Food and Chemical Toxicology 121 (2018) 343–350

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Food and Chemical Toxicology

journal homepage: www.elsevier.com/locate/foodchemtox

Review

Chemical warfare agent NOVICHOK - mini-review of available data T

a b,∗
Eugenie Nepovimova , Kamil Kuca
a
Department of Chemistry, Faculty of Science, University of Hradec Kralove, Rokitanskeho 62, 500 03, Hradec Kralove, Czech Republic
b
Philosophical Faculty, University of Hradec Kralove, Rokitanskeho 62, 500 03, Hradec Kralove, Czech Republic

A R T I C LE I N FO A B S T R A C T

Keywords: The Cold War period is characterized by the infighting between the Western countries and the USSR in diverse
NOVICHOK areas. One of such fields was development of the weapons of mass destruction. Within various programs on both
FOLIANT sides, a wide scale of different agents have been developed. However, information about some of them are still
Nerve agent protected under the designation “top secret”. Notwithstanding, in history several cases are known when such
Chemical warfare agent
information beheld the daylight. One of such cases was the program FOLIANT and NOVICHOK. Both programs
were developed by the USSR as a reaction to English/American invention of VX agent. If at least a part of
available information is truthful, we can allege that these compounds belong among the most toxic synthetic
agents ever. Within this contribution, we have reviewed available Eastern and Western data about the A-agents
and their precursors, so-called NOVICHOKs, including their history, synthesis, physical-chemical properties,
pharmacological characteristics and clinical manifestation.

1. Historical overview patent applications concerning these agents were declared secret
(Szinicz, 2005). Work on these compounds was carefully guarded and
Immediately after the World War I, nobody doubted that the next was realized under the code name Trilon. The discovery of tabun and
war would run without chemical warfare agents (CWAs). The more the sarin was further followed by the revelation of a pinacolyl analogue of
conflict seemed to be realistic, the more intensive were the preparations sarin – soman (3,3-dimethylbutan-2-yl methylphosphonofluoridate,
of potential participants. Not only the experience rising from the pre- GD, Fig. 1) in 1944 by the Nobel laureate Richard Kuhn and Konrad
vious usage of CWAs was thoroughly evaluated, but also searching for Henkel(Tucker, 2006). Tabun, sarin and soman belong to the class of
novel more effective substances gradually escalated(Halámek and nerve agents (NAs) that are collectively termed “G-agents”; the G stands
Kobliha, 2011; Klement, 2011). for German since German researchers discovered this group of com-
In 1934, a project on synthetic insecticides was launched at in- pounds(Ledgard, 2006).
dustrial corporation I. G. Farben (Germany) by Otto Bayer who as- Until April 1945, almost 9000 tonnes of tabun, 1300 tonnes of sarin
signed this research branch to the chemist Gerhard Schräder. In 1936, and 20 tonnes of soman were mass-produced by the Nazi regime,
Schräder's interest turned to organophosphorus compounds. His sys- however, they were never used (Halámek and Kobliha, 2011). There
tematic work on organophosphate (OP) insecticides led to the synthesis has been a considerable debate about why the Germans did not use
of more than 2000 compounds, including highly toxic ethyl dimethyl- their chemical arsenal in the World War II. Several explanations in-
phosphoramidocyanidate (tabun, GA, Trilon 83, Fig. 1)(Szinicz, 2005). clude: i) personal negative experience of Adolf Hitler who was seriously
Subsequently, Schräder and co-workers discovered a more lethal OP affected by mustard gas during the World War I; ii) fear of retaliation,
compound similar to tabun – propan-2-yl methylphosphonofluoridate and iii) underestimation of German superiority in chemical weapons
(GB, Trilon 46, Fig. 1). They named this compound sarin in honour of (Paxman and Harris, 2011; Pitschmann, 2014; Tucker, 2006).
the team members: Schräder, Ambros, Ritter and van der Linde After World War II, another chapter in the history of development
(Coleman, 2005). Since 1935, an official decree required for all in- and production of CWAs began. As Nazi resistance collapsed, the Soviet
ventions possessing potential military application to be reported to the and Allied forces captured the NA production facilities and technologies
German Ministry of War. In 1937, the samples of tabun and sarin were of the Germans' including scientific archives and experts in this field.
sent to the German Army Weapons Office (Wa Prüf 9) where their value Both groups quickly reorganized the military potential of these agents
for military purposes had been immediately recognized and hence all and set to work at developing and stockpiling their own supplies

∗
Corresponding author.
E-mail address: kamil.kuca@uhk.cz (K. Kuca).

https://doi.org/10.1016/j.fct.2018.09.015
Received 13 August 2018; Received in revised form 4 September 2018; Accepted 8 September 2018
Available online 11 September 2018
0278-6915/ © 2018 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/BY-NC-ND/4.0/).
E. Nepovimova, K. Kuca Food and Chemical Toxicology 121 (2018) 343–350

Fig. 1. Chemical structures of G-agents.

Fig. 2. Chemical structures of V-agents.

(Wiener and Hoffman, 2004). In 1949, British chemist Ranajit Ghosh to those handling, transporting or disposing the weapons. The compli-
discovered novel, military important group of OP esters derived from cations associated with BWs involve complicated construction of am-
variously substituted 2-aminoethanethiole. Shortly after this discovery, munition, smaller cartridges where the precursors are imposed and
the existence of these compounds had been reported to the British lower yields of the final step.(Halámek and Kobliha, 2011).
Chemical Warfare Establishment in Porton Down. VX agent (S-{2-[di Initially, the Soviets had very reserved attitude to BWs. They were
(propan-2-yl)amino]ethyl} O-ethyl methylphosphonothioate, Fig. 2) convinced that this type of ammunition is generally less efficient and
has been selected as the most promising substance of the series(Szinicz, due to complicated construction more expensive comparing to weapons
2005). In a deal brokered between the British and the US governments, filled with unitary agents. The Soviets perceived the BW program as a
the British traded the VX technology for the thermonuclear weapons way of the USA how to circumvent the upcoming Chemical Weapons
technology of the United States. Thereafter, full scale production of VX Convention (CWC) (Halámek, 2008). Such negative attitude of the
commenced in 1961 in the USA (Tucker, 2006). During the same USSR was probably determined by huge amounts of stockpiled unitary
period, the Soviet scientists developed independently of the UK and the CWAs but also by relatively low age of chemical arsenal. Not negligible
USA an isomer of VX agent – the so-called Russian VX (VR, RVX, was also the fact that the USSR did not puzzle over the disposal of
Substance 33, S-[2-(diethylamino)ethyl] O-(2-methylpropyl) methyl- expired or defective ammunition. However, with the progress of the
phosphonothioate, Fig. 2) which later became a prototype for the series time all these arguments lost importance since signing of the CWC was
of NOVICHOK agents. Another structural analogue of VX known as drawing nearer as well as the Soviet chemical arsenal was gradually
Chinese VX (CVX, O-butyl S-[2-(diethylamino)ethyl] methylpho- expiring.(Halámek, 2008).
sphonothioate, Fig. 2) was also developed and studied(Romano et al., At the time, when in the US binary weapon program was at its peak,
2007). several long-term research projects took place also in the USSR. Among
VX and its analogues belong to the class NAs that are collectively them, the most important in the field of chemical sciences were FLU-
termed “V-agents”; the V stands for venemous because they are very toxic ORINE (in Russian “FTOR”) and PHOSPHORUS (in Russian “FOSFOR”).
and the symptoms of their intoxication resemble the manifestation of These projects were of high priority not only from the point of view of
snake venom poisoning(Tucker, 2006). These phosphonylated and national economy but also of the military sector. Particular attention
phosphorylated thiocholine derivatives, lacking electron-withdrawing was paid to compounds with strong biocidal effect(Halámek, 2008;
groups such as halogen or cyano group, are very reluctant to nucleo- Vásárhelyi and Földi, 2007).
philic substitution and so to hydrolysis. This fact together with ex- Moreover, it was found that within the storage process Russian VX
tremely low volatility ensure the resorption of these substances through was very sensitive to moisture. The necessity of RVX stabilization to-
unprotected skin into the bloodstream(Ellison, 2007; Wiener and gether with development of its thickened version or even conversion to
Hoffman, 2004). the binary form led to initiation and escalation of the top secret Soviet
Until the 1950′s, the CWAs were unitary, i.e. the toxic agent was project FOLIANT between 1973 and 1976. The knowledge obtained
filled in the ammunition and then stored until its use. The problems within FLUORINE and PHOSPHORUS projects was incorporated into
resulting from the production, stockpiling of unitary CWAs as well as the FOLIANT program as well. The main aim of this project was
from requirement for expensive disposal of defective or expired che- synthesis of the third generation of NAs with higher toxicity compared
mical ammunition, disagreement of local population, activities of var- to V-agents that will be undetectable using NATO standard chemical
ious ecological organisations and last, but not least from the absence of detection equipment(Halámek, 2008; Halámek and Kobliha, 2011;
novel CWAs with desirable stability within the process of storage Vásárhelyi and Földi, 2007). More than 200 chemists and engineers
launched in the USA the project denoted Binary Lethal Weapons were involved in the FOLIANT program. According to available sources,
System. For the purpose of this project two NAs were selected – sarin at least three unitary chemical weapons were synthesized (A 230, A 232
and VX (Fig. 3) (Gupta, 2015; Halámek and Kobliha, 2011). Such and A234, Fig. 4). Structures of so-called A-agents have been never
technology involves two or more non-toxic chemical precursors physi- published. Over the last few years, the information that these com-
cally separated from each other. The final step of synthesis of the toxic pounds are derivatives of dihaloformaldoxime started to appear. This
agent from above-mentioned precursors is performed immediately be- assumption was based on published works of Soviet chemists who
fore or in the process of firing of the ammunition(Wiener and Hoffman, probably participated on the FOLIANT program(Kruglyak et al., 1972a,
2004). Therefore, binary weapons (BWs) prevent unintentional toxicity 1972b; Malekin et al., 1972; Martynov et al., 1969; Petrov et al., 1967;

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Fig. 3. The principle of binary weapons depicted on sarin and VX.

Fig. 4. Plausible chemical structures of A-agents.

Razumova et al., 1968). Interestingly, almost at the same time the fact The author of this publication was Vil Mirzayanov, a researcher of the
that acetylcholinesterase (AChE, E. C. 3.1.1.7) reactivators are able to State Research Institute of Organic Chemistry and Technology.
restore the function of OP-inhibited AChE was published(Hobbiger, Promptly after the article publication, he was arrested for high treason
1957). From the chemical point of view, such agents are oximes. Later, but due to a huge public echo a few months later he was released and
however, it was revealed that phosphorylated/phosphonylated oximes emigrated into the USA (Halámek, 2008; Karev, 2009). In this way, the
are formed immediately after the process of enzyme recovery. Such West first learned about the existence of the novel class of CWAs with a
intermediates, instead of reducing the inhibitory effect of parent OP much stronger toxic effect than the synthetic chemical warfare avail-
towards AChE, inhibit it more strongly(Andersen, 1978; Fossier et al., able at that time. Today, the information on synthesis, physical-che-
1983). mical properties, toxicity and military characteristics of A-agents and
A significant drawback of A-agents was their low stability in the NOVICHOKs is still guarded under the designation “top secret”. A
environment(Karev, 2009). However, this shortcoming seems to be substantial part of revealed data is derived only from the interviews and
negligible when a binary form of afore-mentioned agents is developed publications of Mirzayanov, Uglev and Zheleznyakov, as well as from
passing the requirement of stability on their precursors. Thus, at least 5 the chairman of the Union for Chemical Security Lev Fyodorov(Karev,
types of binary A-agents were prepared. This program bore the code 2009). The latter environmental activist was intensively involved in
name NOVICHOK. Compounds, referred to as NOVICHOKs, were declassification of the Russian chemical weapons program, in its dis-
commonly used intermediates within chemical industry and were not posal, environmental burden elimination as well as health protection of
included in proposed CWC. Therefore, particularly these compounds the population in affected areas. The ending of the project NOVICHOK
were selected as precursors of binary form of A-agents(Karev, 2009). was influenced by the coincidence of the CWC adoption (Table 1)
According to S. L. Hönig, the structural moiety of so-called NOVICHOKs (Halámek, 2008).
could be 2-fluoro-1,3,2-dioxophospholane (Fig. 5) (Hoenig, 2007). In March 2018, the former Russian spy Sergey Skripal and his
Though much of the knowledge about the synthesis and various prop- daughter Yuliya were found unconscious in Salisbury, UK. Later, the
erties of phosphorylated and/or phosponylated oximes, amidates and British authorities classified this incident as poisoning with a nerve
related compounds was published in Russian literature in the 1960′s agent from the Novichok group. On March 12, 2018, British Prime
and 1970′s, (Kruglyak et al., 1972b, 1972a; Malekin et al., 1972; Minister Theresa May stated in the parliament: “Either this was a direct
Martynov et al., 1969; Petrov et al., 1967; Razumova et al., 1968), the action by the Russian state against our country, or the Russian gov-
first exact information on A-agents and NOVICHOKs was posted only ernment lost control over the potentially catastrophically destructive
after the end of the FOLIANT project (after 1992) from the defectors substance and allowed it to fall into hands of others. Russia refused this
(Mirzayanov, 2009). blame. Inasmuch as the investigation of this incident is still under way,
In September 1992, an article describing that the USSR violates the it is currently not possible to certainly identify which country is right
CWC continuing to produce and test the NAs of the third generation, and which is deceiving(Nepovimova and Kuca, 2018).
was published in the newspaper the Moscow News(Mirzayanov, 1992).

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Fig. 5. Plausible chemical structures of precursors –

NOVICHOKs.

Table 1
The results of FOLIANT and NOVICHOK projects.
RVX RVX A 230 A 232 A 232 A 234

Form unitary binary unitary unitary binary binary

Precursor – n.a. – – Novichok 5 Novichok 7
Amount 15 000 tonnes tens of tonnes tens of tonnes several tonnes several tonnes tens of tonnes
Years of Testing n.a. 1988–89 1988–89 n.a. 1989–90 1993
Weaponization n.a. weaponized in 1990 weaponized in 1990 was not weaponized approved in 1989 n.a.

n.a. data not available.

Fig. 6. Possible synthetic approach leading to phosphorylated oximes.

2. Synthesis Novichok 5, Novichok 7) (Fig. 5) by the reaction of phosphorus

trichloride with appropriate diol and subsequent nucleophilic sub-
Since there are discrepancies in exact chemical structure of A- stitution of chlorine atom by fluorine. Resulting 2-fluoro-1,3,2-dioxa-
agents, this section is divided into two parts describing plausible syn- phospholanes readily react with dichloro(fluoro)nitrosomethane, a
thetic routes leading to single structural type. compound structurally similar to choking agents such as chloropicrin
If A-agents belong to the group of phosphorylated oximes (Fig. 4), (trichloronitromethane), fluoropicrin (trifluoronitromethane) or tri-
their synthesis would probably involve three steps(Ellison, 2016; fluoronitrosomethane. The mechanism of such reaction between phos-
Halámek and Kobliha, 2011) (Fig. 6). The first two steps would lead to phites and α-trihalonitrosoalkanes to form oxime esters of phosphates
preparation of A-agents precursors, so-called NOVICHOKs (Novichok ?, was formerly described by Allen(Allen et al., 1956; Allen and Johnson,

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Fig. 7. Synthetic route leading to phosphoroamidates described by V. Mirzayanov.

1955). So-called Allen's reaction is a subtype of Michaelis – Arbuzov data listed above, one can assume that several shortcomings typical for
reaction of trialkyl phosphite and alkyl halide to form phosphonates the first two generations of NAs had been removed.
(Burton and Flynn, 1977). The stability of formed intermediates is
probably temperature-dependent. Under subzero temperatures (- 40 °C)
they are stable but upon being warmed the nucleophilic attack by 4. Mode of action
chloride anion is facilitated resulting in phospholane ring opening
(Halámek and Kobliha, 2011; Hoenig, 2007). The mode of action of A-agents is irreversible inhibition of the AChE
Not only phosphorus chlorides or phosphorus oxychlorides could be (Hoenig, 2007). Under physiological conditions, Ser-His-Glu triad, lo-
used as starting materials, but also many other intermediates of phos- cated in the active site of AChE, hydrolyzes neurotransmitter acet-
phorus chemistry used in pesticide, plasticizer or detergent industry. ylcholine (ACh) reducing thus its concentration at neuronal cholinergic
With regard to the structures of A-agents, several general conclu- synapses and neuromuscular junctions(Dvir et al., 2010). Once the A-
sions should be mentioned: agent reaches the bottom of the active site gorge, the nucleophilic at-
tack of phosphorus atom by the hydroxyl group of serine occurs. This
1) There is no C-P bond in the molecule. Therefore, these compounds attack is accompanied by a simultaneous departure of the fluoride ion
are related to fluorophosphates which are NOT within the scope of and formation of phosphorylated enzyme (Fig. 8). Due to formation of
the CWC. the covalent bond between the phosphorus atom of OP and AChE's
2) The secondary alkoxy side chains, not being methyl, ethyl, propyl or serine, the spontaneous hydrolysis of phosphorylated enzyme is ex-
isopropyl, are also OUTSIDE the CWC scope. tremely slow varying from hours to days(Korabecny et al., 2014).
3) P-O-N linkage is NOT mentioned in the CWC. From the literature it is well-known that phosphorylated oximes are
4) The precursor fluorodichloronitrosomethane is NOT regulated by relatively unstable (Leader et al., 1999; Stenzel et al., 2007). Therefore,
the CWC, although chloropicrin is.(“Organisation for the Prohibition an analogy between phosphorylated oxime and AChE-inhibited by A-
of Chemical Weapons,” n.d.) agent could be assumed (Fig. 9). Rapid hydrolysis of = N-O- bond
within the A-agent-AChE adduct may result in so-called aged form of
According to Mirzayanov, a defector devoted to Soviet chemical the enzyme where the phosphonic oxyanion forms a salt bridge with the
weapons, A-agents belong to the group of phosphoroamidates (Fig. 7). protonated histidine that strongly stabilizes the conjugate (Fig. 8). Once
In his book State Secrets. An Insider´s Chronicle of the Russian Chemical aging occurs, the enzyme is permanently inactivated and no therapy
Weapons Program, he alleges that for the preparation of A 232 a reaction can restore its activity(Kuca and Pohanka, 2010; Sharma et al., 2015).
of methyl phosphorocyanidofluoridate with N,N-diethylethanimida- Such hypothesis is at least partly supported by the well-known fact that
mide had been approved. The latter intermediate is a product of the AChE inhibited by the A-agent ages rapidly. In addition, according to
reaction of diethylamine and acetonitrile(Halámek and Kobliha, 2011; available literature, the aging half-time of A 230 is similar to that ob-
Mirzayanov, 2009). In context of above-mentioned synthesis and served in soman, i.e. 2–4 min(Karev, 2009; Sirin et al., 2012).
character of reactants, a question arises – which of the starting com- From the therapeutic point of view, we should probably part with
pounds act as NOVICHOK precursor? the idea of restoring the function of inhibited AChE by A-agents. Not
only because of the rapid aging process, but also due to debilitated
3. Physical-chemical properties partial positive charge on phosphorus atom. Forasmuch as, the nu-
cleophile represented by currently available reactivators (pralidoxime,
Only very little information is available about physical-chemical obidoxime, HI-6) would attack such phosphorus very hardly (Fig. 8).
properties of A-agents and NOVICHOKs(Halámek and Kobliha, 2011; Therefore, the only effective treatment approaches would be sympto-
Hoenig, 2007; Karev, 2009; Mirzayanov, 2009; Pitschmann, 2014). matic therapy (combination of anticholinergic agent and antic-
Table 2 represents the best known physical-chemical properties of A- onvulsant) or administration of so-called bioscavengers, such as bu-
agents, sarin and VX. tyrylcholinesterase (BChE, E. C. 3.1.1.8). Such exogenously
A significant drawback of currently available NAs is an imbalance administered molecule/enzyme would bind and detoxify free NA en-
between their persistency and volatility. On one side, there are G-agents tering a patient's circulation so that it would not be able to reach the
for which high volatility and low persistency is characteristic. On the tissue AChE and produce clinical symptoms of intoxication(Bajgar
other hand, there are V-agents that at the expense of high persistency et al., 2009). Generally, bioscavengers belong among the prophylactic
display decreased efficacy in live power elimination by a direct action approaches, however, in case of A-agents intoxication when causal
of vapours(Newmark, 2007; Wiener and Hoffman, 2004). The aim of therapy is ineffective, they can be used ex post. Additionally, BChE is
military chemists working on the third generation of NAs was to de- able to detoxify all types of NAs representing thus the universal ap-
velop compounds with balanced physical-chemical properties such as proach since it takes quite long time to determine which NA has been
volatility, density and stability towards light and moisture. Based on the used and select appropriate therapy(Bajgar et al., 2009).

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Table 2
Available data about physical-chemical properties of A-agents, sarin and VX.
A 230 A 232 A 234 sarin VX

Molecular mass 241.95 255.97 270.00 140.09 211.2

Boiling point 61–62 °C 70–71 °C 73–74 °C 147 °C 256 °C
Density 1.612 g/mL 1.515 g/mL 1.414 g/mL 1.102 g/mL 1.062 g/mL
State liquid n.a. n.a. liquid liquid
Behaviour at low solidifies at low does not solidify at low temperature n.a. does not solidify at low does not solidify at low
temperature temperature temperature temperature
Volatility volatile more volatile than A 230 or RVX n.a. volatile not volatile
Moisture stability resistant to moisture less stable against moisture than n.a. resistant to moisture resistant to moisture
A 230 or RVX

n.a. data not available.

Fig. 8. Mechanism of AChE inhibition by A-agent, aging and reactivation by oximes.

5. Toxicity Nevertheless, there are also several differences. Within un-

authorized reports it was mentioned that the symptoms of intoxication
According to unauthorized sources, so-called A-agents exert higher were virtually incurable as well as that people who were exposed to
or at least similar toxicity as VX. Within seminar work of Karev ela- such kind of NA remained disabled invalids(Karev, 2009). Probably
borated at Penza State University, Russia, the exact toxicological data of these reports are describing the so-called delayed neurotoxicity char-
A-agents could be found(Karev, 2009). These parameters are listed in acterized by severe disruption of the nerve system manifesting as pa-
Table 3. ralysis occurring within 1–3 weeks after the intoxication(Gupta, 2015).
As NAs, also A-agents belong to the class of irreversible inhibitors of The above-mentioned scientist Andrey Zheleznyakov was probably one
AChE. These compounds block the action of AChE preventing thus of the persons who was exposed to A 232. Five years after the in-
physiological breakdown of the neurotransmitter ACh. Therefore, ACh toxication, he died suffering from cirrhosis, trigeminal neuritis and
accumulates in the synaptic cleft where it causes cholinergic receptors epilepsy(Karev, 2009).
overstimulation. The severity of the symptoms depends mainly on the
amount of the agent, which entered the body. Generally, the symptoms 6. Treatment management
could be divided into three groups: muscarinic, nicotinic and central.
Overstimulation of muscarinic cholinergic receptors causes pupils General treatment protocol of nerve agent intoxication includes
constriction, glandular hypersecretion, urination, defecation, diaphor- pharmacological as well as non-pharmacological approaches. Among
esis and gastric emesis. Among nicotinic symptoms belong initial de- the non-pharmacological methods belong oxygen supply, resuscitation
fasciculation followed by weakness and flaccid paralysis. Finally, within and decontamination(Sharma et al., 2015). For the decontamination
central nervous system, NA poisoning manifests as irritability, giddi- purposes, different countries are using diverse decontamination means
ness, fatigue, lethargy, seizures, coma and mostly fatal respiratory de- as well as different decontamination approaches based on the national
pression(Korabecny et al., 2014; Kuca et al., 2013). standards. Among them, probably the most preferred seems to be

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Fig. 9. Analogy between the A-agent and phosphorylated

oxime.

Transparency document
Table 3
Toxicological parameters of G-, V- and A-agents. Transparency document related to this article can be found online at
LCt50 (mg*min/m−3) LD50 percutaneous administration (mg/person) https://doi.org/10.1016/j.fct.2018.09.015.

sarin 100 1700 Abbreviations

soman 70 350
VX 50 10
A 232 6–10 1–2 ACh – acetylcholine; AChE – acetylcholinesterase; BChE – butyr-
A 234 7 5 ylcholinesterase; BW – binary weapon; CVX – Chinese VX; CWA –
chemical warfare agent; CWC – Chemical Weapon Convention; GA –
tabun; GB – sarin; GD – soman; NA – nerve agent; OP – organopho-
sphate; RSDL – reactive skin decontamination lotion; RVX – Russian VX;
VR – Russian VX.
personal and scene decontamination. Immediate skin decontamination
can preserve survival of casualties and prevent the progression of References
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