine could increase clearance rate [CL(S)] from (20.8+/-1.5) to (31+/-11) mL/kg/s and reduce AUC of C(+/-)P(-)soman from (2.08+/-0.15) to (1.6+/-0.4) mg/s. Nimodipine (10 mg/kg, ip, 1 h pre-treated) treatment could significantly reduce the distribution amount of bound [3H]soman in plasma, brain, lung, and liver, moreover increased the distribution amount of bound [3H]soman in small intestine during 0-120 min after mice received [3H]soman (0.544 GBq*119 microg/kg, sc) compared to soman control group. CONCLUSION: Nimodipine might alter the distribution of soman and reduce the initial concentration of soman in rabbit blood, then accelerated the metabolic detoxication of soman.
Asunto(s)
Bloqueadores de los Canales de Calcio/farmacología , Nimodipina/farmacología , Soman/farmacocinética , Animales , Área Bajo la Curva , Cromatografía de Gases , Interacciones Farmacológicas , Femenino , Inactivación Metabólica , Masculino , Ratones , Conejos , Soman/sangre , Distribución TisularRESUMEN
The portal vein, liver artery ligation treatment and the portal vein ligation treatment could increase the concentration of P(-) soman in rabbit blood 3.6-19.3 times as compared with soman control group at each time points after soman injection (43.2 microgkg(-1), i.v.). Toxicokinetics parameters showed that portal vein, liver artery ligation treatment and portal vein ligation treatment could reduce the clearance (CL) and distribution volume (V(d)). Meanwhile, they could significantly increase the AUC of soman in rabbits from 2.08+/-0.154 to 18.2+/-2.96 and 22.9+/-3.73 mg s l(-1), respectively. All these data showed that the liver and intestine play a very important role on elimination the free soman in rabbit's blood at high dosing of soman.
Asunto(s)
Inhibidores de la Colinesterasa/farmacocinética , Hígado/irrigación sanguínea , Conejos/metabolismo , Soman/farmacocinética , Animales , Área Bajo la Curva , Inhibidores de la Colinesterasa/sangre , Inhibidores de la Colinesterasa/toxicidad , Femenino , Arteria Hepática/cirugía , Ligadura/veterinaria , Vena Porta/cirugía , Conejos/cirugía , Soman/sangre , Soman/toxicidad , EstereoisomerismoRESUMEN
The potential of obidoxime and other pyridinium-4-aldoximes to reactivate dimethyl- and diethylphosphorylated cholinesterases is markedly restricted by the inevitable formation of rather stable phosphoryl oximes (POXs) with high anticholinesterase activity. This effect is hardly seen with very dilute enzyme preparations, but becomes significant at physiological enzyme concentrations. Human plasma with the butyrylcholinesterase irreversibly blocked by soman was able to stimulate obidoxime-induced reactivation of concentrated erythrocyte acetylcholinesterase (Ery-AChE) to the same extent as was observed with a dilute preparation, suggesting phosphoryl oxime-destroying capacity. The inactivating factor, which was tentatively termed POX-hydrolase, had (1) a molecular weight of >100 kDa; (2) required Ca2+ , which could not be substituted by Zn2+ or Mg2+; and (3) lost its catalytic activity reversibly in the presence of ethylenediamine-tetraacetic acid (EDTA). The enzyme activity varied widely (20-fold) among different subjects and did not follow the activity pattern of human serum paraoxonase (PON1). Rabbit plasma with its particularly high paraoxonase content showed only weak POX-hydrolase activity. These data suggest POX-hydrolase to be a different entity. POX-hydrolase was most active with the putative phosphoryl-obidoxime from paraoxon-ethyl, less with the product from paraoxon-methyl and least with that from diisopropylfluorophosphate. The analogue TMB-4 reacted similarly to obidoxime. The putative phosphonyl oximes arising by the reaction of obidoxime with nerve agents were apparently not cleaved. The variation in POX-hydrolase activity may additionally contribute to the variable response to oxime therapy in patients with organophosphate insecticide poisoning.