毒剂的生化分析技术

本文论述了酶法分析、核酸探针、免疫分析法特别是生物传感器在毒剂检测中的应用，提出了毒剂生化分析的发展方向。     

Liquid chromatography electrospray tandem mass spectrometric and desorption electrospray ionization tandem mass spectrometric analysis of chemical warfare agents in office media typically collected during a forensic investigation

Most prior analytical studies have dealt with the determination of chemical warfare agents in environmental or biological matrices that would typically be collected following battlefield use or in support of the Chemical Weapons Convention. These methods may be useful for some investigations, but may not be practical for indoor forensic investigations where chemical warfare agent use is suspected. There is a need for analytical methods for chemical warfare agent identification in office media, including flooring, wall surfaces, office fabrics and paper products, which would typically be collected in an office environment during forensic investigations. During this study, typical office environment media were spiked at the 4-20microg/g level with either a complex munitions grade sample of tabun (GA) or with a standard containing the three nerve agents, sarin (GB), cyclohexyl methylphosphonofluoridate (GF), soman (GD) and the nerve agent simulant, triethyl phosphate (TEP), to evaluate the potentials of liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) and liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) for forensic purposes. An emerging technique, desorption electrospray ionization (DESI-MS/MS), was also investigated for the direct determination of TEP, GB and GD sampled onto solid phase microextraction (SPME) fibers exposed to spiked office media. The spiked chemical warfare agents were recovered with varying efficiencies during this study, but in all cases sufficient chemical warfare agent was recovered for mass spectrometric identification purposes. Full high resolution mass spectra were acquired for all the chemical warfare agents in the continuum mode, which typically resulted in mass measurement errors of 0.001Da or less.     

Mass spectrometric identification of toxins of biological origin

The chemical/biological (CB) threat spectrum encompasses a wide range of potential agents including chemical warfare agents, biological warfare agents and toxins of biological origin that fall between these two main agent categories. These proteinaceous and non-proteinaceous toxins, commonly referred to as mid-spectrum agents, range in molecular mass from a few hundred to more than a hundred thousand daltons. The large number of potential candidates as well as the structural diversity of possible mid-spectrum agents makes identification of these compounds a challenge. The NATO defense community has recognized these challenges and has a working group that is developing identification protocols and evaluating methods through a series of international analytical exercises. Identification strategies rely heavily on recent advances that have been made in both mass spectrometry (MS) and liquid chromatography (LC), with LC-MS typically being employed as the primary method for separation/identification. While this paper focuses on the application of these and related instrumental analytical techniques for the identification of mid-spectrum agents, the approach described could be applied in the fields of toxicology, forensic science and environmental analysis. Areas for future research have been identified and application of developed mid-spectrum identification methods to the ongoing biological and toxin weapons convention (BTWC) are anticipated.     

Functionalized gold nanoparticle supported sensory mechanisms applied in detection of chemical and biological threat agents: A review

There is a great necessity for development of novel sensory concepts supportive of smart sensing capabilities in defense and homeland security applications for detection of chemical and biological threat agents. A smart sensor is a detection device that can exhibit important features such as speed, sensitivity, selectivity, portability, and more importantly, simplicity in identifying a target analyte. Emerging nanomaterial based sensors, particularly those developed by utilizing functionalized gold nanoparticles (GNPs) as a sensing component potentially offer many desirable features needed for threat agent detection. The sensitiveness of physical properties expressed by GNPs, e.g. color, surface plasmon resonance, electrical conductivity and binding affinity are significantly enhanced when they are subjected to functionalization with an appropriate metal, organic or biomolecular functional groups. This sensitive nature of functionalized GNPs can be potentially exploited in the design of threat agent detection devices with smart sensing capabilities. In the presence of a target analyte (i.e., a chemical or biological threat agent) a change proportional to concentration of the analyte is observed, which can be measured either by colorimetric, fluorimetric, electrochemical or spectroscopic means. This article provides a review of how functionally modified gold colloids are applied in the detection of a broad range of threat agents, including radioactive substances, explosive compounds, chemical warfare agents, biotoxins, and biothreat pathogens through any of the four sensory means mentioned previously.     

Fluorescent sensors for the detection of chemical warfare agents

Along with biological and nuclear threats, chemical warfare agents are some of the most feared weapons of mass destruction. Compared to nuclear weapons they are relatively easy to access and deploy, which makes them in some aspects a greater threat to national and global security. A particularly hazardous class of chemical warfare agents are the nerve agents. Their rapid and severe effects on human health originate in their ability to block the function of acetylcholinesterase, an enzyme that is vital to the central nervous system. This article outlines recent activities regarding the development of molecular sensors that can visualize the presence of nerve agents (and related pesticides) through changes of their fluorescence properties. Three different sensing principles are discussed: enzyme-based sensors, chemically reactive sensors, and supramolecular sensors. Typical examples are presented for each class and different fluorescent sensors for the detection of chemical warfare agents are summarized and compared.     

Mass spectrometric analysis of chemical warfare agents and their degradation products in soil and synthetic samples

A packed capillary liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS) method was developed for the identification of chemical warfare agents, their degradation products and related compounds in synthetic tabun samples and in soil samples collected from a former mustard storage site. A number of organophosphorus and organosulfur compounds that had not been previously characterized were identified, based on acquired high-resolution ESI-MS data. At lower sampling cone voltages, the ESI mass spectra were dominated by protonated, sodiated and protonated acetonitrile adducts and/or their dimers that could be used to confirm the molecular mass of each compound. Structural information was obtained by inducing product ion formation in the ESI interface at higher sampling cone voltages. Representative ESI-MS mass spectra for previously uncharacterized compounds were incorporated into a database as part of an on-going effort in chemical warfare agent detection and identification. The same samples were also analyzed by capillary column gas chromatography (GC)-MS in order to compare an established method with LC-ESI-MS for chemical warfare agent identification. Analysis times and full-scanning sensitivities were similar for both methods, with differences being associated with sample matrix, ease of ionization and compound volatility. GC-MS would be preferred for organic extracts and must be used for the determination of mustard and relatively non-polar organosulfur degradation products, including 1,4- thioxane and 1,4-dithiane, as these compounds do not ionize during ESI-MS. Diols, formed following hydrolysis of mustard and longer-chain sulfur vesicants, may be analyzed using both methods with LC-ESI-MS providing improved chromatographic peak shape. Aqueous samples and extracts would, typically, be analyzed by LC-ESI-MS, since these analyses may be conducted directly without the need for additional sample handling and/or derivatization associated with GC-MS determinations. Organophosphorus compounds, including chemical warfare agents, related compounds and lower volatility hydrolysis products may all be determined during a single LC-ESI- MS analysis. Derivatization of chemical warfare agent hydrolysis products and other compounds with hydroxyl substitution would be required prior to GC-MS analysis, giving LC-ESI-MS a definite advantage over GC-MS for the analysis of samples containing chemical warfare agents and/or their hydrolysis products.     

用于化学毒剂检测的微萃取技术研究进展

对化学毒剂及其降解产物的分析检测是准确鉴别化学沾染的重要手段.由于化学毒剂及其降解产物的样品可能存在于各种基质中,且部分化学毒剂在水体等基质中降解速度过快,所以将痕量样品从复杂基质中快速高效的富集提取出来显得尤其重要.微萃取技术具有装置体积小、使用少量或不使用溶剂、绿色环保、易与色谱分析技术联用等突出优势受到广泛的关注...     

Screening hydrolysis products of sulfur mustard agents by high-performance liquid chromatography with inductively coupled plasma mass spectrometry detection

Sulfur mustard (HD), bis(2-chloroethyl)sulfide, is one of a class of mustard agents which are chemical warfare agents. The main chemical warfare hydrolysis degradation products of sulfur mustards are: thiodiglycol, bis(2-hydroxyethylthio)methane, 1,2-bis(2-hydroxyethylthio)ethane, 1,3-bis(2-hydroxyethylthio)propane, and 1,4-bis(2-hydroxyethylthio)butane. The aim of this study is to identify these five hydrolysis degradation products utilizing reversed-phase high-performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (ICP-MS) for element-specific sulfur detection using a collision/reaction cell and electrospray ionization mass spectrometry to confirm the identification. To date, this is the first study utilizing ICP-MS with ³²S element-specific detection for the analysis of vesicant chemical warfare agent degradation products.     

New method for comprehensive detection of chemical warfare agents using an electron-cyclotron-resonance ion-source mass spectrometer

We developed a detection technology for vapor forms of chemical warfare agents (CWAs) with an element analysis system using an electron cyclotron resonance ion source. After the vapor sample was introduced directly into the ion source, the molecular material was decomposed into elements using electron cyclotron resonance plasma and ionized. The following CWAs and stimulants were examined: diisopropyl fluorophosphonate (DFP), 2-chloroethylethylsulfide (2CEES), cyanogen chloride (CNCl), and hydrogen cyanide (HCN). The type of chemical warfare agents, specifically, whether it was a nerve agent, blister agent, blood agent, or choking agent, could be determined by measuring the quantities of the monatomic ions or CN(+) using mass spectrometry. It was possible to detect gaseous CWAs that could not be detected by a conventional mass spectrometer. The distribution of electron temperature in the plasma could be closely controlled by adjusting the input power of the microwaves used to generate the electron cyclotron resonance plasma, and the target compounds could be detected as molecular ions or fragment ions, enabling identification of the target agents.     

The use of thermospray-liquid chromatography/mass spectrometry for the verification of chemical warfare agents

Summary Thermospray-liquid chromatography mass spectrometry (TSP-LC-MS) is a relatively new analytical technique which proved to be useful for the verification of chemical warfare agents and their polar degradation products in aqueous solutions. The principles of the technique are described and comparisons are made with other forms of mass spectrometric analysis. A survey is presented of the results obtained so far at the Prins Maurits Laboratory TNO. The analysis of organophosphorus nerve agents and their hydrolysis products (organophosphorus acids) in various types of water is described. Special attention is paid to the nerve agent VX. Direct analysis of vesicants in water by TSP-LC-MS is limited. However, analysis of their hydrolysis products, as well as related compounds such as adducts of mustard gas with nucleosides and peptides, is possible. Finally, the use of TSP-LC-MS for the analysis of other compounds of chemical warfare interest (toxins) is indicated.     

荧光法检测化学战剂

化学战剂,包括神经性毒剂(如沙林、VX)和糜烂性毒剂(如芥子气),属于剧毒化学品,能够严重危害国家安全和环境安全.因此,针对各类化学战剂,开发简单、快速、可便携化、高灵敏度和高选择性的荧光检测技术具有重要的意义.本文综述了近年来国内外荧光法检测化学战剂的研究进展,并对该领域所面临的挑战和未来发展方向进行了总结和展望.     

Doubly Protective MOF-Photo-Fabrics: Facile Template-Free Synthesis of PCN-222-Textiles Enables Rapid Hydrolysis,Photo-Hydrolysis and Selective Oxidation of Multiple Chemical Warfare Agents and Simulants

New materials and chemical knowledge for improved personal protection are among the most pressing needs in the international community. Reported attacks using chemical warfare agents (CWAs,) including organophosphate soman (GD) and thioether mustard gas (HD) are driving research in field-deployable catalytic composites for rapid toxin degradation. In this work, we report simple template-free low temperature synthesis that enables for the first time, a deployable-structured catalytic metal-organic framework/polymer textile composite “MOF-fabric” showing rapid hydrolysis and oxidation of multiple active chemical warfare agents, GD and HD, respectively, and their simulants. Our method yields new zirconium–porphyrin based nano-crystalline PCN-222 MOF-fabrics with adjustable MOF loading and robust mechanical adhesion on low-cost nonwoven polypropylene fibers. Importantly, we describe quantitative kinetic analysis confirming that our MOF-fabrics are as effective as or better than analogous MOF powders for agent degradation, especially for oxidation. Faster oxidation using the MOF-fabrics is ascribed to the composite geometry, where active MOF catalysts are uniformly displayed on the MOF-textile enabling better reactant transport and reactive oxidant generation. Furthermore, we note the discovery of visible photo-activation of GD hydrolysis by a MOF-fabric, which is ascribed to oxidation at the active metal node site, significantly increasing the rate over that observed without illumination. These results provide important new insights into the design of future materials and chemical systems to protect military, first-responders, and civilians upon exposure to complex chemical toxins.     

Electrochemical Biosensors for Detection of Biological Warfare Agents

This review discusses current development in electrochemical biosensors for detection of biological warfare agents. This could include bacteria, viruses and toxins that are aerosoled deliberately in air, food or water to spread terrorism and cause disease or death to humans, animals or plants. The rapid and unequivocal detection and identification of biological warfare agents is a major challenge for any government including military, health and other government agents. Reliable, specific characterization and identification of the microorganism from sampling location, either air, water, soil or others is required. This review will survey different types of electrochemical biosensors has been developed based on the following: i) Immunosensors ii) PCR (DNA base Sensor) iii) Bacteria or whole cell sensor and iv) Enzyme sensor. This article gives an overview of electrochemical biosensor for detection of biological warfare agents. Electrochemical biosensors have the advantages of sensitivity, selectivity, to operate in turbid media, and amenable to miniaturization. Recent developments in immunofiltration, flow injection, and flow‐through electrochemical biosensors for bacteria, viruses, and toxin detection are reviewed. The current research and development in biosensors for biological warfare agents detection is of interest to the public as well as to the defense is also discussed.     

Targeting Chemical and Biological Warfare Agents at the Molecular Level

After the September 11 tragedies of 2001, scientists and law‐enforcement agencies have shown increasing concern that terrorist organizations and their “rogue” foreign government‐backers may resort to the use of chemical and/or biological agents against U.S. military or civilian targets. In addition to the right mix of policies, including security measures, intelligence gathering and training for medical personnel on how to recognize symptoms of biochemical warfare agents, the major success in combating terrorism lies in how best to respond to an attack using reliable analytical sensors. The public and regulatory agencies expect sensing methodologies and devices for homeland security to be very reliable. Quality data can only be generated by using analytical sensors that are validated and proven to be under strict design criteria, development and manufacturing controls. Electrochemical devices are ideally suited for obtaining the desired analytical information in a faster, simpler, and cheaper manner compared to traditional (lab‐based) assays and hence for meeting the requirements of decentralized biodefense applications. This articler presents a review of the major trends in monitoring technologies for chemical and biological warfare (CBW) agents. It focuses on research and development of sensors (particularly electrochemical ones), discusses how advances in molecular recognition might be used to design new multimission networked sensors (MULNETS) for homeland security. Decision flow‐charts for choosing particular analytical techniques for CBW agents are presented. Finally, the paths to designing sensors to meet the needs of today's measurement criteria are analyzed.     

Determination of trace level chemical warfare agents in water and slurry samples using hollow fibre-protected liquid-phase microextraction followed by gas chromatography-mass spectrometry

A simple and solvent-minimised sample preparation technique based on hollow fibre-protected liquid-phase microextraction was investigated for the gas chromatography/mass spectrometric analysis of chemical warfare agents in water and slurry samples. The chemical warfare agents included four nerve agents and a blister agent. Parameters such as extraction solvent, salt concentration, stirring speed and extraction time were optimised using spiked deionised water samples. The technique provided a linear range of two orders of magnitude, good repeatability (RSDs < 10%, n = 6), good linearity (r2 >or= 0.995) and limits of detection (LODs) in the range of 0.02-0.09 microg l(-1) (S/N = 3) under full scan mode. The optimised technique was also applied to more complex slurry samples and similar precision (RSD < 15%, n = 3) and limits of detection (0.02-0.2 microgl(-1), S/N = 3) were obtained.     

Investigation of dimethyl methylphosphonate (DMMP) with an Ion mobility spectrometer using a pulsed electron source

Ion mobility spectrometry (IMS) has been an important tool for decades in the field of trace gas analysis of substances such as explosives, drugs of abuse or chemical warfare agents. In recent years, its application has been extended to more complex set ups. In this paper we present the application of a standard IMS device equipped with a novel pulsed electron gun for ionization in the investigation of the chemical warfare agent simulant dimethyl methylphosphonate (DMMP). The signal decay times of the reactant ion peak (RIP), the DMMP monomer and dimer have been investigated. Thus, further information could be obtained of the innovative application of different signal decay times in order to filter out signals of contaminants with focus on the decay dependence on the concentration. Additionally, further details regarding the still not fully understood underlying decay mechanisms have been found.     

Chromatographic analysis of chemical warfare agents

The usefulness and applications of the particular types of chromatography in the analysis of chemical warfare agents have been reviewed. A major problem in the chromatographic analysis of chemical warfare agents is the collection and preparation of the samples. The importance of this problem differs for the various types of chromatography. Significant differences occur in the way in which samples are collected from air, water, soil, vegetables or animal organisms. The analyses are characterized by the main groups of chemical warfare agents, e.g., organophosphorus, vesicants, irritants, etc. Account has been taken of the relationships between their properties and the possibilities of their chromatographic analysis. The advantages and disadvantages of particular types of chromatography in the analysis of the particular groups and individual agents have been considered. The detectability of particular chemical warfare agents has been assessed, together with the separating efficiency for their mixtures. Examples of applications of chromatographic systems and conditions of chromatographing are summarized in tables. It is concluded that chromatography is a very useful tool in the analysis of chemical warfare agents; GC and TLC have the most advantageous properties, HPLC being slightly inferior.     

Determination of sarin, soman and their hydrolysis products in soil by packed capillary liquid chromatography-electrospray mass spectrometry

An analytical method based on aqueous ultrasonic extraction and packed capillary liquid chromatography-electrospray mass spectrometry (LC-ESI-MS) analysis was developed and compared to an existing gas chromatography(GC)-MS based method for the determination of sarin, soman and their hydrolysis products in contaminated soil. Three soils, a red clay, a tan sandy clay and a brown sandy clay loam, were spiked with sarin and soman and their initial hydrolysis products, isopropyl methylphosphonic acid and pinacolyl methylphosphonic acid, at the 10 microg/g level to assess recovery efficiency. Recovery of sarin and soman from the aqueous soil extracts was comparable to the existing analytical method, with a significant improvement in recovery being demonstrated for the chemical warfare agent hydrolysis products. Sarin and soman were recovered in the 20-90% range from the three soil types with aqueous extraction, while the hydrolysis products of these chemical warfare agents were extracted with recoveries in excess of 80%. The developed soil extraction and analysis method appears to be an attractive alternative to the GC-MS based method, since aqueous extracts containing chemical warfare agent hydrolysis products may be analysed directly, eliminating the need for additional sample handling and derivatization steps.     

Fluorescent molecular probes for the detection of chemical warfare agents and their mimics

Owing to their direct toxic effects on human beings, animals, and plants, chemical warfare agents (CWAs) and their mimics have become widespread in chemical warfare and agriculture. The considerable concerns about their entry into biological systems and the residues in environment stimulate the development of rapid and sensitive methods for the detection and analysis of this family of compounds. In the progress of sensitive, selective, and fast responsive detection, fluorescent molecular probes have been widely used in the detection of CWAs in recent years. Here the recent reports on the design of fluorescent molecular probes and their advantages in the detection of CWAs were reviewed. Furthermore, the extensive interests accelerate the development of novel fluorescent molecular probes and detection techniques in this field.     

化学毒剂探测技术发展现状

化学毒剂(CWAs)作为一种大规模杀伤性武器,具有杀伤力强、影响范围广、防护困难、易于制作施放等特征,从诞生之初即常被用于战争冲突,在现代非对称战争、恐怖袭击中也造成了巨大威胁.因此如何及时地探测化学毒剂成为世界各国国土安全的重点研究方向.该文聚焦于化学毒剂现有主流及具有潜力的各类探测技术,对各种检测技术的基本原理及其...