AnalysisofCWA - END Analyza Bojovych Chemickych Latek

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UNIVERSITY OF DEFENCE
Institute of NBC Defence
__________________________________________
S ???
ANALYSIS
OF CHEMICAL WARFARE AGENTS
The Authors:
Prof. Ing. Emil HALÁMEK, CSc.

Prof. Ing. Zbyněk KOBLIHA, CSc.
Doc. Ing. Vladimír PITSCHMANN, CSc.
VYŠKOV
2009
Motto:
The discovery of poison has undoubtedly been one of the first

revolutionary inventions at the very dawn of mankind history.
E.H.
2
___________________________________________________________________________
© Halámek, E., Kobliha, Z., Pitschmann, V. Brno 2009

ISBN
This publication or any part thereof must not be copied, reproduced or otherwise circulated
without the written permission of the publisher.
All rights of the authors are reserved.
3
Reviewers:
Ing. Ladislav Středa, CSc.

State Office for Nuclear Safety, Prague, Czech Republic
Ing. Ivana Tušarová, CSc.

Oritest Ltd, Prague, Czech Republic
PhDr. Alena Langerová

Language Training Centre, University of Defence, Brno, Czech Republic
4
Contents
List of abbreviations 6
Preface 9
1. System of technical devices used in chemical reconnaissance and chemical
check check 10
Objectives and duties of chemical reconnaissance and chemical check 10
Factors affecting the concept of chemical reconnaissance and chemical check 10
Conception of system of technical devices used in chemical reconnaissance and
chemical check 11
System of technical devices used in reconnaissance and chemical check 12
2. Analysis of chemical warfare agents 16
Phosgene 16
Diphosgene 21
Chloropicrin 23
Hydrogen cyanide 26
Cyanogen chloride 33
Sulfur yperite 37
Nitrogen yperites 49
Lewisite 58
Organophosphorus chemical warfare agents 63
Substances of the G-series (sarin-type) 71
Tabun 78
Organophosphorus fluoridates 81
Sarin 85
Soman 87
Substances of the V-series (VX-type) 89
Substance GP 97
Novichok (A-230, A-232, A-234) 100
Supplement to Chapters 2.9-2.17 101
Bromobenzyl cyanide 103
Chloroacetophenone 107
o-Chlorobenzylidenemalononitrile, compound CS 111
Dibenzo[b,f]-1,4-oxazepine, compound CR 118
Diphenylchloroarsane 123
Diphenylcyanoarsane 126
10-Chloro-9,10-dihydrophenarsazine (adamsite) 127
Quinuclidin-3-yl diphenylhydroxyacetate, compound BZ 131
3. Chemical reconnaissance and chemical check devices 140
Detector papers 140
Colorimetric biosensor 141
Chemical detectors 141
Mobile laboratories 147
4. Trends in development within the system of technical devices used in
chemical reconnaissance and check 151
5. Recommended reviews 153
List of abbreviations
A-230 O-chlorofluoroformimino-O-(2-chloroethyl) phosphoro-

fluoridate
A-232 O-chlorofluoroformimino-O-[(2-chloro-1-methyl)ethyl]
phosphorofluoridate
A-234 O-chlorofluoroformimino-O-[(2-chloro-1-methyl)propyl]
phosphorofluoridate
A-3 box trailer
ABC-M8 VGH paper for detection of liquid CWAs (US)
AC hydrogen cyanide
AČR Czech Army
ACHE acetylcholinesterase
AL-1 automobile laboratory
AL-2/ch mobile field chemical laboratory
BTY breakthrough time for sulfur yperite
BuCHE butyrylcholinesterase
BZ compound BZ; quinuclidin-3-yl diphenylhydroxyacetate;
3-quinuclidinyl benzilate
CA bromobenzyl cyanide; α-bromobenzyl cyanide
CALID-3 paper for detection of liquid CWAs (CZ)
CAMAG TLC kit
CG phosgene, carbonyl dichloride
CK cyanogen chloride
CN chloroacetophenone; ω-chloroacetophenone
CR compound CR; dibenzo[b,f]-1,4-oxazepine
CS o-chlorobenzylidenemalononitrile
CWA chemical warfare agent
DA diphenylchloroarsane; Clark I
DB-3 4-(4`-nitrobenzyl)pyridine
DC diphenylcyanoarsane; Clark II
DETEGAS-1 mobile set for analytical checking of industrial harmful agents
DETEHIT nerve paralytical agents detector; colorimetric biosensor
DM adamsite; 10-chloro-9,10-dihydrophenarsazine
DP diphosgene; trichloromethyl chloroformate
DT detector tube; tube detector, indicator tube
GA tabun; ethyl (dimethylphosphoramido)cyanidate
GB sarin; isopropyl methylphosphonofluoridate
GC gas chromatography
GC-MS gas liquid chromatography / mass spectrometry
GD soman; (3,3-dimethylbut-2-yl) methylphosphonofluoridate
GE ethylsarin; isopropyl ethylphosphonofluoridate
GF cyclosarin; cyclosin; cyclohexyl methylphosphonofluoridate
GP [2-(dimethylamino)ethyl] (dimethylphosphoramido)fluoridate
GSP-1 automatic CWA signalizer
GSP-11 automatic CWA signalizer
GSA-12 automatic CWA signalizer
GV substance GP; [2-(dimethylamino)ethyl] (dimethylphosphor-
amido)fluoridate
H sulfur yperite; bis(2-chloroethyl) sulfide
6
HD distilled sulfur yperite; bis(2-chloroethyl) sulfide
HELIOS-β spectrophotometer UV/VIS
HN-1 nitrogen yperite; bis(2-chloroethyl)ethylamine
HN-2 nitrogen yperite; bis(2-chloroethyl)methylamine
HN-3 nitrogen yperite; tris(2-chloroethyl)amine
HPLC high performance liquid chromatography
CHP-5 chemical detector
I50 50% inhibition of enzyme
IMS ion mobility spectrometry;
ISO-1C standard army container
K-7a detector paper for sulfur yperite (CZ)
KNAUER high performance liquid chromatography set;
KLAUS-STEELBRO SB30 container side-loader
KU-1 detector paper for sulfur yperite (CZ)
L lewisite
L-1 α-lewisite; (2-chlorovinyl)dichloroarsane
L-2 β-lewisit; bis(2-chlorovinyl)chloroarsane
L-3 γ-lewisit; tris(2-chlorovinyl)arsane
LIMS laboratory information and methodical system
LR Defender light terrain vehicle
M6 automatic CWA signalizer (US)
M8 automatic CWA signalizer (US)
M43 air contamination detector (US)
M256A1 CWA detector set (US)
NAVD nerve agents vapour detector, colorimetric biosensor (CA,
US)
NA nerve agent
ORI-217 CWA detection kit
PCHL-90 portable chemical laboratory
PP-1 detector paper for substance VX (CZ)
PP-3 detector paper for liquid CWAs (CZ)
PPCHL-AL-2/ch mobile field chemical laboratory T-815
PS chloropicrin; trichloronitromethane
PVC polyvinyl chloride
P-V3S medium terrain truck
Q sesqui-yperite; 1,2-bis[(2-chloroethyl)thio]ethane
R-33 S-[2-(diethylamino)ethyl] O-isobutyl methylphosphonothioate
RAID-1M instrument for rapid detection of chemical compounds by IMS
SA arsane, arsine
SONDA-CH automobile chemical laboratory T-815
SPEKOL-11 spectrophotometer UV/VIS
SRI gas chromatograph set; GC
T oxole yperite; bis{[(2-chloroethyl)thio]ethyl} ether
T-135 reagent for determination of yperites
T-815 heavy terrain truck
TLC thin-layer chromatography
VE S-[2-(diethylamino)ethyl] O-ethyl ethylphosphonothioate
VG S-[2-(diethylamino)ethyl] O,O-diethyl phosphorothioate,
Amiton
7
VM S-[2-(diethylamino)ethyl] O-ethyl methylphosphonothioate
VS S-[2-(diisopropylamino)ethyl] O-ethyl ethylphosphonothioate
VX S-[2-(diisopropylamino)ethyl] O-ethyl methylphosphonothioate
VXC S-[2-(diethylamino)ethyl] O-butyl methylphosphonothioate
VX model S-[2-(dimethylamino)ethyl] O-ethyl methylphosphonothioate
3-WAY paper for detection of liquid CWAs V-G-H (CA, US)
8
Preface
Advances in physico-chemical instrumentation used for analytical checking has long

been diverting the chemists' attention from classical chemical analytical methods. Naturally,
this trend has affected the whole field of military chemistry, and, in particular, the analysis of
chemical warfare agents and other toxic compounds of military importance. The objective of
the present work is to bridge, at least partially, this looming gap in the knowledge of chemical
analysis of warfare agents. However, the methods and procedures of chemical analysis
constitute a basis of not only laboratory chemical checking but also of the many various
means of chemical reconnaissance and checking, and simple means and detection methods of
chemical warfare agents. These means are still being widely utilized by the military, to protect
civilians, and by fire brigades and rescue service as the primary source of information on a
chemical situation. These means and methods are also of irreplaceable significance as
complementary information sources. Of no less importance is also the fact that these means
and methods enable a rapid and flexible response to the changing situation in providing
protection against weapons of mass destruction (chemical weapons), as distinct from the often
complicated specialized appliances and very expensive automatic detectors with rigidly
preprogrammed sensitivity and selectivity. This publication as such should serve as a
complementary source of relevant information mainly for chemical specialists of the Czech
Army, students of military academies, interested persons from member organisations of the
Association of Defence Industry of the Czech Republic, members of the Fire and Rescue
Brigade of the Czech Republic, and for selected staff of the state administration responsible
for the protection of population.
Vyškov, April 4, 2007
The Authors
Preface to the amended and revised second edition
Soon after the Czech edition was published, the demand for an English version has
appeared repeatedly in addition to many mostly positive reactions. This demand was urgent
especially on the part of international students at our university. Therefore, we created the
English translation without any hesitation. Concurrently, we updated the text and added
sources of literature with the aim of improving the monographic value of this specific
textbook.
Vyškov, April 24, 2009
The Authors
9
1. System of technical devices used in chemical reconnaissance and chemical check
The present world situation can be characterized as a dynamic period of great and violent
changes. The end of the Cold War has brought an only seemingly lower risk of a high
intensity global conflict because, at the same time, it has resulted in a series of regional
conflicts of lower intensity but expected to last quite long. These conflicts together with new
forms of international terrorism represent an acute danger because due to the bipolar world
disintegration they are accompanied by the proliferation of weapons of mass destruction and
by the spread of relevant technologies difficult to control. Ominous visions and prospects of a
deideologized world tossed by ethnic and religious conflicts are beginning to take shape at a
horrifying rate. Every day, world events confirm the fact known to specialists that chemical
weapons as a type of mass destruction weapons are technologically accessible. Moreover, it is
evident that chemical weapons may easily be exploited by terrorists for conducting a
psychological war. High standard chemical reconnaissance and chemical check become thus a
permanent prerequisite for effective protection against the effects of chemical weapons, the
consequences of accidents and disasters, acts of sabotage and terrorist attacks accompanied by
the escape of toxic chemicals as well as against the threat of their use. Analytical chemical
check is also an indispensable part of environmental surveillance of the armed forces as well
as the population.
1.1 Objectives and duties of chemical reconnaissance and chemical check
Chemical reconnaissance and chemical check must support many tasks. The most
important task is to detect a chemical attack, identify the chemical weapon (CWA) used and
sound the end of chemical contamination. Early determination of the beginning and end of
contamination is in fact decisive for how long individual and collective protective equipment
is to be used and also for the magnitude of revocable and irrevocable losses of manpower due
to the toxic effects of chemical weapons and possibly for the consequences of long-term
employment of protective equipment. Identification of the type of a CWA or other toxic
compound is important for assessing the losses, duration of contamination of air, material
surface, military equipment and the terrain, for choosing decontamination technologies,
procedures and protocols and possibly also for selecting specific antidotes and drugs. Out of
the numerous demanding and specific tasks for analytical chemical check that can be
performed only by chemical specialists in mobile or stationary laboratories, the main tasks are
to supplement and extend the results obtained by chemical reconnaissance. The aim is to
increase, using chemical analysis, the reliability of information gained from the contaminated
samples, to determine the extent of contamination, i.e. concentration of a CWA or harmful
agents and possibly to determine other militarily significant compounds, such as diverse and
sabotage poisons, industrial poisons, agrochemicals, rocket fuels, etc. In a broader sense,
chemical check ranging from mobile devices to stationary laboratories must provide all
relevant information on any chemical contamination on the territory and provide analytical
check and expertise on all chemical materials, including constitutive analysis of organic
compounds and identification of unknown toxic compounds.
1.2 Factors affecting the concept of chemical reconnaissance and chemical check
The tasks of chemical reconnaissance and chemical check are limited by a number of
factors. They include the military doctrine reflecting the decisive political and regional
aspects, opinions on the use of chemical weapons in combat and operation – whether of
10
friendly forces or a potential enemy, the industrial and technological level of the economy,
proportion of the national budget allocated to defence and the level of the training and
educational system of the army, the alliance and the country. These as well as other factors
influence the concept of chemical reconnaissance and chemical check, which under the army
conditions is expressed as a significant part of the position and role of the Chemical Corps,
being in the materially-technical field manifested as a system of technical devices of chemical
reconnaissance and chemical check.
1.3 Concept of technical devices system used in chemical reconnaissance and chemical
check
In addition to the above-mentioned basic tasks, the system of technical devices of
chemical reconnaissance and chemical check must provide two kinds of protection against
chemical weapons. First, it is the information on the employment of chemical weapons and
other militarily significant toxic substances in the theatre of war, which allows the unit
commands to effectively control the combat activity with stress put on tactical warning and
the elimination of consequences. Second, but by no means less important, it is the necessity of
supporting the survival of individuals under the demanding conditions of using chemical
weapons even in extreme situations with no available communication, in cases of separation
from the unit, not to mention the military professions fulfilling their tasks independently, such
as surveyors, observers, forward air controllers, shot down pilots, etc. Moreover, technical
devices of chemical reconnaissance and chemical check are expected to be operationally
reliable with low operation demands or even working automatically and, at the same time,
being able to rapidly, sensitively and selectively identify all the relevant warfare agents or
possibly other militarily significant toxic compounds. Other requirements also have to be
mentioned, such as the autonomous function independent of power sources, materials such as
fillings, indicator devices, etc. and further, small size and weight, domestic production,
resistance to battleground influences, ability of interconnection with an automated data
collection system and, last but not least, low price. It is thus evident that no single device can
be designed in such a way as to meet all the requirements mentioned. This can be satisfied
only by a whole system of technical devices which, moreover, is continually replenished and
modernized. Logically, the varied and numerous tasks of chemical check rule out the
employment of only one device or equipment. With the exception of some limited and special
tasks, chemical check can be carried out only in a laboratory, i.e. in mobile laboratories of
various sizes and purposes in a field army. According to their size and weight, such
laboratories fall into two categories, i.e. portable laboratories transportable at least over a
short distance by an individual or mobile laboratories which depend on transport vehicles. In
addition to these incorporated mobile chemical laboratories, there is a stationary installation
built on the national territory and designed for army needs in which all chemical check tasks
beyond the possibilities of the deployed field forces would be solved. Such an installation
must be adequately manned as well as materially capable of verification and identification of
extremely toxic organic compounds including the capability of issuing relevant
recommendations and suggestions for protection against toxic effects of any chemical
compound. This is one of the most complicated problems which the army and its chemical
specialists may face during a conflict. In fact, all these chemical check tasks are being solved
even in peacetime as part of the necessary enhancement of protection against the effects of
mass destruction weapons.
11
1.4 System of technical devices used in chemical reconnaissance and chemical check
It is evident that the main tasks of chemical reconnaissance and chemical check will
necessarily be supported using a whole system of technical devices at the corresponding
command levels. The interconnection of systems of technical devices used in chemical
reconnaissance and chemical check is important not only from the viewpoint of early tactical
warning, but advancing to higher levels of command its importance increases even as
concerns the defence doctrine of the country. The required tasks and their scope can be
fulfilled only by applying various principles used by chemical reconnaissance and chemical
check, i.e. physical, chemical, biochemical and physico-chemical principles as well as by
using devices of various design level. Any one-sided preference of a principle or method must
sooner or later end in the loss of effectivity of the whole system. Moreover, the system has to
be systematically complemented and modernized to keep up the pace with the changes in the
chemical warfare. All the current changes in the armed forces, such as their
restructuralization, rearmament, integration into the NATO structures and UN Peace Forces,
participation in foreign missions as well as the fulfilment of the military doctrine itself, must
be supported by a concept of chemical reconnaissance and chemical check materialized in the
existing system of technical devices (see Table 1).
Table 1: System of technical devices used in chemical reconnaissance and check

(a variant)
Type of Task of chemical Command level
the device reconnaissance and chemical
ind sq pn coy bn bde ac
check
A Detection of dangerous NA
concentrations in air, on surface, ⊕
in water and food
B Detection of surface
contamination by liquid CWAs ⊕
C Detection of CWAs in air and on
the surface of military equipment,
material and terrain, ⊕
determination of the type of
CWAs
D Rapid and automatic detection of
CWAs in air ⊕
E Determination of degree of
contamination by CWAs ⊕
F Determination of other toxic
substances ⊕
G Air chemical reconnaissance ⊕
Abbreviations: ind – individual; sq – squad; pn – platoon; coy – company; bn – battalion;
bde - brigade; ac - army corps
12
Type A – Individual detector
This device should support the identification of the most significant CWAs, i.e. at least nerve
agents (NAs) and sulfur yperite in dangerous and tolerable concentrations in air, water, food
and on the surface of weapons, equipment and terrain. It has to be independent of power
sources and other materials simple to handle and has to enable the survival of manpower
under extreme conditions of using chemical weapons. It must support a timely and safe
removal of individual protection equipment, prevent the unnecessary exhaustion of manpower
and thus operability decline of the combat readiness of troops. In an emergency it must enable
checking water and food for the content of NAs and yperite.
Principle used: Biochemical reactions (colorimetric biosensor), chemical chromogenic
reactions.
Type B – Detector of liquid CWAs
A simple detector in the form of paper, foil, chalk or dye. It must rapidly indicate surface
contamination by liquid CWAs. In an optimal case, it should identify the substance or type of
a CWA and optionally enable the assessment of contamination degree. Stress is laid on
persistent CWAs that contaminate the surface for several hours to several days.
Principle used: Physical (visual changes accompanying the dissolution of dyes and indicators
in a liquid CWA) or chemical, chromogenic reactions.
Type C – Chemical detector
A simple, widely used device, usually employing detector tubes (DTs) to identify groups of
CWAs or individual CWAs. Chromogenic agents are predominantly applied on inert carriers
at given doses or they are kept in the ampoules of DTs. The presence of CWAs is indicated by
the corresponding colour change. Apart from DTs, the chemical detector kit usually contains
a manual or electric pump and other accessories. The device represents a classical tool
traditionally used by all armies. Recently, DTs without ampoules for long-time check of air,
water analysis or, if necessary, of capability to assess the concentration of CWAs in a vapour-
air mixture are required. In addition to DTs, recently there has been a trend towards using disc
carriers of chromogenic agents with the corresponding developing agents in ampoules. It is
not necessary to pass air through the detection part. For the passing of air, one can use a pump
or also a modified intake chamber of a protecting mask.
Principle used: Chemical and biochemical colorimetric reactions.
Type D – Fast detector of CWAs
An automatic electronic device with a rapid response to the presence of CWAs in air (in an
order of seconds) that can be integrated into an automated data collection system. Since it has
to work even without being connected to the vehicle’s power network, it must be equipped
with its own battery. The presence of a CWA at a given site is usually signalled acoustically
and also visually by a warning light. Sometimes, the kit comprises a long-range signalling
panel. Instruments which do not require further supporting devices, e.g. indication kits are
preferred. The device is considered to be the main component of the system of technical
devices used in chemical reconnaissance that enables warning of troops, particularly those in
the downwind direction. Rapid monitoring of a momentary chemical situation allows the
qualified assessment of casualties and thus supports effective decision-making process at
higher level of command. These devices linked to the automated data collection network must
13
be available for use at a command level corresponding to a standard target of a chemical
weapon attack, i.e. a company (battalion).
Principle used: Ionization method, ion mobility spectrometry (IMS), flame-ionization
photometry (PFD), gas chromatography-mass spectrometry (GC/MS).
Type E – Portable field chemical laboratory
Equipment which supports the basic task of chemical check, i.e. determination of the
contamination degree caused by all CWAs known. This implies mainly a collection of
samples and their qualitative and, if need be, quantitative analysis. Currently, this task has
been accomplished using a portable chemical laboratory intended for the analysis of collected
samples. Moreover, such equipment enables to confirm and complete information obtained by
chemical reconnaissance. Such equipment is preferred since it is simple to use; further, it uses
fast methods, pre-prepared reagents and materials. Simple devices of A, B and C types are
used as well, e.g. detector papers, tickets and DTs.
Principle used: Chemical and biochemical reactions with visual indication, thin-layer
chromatography (TLC).
Type F – Mobile chemical laboratory
The determination of other important compounds of military interest, such as toxic industrial
harmful agents, agrochemicals, sabotage poisons and rocket fuels, requires the more
complicated analysis of collected and delivered samples and thus a better equipped chemical
laboratory staffed by specially trained university graduates. This laboratory dealing with such
a broad task is often placed on a truck undercarriage or in transport containers. The activities
performed in this laboratory include the analysis of decontamination compositions and their
components, fogging and ignition mixtures, explosives as well as equipment of individual
chemical protection, both own and captured.
Principle used: In addition to the classical methods of chemical analysis, the instrumental
physico-chemical methods, such as UV/VIS spectrophotometry, thin-layer chromatography
(TLC), high performance liquid chromatography (HPLC) and gas-liquid chromatography with
mass spectrometric detection (GC-MS) or optionally with detectors of other types.
Type G – Long-range detector of CWAs
Air chemical reconnaissance using a long-range detector represents not only the only possible
solution of this task, but also the only effective utilization of this toxic agent detection device.
Its high cost, terrain ruggedness, high pollution of the lower layer of the atmosphere at the
battlefield and the long-range capacity of detectors are the reasons usually not allowing the
effective utilization of this device on the ground. By employing air chemical reconnaissance
over the region of interest, the long-range detector can effectively be used without being
affected by the above-mentioned undesirable factors.
Principle used: IR spectroscopy, Raman spectroscopy.
References to chapter 1.
1. ETTEL, V.: Chemická válka. Vědecký vojenský ústav. Praha 1932. 415 s..
2. TOMEČEK, I., MATOUŠEK, J.: Analýza bojových otravných látek. SPN. Praha 1961. 235 s.
3. TOMEČEK, I.: Chemie a analýza bojových otravných látek. VA Brno 1966. 166 s.
4. FRANKE, S.: Lehrbuch der Militärchemie. 2. vydání. Díl 1 a 2. Deutscher Militärverlag der DDR. Berlin
1977. 512/616 s.
14
5. ČEBOTARJEV, O.V.: Vojskovaja indikacija. Izdanie akademii. Moskva 1983.
6. HALÁMEK, E.: Perspektiva vývoje prostředků detekce otravných látek. In: Sborník chemického vojska, 27.
Praha, MNO/SCHV, 1983, s. 73 - 80.
7. HALÁMEK, E.: Chemická kontrola otravných látek a dalších vojensky významných sloučenin. [Habilitační
práce]. Vyškov 1990. 216 s. VVŠ PV.
8. HALÁMEK, E.: Trendy vývoje prostředků chemického průzkumu a kontroly. In: Vojenské rozhledy, 9(41),
2000, zvláštní číslo, s.89 - 94.
9. RYBALČENKO, I.V.: Iděntifikacija toksičnych chimikatov. In: Ros. Chim. Ž., 2002, T. XLVI, No 4, s.64-70.
15
2. Analysis of chemical warfare agents
2.1 Phosgene
Cl
C O
Cl
Phosgene is a typical acylating and halogenating compound easily undergoing

nucleophilic substitution reactions. Practically the oldest method for the detection of phosgene
in air is its reaction with aniline water (about 3% aniline solution in water). The reaction with
phosgene results in white turbidity caused by the sparingly water-soluble diphenylurea.
2.1.1
NH
COCl2 4 NH2 O C 2 NH3 Cl
NH
diphenylurea
difenylmočovina aniline hydrochloride
anilinium chlorid
The procedure has been used for the gravimetric quantitative determination of phosgene;
however, it is not sensitive enough for its detection [1].
In the presence of some metal and non-metal chlorides (LEWIS acids), phosgene acylates
tertiary aromatic amines to give products typical for FRIEDEL-CRAFTS reactions. Thus, e.g.
phosgene reacts with N,N-dimethylaniline in the presence of ZnCl2 under the formation of
MICHLER’s ketone or, in the presence of AsCl3 or PCl3, it gives a carbinol, Crystal Violet.
2.1.2
O
ZnCl2
COCl2 4 N(CH3)2 (H3C)2N C N(CH3)2
bis(p-dimethylaminofenyl)keton
bis(p-dimethylaminophenyl)ketone
2 NH(CH3)2 Cl
N,N-dimethylaniliniumchloride
N,N-dimethylanilinium chlorid
AsCl3 2.1.3
COCl2 5 N(CH3)2
OH
(H3C)2N C N(CH3)2 2 NH(CH3)2 Cl
N(CH3)2
tris(p-dimethylaminofenyl)karbinol
tris(p-dimethylaminophenyl)carbinol
16
These products are also chromogenic analytical reagents that can be employed for the
construction of simple detecting devices. Chromogenic products also arise in the reaction of
phosgene with p-dimethylaminobenzaldehyde and aromatic amines [2,3].
The most frequently used method is based on the reaction of phosgene with HARRISON
reagent, p-dimethylaminobenzaldehyde and N,N-dimethylaniline (2.1.4). In accordance with
the reaction mechanism, the blue-green coloration of the diphenylmethine dye and of the
oxidized form of Crystal Violet develops gradually. The reaction is selective for phosgene and
diphosgene. Acid fumes, hydrogen chloride, nitrogen oxides, adamsite and chloropicrin cause
a yellow coloration. The sensitivity of a detector tube based on this reaction is at least 5 µg.l-1
in air. The use of other aromatic amines instead of N,N-dimethylaniline results in different
colorations (different dyes). Detector papers and detector tubes traditionally make use of
diphenylamine which gives yellow coloration. m-Dimethylaminophenol gives a blue-violet
pyronine-type dye [4]. In the HARRISON reagent, p-dimethylamino-benzaldehyde can be
replaced by e.g. MICHLER’s ketone [5].
2.1.4
COCl2 (H3C)2N CHO (H3C)2N CHCl2 CO2
(H3C)2N CH N(CH3)2 N(CH3)2

- HCl
Cl
(H3C)2N (H3C)2N CH N(CH3)2 Cl
tautomerizacígives
tautomerization vzniká
rise modrozelené zbarvení of
to blue-green coloration
difenylmethanového
diphenylmethane (methine)(methinového)
dye barviva
(H3C)2N CH NH(CH3)2 Cl
bezbarváleuco-base
colourless leukobázeofkrystalové violeti
Crystal Violet
N(CH3)2
+ 1/2 O2
(H3C)2N C N(CH3)2 Cl H2O
violet oxidized
fialová formforma
oxidovaná of Crystal Violet violeti
krystalové
N(CH3)2
17
Another frequently employed reaction for the detection of acyl halides and optionally also
alkyl halides is the reaction with 4-(4'-nitrobenzyl)pyridine [3]. Phosgene with this reagent
gives rise to a yellow to orange coloration.
2.1.5
COCl2 2 O2N CH2 N
O2N CH2 N C N CH2 NO2 2 Cl
2 Cl
O2N CH NH C NH CH NO2
The formed condensation product of substituted urea character is converted by

tautomerization into a coloured (red) mesoquinoid form of methine dye. The selectivity of the
reaction is not high; similar reaction also occurs with many acyl and alkyl halides. Higher
selectivity, sensitivity and colour deepening to red can be achieved by adding
N-phenyl(benzyl)amine. The reaction is employed for the construction of detector papers [6-
9], chalks [10], pencils and DTs [11-21]. Similarly to 4-(4'-nitrobenzyl)pyridine, anabasine
also reacts with phosgene under the formation of a red coloration. The reaction has been
recommended for the preparation of detector papers.
The aminoperoxide or hydrogen peroxide reaction, usually named the SCHÖNEMANN
reaction or the ALEXEJEV reaction in the Russian literature, is also a group reaction for the
determination of alkyl halides. It is based on nucleophilic substitution of halogen atom with
hydrogen peroxide ion in a slightly alkaline medium of pH 7-8 leading to the corresponding
peroxy acid of high oxidation potential. This peroxy acid then reacts with the reduced form of
a redox indicator to give its coloured oxidized form.
2.1.6
pH 7 - 8
COCl2 2 OOH CO(OOH)2 2 Cl
2 H2N NH2
benzidine
benzidin
HN NH H2N NO benzidine
benzidin
2
3 H2O CO2 2 H2O CO2

- H2O
H2N N N NH2
18
With benzidine as a redox indicator, the oxidized product has a yellow-green colour.
According to the literature, the product is an azo-dye arising by the condensation of
mesoquinoid form of benzidine. However, the formation of coloured oxidized forms of
aromatic amine such as nitroso or nitro derivatives seems to be more probable. Because of the
suspected carcinogeneity of benzidine the redox indicators commonly used are its methyl
derivatives, traditionally o-tolidine, o-dianisidine and others. The coloration obtained is also
brighter, being red, violet or blue. The most often employed reagent is o-dianisidine. Besides
detector papers and DTs, this chromogenic reaction has also been applied to automatic CWA
signalizers (GSP-1 or M6). For phosgene as well as diphosgene, this method is not very
sensitive (0.1 mg.l-1 in air) probably due to the low chemical stability of the arising
peroxycarbonic acid. In addition to its low sensitivity, the method does not afford even
reproducible results in quantitative determinations.
Acylation of phenylhydrazine, leading to diphenylcarbazide, represents one of other reactions
for the detection of phosgene. As a metallochromic indicator, this compound forms
characteristically coloured complexes with heavy metals. The reaction with Cu2+ leading to a
benzene-extractable red chelate is usually employed [22].
2.1.7
HN NH
COCl2 4 NH NH2 O C 2 NH NH3 Cl
HN NH
HN NH N NH
2+
O C Cu Cu C O 2 H+
HN NH N NH
In an acid medium, diphenylcarbazide affords complexes with Hg2+ ions of an intensive blue
colour. Besides for determination in a solution, the reaction is used for the construction of
detector papers. The detection is selective and its sensitivity amounts to 20 µg .l-1 in air [23].
Phosgene can be quantitatively determined by titrimetric methods. Acid-base titration is based
on the reaction of phosgene as an acyl halide with an excess hydroxide solution which then is
back titrated with a volumetric acid solution, usually to an acid-base indicator or using a glass
electrode potentiometric indication.
2.1.8
COCl2 2 NaOH CO2 2 NaCl H2O

2.1.9
COCl2 3 NaOH Na HCO3 2 NaCl H2O
2.1.10
COCl2 4 NaOH Na2CO3 2 NaCl 2 H 2O
Analogously, the argentometric determination of formed chlorides is possible by titration with

AgNO3 which forms white insoluble AgCl. The equivalence point can be indicated by a silver
metal electrode or a silver halide membrane ion-selective electrode. The titrimetric
determination of chlorides by mercurimetry is also possible, usually with a mercury(II) nitrate
19
solution and the visual indication using metallochromic indicators or ion-selective chloride
electrodes because the reaction of chlorides with mercury(II) ions gives rise to undissociated
mercury(II) chloride [24-28].
As a chlorinating agent, phosgene reacts with a potassium iodide solution in anhydrous
acetone with the liberation of iodine.
2.1.11
COCl2 KI 2 KCl I2 CO
This reaction can be employed for the iodometric determination of phosgene by sodium
thiosulfate. The reaction also takes place stoichiometrically in acetone with diphosgene and
can be utilized for the spectrophotometric determination of phosgene at 365 nm.
2.1.12
N N
COCl2 COCl2
N N N N
N N
The direct alkalimetric determination of phosgene is best performed after its reaction
with hexamethylenetetramine (urotropine) with which it forms a stable addition product [29].
20
2.2 Diphosgene
Cl
O O
C C
Cl
Cl Cl
Diphosgene as well as its chemical properties is related to phosgene. From the

viewpoint of particular detection devices and the reaction principles used in them, there is no
difference between these two compounds. The similarity is reflected not only by their trivial
names, but also by the thermal cleavage of diphosgene to phosgene which is quantitative at
temperatures of 300-350 °C.
2.2.1
Cl O
t
Cl C O C Cl 2 COCl2
Cl
In comparison with phosgene, the only difference is the so-called dimethylaniline test in
which diphosgene on heating affords an unidentified violet product [30].
From the viewpoint of simple reconnaissance instruments, there is a fundamental difference
in the physical properties of both compounds, mainly in their physical state under normal
conditions. An analogous situation is also with triphosgene (perchlorodimethyl carbonate).
References to chapter 2.1-2.2

1. ALEXANDROV, V.N., JEMELJANOV, V.I.: Otravljajuščije věščestva. 2. vydání. Vojennoje izdatělstvo.
Moskva 1990. 272 s.
2. MOUREU, H., CHOVIN, P., TRUFFERT, L.: Sur la transformation photochimique de la loropicrine en
phosgene et la caracterisation de ces deux corps. Compt. Rend. 228, 1954-1956 (1949).
3. LINCH, A.L., LORD, S.S., KUBITZ, K.A., DE BRUNNER, M.R.: Phosgene in air – development of
improved detection procedures. Am. Ind. Hyg. Assoc. J. 26(5), 465-474 (1965).
4. SPĚVÁK, A., KRATOCHVÍL, V.: Rychlé semikvantitativní stanovení koncentrace fosgenu v ovzduší. Chem.
Prům. 15(11), 682-685 (1965).
5. GILMAN, H., SWEENEY, O.R., HECK, L.L.: A delicate color test for Michler's ketone and a less sensitive
test for phosgene and dialkylanilines. J. Am. Chem. Soc. 52(4), 1604-1607 (1930).
6. DIXON, B.E., HANDS, G.C.: A field method for the determination of phosgene. Analyst 84, 463-464 (1959).
7. NAKANO, N., YAMAMOTO, A., KOBAYASHI, Y., NAGASHIMA, K.: Development of a monitoring tape
for phosgene in air. Talanta 42(4), 641-645 (1995).
8. PITSCHMANN, V., KOBLIHA, Z., HALÁMEK, E., TUŠAROVÁ, I.: A simple in situ visual and
tristimulus colorimetric method for the determination of diphosgene in air. J. Serb.Chem.Soc., 72(10), 1031-
1037 (2007).
9. ORITEST. Indikační roztok pro monitorovací pásku ke stanovení fosgenu. Původce: Pitschmann, V.,
Tušarová, I., Veverka, V. Int. G01N31/22. CZ 14647. Zapsáno 24. 8. 2004.
10. WITTEN, B., PROSTAK, A.: Sensitive detector crayons for phosgene, hydrogen cyanide, cyanogen
chloride, and lewisite. Anal. Chem. 29(6), 885-887 (1957).
11. KOVÁŘ,V., HORÁK, O., MATOUŠEK, J.: Trubičkové detektory průmyslových škodlivin pro kontinuální
kontrolu ovzduší. Chem. Prům. 14(12), 663-665 (1964).
21
12. PITSCHMANN, V., KOBLIHA, Z., HALÁMEK, E., TUŠAROVÁ, I.: New detector tube for phosgene,
hydrogen cyanide and cyanogen chloride and its evaluation by tristimulus colorimetry. Chem. Anal. (Warsaw),
2007, ročník 53, číslo 2, s. 47-57.
13. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z.: Trubička na určení koncentrace fosgenu a difosgenu.
Sborník VVŠ PV Vyškov, 2003, číslo 1, s. 19-25.
14. ORITEST. Způsob analytické kontroly průmyslových škodlivin.
Původce vynálezu: Pitschmann V., Halámek E., Tušarová I. Int. Cl. G01N21/78. CZ 284 693. Udělení
oznámeno ve Věstníku č. 2/99, 17.2.1999.
15. ORITEST. Detekční trubička pro zjišťování dusivých a krevních jedů ve vzduchu. Původce vynálezu:
Pitschmann V., Halámek E. Int. Cl. G01N31/22. CZ 288 125. Udělení oznámeno ve Věstníku č. 4/2001,
11.4.2001.
16. ORITEST. Detekční trubička pro zjišťování yperitu a fosgenu či difosgenu ve vzduchu. Původce:
Pitschmann V., Halámek E. Int. Cl. G01N31/22. CZ 8222. Zapsáno 1.2.1999.
17. ORITEST. Indikační náplň detekční trubičky na fosgen a difosgen. Původce: Pitschmann V., Halámek E.,
Kobliha Z. Int. Cl. G01N31/22. CZ 12413. Zapsáno 8.7. 2002.
18. ORITEST. Detekční trubička pro zjišťování bojových chemických látek. Původce: Pitschmann V., Orel J.
Int. Cl. G01N31/22. CZ 12840. Zapsáno 2.12.2002.
19. ORITEST. Detekční trubička na fosgen, difosgen, kyanovodík a chlorkyan, s kapalným chloračním činidlem.
Původce: Pitschmann V., Halámek E. Int. Cl. G01N31/22. CZ 17643. Zapsáno 2. 7. 2007.
20. ORITEST. Indikační náplň ke stanovení nízkých koncentrací fosgenu v ovzduší. Původce: Pitschmann V.
PV 2007-802 (přihlášeno 19.11.2007)
21. OLTHOFF, U., EICHARDT, A., PEIZING, K., CZEPUCK, A.: Prüfröhrchen zum Nachweis toxischer
Substanzen in der Luft. Int. Cl. G01N31/22. DD 300 836 A7. (13.8.1992). Bundesamtes für wehrtechnik und
beschaffung, Koblenz.
24. ČIHALÍK, J.: Potenciometrie. Nakladatelství ČSAV. Praha. 1961. 772 s.
25. KORYTA, J., ŠTULÍK, K.: Iontově selektivní elektrody. Akademia. Praha . 1984. 208 s.
26. HALÁMEK, E.: Příspěvek k elektrochemickému stanovení některých bojových otravných látek.
[Kandidátská disertace]. Brno 1979. 118 s. - Vojenská akademie.
27. ČAPOUN, T., KOBLIHA, Z., HALÁMEK, E.: CO-3-16 Analýza vysoce toxických látek v chemických
laboratořích Civilní ochrany ČR. Příloha 6: Potenciometrické titrace. MO Praha. 23 s.; MV ČR, Praha: Příloha
6 k PO-28/28-164/ICO-2002. Potenciometrické titrace. 2002. 28 s.
28. HALÁMEK, E., SOUČEK, J.: Možnosti iontově selektivních elektrod pro stanovení otravných látek. Civilní
obrana, 28, 1986, 2, s . 75-87.
1977. 512/616 s.
30. TOMEČEK, I., MATOUŠEK, J.:Analýza bojových otravných látek. SPN. Praha. 1961. 235 s.
22
2.3 Chloropicrin
Cl
Cl C Cl
NO2
Chloropicrin (trichloronitromethane) reacts readily with aqueous-alcoholic solutions

of alkali metal sulfides, liberating free sulfur which makes the solution turbid. The products
and their proportions depend on the reaction ratio.
2.3.1
Cl3C NO2 Na2S S CO NOCl 2 NaCl
2.3.2
2 Cl3C NO2 3 Na 2S 3S N2 2 CO2 6 NaCl
2.3.3
7 Cl3C NO2 11 Na2S 7S 2 CO 2 CO2 2 COS CS2 N2
4 NO NaNO2 21 NaCl
Rather than for detection, the reaction of chloropicrin with alkali metal sulfides is used for the
disposal of this compound that otherwise is very resistant to common nucleophilic
substitution-based decontamination procedures.
The ethanolic potassium iodide solution reacts with chloropicrine to afford tetraiodomethane,
potassium chloride and potassium nitrite. Subsequently, the nitrite, a strong oxidation reagent,
converts the iodide to free iodine.
2.3.4
I
Cl3C NO2 4 KI I C I 3 KCl KNO2
It is essential to perform the reaction with KI in an acid medium to liberate iodine because
under neutral conditions it leads to the formation of colourless hypoiodites.
2.3.5
2 KNO2 6 KI 8 H+ 8 K+ 3 I2 N2 4 H 2O
2.3.6
2 KNO2 3 KI H2O N2 3 KIO 2 KOH
Traces of free iodine can be visualized by adding a starch or dextran solution which forms a
blue clathrate complex with iodine.
23
2.3.7
I2 (C 6H 10O 5)n [(C 6H 10O 5)n . I2]
At a normal temperature, chloropicrin is hydrolyzed very slowly with water and strong
alkalies. It is, however, decomposed by the nucleophilic substitution reaction with alcoholate
ions, usually with sodium or potassium ethoxide in ethanol, which proceeds readily and
quantitatively [1].
2.3.8
Cl3C NO2 4 OR C(OR)4 3 Cl NO2
Under acid conditions, nitrites are converted into nitrous acid which, as a diazotation reagent,
converts primary aromatic amines into highly reactive diazonium salts.
2.3.9
NO2 HCl HNO2 Cl

2.3.10
NH2
N N
HNO2 2 H2O
H
SO3H
SO3H
NH2
H HO3S N N NH2
The diazonium salt, formed by sulfanilic acid (p-aminobenzenesulfonic acid), couples with
the so-called passive component (1-naphthylamine) under the formation of a red azo-dye.
This GRIESS reaction is used for the selective and sensitive detection of chloropicrin as well
as for its UV-VIS spectrophotometric quantitative determination. (Reagent containing
sulfanilic acid and 1-naphthylamine in acetic acid is called the GRIESS reagent) [2]. The
method offers many modifications by replacing sulfanilic acid with a series of primary
aromatic amines and using great number of passive components that couple with diazonium
salts in acid as well as a neutral or alkaline medium affording many azo-dyes. At the same
time, some aromatic amines, e.g. benzidine derivatives may function as passive components.
Chloropicrin is prepared by oxochlorination of picric acid in an alkaline medium. As a
compound of high oxidation potential, it is reduced very easily. One of the possible reducing
agents is tin(II) chloride which converts chloropicrin to cyanogen chloride.
24
Cl3C NO2 3 SnCl2 4 HCl Cl CN 3 SnCl4 2 H2O
2.3.11
The chlorinating effects of chloropicrin are shown in its reaction with potassium cyanide that
leads to cyanogen chloride as well [1].
2.3.12
Cl Cl
2 Cl3C NO2 KCN O2N C C NO2 Cl CN KCl
Cl Cl
The arising cyanogen chloride is determined by some of the modifications of the KÖNIG-
ZINCKE reaction (see Chapter 2.5, reaction 2.5.5) [3] or as an acyl halide by the
SCHÖNEMANN aminoperoxide reaction. Interestingly enough, this reaction principle has
been used for the construction of a detector tube containing an ampoule with the pyridine-
dimedone reagent and another ampoule with crystalline potassium cyanide. The detection
limit is 6 µg.l-1 in air [4]. Currently, detector tubes for chloropicrin contain a layer with a
strong oxidation reagent such as Cr6+ in oleum (sulfur trioxide solution in sulfuric acid) which
converts chloropicrin to phosgene and nitrosyl chloride.
2.3.13
6+
Cr , H2SO4
Cl3C NO2 COCl2 NOCl
Phosgene is then detected by known chromogenic reactions, e.g. by the reaction with
4-(4'-nitrobenzyl)pyridine.
Simple quantitative determination is usually carried out by the argentometric titration of
chlorides with silver nitrate indicated by a silver electrode or an Ag2S or a silver halide
membrane ion-selective electrode. The potentiometric titration is suitable because the reaction
of chloropicrin with sodium ethoxide leads to coloured products [5-8]. The iodometric
titration of the nitrite arising in the reaction with potassium iodide (see reaction 2.3.5) is
another option. The titration is performed with sodium thiosulfate with the visual indication of
equivalence point.
References to chapter 2.3

1. FRANKE, S.: Lehrbuch der Militärchemie. 2. vydání. Díl 1 a 2. Deutscher
Militärverlag der DDR. Berlin 1977. 512/616 s.
2. GRIESS P.: Bemerkungen zu der Abhandlung der H.H. Veselsky und Benedikt „Ueber einige
Azoverbindungen“. Ber. 12, 426-428 (1879).
3. MOUREU, H., CHOVIN, P., TRUFFERT, L.: Sur la transformation photochimique de la loropicrine en
phosgene et la caracterisation de ces deux corps. Compt. Rend. 228, 1954-1956 (1949).
4. Chem 51-3: Určování OL. MNO Praha. 1957. 143 s.
5. HALÁMEK, E.: Příspěvek k elektrochemickému stanovení některých bojových otravných látek. [Kandidátská
disertace]. Brno 1979. 118 s. - Vojenská akademie.
obrana, 28, 1986, 2, s . 75-87.
25
2.4 Hydrogen cyanide
H C N
In an aqueous solution, hydrogen cyanide behaves as a weak acid. Its salts, cyanides, are
relatively stable compounds markedly tending to form insoluble or soluble, but little
dissociated, compounds with transition metal ions. Some of them are characteristically
coloured. Free cyanides readily occupy the ligand field around the central metal cation and
thus form stable cyano complexes of which some are of importance as analytical reagents.
The formation of cyano complexes is utilized for the determination of cyanides and thus also
of hydrogen cyanide. The best-known and most used reaction is the formation of the so-called
Berlin Blue (Prussian Blue). A soluble ferrous salt is added to an alkaline sample in which
cyanide ions are able to exist. A sparingly soluble lightly green iron(II) cyanide is formed first
and then it dissolves in the excess cyanide to give a yellowish hexacyanoferrate(II).
Acidification, e.g. with sulfuric acid, followed by addition of an iron(III) salt solution, results
in a characteristically coloured blue solution. At higher concentrations, blue amorphous
iron(III) hexacyanoferrate(II) precipitates that readily forms colloid solutions [1,2]
2.4.1
2 CN - Fe 2+ Fe(CN)2
4 CN - Fe(CN)2 [Fe(CN)6] 4 -
2.4.2
(H2SO4)
3 [Fe(CN)6] 4 - 4 Fe 3+ Fe4[Fe(CN)6]3
Since solutions of iron(II) salts are oxidized spontaneously and rapidly with atmospheric
oxygen to give iron(III) salts, the blue coloration often appears already after the acidification
with sulfuric acid. The reaction is very selective and when carried out on a dropping plate or
paper, it is relatively sensitive (3 µg.ml−1) [3,4].
The reaction of cyanides is also highly sensitive, especially with sulfur, ammonium
polysulfide or thiosulfate leading to thiocyanates which form a red complex with trivalent
iron [5].
2.4.3
CN - (NH4)2Sn (NH4)2Sn-1 SCN -
SCN - Fe 3+ [FeSCN] 2+
SCN - [FeSCN] 2+ [Fe(SCN)2] +
SCN - [Fe(SCN)2] + Fe(SCN)3
Its sensitivity is 10 µg.ml−1 in a sample or up to 5 µg.l−1in air.

The ability of cyanide ions to form stable complexes as a result of strong donor-acceptor bond
is utilized for their indirect proof by decomposing less stable chelates [6]. The liberated
multifunctional ligand of chelate is then converted to a markedly coloured complex, called
lake pigment. Thus, for example the TSCHUGAEV reagent (dimethylglyoxime) with
26
palladium(II) ions forms a crystalline yellow complex that is decomposed by cyanides
affording dimethylglyoxime and a yellowish tetracyanopaladate. Addition of nickel(II) ions
results in the formation of red amorphous precipitate of nickel(II) dimethylglyoximate.
2.4.4
H
O O
H3C N N CH3
H3C C N OH
Pd 4 CN - [Pd(CN)4] 2- 2
žlutý
žlutý H3C C N O
H3C N N CH3 yellow
O O
H
žlutý
žlutý H
yellow
O O
H3C N N CH3
H3C C N OH Ni
2 Ni 2+
H3C C N O H3C N N CH3
O O
H
red
červený
červený
The sensitivity of this indirect detection reaction amounts to 1 µg.ml−1 in a sample. Several
indirect analytical detection methods for cyanide ions have been described and employed.
The detection using cupric ions is based on the reaction of hydrogen cyanide and cyanides
with metal ions. The basis is a redox disproportionation reaction of the arising copper(II)
cyanide affording copper(I) cyanide with the liberation of dicyanogen, a compound of marked
oxidation potential.
2.4.5
-
Cu(CH3COO)2 2 CN - Cu(CN)2 2 CH3COO
2 Cu(CN)2 2 CuCN (CN)2
Dicyanogen itself, when reacting with water, yields back hydrogen cyanide and active oxygen
in statu nascendi.
2.4.6
NC CN H2O 2 HCN O
The chromogenic effect is then produced by the reaction with reduced, usually colourless,
forms of redox indicators, such as benzidine (benzidine blue) [7], o-tolidine (yellow-orange
coloration), bis(N,N’-dimethylaminophenyl)methane (blue coloration) [8,9] and many other
reactions, such as oxidation of phenolphthalin to phenolphthalein [10-12], o-cresolphthalin to
27
o-cresolphthalein [11], fluorescin to the fluorescing fluorescein or the appearance of
luminiscence with luminal [13].
2.4.7
2 (H3C)2N CH2 N(CH3)2 (CN)2
(H3C)2N CH N(CH3)2
2 HCN
(H3C)2N CH N(CH3)2
The arising products of mesoquinoid character are stabilized in charge-transfer complexes.

The sensitivity of detection is high, in a sample up to 0.05 µg.ml-1; however, the reaction is
not very selective. It is used in detector papers and detector tubes. The detection is interfered
with many oxidation reagents.
In some important detection reactions of hydrogen cyanide the analyte acts as a catalyst. The
otherwise weak reducing properties of hydrogen cyanide are enhanced by conversion to
cyanohydrines in the reaction with aldehydes and ketones. Hydrogen cyanide, a weak
reducing agent, catalyzes reactions of chromogenic reagents and this property is utilized for
the analysis of air and solutions using detector papers as well as detector tubes, despite that
sometimes the reactions are not studied and described enough. The reaction of
p-nitrobenzaldehyde may serve as an example.
2.4.8
O HO
HCN C NO2 NC C NO2
H H
HO O
NC C NO2 C NO2
H H
HCN O2N CH C NO2
OH O
OH - - H2O
N C C NO2
O
OH O
28
The red coloration of the product is due to the mesoquinoid form of 4,4'-dinitrobenzoin. The
detector paper for hydrogen cyanide and tabun was based on this reaction. The so-called
CANNIZZARO reaction is the acyloin condensation of p-nitrobenzaldehyde to
4,4'-dinitrobenzoin, catalyzed by cyanide ions or hydrogen cyanide [14,15]. To increase the
sensitivity and selectivity of the detection, o-dinitrobenzene [16-19], triphenyltetrazolium
chloride [18] or Tetrazolium Blue [20] (the reduction products of which are also coloured) are
added to the reaction mixture. The published detection limit amounts to 10 µg.l-1 in air.
Another example of a catalytic reaction is the detection of hydrogen cyanide by ninhydrin
[21,22].
2.4.9
O O O CN
OH OH
+ CN
O OH -
O O
O O O
- OCN
O O O
OH OH
+ HCl + CN
O
CN - Cl CN
OH O OH
red colour pH < 10

červené zbarvení pH <10
blue colour pH > 10
modré zbarvení pH >10
+ HCl - Cl OH O O
OH OH OH
+ 1/2 O2 + H2O H
CN
- H2O COOH
CN CN
OH O
- HCN
light pinkrůžové
světle colour
zbarvení O O
CHO
O
OH
- H2O
O O
The sensitivity of catalytic reactions is very high, being up to 0.05 µg.ml−1 in a sample; their
selectivity, however, is not sufficient since many reducing compounds afford a similar
coloration.
The detection of hydrogen cyanide using the ELLMAN reagent which affords yellow-orange
products is also based on its reducing properties [23,24]. The reaction of hydrogen cyanide
with 5,5'-dithiobis(2-nitrobenzoic acid) proceeds probably in the following way:
29
2.4.10
O2N S S NO2 HCN H2O HOCN 2 HS NO2
HOOC COOH COOH
Another reaction employed for the detection of hydrogen cyanide is the formation of orange
to brown sodium isopurpurate from picric acid or sodium picrate [25].
2.4.11
ONa ONa
O2N NO2 O2N NH OH O
3 HCN HC NH2
NC CN
NO2 NO2
O
O2N NH OH
NC CN
NaO N O
The reaction is performed in a solution, on a paper [26] and in a detector tube [26,27]. The
sensitivity in a DT is up to 1 µg.l−1 in air or 10 µg.ml−1 in a sample. As suggested by the
present authors, the reaction can be accelerated by addition of aprotic solvents, such as
dimethyl sufoxide.
Besides the legendary formation of Berlin (Prussian) Blue, probably the most often applied
method for the detection of hydrogen cyanide and cyanides is halogenation and the
subsequent formation of cyanogen halide. The halogenation, usually chlorination, is
performed with the so-called chloroamines, actually chloroamides of sulfonic acids or
hypochlorites [29] or bromine water (in this case the product is cyanogen bromide).
2.4.12
O O
Na Na
H3C S N NaCN H3C S N ClCN
Cl Na
O O
Further, the arising cyanogen chloride is subjected to the KÖNIG-ZINCKE reaction (see
Chapter 2.5, Cyanogen chloride).
The detection of hydrogen chloride formed in the reaction with mercury(II) chloride
represents a classical very simple principle still utilized in detector tubes for hydrogen
cyanide.
30
2.4.13
HCN HgCl2 Hg(CN)2 2 HCl
Hydrogen chloride is then detected with an acid-base indicator, usually Methyl Orange or
Bromophenol Blue.
The quantitative determination of hydrogen cyanide and cyanides is carried out by titrimetric
methods. The titration with a silver nitrate solution is employed most often. The classical
LIEBIG argentometric titration proceeds according to the following scheme:
2.4.14
+ - - + 2 AgCN
Ag 2 CN [Ag(CN)2] Ag
The titration to the first degree (dicyanoargentate anion) can be followed visually, for further
titration to the white insoluble silver cyanide a potentiometric titration with a silver metal
electrode or with a silver iodide, chloride, bromide or preferably a sulfide membrane ion-
selective electrode is recommended. Cyanides can also be titrated mercurimetrically, e.g. with
mercury(II) nitrate. The reaction is analogous to the argentometric one, but mercury(II)
cyanide is a soluble non-dissociated salt. The equivalence point may also be indicated by
metallochromic agents or better potentiometrically using a sulfide ion-selective electrode [30-
33].
2.4.15
Hg 2+ 4 CN - [Hg(CN)4] 2- Hg 2+ 2 Hg(CN)2
Hydrogen cyanide and cyanides can also be determined iodometrically by an iodine solution
under the formation of cyanogen iodide, the excess iodine being titrated with thiosulfate to
starch.
2.4.16
HCN I2 ICN HI
Another possibility is the titration with a nickel(II) salt solution to dimethylglyoxime as a

metallochromic indicator.
2.4.17
Ni 2+ 4 CN -
[Ni(CN)4] 2- Ni 2+ 2 Ni(CN)2
The nickel salt solution is usually standardized chelatometrically [34].

1. OKÁČ, A.: Analytická chemie kvalitativní. Akademie. Praha. 3. vydání. 1966.576 s.
2. SVEHLA, G.: Vogel´s Quantitative Inorganic Analysis. Longman Ltd. Harlow UK. 2003. 347 s.
3. WILLEKENS, G.J., BULCKE, A.: Spectrophotometric determination of trace amounts of free cyanide in
Prussian Blue. Analyst 104, 525-530 (1979).
4. DIXON, B.E., HANDS, G.C., BARTLETT, A.F.F.: A field method for the rapid determination of hydrogen
cyanide in air. Analyst 83, 199-202 (1958).
5. FRANCIS, C.K., CONNELL, W.B.: The colorimetric method for determining hydrocyanic acid in the plants
with special reference to kafir corn. J. Am. Chem. Soc 35(10), 1624-1628 (1913).
31
6. ISHII, H., KOHATA, K.: Indirect spectrophotometric determination of trace cyanide with cationic porphyrins.
Talanta 38(5), 511-514 (1991).
7. SIEVERTS, A., HERMSDORF, A.: Der Nachweis gasförmiger Blausaure in Luft. Angew. Chem. 34, 3-5
(1921).
8. FEIGL, F., ANGER, V.: Replacememnt of benzidine by copper ethylacetate and tetra base as spot-test reagent
for hydrogen cyanide and cyanogen. Analyst 91, 282-284 (1966).
9. NAKANO, N., YAMAMOTO, A., KOBAYASHI, Y., NAGASHIMA, K.: An automatic measurement of
hydrogen cyanide in air by a monitoring tape method. Anal. Chim. Acta 398 (2-3), 305-310 (1999).
10. CACACE, D., ASHBAUGH, H., KOURI, N., BLEDSOE, S., LANCASTER, S., CHALK, S.:
Spectrophotometric determination of aqueous cyanide using a revised phenolphtalin method. Anal. Chim. Acta
589(1), 137-141 (2007).
11. NICHOLSON R.I.: The estimation of hydroen cyanide by the phtalin method. Analyst 66, 189-192 (1941).
12. HAJ-HUSSEIN A.T.: Flow injection spectrophotometric determination of cyanide by the phenolphtalin
method. Talanta 44(4), 545-551 (1997).
13. LU, J., QIN, W., ZHANG, Z., FENG, M., WANG, Y.: A flow-injection type chemiluminiscence-based
sansor for cyanide. Anal. Chim. Acta 304(3), 369-373 (1995).
14. KOVÁŘ, V., HORÁK, O., MATOUŠEK, J.: Trubičkové detektory průmyslových škodlivin pro kontinuální
kontrolu ovzduší. Chem. Prům. 14(12), 663-665 (1964).
15. KRATOCHVÍL V.: Rychlé semikvantitativní stanovení kyanovodíku v ovzduší. Chem. Prům. 37(5), 265-
267 (1987).
16. FEIGL, F., CALDAS, A.: Catalytic detection of cyanide through the benzoin condensation. Microchim. Acta
1955, 992-995.
17. GUILBAULT, G.G., KRAMER, D.N.: Ultra sensitive, specific method for cyanide using p-
nitrobenzaldehyde and o-dinitrobenzene. Anal. Chem. 38(7), 834-836 (1966).
18. KRAMER, D.N., GUILBAULT, G.G.: The reduction on 1,2-dinitrobenzene by the cyanohydrin of p-
nitrobenzaldehyde. J. Org. Chem. 31, 1103-1106 (1966).
19. FÁVERO, J.A.D., TUBINO, M.: Semi-quantitative „spot-test of cyanide. Anal. Sci. 19, 1139-1142 (2003).
20. PITSCHMANN, V.: Spektrofotometrické stanovení kyanidů tetrazoliovou modří. Vodní hospodářství 12,
395 (1997).
21. DROCHIOIU, G., MANGALAGIU, I., AVRAM, E., POPA, K., DIRTU, A.C., DRUTA, I.: Cyanide
reaction with ninhydrin: elucidation of reaction and interference mechanism. Anal. Sci. 20, 1443-1447 (2004).
22. NAGARAJA, P., HEMANTHA KUMAR, M.S., YATHIRAJAN, H.S., PRAKASH, J.S.: Novel sensitive
spectrophotometric method for the trace determination of cyanide in industrial effluent. Anal. Sci. 18, 1027-
1030 (2002).
23. HUMPHREY, R.E., HINZE, W.: Spectrophotometric determination of cyanide with organic disulphides.
Talanta 18, 491-497 (1971).
24. SPURLIN, S., HINZE, W., ARMSTRONG, D.W.: Use of an aqueous micellar medium to improve the
spectrophotometric determination of cyanide ion with 5,5'-dithiobis(2-nitro-benzoic acid). Anal. Lett. 10(12),
997-1008 (1977).
25. DROCHIOIU, G., MANGALAGIU I., TATARU, V.: Specific spectrophotometric determination of
hydrocyanic acid in the environment. Analyst 125(5), 939-941 (2000).
26. STEC, H., CZERNIEC J., GREGOROWICZ Z.: Wskaznik do pólilosciowego oznaczania jonów
cyjankowych i cyjanowodorodu. Int. Cl. G01N31/22. PL 101439. (20.4.1979). Polytechnika slaska.
11.4.2001.
28. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z.: Trubičkový detektor s kyselinou pikrovou pro určení
kyanovodíku. Sborník VVŠ PV, Vyškov, 2001, číslo 1, s.17-22.
29. PITSCHMANN, V., KOBLIHA, Z., HALÁMEK, E.: Použití chlornanů ke stanovení kyanidů podle Königa
a Zinckeho. Sborník VVŠ PV, Vyškov, 2003, číslo 2, s. 113-119.
30. TOMÍČEK, O.: Odměrná analýza. Československá chemická společnost pro vědu a průmysl. Praha 1949.
380 s.
32.ČAPOUN, T., KOBLIHA, Z., HALÁMEK, E.: CO-3-16 Analýza vysoce toxických látek v chemických
obrana, 28, 1986, 2, s . 75-87.
32
2.5 Cyanogen chloride
Cl C N
Although cyanogen chloride is genetically related to hydrogen cyanide, formally it is

the chloride of cyanic acid as manifested by the reaction of both compounds with alkaline
solutions. This procedure also enables the separation of hydrogen cyanide from cyanogen
chloride.
2.5.1
ClCN 2 NaOH NaCl NaOCN H2O
Cyanates do not undergo reactions characteristic of cyanides. Similarly to other halides,

cyanogen chloride reacts with silver salts in an ammonia solution precipitating a yellow silver
salt of cyanamide [1].
2.5.2
ClCN 2 AgNO3 4 NH4OH Ag2NCN NH4Cl 2 NH 4NO3 4 H 2O
With sulfides, cyanogen chloride gives thiocyanates, easily and sensitively detectable as red
iron(III) thiocyanate [2].
2.5.3
ClCN Na2S NaSCN NaCl
3 NaSCN FeCl3 Fe(SCN)3 3 NaCl
As an acylation reagent, cyanogen chloride affords reactions typical for acyl halides.
Similarly to phosgene, it undergoes the SCHÖNEMANN aminoperoxide reaction.
2.5.4
pH 7-8
ClCN - - [HOOCN]
OOH Cl
H2N NH2
3 [HOOCN] 5 H2O
H3CO OCH3
H2N NO2 3 NH4HCO3
H3CO OCH3
H2N N
H3CO OCH3
33
o-Dianisidine is oxidized with peroxycyanic acid to give the nitro derivative which in an
alkaline medium forms a red quinoid form of the Na salt. Due to the unstability of cyanic acid
and even peroxycyanic acid, the sensitivity of cyanogen chloride detection is not high, being
about 50 µg.l-1 in air.
The most often employed procedures for the determination of cyanogen chloride (cyanogen
halides) use the KÖNIG-ZINCKE reaction [3,4] and a number of such procedures have been
published [5-9]. They differ in a reagent used for replacing the unpleasant pyridine. They are
usually pyridine derivatives, such as picoline (methylpyridine), nicotinamide,
isonicotinamide, isonicotinic acid, 4-phenylpyridine, 4-(4'-nitrobenzyl)pyridine or
4-benzylpyridine. Several compounds are also employed as passive components for the
preparation of Schiff’s bases. Resorcinol, floroglucinol, dimedone [10-15], barbituric acid,
1,3-dimethylbarbituric acid [16,17], 2,4-quinolinediol, ethyl benzoylacetate or heterocyclic
compounds such as 3-methyl-1-phenyl-5-pyrazolone are very often used in addition to
common primary amines, such as aniline, sulfanilic acid, anthranilic acid, benzidine, o-
tolidine or p-phenylenediamine.
The mechanism of the whole reaction is based on the condensation of acyl halides and some
alkyl halides with pyridine. In a polar medium, the adduct undergoes the von BRAUN
rearrangement under the formation of glutacondialdehyde, a reactive polymethine compound.
This is a special case of the von BRAUN rearrangement [18] of tertiary amines leading to
secondary amines. Glutaconaldehyde-cyanamide is then cleaved in a polar medium to give
glutacondialdehyde and cyanamide.
2.5.5
ClCN Cl -
N N
+ H2O
CN
C CH CH CHO HCl
N
N NH CH CH
NC OH
C CH CH CH OH
N N CH CH
+ H2O glutaconaldehyde-cyanamide
glutakonaldehyd-kyanamid
CH CH CH
NH2CN
O CH CH OH
glutacondialdehyde
glutakondialdehyd
34
O O
CH CH CH
O CH CH2 O
H3C CH3
O
CH3
CH CH CH H2O
O CH CH
CH3
HO
O O
- H2O
O O
H3C CH3
H3C CH3
CH CH CH
CH CH
H3C CH3
OH O
červené polymethinové
red polymethine dye barvivo
The reaction is very sensitive, up to 0.2 µg.ml−1 in a sample and 0.5 µg. l−1 in air. Apart from
its use in detector tubes, it is also performed in a solution and also as the quantitative
photometric determination. It can be carried out even on a paper strip, saturated with a
combined reagent. The same coloration is produced by phosgene oxime. A detection chalk for
cyanogen chloride has also been described which comprises 4-benzylpyridine instead of the
volatile pyridine and barbituric acid as a passive component [19]. Cyanogen chloride itself
affords coloured products with some pyridine derivatives. Thus, e.g. it gives a yellow
coloration with green UV-fluorescence with nicotinamide. A yellow product also arises in the
reaction of cyanogen chloride with 4-(4'-nitrobenzyl)pyridine. However, the sensitivity of
these reactions is not sufficient.
The simple quantitative determination of cyanogen chloride is based on the titration of
chlorides, arising by hydrolysis with the volumetric solution of silver nitrate or mercury(II)
nitrate.

3. KÖNIG, W.: Über eine neue von Pyridin derivierende Klasse von Farbstoffen. J. Prakt. Chem. 69, 105-137
(1904).
4. KÖNIG, W., BAYER, R.: Über die Aufspaltung des Pyridins. J. Prakt. Chem. 83, 325-328 (1911).
35
5. WITTEN, B., PROSTAK, A.: Sensitive detector crayons for phosgene, hydrogen cyanide, cyanogen chloride,
and lewisite. Anal. Chem. 29(6), 885-887 (1957).
6. LAMBERT, J.L., RAMASAMY, J., PAUKSTELIS, J.V.: Stable reagents for the colorimetric determination
of cyanide by modified König reactions. Anal. Chem. 47(6), 916-918 (1975).
7. MARKLEY, B., MELOAN, C.E., LAMBERT, J.L., CHIANG, Y.C.: A sensitive, and rapid colorimetric
method to determine cyanide contamination in containers, capsules, and liquids. Anal. Lett. 20(8), 1225-1236
(1987).
8. EPSTEIN, J.: Estimation of microquantities of cyanide. Anal. Chem. 19(4), 272-274 (1947).
9. KRYŃSKA, A.: Rurka wskaźnikowa do oznaczania śladowych ilości cyjanowodorodu w powietrzu. Prace
Centr. Ind. Ochr. Pracy 15(47), 223-131 (1965).
11.4.2001.
11. ORITEST. Detekční trubička pro zjišťování bojových chemických látek. Původce: Pitschmann V., Orel J.
Int. Cl. G01N31/22. CZ 12840. Zapsáno 2.12.2002.
12. ORITEST. Detekční trubička na fosgen, difosgen, kyanovodík a chlorkyan, s kapalným chloračním činidlem.
Původce: Pitschmann V., Halámek E. Int. Cl. G01N31/22. CZ 17643. Zapsáno 2. 7. 2007.
13. ORITEST. Detekční trubička pro zjišťování kyanovodíku a chlorkyanu ve vzduchu. Původce:
14. ORITEST. Detekční trubička ke stanovení kyanidů ve vodě. Původce: Pitschmann V. Int. G01N31/22. CZ
14305 U1. Zapsáno 3.5.2004.
15. ORITEST. Detekční trubička ke zjišťování chlorkyanu a chlóru. Původce: Pitschmann V. Int. Cl.
G01N31/22. CZ 14952. Zapsáno 29. 11. 2004.
16. ORITEST. Způsob analytické kontroly průmyslových škodlivin.
Původce vynálezu: Pitschmann V., Halámek E., Tušarová I. Int. Cl. G01N21/78. CZ 284 693. Udělení
oznámeno ve Věstníku č. 2/99, 17.2.1999.
17. ORITEST. Detekční trubička ke zjišťování chlorkyanu a chlóru. Původce: Pitschmann V. Int. Cl.
G01N31/22. CZ 14952. Zapsáno 29. 11. 2004.
18. BRAUN, J.: Die Einwirkung von Bromcyan auf tertiäre Amine. Ber. 33, 1438-1452 (1900).
19. WITTEN, B., PROSTAK, A.: Sensitive detector crayons for phosgene, hydrogen cyanide, cyanogen chloride
36
2.6 Sulfur yperite
CH2 Cl
CH2 CH2 Cl
S CH2
Besides yperite (bis(2-chloroethyl) sulfide), some of its known analogues, such as

sesquic yperite (Q) and oxole yperite (T), also contain sulfur. These compounds behave
analogously and their chromogenic reactions in simple detection devices are similar to those
of yperite. Historically, the German DRÄGER-SCHRÖTER tube comprising silica gel and an
ampoule with a violet solution of potassium permanganate which on the reaction with yperite
turned yellow is the first among the detector tubes for yperite. Up to now, this reaction is used
for the detection of easily oxidizable compounds [1].
2.6.1
O
3 S(CH2CH2Cl)2 4 KMnO4 6 H2O 3 S(CH2CH2Cl)2 4 MnO(OH)2 4 KOH
O
One of the first American detectors for sulfur yperite used the reduction of selenic acid in
sulfuric acid yielding red-orange elemental selenium [2].
2.6.2
H2SO4 O
H 2SeO3 S(CH 2CH 2Cl)2 Se S(CH2CH2Cl)2 H2O
O
Besides electrophilic reactions involving free electron pairs of sulfur as well as coordination
bonds, yperite is, first of all, a strong alkylation agent. Some of the oldest reactions of yperite
used are its precipitation reactions affording sparingly soluble heavy metal complexes. The
reaction with the GRIGNARD reagent, tetraiododicuprate, may be an example. The reagent is
prepared from copper sulfate and the excess of sodium iodide.
2.6.3
CuSO4 2 NaI CuI2 Na2SO4
2 CuI2 Cu2I2 I2
The black copper(II) iodide disproportionates to white copper(I) iodide and free iodine that
may be removed by thiosulfate or by extraction into an organic water-immiscible solvent.
2.6.4
Cu 2I2 2 NaI Na2[Cu2I4]
The insoluble copper(I) iodide dissolves in the excess of iodide to give tetraiododicuprate(I)
which with yperite in an aqueous solution affords a yellow insoluble complex, more
accurately a complex of copper(I) iodide with bis(2-iodoethyl) sulphide [3].
2.6.5
Na2[Cu2I4] S(CH2CH2Cl)2 [Cu2I2 .S(CH2CH 2l)2] 2 NaCI
37
The sensitivity of the reaction is 30 µg.ml-1. The NĚKRASOV reagent
(tetraiodomercurate(II)) or the related NESSLER reagent (alkalized solution of
tetraiodomercurate(II) for the detection of ammonia) also behaves very similarly. The
procedure for yperite has been described by DESGREZ [4].
2.6.6
HgCI2 4KI K2[HgI4] 2 KCl
K2[HgI4] S(CH2CH2Cl)2 [HgI2 .S(CH2CH2l)2] 2 KCl
The reaction gives a yellow precipitate of a complex compound. The sensitivity is 25 µg.ml-1.
The reaction of sulfur yperite with gold(III) chloride is very widespread and still used even
with detector tube design. The yellow reagent precipitates a yellow crystalline complex with
yperite from a solution.
2.6.7
AuCl3 S(CH2CH2Cl)2 [AuCl3 .S(CH 2CH2Cl)2]
This OBERMILLER-SCHRÖTER reaction [5,6], often used as a spot reaction on paper, may
be accentuated by auxiliary reagents enhancing the colour of the precipitate, e.g. by
monochloroamine T which affords a red-brown colour. A detector tube, comprising
tetrachloroauric(III) acid on a support and an ampoule with 0.5% aqueous monochloroamine,
is based on a similar principle. After sucking air and breaking the ampoule, an orange zone
appears in the presence of yperite. It is supposed that the compound of yperite with trivalent
gold is decomposed under the formation of its complex salt with benzenesulfochloroamide.
The selectivity is very good, but a similar reaction also occurs with nitrogen yperite. The
sensitivity is good: it is possible to detect 1 µg.l-1 in air.
2.6.8
H[AuCl4] S(CH2CH2Cl)2 [AuCl3 .S(CH2CH2Cl)2] HCl
žlutý
yellow
Na
[AuCl3 .S(CH2CH2Cl)2] 4 SO2N
Cl
Cl
SO2N Au SO2N S(CH2CH2Cl)2 4 NaCl
3
oranžový
orange
Modifications of this method use heteropolyacids, such as phosphotungstic or

phosphomolybdic acids. In addition to the detection of yperite, the reaction is also widely
used for the determination of other sulfur compounds. It is not very sensitive (100 µg.ml-1)
[7]. These precipitation reactions leading to complex ion pairs are used for the standard
detection of aliphatic as well as heterocyclic amines. It is necessary to keep this in mind when
carrying out the detection of yperite or, in turn, amines.
38
Protecting properties of materials (breakthrough time of yperite; BTY) can be assessed by a
precipitation, turbidity test using sodium diethyldithiocarbamate, the so-called cupral, as a
reagent. Hence, the method is called cupral method for the determination of BTY.
2.6.9
C2H5 C2H5
N C SNa N C SCH2CH2SCH2CH2Cl
S(CH2CH2Cl)2
C2H5 C2H5
- NaCI
S S
The cupral reaction also is not very sensitive, 100 µg.ml-1 in a sample, however, for the given
purpose it is sufficient.
A very sensitive and, at the same time, selective reaction, employed for the preparation of
detector papers (KU-1), detector tubes [8-10] and, possibly, even for the determination in a
solution, is the reaction with ethyl derivative of MICHLER’s ketone, bis(p-
diethylaminophenyl) ketone which gives a red-brown or orange coloration. MICHLER’s
thioketone was also used. Other components of the detection protocol are mercury(II) or
copper(II) chloride or bromide. The detailed reaction mechanism has not been examined
thoroughly. According to the literature, the reaction involves the formation of a three-
component complex comprising yperite, S-bonded to the mercury salt where the ketone only
deepens the coloration to red-brown.
2.6.10
O
S(CH2CH2Cl)2 HgCI2 (C2H5)2N C N(C2H5)2
(C2H5)2N C N(C2H5)2 HgCI2 S(CH2CH2Cl)2
However, the reaction does not apparently depend on the formation of a heavy metal complex
because after the replacement of mercury(II) chloride by magnesium perchlorate, the
indication effect is unchanged. The coloration becomes brilliant red and a detector tube based
on this principle functions as a linear one. The minor 2-3 µg.l-1 in air and on warming it may
increase by at least one order of magnitude [11-13]. The selectivity is very high. In its original
version with Hg2+, the reaction was utilized even for the construction of a yellow paper for the
detection of liquid sulfur yperite that made deep red spots.
Detector papers (K-7a) utilize the ability of liquid yperite to decompose the light blue
complex of copper(II) chloride and mercury(II) chloride with Crystal Violet. Yperite
dissolves this dye to give a violet solution and forms blue spots on the detector paper [14].
39
2.6.11
CuCI2 (H3C)2N CH HgCI2 2 S(CH2CH2Cl)2

3
1
/2 O2
[CuCI2 .S(CH2CH2Cl)2] [HgCI2 .S(CH2CH2Cl)2]
(H3C)2N C N(CH3)2 OH -
N(CH3)2
The reaction is not selective enough: a similar effect is observed with some organic solvents.
Liquid lewisite produces a green spot on a detector paper.
The extraordinary lipophilic properties of liquid yperite (ability to dissolve oleophilic dyes)
are often utilized for its detection. One of the most often used simple devices to detect liquid
CWAs visualizes yperite droplets using a dye powder (Dye Red E) incorporated in a paper or
paper material. The liquid compound produces red spots on detector papers (PP-3, 3-WAY,
CALID, ABC-M8). The selectivity of this version is satisfactory; the same effect is observed
with lewisite and several organic solvents.
2.6.12
H3C CH3
CH
N CH3 H3C N
N N CAS: 60033-00-3
OH HO
Dye Red E
O O
C C
NH NH
The same principle is also applied to various detection pencils, chalks or paints directly on
military equipment and military material. Previously, other dyes soluble in oils and fats were
also used for the preparation of powders detecting liquid yperite and, possibly, other liquid
40
stable CWAs, such as tabun, nitrogen yperites or lewisite. Sudan IV (IP-1) or Methyl Red (IP-
2) dyes may serve as examples. However, these powders are not selective and give coloration
even with motor fuels, lubricating oils and fats and with organic solvents [15].
2.6.13
CH3 CH3 HO
HOOC
H3C
N N N N
N N N
H3C
CAS: 85-83-6 CAS: 493-52-7
SudanIVIV
Sudan Methyl Red červeň
Methylová
Another method to detect yperite used for the determination of protective materials (BTY) is
more ingenious. The MIKROTEST method is based on the reaction of yperite with
chloroamide [CNITI-8; N-chloro-N-(2-tolyl)benzamide]. The reaction liberates hydrogen
chloride which converts the alkaline form of an acid-base indicator to the acidic one (here the
red form of Kongo Red to the blue form) using azo-hydrazone tautomerism.
2.6.14
CH3
O Cl
C N
S(CH2CH2Cl)2
Cl CH3
O
CH CH2 Cl
S C NH
CH2 CH2 Cl
CH CH Cl
S HCl
CH2 CH2 Cl
41
NH2 NH2
N N N N
2 H2O
red
červeň
SO3Na SO3Na
+2H + 2 OH
NH2 NH2
N NH NH N
2 Na
modř
blue
SO3 SO3
In field laboratories, yperite is determined using the so-called pyridine reaction that,
analogously to the KÖNIG-ZINCKE reaction, is based on the determination of cyanogen
halide by the von BRAUN cleavage of pyridine and rearrangement to a polymethine
SCHIFF’s base. However, in the case of alkyl halides, the rearrangement requires an alkaline
medium and heating to a higher temperature. The reaction with p-aminoacetophenone affords
a raspberry-red coloration. The selectivity is not sufficient. On the other hand, the sensitivity
is good, being 1 µg.ml-1.
2.6.15
t
S(CH2CH2Cl)2 Cl
N N
CH2CH2SCH2CH2Cl + 2 OH
HO CH2CH2 S CH2CH2 N CH CH CH CH CH OH 2 Cl
HO CH2CH2 S CH2CH2 N CH CH CH CH CH OH H2N C
CH3
C NH CH CH CH CH CH N CH2CH2 S CH2CH2 OH H2O

H3C
red SCHIFF's
červená base
SCHIFFova báze
O
C N CH CH CH CH CH NH CH2CH2 S CH2CH2 OH H2O
H3C
42
The reaction of yperite with 4-(4'-nitrobenzyl)pyridine [16] (reagent DB-3), used in detector
tubes [17-19] as well as in other simple devices, is an analogy of the reaction with pyridine
[20,21].
2.6.16
S(CH2CH2Cl)2 2 O2N CH2 N
O2N CH2 N CH2 CH2
S 2 Cl
O2N CH2 N CH2 CH2
O2N CH NH CH2 CH2
S 2 Cl
O2N CH NH CH2 CH2
+ 2 OH
O
N CH N CH2 CH2
O
S 2 Cl 2 H2O
O
N CH N CH2 CH2
The addition of hydroxide results in a blue coloration. Although the sensitivity is good, the
selectivity is low. In addition to yperites, the reaction is positive with many compounds of
acyl halide and alkyl halide type. Concerning CWAs, a positive reaction is observed with
sarin, soman, nitrogen yperites, lewisite, diphenylchloroarsane, chloroacetophenone,
phosgene, diphosgene and others. The sensitivity to yperite can be enhanced up to 1 µg. l-1 in
air by the addition of sodium perchlorate. Further sensitivity enhancement (more than by one
order of magnitude) may be achieved by the modification of packing and by warming.
A classical and interesting method of yperite detection consists in its reaction with thiourea
and an ammonia solution of nickel(II) sulphate [22]. Alkyl halides and yperite as well react
with thiourea to afford the corresponding quaternary compounds, in this case
S,S`-[(3-thiapent-1.5-diyl)]-dithiouronium dichloride.
2.6.17
NH3
NH2 CH2CH2 S C
NH
S(CH2CH2Cl)2 2 S C S 2 Cl
NH
CH2CH2 S C
NH2
NH3
43
When treated with sodium hydroxide, this bis-quaternaty salt is cleaved to give
bis(2-sulfanylethyl) sulfide which in an ammonia medium gives rise to red chelate with
nickel(II) salt.
2.6.18
NH3
CH2CH2 S C
NH
S 2 Cl 6 NaOH 2 NaCl 2 Na2CO3 4 NH3
NH
CH2CH2 S C
S(CH2CH2SH)2
NH3
S
CH2CH2 SH S S
2 S 2 Ni 2+
4 NH3 Ni Ni 4 NH4
S
CH2CH2 SH S
S
In the past, field laboratories used the so-called reagent T-135 widely. The detection is based
on the condensation of yperite with an alkaline (coloured) form of phthalein, in this case
thymolphthalein, under the formation of ester which in an acid medium cannot form lactone,
phthaleine. Heating in an alkaline aqueous-ethanolic medium affords yellow to orange ester
of yperite and thymolphthalein. The cooling and acidification of the reaction mixture removes
the excess of a blue unreacted reagent and the yellow or orange product itself is visualized
[23]. The reaction is very sensitive (up to 0.5 µg.ml-1 in a sample solution), but it is not
reproducible enough for the quantitative determination of yperite. Its selectivity is
insufficient; the same reaction occurs not only with all yperites, but also with other CWAs
and components of decontamination compositions of alkyl halide character.
2.6.19
H3C CH3 H3C CH3
H 3C CH3 H3C CH3
CH CH
CH CH
HO O
HO OH
+ OH
H2O
-H C
C
O CH 3 CH3
CH3 CH3
C O
C O
O
H3C CH3 H3C CH3
CH CH
HO O S(CH2CH2Cl)2 OH - 2 Cl
CH3 CH3
C O CH2CH2SCH2CH2OH
44
To confirm a positive reaction for sulfur yperite and to enhance the selectivity of the reaction
with reagent T-135, the sulfur yperite-thymolphthalein ester reacts with palladium(II) salts.
The formation of a red complex compound represents a highly selective reaction for sulfur
yperite.
2.6.20
H3C CH3 H3C CH3
CH CH
HO O
CH3 CH3
C O CH2CH2SCH2CH2OH
O
PdCl2
This yperite determination using phthaleines has principally been optimized by the present
authors as an ion-pair extraction method [24-27]. The reaction is carried out in a polar
aqueous-methanolic medium with carbonate buffer-adjusted pH, o-cresolphthalein being
selected as suitable phthalein. In an aqueous solution, yperite is converted slowly to reactive
intermediate, S-(2-chloroethyl)thiiranium chloride.
2.6.21
S(CH2CH2Cl)2 S CH2CH2Cl Cl
In an alkaline medium, this cation becomes a counterion of the so-called keto form of
o-cresolphthaleine.
2.6.22
CH3 CH3 CH3 CH3
HO OH HO O
+ OH
C - H2O
C
O
C O
C O
O
leukoforma
leuco form ketoketoforma
form
45
S CH2CH2Cl
CH3 CH3
HO O
S CH2CH2Cl C
C O
O
w
CH3 CH3
extrakce HO O
extraction
C O CH2CH2SCH2CH2Cl
O
org
The arising ion pair reacts gradually to give an ester and is extracted from the aqueous
medium (w) into a water-immiscible organic solvent (org), such as benzene, toluene, but also
chloroform, dichloroethane or tetrachloromethane. Unlike the method that makes use of
reagent T-135, this procedure is highly selective with simultaneous retention of sensitivity.
o-Cresolphthalein reacts only with yperites, moreover, it is not necessary to warm the reaction
mixture and to remove the excess alkaline form of phthalein by acidification. The results of
quantitative determination are fully reproducible. Besides o-cresolphthalein chosen for its
brilliant colour, other phthaleins may also be employed as reagents.
The quantitative determination may also be performed advantageously using volumetric
titration methods. After the decomposition of yperite to chlorides they can be determined
argentometrically or mercurimetrically with the visual or potentiometric indication [28-30].
2.6.23
S(CH2CH2Cl)2 2 KOH S(CH2CH2OH)2 2 KCl
KCl AgNO3 AgCl KNO3
2 KCl Hg(NO3)2 HgCl2 2 KNO3
46
Volumetric redox methods are preferred. The bromatometric determination of yperite is
based on its oxidation to sulfoxide.
2.6.24
S(CH2CH2Cl)2 Br2 H 2O O S(CH2CH2Cl)2 2 HBr
A mixture of bromide and bromate, used as a source of bromine, liberates an equimolar

amount of bromine. The excess of the unused bromine is converted to free iodine with
potassium iodide and this is titrated with sodium thiosulfate. The bromatometric titration can
also be performed directly with a redox indicator, such as Methylene Blue or
potentiometrically with a platinum electrode indication. The method is suitable only for pure
yperite because other accompanying sulfur impurities are also oxidized.
Another redox titration is based on condensation with monochloroamine T.
2.6.25
Cl
S(CH2CH2Cl)2 H3C SO2N H3C SO2N=S(CH2CH2)Cl2

Na - NaCl
The sample is mixed with an excess of iodometrically standardized aqueous solution of

monochloroamine T and the mixture is acidified with acetic acid. After 10 minutes, potassium
iodide is added and liberated iodine is titrated with a thiosulfate solution. The difference
serves for the calculation of the yperite content.
Another iodometric titration method is based on the reaction of yperite with hexaiodoplatinate
which leads to a complex compound of yperite with iodoplatinate liberating an equimolar
amount of iodine.
2.6.26
2 S(CH 2CH2Cl)2 [PtI6]2- [Pt{S(CH2CH2Cl)2}2I2] 2I- I2
The free iodine is titrated using a sodium thiosulfate solution with starch as a visual indicator.
Sulfur yperite can also be determined by its reaction with thiosulfates.
2.6.27
CH2CH2 Cl CH2CH2 S SO2 ONa
S 2 Na2S2O3 S 2 NaCl
CH2CH2 Cl CH2CH2 S SO2 ONa
The reaction is quantitative and rapid and is used for checking the synthesized yperite directly
in manufacturing plants [31].

1. PITSCHMANN, V.: Analýza toxických látek detekčními trubičkami. Econt Colsunting. Drahelčice. 2005.
194 s.
2. YABLICK, M., PERROTT, G.ST.J., FURMAN, N.H.: Quantitative determination of traces of dichloro-
ethylsulfide (mustard gas) in air. J. Am. Chem. Soc. 42(2), 266-274 (1920).
3. GRIGNARD, V., et al., Ann. Chim., 15, 1921, 5.
47
5. OBERMILLER, M.: Ein spezifischer Nachweis von Dichlordiäthylsulfid neben anderen Kampfstoffen.
Angew. Chem. 49, 162-164 (1936).
6. SCHRÖTER, G.A.: Der spezifische Lostnachweis mit dem Adsorptionsverfahren (Gaspürgerät). Angew.
Chem. 49, 164-165 (1936).
1977. 512/616 s.
8. OLTHOFF, U., EICHARDT, A.: Indikatorrörchen. Int. Cl. G01N31/22. DD 299 765 A7. (7.5.1992).
JENAPHARM.
9. BITTER, R., NOLDE, M., HOFFMANN, J., SEIDEL, D.: Indikatorrörchen zur mehrmaligen Verwendung.
Int. Cl. G01N31/22. DD 288 954 A7. (18.4.1991). LABORCHEMIE APOLDA.
10. BITTER,R., NOLDE, M., HUTZSCHENREUTER, J., FRANZ, P.: Indikatorrörchen zum Nachweis von
Yperit in der Luft. Int. Cl. G01N31/22. DD 288 728 A7. (11.4.1991). LABORCHEMIE APOLDA.
11. ORITEST. Detekční trubička pro zjišťování yperitu a fosgenu či difosgenu ve vzduchu. Původce:
12. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z., VEVERKA, V.: Zjišťování nízkých koncentrací
sulfidického yperitu ve vzduchu trubičkovým detektorem. Sborník VVŠ PV Vyškov, 2001, číslo 1, s.23-28.
13. ORITEST. Detekční trubička pro zjišťování yperitu ve vzduchu.
Původce vynálezu: Pitschmann V., Halámek E. Int. Cl. G01N31/22. CZ 285 895. Udělení oznámeno ve
Věstníku č. 11/99, 17.11.1999.
15. Chem 51-3: Určování OL. MNO Praha. 1957. 143 s.
16. EPSTEIN, J., ROSENTHAL, R.W., ESS, R.: Use of γ-(4-nitrobenzyl)pyridine as analytical reagent for
ethylenimines and alkylating agents. Anal. Chem. 27(9), 1435-1439 (1955).
17. KRATOCHVÍL, V., MARTINEK, J.: Neues Verfahren zum Nachweis von Yperit. Chem. Zvesti 23, 282-390
(1969).
18. ORITEST. Detekční trubička ke zjišťování zpuchýřujících látek.
Původce: Pitschmann V. Int. Cl. G01N31/22. CZ 9949. Zapsáno 2.5.2000.
19. ORITEST. Trubička ke zjišťování sulfidických a dusíkových yperitů v ovzduší.Původce: Pitschmann V.
Int. Cl. G01N31/22. CZ 15979. Zapsáno 31.10. 2005.
20. PITSCHMANN, V.: Jednoduché stanovení bis(2-chlorethyl-sulfidu. Chemické listy 92, 149-150 (1998).
21. AGNEW, F.R., BYRNE, F.P., KACZMAREK, T.G., KRIEGE, O.H., GAINER, G.C., LUCK, R.M.:
Chemical contaminant detection samolet. Int. Cl. G01N31/00. US 3 689 224. (5. 9. 1972).
22. MASON-HARLEY, J.: Some aliphatic thiols and their derivatives. Part II. Thiols derived from 2,2'-
dichlorodiethyl sulphide and its analoques. Nickel complexes from 1,5-dithiols. J. Chem. Soc. 1952, 146-149.
23. ISSA, F.M., YOUSSEF, H.Z., ISSA, R.M.: Spectrophotometric determination of bis(2-chloroethyl)sulfide
(sulfur mustard HD) and tris(2-chlorethyl)amine (nitrogen mustard HN) using thymolphtalein. Egypt. J. Chem.
18(2), 257-64 (1975).
24. KOBLIHA, Z., HALÁMEK, E,: Extrakčně spektrofotometrické stanovení bis(2-chlorethyl)sulfidu ftaleiny.
In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1987, s. 2-24.
25. HALÁMEK, E., KOBLIHA, Z.: Extraction photometric detetermination of yperite by phthaleins. Talanta,
48, 1999, p. 163-171.
26. KOBLIHA, Z., HALÁMEK, E., ČAPOUN, T., KRYKORKOVÁ, J: CO-3-16 Analýza vysoce toxických
látek v chemických laboratořích Civilní ochrany ČR. Příloha 5: Extrakčně spektrofotometrické metody. MO
Praha. 28 s. MV ČR, Praha: Příloha 5 k PO-28/28-164/ICO-2002 Extrakčně spektrofotometrické metody.
2002. 28 s.
27. ČAPOUN, T., KRYKORKOVÁ, J., KOBLIHA, Z., HALÁMEK, E.: CO-3-16 Analýza vysoce toxických
látek v chemických laboratořích Civilní ochrany ČR. Příloha 12: Stanovení čistoty vzorků otravných látek.
MO Praha. 46s. MV ČR, Praha: Příloha 12 k PO-28/28-164/ICO-2002 Stanovení čistoty vzorků otravných
látek. 2002. 25 s.
laboratořích Civilní ochrany ČR. Příloha 6: Potenciometrické titrace. MO Praha. 23 s.; MV ČR, Praha:
Příloha 6 k PO-28/28-164/ICO-2002. Potenciometrické titrace. 2002. 28 s.
obrana, 28, 1986, 2, s . 75-87,
48
2.7 Nitrogen yperites
CH2CH2Cl R:
C2H5 [HN-1]
R N
CH3 [HN-2]
CH2CH2Cl
Cl C2H4 [HN-3]
Thanks to the two reactive 2-chloroethyl groups of strong alkylation potency and to
isosterism of the nitrogen and sulfur atoms, nitrogen yperites [ethyl-, methyl- or
2-chloroethyl-bis(2-chloroethyl)amine] afford reactions analogous to those of sulfur yperite,
including reactions used for the determination under conditions of mobile analysis. Nitrogen
yperites differ from sulfur yperite in the sense that they form very stable ammonium salts
derived from the tertiary amine nitrogen and they are able to form a cation in ion pairs with
suitable anions [1].
2.7.1
CH2CH2Cl R CH2CH2Cl
R N HX N X
CH2CH2Cl H CH2CH2Cl
The so-called reagents for tertiary amines and alkaloids are various compositions that in an
acid medium afford anions forming sparingly soluble ion pairs, precipitates or turbidity.
Basically, it concerns group reactions for organic compounds of tertiary aliphatic or
heterocyclic amine character. Sometimes, even secondary and primary amines enter the
reactions, but only in an acid medium, i.e. in their quaternized state.
More common precipitation agents for amines are iodo complexes, a legendary reagent being
the most often modified and for the longest time used DRAGENDORFF reagent [2]. It is a
yellow solution prepared from bismuth salt and the excess of potassium iodide with the
addition of acid. The DRAGENDORFF reagent precipitates a great number of amines,
including the heterocyclic ones in the form of orange and deep red crystalline precipitates.
2.7.2
R-N(CH 2CH2Cl)2] H[BiI4] [R-NH(CH2CHCl)2] . [BiI4]
The reaction is carried out in a solution, on a spot plate, a paper and thin layers as well as on
supports in detector tubes [3]. In some cases, such as the detection on cellulose or silica gel,
free iodine is liberated that partly obscures the reaction itself. If needed, the free iodine is
removed using a suitable reducing agent. In addition to the DRAGENDORFF reagent
(tetraiodobismuthic acid), other precipitation agents are often used, such as tetraiodocadmate,
tetraiododicuprate or tetraiodomercurate. The colour of the precipitate invariably corresponds
to that of an anion solution.
49
2.7.3
2 R-N(CH2CH2Cl)2] 2 H +
2 R-NH(CH2CH2Cl)2
[CdI4] 2- [Cu2I4] 2-
[HgI4] 2-
R-NH(CH2CH2Cl)2 [CdI4] R-NH(CH2CH2Cl)2 [Cu2I4]

2 2
R-NH(CH2CH2Cl)2 [HgI4]
2
With colourless precipitating agents the precipitate is white, with yellowish ones it is yellow,
etc. In some cases, however, coloured compounds in a solution may be adsorbed on the
surface of a light-coloured precipitate. The sensitivity of precipitation reactions is not very
high and in a solution it ranges between 10 and 100 µg.ml-1. The detection limit also fairly
depends on the mode of execution or its size. The precipitates of tertiary and secondary
amines of a defined composition are also formed as complexes with thiocyanate ligands.
Thus, e.g. potassium hexathiocyanatochromate and the so-called REINECKE salt (ammonium
tetrathiocyanato(diammine)chromate(III)) give violet and red crystalline precipitates [4].
2.7.4
3 [R-NH(CH2CH2Cl)2] [Cr(SCN)6] 3- [R-NH(CH2CHCl)2]3 . [Cr(SCN)6]
R-NH(CH2CH2Cl)2 NH4[Cr(NH3)2(SCN)4] [R-NH(CH2CHCl)2] . [Cr(NH3)2(SCN)4] NH4 +
The sensitivity in solution is up to 5 µg.ml-1. Similar reactions are also observed with a
number of other complex compounds containing various ligands, often cyano and nitro.
The so-called heteropolyacids belong to an important group of precipitating agents for
amines. They are most often derived from phosphorus(V), arsenic(V), silicium(IV) and
boron(III) in which oxygen is replaced with the residues of some isopolyacids of hexavalent
tungsten, molybdenum or of tetravalent silicium. Examples are phosphotungstic acid
[trihydrogen-tetrakis(tritungstato)-phosphoric acid], phosphomolybdic acid [trihydrogen-
tetrakis(trimolybdato)phosphoric acid] or germanosilicic acid [tetrahydrogen-
tetrakis(trisilicato)germanic acid].
2.7.5
3 R-N(CH2CH2Cl) 2 H3[P(W 3O 10) 4] [R-NH(CH 2CHCl) 2]3 . [P(W 3O10) 4]
3 R-N(CH2CH2Cl) 2 H3[P(Mo3O 10) 4] [R-NH(CH 2CHCl) 2]3 . [P(Mo3O10) 4]
4 R-N(CH2CH2Cl) 2 H4[Ge(Si 3O7) 4] [R-NH(CH 2CHCl) 2]4 . [Ge(Si 3O 7) 4]
The sensitivity of the reactions is not high, about 10-100 µg.ml-1 in solution. The precipitates
usually contain various amounts of crystal water and not all are suitable for quantitative, e.g.
gravimetric determination [5].
A slightly different precipitation reagent for amines is a complex of trivalent boron with
organic ligands - sodium tetraphenylborate – a reagent usually employed for the precipitation
of potassium ions.
2.7.6
[R-NH(CH2CH2Cl)2] Na B [R-NH(CH2CH2Cl)2] B Na +
4 4
50
The sensitivity is 10 µg.ml-1 in a sample. Crystalline precipitates are also obtained with
ammonium, rubidium and cesium ions. The reagent is suitable for gravimetric or titrimetric
determination [6].
Picric acid (2,4,6-trinitrophenol) and picrolonic acid [2,4-dihydro-5-methyl-4-nitro-2-(4-
nitrophenyl)-3H-pyrazol-3-one] are also used for the precipitation of amines. They afford
yellow crystalline precipitates [7].
2.7.7
OH O
O2N NO2 O2N NO2
R-N(CH2CH2Cl)2 [R-NH(CH2CH2Cl)2]
NO2 NO2
2.7.8
H3C N
R-N(CH2CH2Cl)2 N NO2
O2N
O
H3C N
[R-NH(CH2CH2Cl)2] N NO2
O
N
O O
Since precipitates of amines with picric and picrolonic acids have sharp and defined melting
points, they are used for the identification of individual nitrogen compounds. The ion pair
formation underlies another method of the group determination of tertiary amines and
alkaloids. Ion pairs are formed with acidic dyes and are extracted into a water-immiscible
solvent. An example may be the ion pair formation from nitrogen yperite and Bromoxylene
Blue.
Anions of acidic dyes (or cations of quaternized amines) are polar enough to prevent the
precipitation of the ion pair and their non-dissociated part can be extracted into water-
immiscible solvents. Solvents which dissolve water to a very limited extent are preferred.
Examples of such solvents are chloroform, dichloromethane or 1,1,2,2-tetrachloroethane.
Although the reaction is a group reaction, the required selectivity can be achieved by choice
of reaction conditions and the dye. The sensitivity usually ranges between 1 and 10 µg.ml-1 in
solution. The advantage lies in the high rate of counterion association. The detection limit as
well as the determination limit can be increased by alkalization of the extracted associate; this
increases the molar absorption coefficient which is typical for the majority of acidic dyes.
51
2.7.9
Br Br
HO O
R-N(CH2CH2Cl)2
H3C CH3
SO3H
Br Br
HO O
R-NH(CH2CH2Cl)2
H3C CH3
SO3
extraction
extrakce
Br Br
HO O
R-NH(CH2CH2Cl)2
H3C CH3
SO3
org
Similarly to many alkyl and acyl halides, nitrogen yperite reacts with
4-(4'-nitrobenzyl)pyridine [8-10]. The reaction requires the warming of the reaction mixture.
The sensitivity depends on the mode of execution and is 0.5-10 µg.ml-1 in solution. Warming
as well as the addition of sodium perchlorate and other components increases the sensitivity
of the test and shortens its duration.
52
2.7.10
R-N(CH2CH2Cl)2 2 N CH2 NO2
H2CH2C N CH NO2
H
R N 2 Cl
H2CH2C N CH NO2
O
2 NaOH H2CH2C N CH N
O
R N 2 NaCl 2 H2O
O
H2CH2C N CH N
A blue coloration appears in an alkaline medium. Out of CWAs, fluorophosphonates sarin

and soman, sulfur yperites, mono- and disubstituted monochloro- and dichloroarsanes and
chloroacetophenone also give the same coloration.
Nitrogen yperites can also be detected by the reaction with chloranil
(tetrachlorobenzoquinone) which affords red products with primary amines, a violet
coloration with secondary amines, whereas tertiary amines give a green coloration [11].
2.7.11
O O
Cl Cl Cl Cl
R-N(CH2CH2Cl)2
CH2CH2Cl Cl
Cl Cl Cl N
O O CH2CH2Cl
R
Both the selectivity and sensitivity of the reaction are not high. About 50 µg.ml-1 in a solution
can be detected.
Nitrogen yperites are also determined by treatment with agent T-135 (alkalized aqueous-
ethanolic thymolphthalein solution). The reaction and procedure are analogous to those for
sulfur yperite.
53
2.7.12
H3C CH3 H3C CH3

H3C CH3 H3C CH3
CH CH
CH CH
HO O
HO OH
+ OH
H2O
-H C
C
O CH3 CH3
CH3 CH3
C O
C O
H3C CH3 H3C CH3
CH CH
R-N(CH2CH2Cl)2 OH - 2 Cl
HO O
CH3 CH3
C O CH2CH2NCH2CH2OH
O R
Upon the warming and removal of the blue form of thymolphthalein by acidification, the
formed ester remains as an orange or yellow coloured solution. The sensitivity of
determination is 1 µg.ml-1. The selectivity is not high because a similar reaction is observed
with many alkyl and acyl halides. Reagent T-135 reacts even with some CWAs, such as β-
lewisite [bis(2-chlorovinyl)chloroarsane], bromobenzyl cyanide and chloroacetophenone. The
reaction is thus a group reaction. The product which retains properties of an amine can
undergo precipitation reactions with reagents for tertiary amines and alkaloids, thus being
distinguished from sulfur yperites. Phosphotungstic acid behaves slightly differently reacting
with the ester of thymolphthalein and nitrogen yperite under the formation of a lilac product.
2.7.13
H3C CH3 H3C CH3

CH CH
HO O
[P(W3O10)4]
C
CH3 H
CH3
C O CH2CH2NCH2CH2OH
R
O
3
54
By optimization of conditions, the phthalein reaction has become highly selective for the
determination of yperites including nitrogen yperites [12]. The method is based on the
formation of an ion associate: the reaction product of nitrogen yperite in water associates with
phthalein (in this case o-cresolphthalein) anion in a mildly alkaline aqueous-methanolic
medium and the associate is then extracted into an organic water-immiscible solvent (2.7.14).
In the organic phase the ion associate is converted into an ester of nitrogen yperite with
o-cresolphthalein of pink to violet colour. This modified phthalein reaction is very selective
and occurs only with highly reactive alkyl halides such as yperites. The selectivity is
illustrated by the fact that the extraction can be performed with chlorinated hydrocarbons such
as chloroform. Even without warming and acidification, the method substantially retains its
sensitivity of 1 µg.ml-1. As optimized by the present authors, this phthalein reaction enables,
besides the highly selective determination of yperites, the use of other phthaleins as sensitive
reagents, e.g. α-naphtholphthalein, p-xylenolphthalein, thymolphthalein or erythrosin.
2.7.14
R
R-N(CH2CH2Cl)2] N Cl
CH2CH2Cl
CH3 CH3 CH3 CH3
HO OH HO O
+ OH
C - H2O
C
O
C O
C O
R
N
CH2CH2Cl
CH3 CH3
HO O
R
N C
CH2CH2Cl
C O
O
w
55
CH3 CH3
extrakce HO O
R
N C
extraction CH2CH2Cl
C O
org
CH3 CH3
HO O
C O CH2CH2NCH2CH2Cl
O R
org
The detection of liquid nitrogen yperite is based on its ability to decompose yellow pinaverdol
picrate to free pinaverdol which turns red when exposed to air.
2.7.15
O
H 3C
O 2N NO2
N I
C 2H5
N
C 2 H5
H
NO2
R-N(CH2CH2Cl)2
H3C O
O 2N NO2
N I R-NH(CH 2CH 2Cl)2
N
C2 H5
C 2H 5
NO2
56
This reaction has been utilized for the production of detector papers (paper No. 570). Red
spots are also produced by liquid tabun. The detection is interfered by alkaline compounds,
such as amine bases and hydroxide solutions.
Nitrogen yperites can be quantitatively determined by titration. Upon reaction with alcoholate
ions the liberated chlorides can be quantitatively determined. More advantageous is the direct
potentiometric titration of nitrogen yperite with sodium tetraphenylborate in an acid medium
indicated by a softened PVC membrane ion-selective electrode activated in trialkylammonium
tetraphenylborate solution [13].The acid-base titration with perchloric acid in a non-aqueous
medium of glacial acetic acid is more demanding and is indicated visually or
potentiometrically using a glass electrode.

1. Pat. ČSFR 277616. KOBLIHA, Z., SOUČEK, J., HALÁMEK, E.: Rychlý skupinový test k určení aminických
sloučenin. 7.2.1990.
2. CRUIKSHANK, A.J., BEWICK, H.A., CURRAH, J.E., BEAMISH, F.A.: Detection of certain chlorinated
tertiary aliphatic amines. Anal. Chem. 19(11), 849-850 (1947).
3. ORITEST. Indikační náplň do detekční trubičky ke zjišťování dusíkových yperitů.. Původce: Pitschmann V.,
Halámek E. Int. Cl. G01N31/22. CZ 14951. Zapsáno 29. 11. 2004.
5. KELLNER, J., HALÁMEK, E., PŘIKRYL, F.: Potenciometrická titrace organických dusíkatých bazí
s heteropolykyselinami. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1984, s. 10 - 23.
6. HALÁMEK, E., ČAPOUN, T.: Použití skleněné elektrody k indikaci titrací založených na tvorbě iontových
asociátů. Chemické listy, 83, 1989, s. 431 - 434.
s kyselinou pikrovou. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1984, s. 18 - 25.
8. EPSTEIN, J., ROSENTHAL, R.W., ESS, R.: Use of γ-(4-nitrobenzyl)pyridine as analytical reagent for
ethylenimines and alkylating agents. Anal. Chem. 27(9), 1435-1439 (1955).
9. FRIEDMAN, O.M., BOGER, E.: Colorimetric estimation of nitrogen mustard in aqueous media. Anal. Chem.
33(7), 906-910 (1961).
10. KOBLIN A.: Field sampling and analysis of micro quantities of sesquimustard in presence of mustard. Anal.
Chem. 30(3), 430-432 (1958).
11. SASS, S., KAUFMAN, J.J., CARDENAS, A.A., MARTIN, J.J.: Colorimetric estimation of tertiary and
quarternary amines. Anal. Chem. 30(4), 529-531 (1958).
12. Pat. ČSSR 270 370. HALÁMEK, E., KOBLIHA, Z.: Způsob extrakčně spektrofotometrického stanovení
reaktivních alkylhalogenidů ftaleiny. 25.11.1987.
13. Pat. ČSSR 267 737. ČAPOUN, T., HALÁMEK, E.: Iontově selektivní elektroda pro stanovení organických
iontů. 27.1.1988.
57
2.8 Lewisite
Cl AsCl2 Cl H AsCl2
C C Cl CH CH As C C
H H Cl Cl H
Lewisite [(2-chlorovinyl)dichloroarsane] is an organoarsenic compound of acylating

and reducing properties. It is the most effective representative of the large group of toxic
organic arsenic compounds. Its precipitation with sulfane is a group reaction for organic
haloarsanes but with advantage they can also be precipitated with reagents liberating sulfane
in a solution, e.g. with sulfane-hydroquinone adduct.
2.8.1
OH OH
H2S RAsCl2 RAs S 2 HCl
OH OH
The reaction affords a white, water-insoluble alkylarsane sulfide. The sensitivity of the
reaction is 7-10 µg.ml-1 and its selectivity corresponds to that of a group reaction. Inorganic
arsenic compounds afford a yellow precipitate. Heavy metal salts afford a brown or black
precipitate. The detection is interfered by oxidation reagents that oxidize sulfane to colloidal
sulfur. One of very valuable detection reactions for lewisite is the reaction with MICHLER’s
thioketone which is often applied to detector chalks [1] and papers for lewisite (M256A1).
The lewisite arsenic atom is bonded to the sulfur atom under the formation of a green product
arising from yellow thioketone.
2.8.2
Cl
(H3C)2N C N(CH3)2 Cl CH CH As
Cl
S
(H3C)2N C N(CH3)2 Cl
Cl As CH CH Cl
A very similar reaction of lewisite and other monosubstituted dihaloarsanes with

diphenylthiocarbazone affords a blue to violet product [2].
2.8.3
N N
C S RAsCl2 N N C N N 2 HCl
NH NH
S As
R
58
Without mentioning the mechanism, a colour test for lewisite has been described with
aromatic compounds, preferably with 2-chloro-3,5-dinitrobenzotrifluoride which gives a
green coloration [3]. The present authors have suggested a detector tube, containing silica gel
impregnated with 7-chloro-4-nitrobenzofurazane and sodium hydroxide in an ampoule. In this
case, the positive reaction affords a blue coloration.
The reduction of ergosterol with lewisite affording green coloration belongs to historical
procedures [4]. The reducing properties of lewisite are often used for the construction of
detector tubes. As an example, we may mention a detector tube with colourless osmium(VIII)
oxide that turns into brown to black osmium(IV) oxide.
2.8.4
OH
Cl
2 Cl CH CH As OsO4 4 H2O OsO2 2 Cl CH CH As O
Cl - 4 HCl
OH
To enhance the detection layer coloration and to increase sensitivity, a redox indicator of
benzidine type may be employed.
Good selectivity is shown by the reaction of lewisite and other primary dihaloarsanes with
potassium iodate resulting in the liberation of iodine. The reaction is also utilized for the
construction of detector tubes.
2.8.5
OH
Cl
5 Cl CH CH As 2 KIO3 9 H2O 5 Cl CH CH As O 2 KCl I2
Cl - 8 HCl
OH
The detection limit is 3 µg.l-1 in air.

The reducing properties of lewisite are also used in the construction of other types of detector
tubes. They are based on the reduction of molybdic acid to Molybdenum Blue. Molybdenum
Blue is an as yet insufficiently identified product of Mo(V) and Mo(VI). The sensitivity of the
reaction is good, but its selectivity is poor. The same holds for a detector tube containing
silica gel saturated with tin(II) chloride solution; here the presence of lewisite and primary
dichloroarsanes is manifested by a yellow ring. The group reaction for organoarsenic CWAs,
and thus also for lewisite, is their decomposition with a concentrated solution of hydroxide or
alcoholate ions under the formation of arsenites or arsenates.
2.8.6
Cl
- -
Cl CH CH As 6 OH HC CH 3 Cl AsO33 - 3 H 2O
Cl
In an acid medium, these inorganic arsenic compounds are reduced with hydrogen in statu
nascendi, usually generated by dissolving zinc (arsenic-free) in hydrochloric acid to give
gaseous arsane.
59
2.8.7
AsO33 - 3 Zn 9 HCl H3As 3 ZnCl2 3 H2O 3 Cl -
Arsane is usually sucked through a detector paper saturated with a reagent based on heavy
metals. The most often used reaction is the so-called GUTZEIT reaction with silver nitrate
which gives a yellow, gradually darkening precipitate of silver arsenide. Similarly, the
reaction with lead nitrate affords brown-black lead arsenide.
2.8.8
H3As 4 AgNO3 Ag3As AgNO3 3 HNO3
2 H3As 3 Pb(NO3)2 Pb3As2 6 HNO3
Arsane has also been detected using a tube containing silica gel saturated with a mercury(II)
bromide solution. The colour of the tube changes from white to yellow-brown depending on
arsane concentration [5,6]. The sensitivity is good, 5 µg.l-1 in air. However, phosphane and
sulfane give the same coloration.
2.8.9
2 H3As 3 HgBr2 Hg3As2 6 HBr
The WINKLER reaction with gold(III) chloride that gives a characteristic violet product
containing colloidal gold is regarded as the most sensitive reaction of arsane, both on a
detector paper and in a detector tube [7].
2.8.10
H3As 2 AuCl3 3 H2O 2 Au H3AsO3 6 HCl
On the whole, the hitherto most sensitive selective reaction described is the reaction according
to VAŠÁK [8-10] in which silver diethyl dithiocarbamate forms a complex with arsane in
pyridine or dimethyl sulfoxide giving a deep red colour. Its sensitivity is up to 0.1 µg.ml-1
exhibiting good selectivity.
2.8.11
S
S
H3As 3 (C2H5)2N C SAg H3 As
(C2H5)2N C SAg
3
The decomposition of lewisite by base and its reduction to arsane can be carried out,
particularly in detector tubes using complex hydrides, such as potassium borohydride,
K[BH4].
2.8.12
Cl
-
Cl CH CH As K[BH4] 3 OH H3As HC CH 3 CI - H 2O KBO2
Cl
60
The detection reaction of arsane itself is then based on the reduction of selenium(IV) oxide to
orange elemental selenium.
2.8.13
4 H3As 3 SeO2 3 Se 4 As 6 H2O
Unequivocally, most often used detection reaction for lewisite is that with the ILOSVAY
reagent [11-13]. The reaction is very selective and also of good sensitivity. It is based on the
precipitation of acetylene formed by alkaline decomposition of trans-lewisite with
monovalent copper under the formation of red copper(I) acetylide.
2.8.14
Cl
CH CH Cl 6 OH - HC CH 3 Cl - AsO33 - 3 H2O
As
Cl
trans lewisit
trans-lewisite
HC CH 2 [Cu(NH3)2] + Cu C C Cu 2 NH4 + 2 NH3
The ILOSVAY reagent is prepared by reducing a solution of copper(II) nitrate in aqueous

ammonia with hydroxylamine or hydrazine to give colourless diammine-copper(I) nitrate.
2.8.15
2+
NH2OH [Cu(NH3)4] OH - [Cu(NH3)2] + 1
/2 N 2 2 H2 O 2 NH 3
This highly selective reaction exhibiting good sensitivity (1 µg.l-1 in air) has been used in a
detector tube by the present authors [14]. The modified ILOSVAY reagent is applied to silica
gel and a concentrated hydroxide solution for lewisite decomposition is contained in an
ampoule. Upon passing contaminated air and breaking the ampoule, the presence of lewisite
is revealed by pink coloration.
The modification of the ILLOSVAY reagent for the construction of detector tubes comprises
saturation of the silica gel layer with diammine-copper(II) salt that after the decomposition of
lewisite with hydroxide is reduced with the liberated arsenite to the active component, i.e. the
diammine-copper(I) salt which reacts with acetylene to red acetylide.
2.8.16
Cl
Cl CH CH As 2 [Cu(NH 3)2]Cl2 10 NaOH Cu2C 2 Na3AsO4 7 NaCl 6 H 2O 4 NH3
Cl
Similar to sulfur yperite, lewisite dissolves lipophilic dyes to characteristically coloured

solutions. In the case of Dye Red E (see 2.6.12) a red to purple coloration appears.
For quantitative analysis of lewisite, the titrimetric determination of decomposition products
after alkaline hydrolysis appears to be the method of choice. Since the reaction affords As(III)
species (arsenites), the simplest solution is the redox titration with potassium permanganate or
iodine solution.
2.8.17
AsO33 - I2 H2O AsO43 - 2I- 2H+
The titration of chlorides after alkaline hydrolysis is not suitable for quantitative
determination because cis-lewisite partly decomposes to vinyl chloride.
61
1. WITTEN, B., PROSTAK, A.: Sensitive detector crayons for phosgene, hydrogen cyanide, cyanogen chloride,
2. TARBELL, D.S., BUNNETT, J.F.: The reaction betwen arsenicals and di-(p-biphenyl)-thiocarbazone. J. Am.
Chem. Soc. 69, 263-265 (1947).
3. YOE, J.H., COGBILL, E.C.: Organic reagens for the identification of certain vesicants. Microchim. Acta 38,
492-497 (1951).
4. MASON, H.S.: Note on an new color reaction of β-chloro-vinyldichloroarsine. J. Am. Chem. Soc. 67(12),
2267-2268 (1945).
5. SANGER, C.R., BLACK, O.F.: Quantitative estimation of arsenic by the Gutzeit method. J. Soc. Chem. Ind.
26, 1115-1123 (1907).
6. RAHMAN, M.M., FUJINAGA, K., SEIKE, Y., OKUMURA, M.: A simple in site visual and tristimulus
colorimetric method for the determination of trace arsenic in environmental water after its collection on a
Mercury(II)-impregnated paper. Anal. Sci. 20, 165-170 (2004).
7. BAGHEL, A., SINGH, B., PANDEY, P., SELHAT, K.: A rapid field detection method for arsenic in drinking
water. Anal. Sci. 23, 135-137 (2007).
8. VAŠÁK,V., ŠEDIVEC, V.: Kolorimetrické stanovení arsenu. Chem. Listy 46, 341-344 (1952).
9. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z.: Trubička pro detekci arsanu a lewisitu. Sborník VVŠ PV
Vyškov, 2003, číslo 1, s. 27-33.
10. ORITEST. Detekční trubička pro zjišťování toxických sloučenin arsenu. Původce: Pitschmann V.,
Halámek E., Kobliha Z. Int. Cl. G01N31/22. CZ 12292. Zapsáno 4.6.2002.
11. ILOSVAY L. VON NAGY ILOSVA: Ueber die Bereitung ammoniakalischer Cuprolösung mit
Hydroxylamin zum Nachweise des Acetylens. Ber. 32, 2697-2699 (1899).
12. GREEN, S.J., PRICE, T.S.: LVI. – The chlorovinylchloroarsines. J. Chem. Soc., Trans. 119, 448-453 (1921).
13. PITSCHMANN, V.: Jednoduché stanovení lewisitu. Chemické listy 93, 207-209 (1999).
14. ORITEST. Detekční trubička ke zjišťování lewisitu. Původce: Pitschmann V., Halámek E. Int. Cl.
G01N31/22. CZ 8867.Zapsáno 13.7.1999.
62
2.9 Organophosphorus chemical warfare agents
The general formulae of current organophosphorus CWAs are as follows:
X: R2:
SCH2CH2N(iC3H7)2 [VX]
C2H5 [VX]
SCH2CH2N(C2H5)2 [R-33]
CH3
[GB]
[GB,GD,GF,GP, A-230/2/4] CH
F
CH3
CN [GA]
CH3
CHCH2 [R-33]
CH3
X OR2
CH3
P NCH2CH2 [GP]
R1: CH3
R1 O CH3
CH3 [VX,R-33,GB,GD,GF]
H3C C CH [GD]
CH3
N [GA,GP]
CH3 CH3
CH3
H [GF]
Cl R: R':
C N O [A-230/2/4]
F H H [A-230]
CH3 H [A-232] Cl CH CH [A-230/2/4]
CH3 CH3 [A-234] R' R
The main toxic effect of organophosphorus CWAs is their nerve paralytic inhibition of
acetylcholinesterase. The fact that they inhibit the hydrolytic effectivity of cholinesterase
enzymes already in very low concentrations is also utilised in their selective analytical
determination and selective detection. The sensitivity of the cholinesterase reaction is very
high and corresponds to the toxic effect of organophosphorus CWAs on human organisms.
Then this is the reason for using this biochemical reaction unlike the detection reactions based
on a chemical reaction or a physical principle. The limit of determination of
organophosphorus compounds by the cholinesterase reaction is 1.10-5-1.10-7 mg.ml-1(or litre
of air), depending on the inhibitor. Compared with the sensitivity of common chemical
methods, ranging from 1.10-4 to 1.10-2 mg.ml-1 in a sample, this represents a difference of 1 -
5 orders of magnitude in the analyte concentration. This high sensitivity makes the
cholinesterase reaction an ultratrace analytical method and enables the detection of
organophosphorus CWAs even in not acutely dangerous concentrations.
The cholinesterase reaction is utilized within the whole range of technical devices used in
chemical reconnaissance and check. It is based on the use of a suitable hydrolase,
cholinesterase, enzyme that catalyzes the hydrolysis of a physiological substrate efficiently,
acetylcholine (acetic acid ester of amino alcohol choline) [1].
The choice of suitable hydrolases obtainable from living organisms is very wide. However, in
principal, only two basic enzyme types come into consideration: acetylcholinesterase (ACHE,
specific esterase) and butyrylcholinesterase (BuCHE, so-called nonspecific cholinesterase). In
all cases, the enzyme is a protein of molecular weight of the order 105 D which, depending on
conditions, can form larger entities containing usually two or four subunits. Regardless of the
enzyme etiology, the main difference between the two enzyme types consists in their ability
63
to catalyze, in addition to their physiological (so-called specific) substrate, also the hydrolysis
of other esters. As a specific neuronal enzyme, acetylcholinesterase readily hydrolyzes the
neurotransmitter (here the substrate is acetylcholine or its sulfur isoster acetylthiocholine). As
its name suggests, for butyrylcholinesterase, present to a greater extent in blood serum, the
most acceptable partner is the ester of butyric acid with choline, butyrylcholine or its thio
isoster, butyrylthiocholine. Unlike acetylcholinesterase, butyrylcholinesterase relatively
readily catalyzes hydrolysis of many other substrates that only very little resemble
acetylcholine or butyrylcholine, usually containing only an acetyl, or even an ester group, in
their molecule. The affinity of individual inhibitors toward the enzymes (their active sites) is
different.
Since the cholinesterase reaction is mainly used as a kinetic method, the enzyme activity, and
thus its concentration, is derived from its ability to catalyze the hydrolytic conversion of
substrate during a time interval. The originally used international unit was 1 µmol per minute.
The international system of units has introduced an enzyme activity unit, 1 katal (kat), which
is the amount of the converted substrate (in mol) in 1 second. Because 1 kat is too large for
practical use, the commonly used unit is µkat or mkat. Enzyme E (hydrolase) reacts with
substrate S under formation of an enzyme-substrate complex [E.S] which is then hydrolyzed
with water to give products P [2].
2.9.1
H 2O
E S [E.S] E P
If an acetylcholinesterase inhibitor is present (in this case an organophosphorus CWA), it

binds, instead of the substrate, to the active centre of the enzyme (competitive inhibition)
forming thus a stable enzyme-inhibitor complex that hinders further enzymatic hydrolysis of
the substrate.
2.9.2
E I [E.I]
The reaction course can be monitored using either the decreasing concentration of a substrate
or the increasing concentration of products. For several practical reasons, the increase in
product concentration is usually observed. The original AMMON method for the
determination of cholinesterase activity was carried out in the VARBURG apparatus where
the arising acid reacted with a hydrogen carbonate buffer under the evolution of carbon
dioxide, the volume of which was measured [3]. Until recently, cholinesterase methods based
on the measurement of concentration of the arising acid (pH) were very widespread. These so-
called pH-metric methods made use of acid-base indicators in combination with buffers (pH-
regulators) of low capacity, or better, the arising acid was continuously titrated, usually with
glass electrode potentiometric indication. The acid-base indicator method has also been used
in detector tubes [4,5], detector papers as well as in automatic alarms. Butyrylcholinesterase
or its purified concentrate obtained from horse blood plasma was employed as a suitable
enzyme. Butyrylcholine iodide served as the substrate.
2.9.3
O
BuCHE CH3CH2CH2COCH2CH2N(CH3)3 BuCHE CH3CH2CH2COOH (CH3)3NCH2CH2OH

butyrylcholine
butyrylcholin butyric acid
kyselina máselná choline
cholin
64
Phenol Red and Bromothymol Blue have often been used as acid-base indicators. The
already mentioned manometric method, colorimetric (visual), spectrophotometric or
electrochemical method (potentiometry, amperometry, voltametry) can be used as a
pH-metric method. A drawback of these methods is that they are interfered by acid and
alkaline substances and thus depend on the buffering capacity of the measured solution; the
enzyme also has to be quite pure. Accompanying ballast substances in the aqueous solution
behave as active buffer components.
Another group of modified cholinesterase reactions are methods utilizing the so-called
chromogenic or fluorogenic substrates. The methods sometimes use even very ingenious
compounds as substrates that change colour after the hydrolysis of their ester bond. Then this
colour change shows absence of a cholinesterase inhibitor (organophosphorus CWA). As
follows from the principle, the evaluation is done visually or spectrophotometrically. Thus,
e.g. 3-indolyl acetate as a substrate is hydrolyzed to blue-green fluorescing 3-hydroxyindole
which is further oxidized to give blue indigo [6].
2.9.4
O
OCCH3 OH
BuCHE
2 2 H2O 2 2 CH3COOH
N N
H H
O
H 2 O
N
2 H2O
N
H
O
One of the best-known chromogenic substrates used in the construction of detector set NAVD
(Nerve Agents Vapour Detector) is 2,6-dichloroindophenol acetate. In the absence of a
cholinesterase inhibitor this lightly red compound is hydrolyzed to blue
2,6-dichloroindophenol [7].
2.9.5
Cl O
O N O C CH3 H2O
Cl
Cl BuCHE
CH3COOH O N OH
Cl
Cl
HO N O
Cl
65
2-azobenzene-1-naphthyl acetate and p-nitrophenyl acetate are also well-known chromogenic
substrates [8].
2.9.6
O
O C CH3
N N H2O
žlutý
yellow
OH BuCHE
N N
CH3COOH
red
červený
2.9.7
O
BuCHE
H3C C O NO2 H2O HO NO2 CH3COOH
yellow
žlutý
OH
O
O N H2O
O
red
červený
Advantageously, cholinesterase methods with chromogenic substrates can use buffers of

higher capacity than the pH-metric method and thus eliminate the interfering influence of
acidic or basic compounds. A drawback of the methods, however, is a lower rate of the
hydrolytic reaction, an overall lower stability of the substrates, their high price and limitation
exclusively to the non-specific enzyme butyrylcholinesterase which overcomes great
structural differences of substrates from the choline ester more easily. Although the specific
enzyme acetylcholinesterase also cleaves such substrates, the hydrolysis proceeds only very
slowly and, moreover, the enzyme is inhibited easily by substrate solutions already diluted.
Most of the drawbacks of pH-metric methods and methods with chromogenic substrates have
been overcome by using thiocholine substrates: acetylthiocholine for acetylcholinesterase or
butyrylthiocholine for butyrylcholinesterase. The hydrolysis of these substrates affords thiol
(thiocholine) that is very reactive and is oxidized to the corresponding disulfide readily
enabling thus the electrochemical (voltametric, amperometric, coulometric) determination of
the cholinesterase reaction course. This has been utilized also in the construction of automatic
alarms signalling nerve agents [9].
The use of oxidized forms of redox indicators enables not only spectrophotometric
determination of the cholinesterase reaction course, but also the visual colorimetric
determination and thus the construction of detector sets, papers and tubes.
2.9.8
66
O
ACHE
CH3CSCH2CH2N(CH3)3 H2O CH3COOH
acetylthiocholin
acetylthiocholine
HSCH2CH2N(CH3)3
O thiocholine
thiocholin
BuCHE
CH3CH2CH2CSCH2CH2N(CH3)3 H2O CH3CH2CH2COOH
butyrylthiocholine
butyrylthiocholin
2.9.9
2 (CH3)3NCH2CH2SH 2 Ind(ox) (CH3)3NCH2CH2 S S CH2CH2N(CH3)3 2 HInd(red)
It is worth mentioning that most of the oxidized forms of redox indicators Ind(ox) are markedly
coloured (deep colour, high molar absorption coefficient), whereas the reduced forms
HInd(red) are usually colourless (leuco base). Frequently employed redox indicators for
chromogenic reaction with thiocholine include the Guinea Green B (Acid Green 3) dye,
carbinols of MICHLER’s ketone and Malachite Green [10-12].
2.9.10
2 CH2 N C N CH2
C2H5 C2H5
NaO3S SO3
green
zelená
2 (CH3)3NCH2CH2SH
2 CH2 N CH N CH2
C2H5 C2H5
NaO3S SO3H
colourless
bezbarvá
(CH3)3NCH2CH2 S S CH2CH2N(CH3)3
The mentioned chromogenic detection of thiocholine with Guinea Green B is applied as a

remission photometric method for the reaction of BuCHE and butyrylthiocholine in the GSA-
12 automatic alarm. The advantage of cholinesterase methods with acylthiocholine-based
substrates consists in their independence of an accurate pH value and of low buffering
capacity of the medium. This enables the use of smaller amounts of enzyme which, moreover,
need not be so highly purified. The method is interfered by reducing and strongly oxidizing
substances. A certain drawback is that the method evaluates the decrease in coloration
67
intensity; this means that in visual evaluation one has to wait till the indicator colour changes
(here till the whole redox indicator is decolourized). On the other hand, a photometric
measurement has no such drawback. This is probably the reason of the high reputation of the
ELLMAN reagent, 5,5`-dithiobis(2-nitrobenzoic acid). The reagent is exceptional in that its
oxidized form has only an inexpressive, yellowish colour, in contrast to its deep yellow to
orange reduced form [13]. Thus, it is possible to quickly check visually whether the substrate
liberates thiocholine and the reaction mixture (reaction zone) turns yellow or whether the
enzyme is inhibited and the mixture remains colourless, white or only slightly yellowish.
2.9.11
2 (CH2)3NCH2CH2S O2N S S NO2
thiocholinát
thiocholinate HOOC COOH
Ellman reagent
Ellmanovo činidlo
(CH3)3NCH2CH2 S S CH2CH2N(CH3)3 2 O2N S
HOOC
O
N S
O
HOOC
Not only sets for the determination of cholinesterase activity in clinical biochemistry, but also
the colorimetric biosensor called DETEHIT [14-17] are based on the mentioned colorimetric
and spectrophotometric principle. The same principle is also used in the biochemical detector
tube with three red stripes used e.g. in CHP-71 and CHP-5 [18,19] chemical detectors. The
DETEHIT biosensor is included in a number of devices, sets and mobile laboratories of the
Czech Army. Unlike all other similar devices of chemical reconnaissance and check used in
other armies, this device enables checking not only air, but also the surface of military
equipment and material, water and food. This fundamental difference rests in the
immobilization of acetylcholinesterase as a stable enzyme chimera with polysaccharide
cellulose. Thus, the enzyme remains in the solid phase and its use is polyvalent. Since the
biosensor is very sensitive and reaches detection limits for the cholinesterase reaction, it can
be employed for a simple and rapid check of the completeness of decontamination of military
equipment and material as well as the check of the absence of organophosphorus CWAs or
other strong cholinesterase inhibitors in water and food.
The mentioned detector tube also contains the enzyme in its immobilized form and the tube
can be used for the improvised check of water contamination.
Although the cholinesterase reaction is widely used exclusively as a group method for the
detection of organophosphorus CWAs and, if need be, other potent cholinesterase inhibitors
or for the quantitative determination of a specific organophosphate, it can be modified into a
simple procedure that differentiates organophosphates of the G-series (sarin-type) from those
of the V-series (VX-type), thus increasing the accuracy of concentration determination in an
68
otherwise unknown sample. This procedure is based on the finding that ions of alkali metals
and alkali earth metals, probably as effectors, enter the quaternary cholinesterase structures.
They also change the configuration and charge distribution at the active site, facilitating the
entry of organophosphates of the G-series into the active site and thus increasing, or at least
preserving, their inhibitory activity [20-22]. Unlike the compounds of the G-series, the
inhibitory activity of the compounds of the series V is lowered only. In practical execution of
the test, a sample of the organophosphate is diluted with water to achieve an only partial
inhibition of cholinesterase, i.e. the substrate hydrolysis still takes place, though more slowly
than in the control experiment without the inhibitor. If the hydrolysis does not proceed at all,
the entire enzyme is inhibited (I100) and the sample must be further diluted. The achieved
dilution of the inhibitor is compared to the blank experiment without the inhibitor and the
percentage of cholinesterase inhibition is calculated using the relationship
% I = [(ti – tSE)/ti].100
where ti is the time after which the coloration of the diluted sample is changed, and tSE is the
time in the blank experiment without the inhibitor. Thus the value I50 is the dilution at which
the colour change appears after the twofold prolongation of time in comparison with the blank
experiment. After the determination of the degree of cholinesterase inhibition in %, the series
of G and V are defined. The last dilution of the inhibitor sample is repeated in two test tubes:
in one test tube the sample is diluted with water and in the other with 1M aqueous magnesium
sulfate or calcium chloride solution. Saturated sodium or potassium chloride solution can also
be used. After determining the inhibitor kind, the concentration can be assessed according to
% I of butyrylcholinesterase (Table 2).
Table 2: Determination of organophosphate agent concentration according to %I of

butyrylcholinesterase
Agent concentration, mg.ml -1

%I
BuCHE GB VX
20 1.1.10-6 3.0.10-7
30 2.9.10-6 8.6.10-7
40 5.0.10-6 1.5.10-6
50 7.1.10-6 2.3.10-6
60 9.5.10-6 3.4.10-6
70 1.4.10-5 4.8.10-6
80 2.0.10-5 7.1.10-6
The error in method is about ± 25 %.

To merely distinguish between the G- and V-type compounds, the last dilution with water is
simply repeated with the solution of one of the above-mentioned salts and the inhibitory
activity is evaluated as follows: increase (G), decrease (V), unchanged (G) [23].

1. VOET, D., VOETOVÁ, J. G.: Biochemie. Victoria Publishing, a.s., Praha, 1995, 1.vydání, 1349 s.
2. KARLSON, P.: Biochemie. Academia, Praha, 1981, 3.vydání, 504 s.
3. AMONN, R., VOSS, G. Plugers Arch.ges. Physiol. 235, (1935), s.393.
69
4. OLTHOFF, U., KÖNIG, H.: Prüfröhrchen zum Nachweis biologisch aktiver Phosphorsäureester in Luft.
Int. Cl. G01N31/22. DD 301 983 A7. (13.10.1994).
5. BITTER, R., KRAL, E.: Indikatorrörchen zum Nahweis von organischen Phosphorsäureestern. Int. Cl.
G01N31/22. DD 301 363 A7. (10.12.1992).
6. BARNET, R. J., SELIGMAN, A. M. Science, 114, (1951), s. 579.
7. IVARSSON, U, NILSSON, H., SANTESSON, J.: Chemical weapons. FOA Sveden, 1992, 77 p.
1977. 512/616 s.
10. Užitný vzor ČR. 7004. HALÁMEK E., KOBLIHA, Z., OREL, J., TUŠAROVÁ I.: Souprava indikačních
prostředků pro automatický signalizátor GSP 11. 23.1.1998.
11. Užitný vzor ČR. 7006. HALÁMEK E., KOBLIHA, Z., OREL, J., TUŠAROVÁ I.: Souprava indikačních
prostředků pro automatický signalizátor GSA 12. 23.1.1998.
12. HALÁMEK, E., KOBLIHA, Z., SKALIČAN, Z.: Spektrofotometrické stanovení 2-dimethylaminoethan-
thiolu s Guinea Green B a Malachite Green Carbinol Hydrochlorid. In: Sborník VVŠ PV, 3, Vyškov, VVŠ PV,
1998, s. 237 - 247.
13. ELLMAN, G.L., COURTNEY, D.K., ANDRES, V., FEATHERSTONE, R.M.: A new and rapid
colorimetric determination of acetylcholinesterase aktivity. Biochem Pharmacol. 7, 1961, p.88-95.
14. HALÁMEK, E., TUŠAROVÁ, I.: Jednoduchý detektor nervově paralytických látek. In: Zborník
vojenskovedeckých prác 4, 2, Liptovský Mikuláš, VVTŠ, 1989, s. 134 - 150.
15. TUŠAROVÁ, I., HALÁMEK, E., KOBLIHA, Z.: Study on reactivation of enzyme-inhibitor complexes by
oximes using acetylcholine esterase inhibited by organophosphate chemical warfare agents. Enzyme and
Mikrobiological Technology 25, 1999, p. 400 – 403.
16. Pat. ČR 288 576 TUŠAROVÁ I., HALÁMEK E.: Biosenzor pro detekci a rozlišení inhibitorů cholinesteráz,
způsob přípravy zóny biosenzoru s imobilizovanou cholinesterázou, způsob detekce inhibitorů cholinesteráz a
způsob rozlišení inhibitorů cholinesteráz. 22. 5. 2001.
17. Pat. SR 281 395 TUŠAROVÁ, I., HALÁMEK, E.: Biosenzor, spôsob prípravy zóny s imobilizovanou
cholínesterázou, spôsob detekcie a rozlíšenia inhibítorov cholínesteráz biosenzorom. 12. 3. 2001.
18. Pat. ČR 285 242. TUŠAROVÁ I., HALÁMEK E., OREL, J.: Detekční trubička inhibitorů cholinesteráz ve
vzduchu a ve vodě. 16.6. 1999.
19. Pat. SR 281 547. TUŠAROVÁ I., HALÁMEK E., OREL, J.: Detekčná rúrka inhibítorov cholinesteráz vo
vzduchu a vo vode. 9.4. 2001.
20. TUŠAROVÁ, I., HALÁMEK, E., SOUČEK, J.: Rozlišení nervově paralytických látek cholinesterasovou
reakcí . Civilní obrana 31,1989, 5, s. 45 - 48.
21. TUŠAROVÁ, I., HALÁMEK, E.: Použití jednoduchého detektoru inhibitorů acetylcholinesterasy k určení
typu nervově paralytické látky. In: Sborník VVŠ PV, řada B, č. 1, Vyškov, VVŠ PV, 1991, s. 14-21.
22. Pat. ČSSR 2699. TUŠAROVÁ, I., HALÁMEK, E., SOUČEK, J.: Způsob rozlišení organofosforových
inhibitorů cholinesterasovou reakcí. 25.1.1988.
70
2.10 Compounds of the G-series (sarin type)
Compounds of the G-series have the following formula:
X: R2:
F [GB,GD,GF] C2H5 [GA]
CN [GA] CH3
X OR2 CH [GB]
CH3
P
1 CH3
R O
1
R : H3C C CH [GD]
CH3 [GB,GD,GF] CH3 CH3
CH3
[GA] H [GF]
N
CH3
Unlike the V-series compounds, the G-series compounds markedly tend to enter
nucleophilic substitution reactions. This ability is utilized in the analysis of unknown samples.
The presence of sarin is most often detected using the so-called SCHÖNEMANN
aminoperoxide reaction [1]. After confirming the presence of an acetylcholinesterase
inhibitor, this group reaction for acyl halides is used as a sensitive and selective reaction for
sarin-type compounds. The condition for its successful course is a higher rate of the
nucleophilic substitution reaction of the organophosphate with hydroperoxide ions than with
hydroxide ions. Therefore, this hydrogen peroxide reaction is carried out in a very weakly
alkaline medium of pH 7- 8. The reaction with hydroxide ions is a competitive reaction that
lowers the sensitivity, or even makes the determination impossible.
2.10.1
R1 O R1 O
P OOH P X
2 2
R O X R O OOH
The arising peroxy acid is a very strong oxidation reagent which readily converts reduced
leuco forms of redox indicators into their coloured oxidized forms.
2.10.2
R1 O R1 O
P 2 HInd(red) P 2 Ind(ox) H2O
R2O OOH R2O OH
Benzidine-derived indicators, such as o-dianisidine and o-tolidine, are usually employed;

however, the use of triphenylmethane dyes is also described in the literature [2,3].
71
2.10.3
R1 O
2 P H2N NH2
R2O OOH
H3CO OCH3
R1 O
H2N NO 2 P H2O
R2O OH
H3CO OCH3
H2N NH2
H3CO OCH3
H2N N N NH2 H2O
H3CO OCH3 H3CO OCH3
The reaction affords a yellow-green product with benzidine, a yellow-orange coloration with
o-tolidine and a red coloration with o-dianisidine. Descriptions of the course of the
SCHÖNEMANN reaction vary from intermediates of the iminomesoquinone type to nitroso
derivatives, coupling with the starting reduced compound to give an azo dye.
2.10.4
R1 O
P H2N NH2
R2O OOH
H3CO OCH3
R1 O
HN NH P H2O
R2O OH
H3CO OCH3
72
2 HN NH
H3CO OCH3
H2N N N NH2
H3CO OCH3 H3CO OCH3
This reaction has been used for the production of detector tubes and automatic alarms [4-6].
In the determination of sarin-type compounds the sensitivity is usually up to 1 µg.ml-1 in a
sample, however, the reproducibility is not very high. On the other hand, if the reaction
mixture is allowed to react for 20 minutes and the formed coloured substance is subsequently
extracted with chloroform, the results are fully reproducible. The present authors optimized
this procedure for application in field chemical laboratories [7]. Apart from the
spectrophotometric and colorimetric methods, fluorescence- and chemiluminiscence-based
modifications have also been used. The fluorescence method employs indole as an indicator;
this compound is oxidized with the peroxy acid to afford an indoxyl (3-hydroxyindole) of
blue-green fluorescence [8].
2.10.5
R1 O R1 O OH O
P P
N N
R 2O OOH H R 2O OH H
N
H
The reaction is very sensitive, up to 0.02 µg.ml-1 in a sample, and, therefore, this modification
has also been used in automatic alarms. Another modification of the SCHÖNEMANN
reaction is a chemiluminiscence reaction of the peroxy acid with luminal [9,10].
2.10.6
NH2 O NH2 OH
R1 O R1 O
NH N
2 P 2 P O ν
hν
NH O
N
2 2
R O OOH R O OH
O OH
73
The arising luminol peroxide is decomposed with blue-green luminiscence. The sensitivity,
0.5 µg.ml-1, is slightly higher than that of the photometric method.
Another reaction of the sarin-type compounds is their reaction with oximes, particularly with
aldoximes, in a weakly alkaline medium.
2.10.7
R1 O R1 O
P R3HC N OH P HX
R2O X R2O O N CHR3
R1 O
P N CHR3 R3CN H
2
R O O
If the aldoxime has an oxo or hydroxy group in position α, the reaction, instead of alkyl
nitrile, gives hydrogen cyanide which is then detected by usual chromogenic reactions.
2.10.8
R1 O O R1 O
O
P R3C CH N OH P HX
2 2 3
R O X R O O N CH CR
R1 O O
O O
P
3 3 3
N CH CR H R C CN R C O CN H2O
R2O O
2 OH
Oximes are employed in detector chalks, detector tubes as well as in automatic alarms
[11,12]. For example, isonitrosobenzoylacetone is used in the M43/M8 detector. Another
frequently used oxime is diisonitrosoacetone which not only liberates hydrogen cyanide, but
also affords a red product of the hitherto unidentified composition upon the reaction with a
sarine-type compound.
A similar reaction with hydroxamic acids, usually with potassium benzhydroxamate, is also
very widespread [13].
74
2.10.9
O
R1 O R1 O
O
P C NH OH P HX
2
R O X R2O O NH C
O
R1 O
P C NH N C O H2O
2
R O O
-H
NH2 CO2
The product of the nucleophilic reaction of benzhydroxamate with a sarin-type compound

undergoes cleavage and is converted via LOSSEN rearrangement into phenyl isocyanate
which in an alkaline medium is further hydrolyzed to aniline. This can be detected by many
chromogenic reactions, e.g. by facile conversion into an azomethine dye by condensation
with p-dimethylaminobenzaldehyde in an acid medium.
2.10.10
N CH
H3C
NH2 N CHO H2O
H3C
N CH3
H3C
The sensitivity of the reaction ranges from 0.1 to1 µg.ml-1, depending on the method
modification. The low selectivity reflects the fact that it is a group reaction for acyl halides; a
positive reaction is also observed with alkyl halides at high concentrations. The so-called
pyridine reaction with 4-(4'-nitrobenzyl)pyridine in an alkaline medium, affording a blue
coloration is even less selective.
2.10.11
R1 O
P O2N CH2 N
2
R O X
75
R1
O2N CH2 N P OR2 X
OH
R1
O2N CH N P OR2 X H2O
R1
O
N CH N P OR2
O
O
This group reaction is often employed in detector devices. Its sensitivity for sarin-type
compounds is up to 25 µg.ml-1.

1. SCHÖNEMANN, R.B.R.: Eine neue Reaktion für den Nachweis von labilen Nichtmetal-Halogen-Bindungen,
Gasabweher-Laboratorien der deutschen Wehrmacht, Berlin 1944, enhalten im P.B. 119887 des U.S.
Department of Commerce.
2. GRANT, G.A., BLANCHFIELD, R., SMITH, D.M.: Triarylmethane compounds as redox indicators in the
Schoenemann reaction: I. Mechanism of the Schoenemann reaction. Can. J. Chem./Rev. Can. Chim. 35(1),
42-49 (1957).
3. SMITH, D.M., BLANCHFIELD, R., THOMPSON, J.J., GRANT, G.A.: Triarylmethane compounds as redox
indicators in the Schoenemann reaction: II. Syntheses of the triarylmethanes. Can. J. Chem./Rev. Can. Chim.
35(2), 156-162 (1957).
4. SMITH, S.J.: Detection methods for highly toxic organophosphonates. Talanta 30(10), 725-739 (1983).
5. WELCH, C.M., WEST, P.W.: Qualitative microdetermination of organic phosphorus compounds. Anal.
Chem. 29(6), 874-877 (1957).
6. GEHAUF, B., EPSTEIN, J., WILSON, G.B., WITTEN, B., SASS, S., BAUER, V.E., RUEGGEBERG,
W.H.C.: Reaction for colorimetric estimation of some phosphorus compounds. Anal. Chem. 29(2), 278-281
(1957).
7. HALÁMEK, E., KOBLIHA, Z.: Extrakčně spektrofotometrické stanovení sarinu amino-peroxidovou reakcí.
In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1988, s. 49 - 62.
8. GEHAUF, B., GOLDENSON, J.: Detection and estimation of nerve gases by fluorescence reaction. Anal.
Chem. 29(2), 276-278 (1957).
9. GOLDENSON J.: Detection of nerve gases by chemiluminiscence. Anal. Chem. 29(6), 877-879 (1957).
10. CHERRY, R.H., FOLEY, G.M., BADGETT, C.O., EANES, R.D.: Alarms and analyzers for nerve gas
vapors. Anal. Chem. 30(7), 1239-1247 (1958).
76
11. SASS, S., LUDEMANN, W.D., WITTEN, B., FISCHER, V., SISTI, A.J., MILLER, J.I.: Colorimetric
determination of certain organophosphorus compounds and acylating agents. Anal. Chem. 29(9), 1346-1349
(1957).
12. SEVILLE B.: A new approach to the determination of microgram quantities of phosphorylating or acylating
agents. Analyst 82, 269-274 (1957).
13. SCHWIDLER, R., STEINBERG, G.M.: The kinetice the reaction of isopropyl methylphosphonofluoridate
(sarin) with benzohydroxamic acid. J. Am. Chem. Soc. 78(15), 3594-3598 (1956).
77
2.11 Tabun
CH3
H3C N
O
P
C2H5O
CN
Among organophosphorus CWAs of the G-series, the structure of tabun [ethyl

(dimethylphosphoramido)cyanidate] has a special position because the electronegative group
in its molecule is a pseudohalide – cyanide. The dimethylamide group is less usual as well.
The presence of tabun is confirmed by positive cholinesterase and aminoperoxide reactions.
Further, its determination is usually based on the detection of cyanides liberated by alkaline
hydrolysis. It is interesting that, in its original version, the SCHÖNEMANN reaction
consisted directly in a test for the hydrolytically liberated cyanide that affords yellow
copper(II) cyanide upon the reaction with copper(II) salts [1]. This disproportionates into
white copper(I) cyanide and dicyanogen which reacts with water to give hydrogen cyanide
and oxygen in statu nascendi. The atomary oxygen reacts with luminol under formation of the
chemiluminiscent peroxide.
2.11.1
H3C H3C
N O N O
H3C P 2 OH H3C P CN H2O
C2H5O CN C2H5O O
2.11.2
2 CN - Cu 2+ Cu(CN)2
2 Cu(CN)2 2 CuCN (CN)2
(CN)2 H2O 2 HCN O
NH2 O NH2 OH
NH N
2O O
O
N ν
hν
NH
O OH
The sensitivity of the reaction is very good: 0.5 µg.ml-1 in a sample. Its selectivity was so
good that it gave positive results only with tabun but not with sarin discovered later.
SCHÖNEMANN modified the procedure and replaced copper ions by hydrogen peroxide
and converted almost a specific reaction to a group reaction for acyl halides.
78
Cyanide ions are usually detected as Berlin Blue (see Chapter 2.4 Hydrogen cyanide) or
by the KÖNIG-ZINCKE reaction after conversion to cyanogen chloride (see Chapter 2.5
Cyanogen chloride). A detector paper for liquid tabun was based on the cyanide-catalyzed
reaction of p-nitrobenzaldehyde, affording red bis(p-nitro)benzoin. In a solution, the
detection limit is 3 µg.ml-1 in a sample (see 2.4.8).
The tabun molecule can lose dimethylamine which is then detected by group reactions for
amines and alkaloids, but only by the reactions which also precipitate secondary amines,
such as the universal DRAGENDORFF reagent that forms orange precipitates.
2.11.3
H3C H3C H
N O N O HO O H3C
H3C P H H3C P H2O P NH2
C2H5O CN C2H5O CN C2H5O CN H3C
H3C H3C
NH2 [BiI4] NH2 [BiI4]
H3C H3C
An acid hydrolysis product, ethyl phosphorocyanidate, is saponified using concentrated

hydroxides, advantageously with warming, and in this way tabun and its hydrolytic
products afford phosphates.
2.11.4
HO O O O
P P
3 OH OH PO4 3- C2H5OH
-2 H2O -
C 2H 5 O CN - CN C 2H 5 O O
Subsequently, phosphates can be detected by many selective reactions. Thus, e.g.

phosphates are precipitated by silver nitrate as yellow salt.
2.11.5
PO4 3- 3 Ag + Ag3PO4 (žlutá sraženina)
(yellow precipitate)
Zirconium nitrate precipitates white zirconium phosphate insoluble in concentrated

hydrochloric acid.
2.11.6
4 PO4 3- 3 Zr 4+
Zr3(PO4)4 (whitesraženina)
(bílá precipitate)
Molybdenum solution (a solution of ammonium molybdate in nitric acid) precipitates

phosphates to give a yellow precipitate of the composition (NH4)3[P(Mo3O10)4].H2O. The
volume of this precipitate can be increased by addition of an organic nitrogen base.
Molybdatophosphate with benzidine gives deep blue coloration to the detriment of the
arising benzidine and molybdenum blue. The determination is interfered by arsenates and
silicates; these, however, can be masked by an excess of sulfuric acid.
The argentometric or mercurimetric titration of cyanides indicated visually or
potentiometrically is suitable for the quantitative determination of tabun (see Chapter 2.4
Hydrogen cyanide) [2-4].
79
1. SCHÖNEMANN, R.B.R.: Eine neue Reaktion für den Nachweis von labilen Nichtmetal-Halogen-Bindungen,
Gasabweher-Laboratorien der deutschen Wehrmacht, Berlin 1944, enhalten im P.B. 119887 des U.S.
Department of Commerce.
obrana, 28, 1986, 2, s . 75-87.
80
2.12 Organophosphorus fluoridates
R1: R2:
CH3 [GB,GD,GF] CH3

CH [GB]
CH3
CH3
N [GP] R1 OR2
CH3
CH3
P
Cl H3C C CH [GD]
C N O [A-230/2/4] O F
F CH3 CH3
H [GF]
CH3
NCH2CH2 [GP]
R: R': CH3
H H [A-230]
CH3 H [A-232] Cl CH CH [A-230/2/4]
CH3 CH3 [A-234] R' R
The selective determination of organophosphorus fluoridates is based on the detection

of fluorides which usually arise by a suitable nucleophilic substitution reaction, e.g. by
alkaline hydrolysis or by the reaction with alcoholates. Fluorides can be detected as
lanthanum, calcium or thorium fluorides or as lead chlorofluoride precipitated using some
suitable reagents [1].
2.12.1
3F- La 3+ LaF3
2F- Ca 2+ CaF2
4+
4F- Th ThF4
F- Cl - Pb 2+ PbClF
An interesting precipitation reagent for fluorides, triphenyltin chloride, precipitates fluorides

from an aqueous alcoholic medium at pH 7-9 as white triphenyltin fluoride [2].
2.12.2
Sn Cl F- Sn F Cl -
81
However, for the detection of organophosphorus compounds, these fluoride precipitating
reactions are of limited importance, not only due to the low sensitivity of precipitation
reactions which contrasts with the high toxicity of warfare organophosphorus compounds.
Alkyl phosphonic acid esters and phosphoric acid esters are reagents which form very readily
stable complexes with heavy metal cations and thus mask them from fluoride precipitation.
The sensitivity of fluoride precipitation reactions does not exceed 0.5 mg of a warfare agent
per sample [3,4]. On the other hand, a more sensitive and very widespread analytical
procedure for the detection of fluorides is their ability to form highly stable fluoro complexes
of heavy metals. As a source of the central cation, its coloured less stable complexes, the so-
called lake pigments, are used together with some organic ligands, often hydroquinone-type
compounds substituted in the ortho- or peri-positions.
Fluorides, as an indirect evidence, are most often detected using reagents based on zirconium,
thorium, iron(III) or aluminium complexes. By the action of fluorides, red complexes of
zirconium and thorium with alizarinsulfonate (Alizarin Red S) turn yellow due to the
liberation of alizarinsulfonate. The hexafluorozirconium complex and thorium fluoride are
colourless or white compounds.
2.12.3
4-
O O O OH
OH OH
6F- 4H+
Zr 4+ 4
SO3 - [ZrF6] 2- SO3
O O
4
4-
O O O OH
OH OH
4F- 4H+
Th 4+ 4
SO3 - ThF4 SO3
O O
4
The reaction is performed in an acid or neutral medium. In an alkaline medium the coloured
lake pigments are decomposed and the alizarin itself acquires a red-violet colour. The
detection limit for sarin is 0.5 µg.ml-1 in a sample. Analogously, the deep red solution of
iron(III) thiocyanate or hexathiocyanatoferrate(III) is decolourized in the presence of excess
thiocyanate ions.
2.12.4
3- - -
[Fe(SCN)6] 5F 2H+ H 2[FeF5] 6 SCN
This indirect evidence of fluorides must also be performed in an acid medium. In an alkaline
medium, iron(III) thiocyanate decomposes with the formation of iron(III) hydroxide. Unlike
the above-mentioned reagents, aluminium complexes very often exhibit green fluorescence
that is quenched due to the decomposition of the starting complex, usually a morin-aluminium
or quercetin-aluminium lake, in a weakly alkaline medium (pH 4.5).
82
2.12.5
O O
OH
OH Al 3+ 6F- 3H+
HO O
HO
3
OH O
OH
[AlF6] 3- 3
OH
HO O
morin HO
O O
OH
OH Al 3+ 6F- 3H+
HO O
OH
3
OH O
OH
[AlF6] 3- 3
OH
HO O
kvercetin OH
quercetin
The sensitivity of these indirect proofs of fluoride ions is mostly high, being up to 0.1 µg.ml-1
in a sample, but its low selectivity represents a problem. The reaction is interfered by
phosphates, sulfites, thiosulfates, and sulfates, and strong oxidation reagents as well. If
fluorides cannot be separated from the interfering ions, e.g. by precipitating their silver salts,
it is necessary to perform a parallel test with a blind sample of representative composition, or
possibly with various concentrations of a lake-based reagent. Heating a sample up to 80 °C is
recommendable as well. In order to increase the selectivity of the indirect evidence of
fluorides, it is possible to carry out the reaction itself using a detector paper saturated with a
zirconium complex with 1-(4-dimethylaminophenylazo)-4-benzenearsonic acid (azo dye).
The complex dyes the paper brown. If the sample contains other compounds which interfere
the proof of fluorides, it is acidified with sulfuric acid and heated. The arising hydrogen
fluoride decomposes the brown complex on a moistened detector paper under formation of a
red-pink 1-(4-dimethylaminophenylazo)-4-benzenearsonic acid.
83
2.12.6
O
H3C
N N N As O Zr 6 HF
H3C
O
2
CH3 O
2 H3C N N N As OH [ZrF6] 2-
H OH
The sensitivity is up to 0.1 µg.ml-1 in a sample. The indirect proof of fluorine is performed
only at the end of systematic analysis of the sample [5].
Quantitative determination of fluorides in a sample can be advantageously carried out using a
gravimetric method (precipitation of PbClF) or a volumetric determination with
potentiometric indication by a lanthanum fluoride membrane ion-selective electrode or
titration with a lanthanum or thorium nitrate solution [6,7] or, possibly, with ethanolic
triphenyltin chloride.

4. PŘIKRYL, F., SOUČEK, J., KELLNER, J., HALÁMEK, E.: Potenciometrické stanovení fluoru v bojových
otravných látkách série G. In: Sborník VVŠ PV, 4, Vyškov, VVŠ PV, 1980, s. 3 - 15.
obrana, 28, 1986, 2, s . 75-87.
84
2.13 Sarin
O
H3C O
CH P
F
CH3 H3C
After a positive cholinesterase reaction and a nucleophilic reaction, e.g. the

SCHÖNEMANN aminoperoxide reaction and the detection of fluoride ion, it only remains to
confirm the presence of an alkoxy group (here an isopropoxy group) for the identification of
sarin. To detect the sarin isopropoxy group, the sample is extracted into 1,2-dichloroethane.
The organic extract is mixed with a solution of p-dimethylaminobenzaldehyde in
1,2-dichloroethane and the mixture is cautiously underlayered with chemically pure sulfuric
acid. The presence of an isopropoxy group is indicated by a deep red circle at the interface. At
high concentrations of the analyte, the whole lower sulfuric acid phase gradually turns red.
The detection reaction should be preferably compared with a blank. The action of
concentrated sulfuric acid on organic compounds leads to brown to black coloration due to
carbon arising in dehydrogenation and dehydration reactions.
Aromatic compounds in H2SO4 give a brown-red coloration; out of CWAs it is, e.g.
chloroacetophenone, adamsite or compound BZ. The detection reaction is carried out with
warming to 60 °C on a water bath. In the first step of the detection of isopropoxy group, sarin
is re-esterified with concentrated sulfuric acid.
2.13.1
H3C O H3C H3C O
H3C P H2SO4 CH O SO2OH P
CH O F H3C HO F
H3C
The arising isopropyl hydrogen sulfate (isopropylsulfuric acid) couples with

p-dimethylaminobenzaldehyde to give a red polymethine dye [1,2].
2.13.2
H3C H3C
N CHO CH O SO2OH
H3C H3C
H3C
N CH CH2 CH CH3
H3C
OH O SO2OH
- H2O
85
H3C
NH CH CH CH CH2 HSO4
H3C
H3C
N CH CH CH CH3 HSO4
H3C
The detection reaction is not selective for sarin, but it is selective at least for all isopropoxy
derivatives and, therefore, it has to be carried out only as a last confirming test. Its sensitivity
is good, 0.3-0.5 µg.ml-1 in a sample.
Compounds of the sarin type (G) are detected as liquids using detector papers containing the
pertinent dyes, soluble in CWAs of the G-series and exhibiting an acid reaction. The detector
paper PP-3 (ABC-M8), containing Disperse Yellow DY 23, may serve as an example.
2.13.3
CAS: 6250-23-3
N N N N OH
Disperse Yellow 23

1. KARAKČIJEV, N.I.: Toxikologija OV i zašita ot jaděrnovo oružija.Taškent. 1978. Medicina. 3. vydání. 440 s.
86
2.14 Soman
CH3
H3C F
C O
CH P
H3C O
CH3 H3C
After a positive cholinesterase reaction, a nucleophilic reaction for warfare

organophosphates of the G-series and the exclusion of tabun and sarin using selective tests, it
remains to confirm the presence of the pinacolyloxy (3,3-dimethyl-2-butyloxy) group in
soman by a chromogenic reaction. Since the pinacolyloxy group is chemically very similar to
the isopropoxy group (both being alkoxy groups), the reactions of both groups are also
similar. The detection is carried out using a 1,2-dichloroethane extract of the sample, the
vanillin detection reagent (3-ethoxy-4-hydroxybenzaldehyde) dissolved in concentrated
sulfuric acid. The test comprises gradual addition of 3-5 drops of a dichloroethane extract of
the sample to 2 ml of a vanillin solution (10 mg of vanillin in 2 ml of concentrated sulfuric
acid) till a red-brown or red-violet coloration appears. Sulfuric acid reacts with soman in a
reesterification reaction under formation of pinacolyl hydrogen sulfate (pinacolylsulfuric acid)
which couples with the aldehyde group of vanillin. The product rearranges to a polymethine
dye, capable of transition into the tautomeric mesoquinoid form [1,2].
2.14.1
H3C O H3C O
CH3
P H2SO4 (CH3)3C CH OSO2OH P
H3C C CH O F HO F
CH3
CH3 CH3
OHC OH (CH3)3C CH OSO2OH
CH3
OCH3
CH3 OH
H3C C CH CH2 CH OH
CH3 O SO2OH
OCH3
87
H3C
C CH CH CH OH CH3OSO3H H2O
H3C
OCH3
H3C
CH CH CH CH O
H3C
OCH3
The selectivity of the reaction is not high because many branched alkoxy derivatives react
similarly. Its sensitivity is stated to be about 10 µg in 1 ml of a dichloroethane sample extract.

1. KARAKČIJEV, N.I.: Toxikologija OV i zašita ot jaděrnovo oružija.Taškent. 1978. Medicina. 3. vydání. 440 s.
88
2.15 Compounds of the V-series (VX type)
R1: CH3 [VX, R-33, VM, model VX]
C 2H 5 [VE, VS]
C 2H 5O [VG]
R1 O R3 CH3 [model VX]
3
P R C 2H 5 [VG, VM, VE, R-33]
2
R O SCH2CH2N CH3
3 CH [VX, VS]
R CH3
R2: C2H5 [VX, VE, VG, VM, VS, model VX]
H3C
CHCH2 [R-33]
H3C
A positive cholinesterase test [1-5] and a negative nucleophilic detection reaction, for
example the SCHÖNEMANN reaction, indicate the presence of CWAs of the V-series (type
VX). This is a group of compounds, the structure of which differs only in the length of the
alkyl groups, most of them being homologues or isomers to each other, or even both (see VX
compound and R-33). Therefore, these compounds are usually determined as a whole because
common detection reactions are not able to distinguish the alkyl groups in detail. Without
using physico-chemical methods, only Amiton (compound VG) can be simply distinguished
because, unlike the other compounds, it is derived from phosphoric and not from
alkylphosphonic acid and its total saponification should afford phosphates. The cholinesterase
method in the so-called hydrolytic test modification enables to distinguish compounds of the
V-series from those of the G-series. The procedure is again based on the ability of the G-
series compounds to enter nucleophilic substitution reactions (here with hydroxide ions). The
tested sample (1 ml) is added with caution into 5 ml of 0.1M aqueous sodium hydroxide
solution and after 10 minutes the excess hydroxide is titrated with 0.1M sulfuric acid to an
acid-base indicator (Phenol Red). The neutralized solution is then checked again by the
cholinesterase method for the presence of cholinesterase inhibitors. If the sample after the
hydroxide treatment does not inhibit cholinesterase, it contained compound(s) of the G-series;
however, if it still shows inhibitory activity, it contained a compound of the V-series.
Chemical warfare agents of the V-series are basic compounds of tertiary amine character.
They readily react with electrophilic reagents, particularly with hydroxonium ions, under
formation of a bulky cation.
2.15.1
R1 O R1 O
P H3O P H2O
2 3
R2O SCH2CH2N(R3)2 R O SCH2CH2NH(R )2
Then these cations can form ionic compounds that precipitate from aqueous solutions.
Similarly as in the case of nitrogen yperites (see Chapter 2.7), the counterions are typical
group precipitating agents for tertiary amines and alkaloids, such as some heteropolyacids,
with iodo, thiocynato, chloro, nitro or cyano groups and several other complex compounds
89
with a transition metal cation as a central ion. The DRAGENDORFF reagent
(tetraiodobismuthate), phosphomolybdic acid, phosphotungstic acid, potassium
hexathiocyanatochromate and tetraiodomercurate(II) and many others are known and often
employed precipitation reagents. The precipitation is carried out in an aqueous acid. The
precipitates are white in case the reagent is colourless, if it is not, they acquire the colour of
the anion.
2.15.2
R1 O R1 O
n-
n P B
P B
2 3 2 3
R O SCH2CH2NH(R )2 R O SCH2CH2NH(R )2
n
kdeBBisje [BiI4] -, [P(Mo3O10)4]

where 3-
, [P(W3O10)4] 3-, [Cr(SCN)6] 3-, [HgI4] 2-, ...
The palladium(II) chloride solution is a specific reagent. Unlike chloro and iodo complexes of
platinum metals, it does not precipitate compounds of the V-series, but decomposes them to
thiol chloropalladium(II) salts. The reaction is used for the spectrophotometric determination
of the V-series compounds.
2.15.3
R1 O R1 O
P PdCl2 P ClPdSCH2CH2N(R3)2
R2O SCH2CH2N(R3)2 R2O Cl
The precipitation reactions are not selective because other aliphatic and heterocyclic tertiary
and some secondary amines also behave similarly. The sensitivity of the precipitation
reactions is not high and depends on the precipitation agent and the execution of the reaction;
it is about 10-100 µg.ml-1. The reactions are usually performed only in mobile laboratories.
Palladium or gold salts are rarely used for the detection of amine and sulfur compounds in
detector tubes. A traditional tube contains a DRAGENDORFF reagent-based filling.
The ion pair formation of the V-series compounds with acidic dyes and their immediate
extraction into a water-immiscible solvent can also be used as a group reaction for tertiary
amines. Depending on the anionic acidic dye, reaction conditions (pH, ion strength), and an
extraction solvent, it is possible to increase the selectivity, especially for a certain group of
compounds [6-10]. For the determination of VX compound in a mobile laboratory we selected
the Amido Black 10B dye dissolved in a buffer, pH 3-4, with chloroform as an extraction
solvent [11].
2.15.4
H3C O
OH NH2
2 P O2N N N N N NH2
C2H5O SCH2CH2NH(iC3H7)2
O3S SO3
90
H3C O
OH NH2
P O2N N N N N NH2
C2H5O SCH2CH2NH(iC3H7)2
2
O3S SO3 w
extrakce CHCl
extraction with3CHCl3
OH NH2
O2N N N N N NH2
H O3S SO3 H
C2H5O SCH2CH2N(iC3H7)2 (C3H7i)2NCH2CH2S OC2H5
P P
H3C O O CH3
org
The extracted associate of VX compound is blue and is formed in the 2:1 ratio. The sensitivity
is 5 µg of VX compound in a sample. A semiquantitative determination is possible by
comparison of coloration intensity with a standard colour chart, quantitative determination
can be performed spectrophotometrically. The same coloration arises with BZ and GP
compounds, alkaloids, and some other amine compounds.
Most methods for the identification of the V-series compounds are based on their nucleophilic
substitution reactions with e.g. alcoholate or hydroxide ions leading to
2-dialkylaminoethanethiol. The thiol formation together with positive cholinesterase reaction
confirms the presence of the V-series compound in a sample. Then thiols can be determined
by many analytical procedures [12,13].
2.15.5
R1 O R1 O
P OH P HSCH2CH2N(R3)2
R2O SCH2CH2N(R3)2 R2O O
Thiols are readily oxidized to disulfides and thus in a neutral and acid medium they
decolourize iodine and potassium permanganate solutions.
2.15.6
2 (R3)2NCH2CH2SH I2 (R3)2NCH2CH2S SCH2CH2N(R3)2 2 HI
H H
3 3
10 (R )2NCH2CH2SH 2 KMnO4 16 H + 5 (R )2NCH2CH2S SCH2CH2N(R3)2
2 Mn 2+ 2 K+ 8 H2O
6 (R3)2NCH2CH2SH 2 KMnO4 3 (R3)2NCH2CH2S SCH2CH2N(R3)2
2 MnO(OH)2 2 KOH
91
The fact that thiols are also oxidized with air oxygen dissolved in a sample may negatively
influence the detection of traces of the analyte or its quantitative determination [14]. Thiol can
be detected using oxidized forms of redox indicators as chromogenic reagents. In the presence
of thiol the coloured oxidized form of the indicator is decolourized under formation of the
colourless (leuco) form [15].
2.15.7
2 (R3)2NCH2CH2SH Indox (R3)2NCH2CH2S SCH2CH2N(R3)2 H2Indred
The reaction is similar to that used in the thiocholine detection (see the cholinesterase
reaction, Chapter 2.9). One of the reagents for thiols, bis(p-nitrophenyl) disulfide, reacts with
the analyte to give p-nitrobenzenethiol which is yellow in an alkaline medium. The reaction is
executed in acetone because the reagent is insoluble in water.
2.15.8
S NO2
3
2 (R )2NCH2CH2SH (R3)2NCH2CH2S SCH2CH2N(R3)2
S NO2
2 HS NO2
OH -
S N H2O
This chromogenic reagent, employed for the detection of the V-series compounds in field
laboratories, is analogous to the ELLMAN reagent for the detection of thiol or thiocholine in
the determination of warfare organophosphates by cholinesterase reaction. The reaction is
relatively sensitive: 5 µg.ml-1 in a sample. Its selectivity is good, only reducing agents are
interfered. The application of the ELLMAN reagent is more advantageous and universal,
especially because of its solubility in water.
Sulfur compounds, particularly thiols, readily afford mercury-containing compounds. With
mercury(II) compounds, thiols are precipitated under formation of sparingly soluble
precipitates, often via soluble complexes.
2.15.9
4 (R3)2NCH2CH2SH Hg 2+
{[(R3)2NCH2CH2S]4Hg} 2-
4H +
{[(R3)2NCH2CH2S]4Hg} 2-
Hg 2+ 2 [(R3)2NCH2CH2S]2Hg
An interesting reagent for thiols is dihydroxymercurifluorescein which exhibits a markedly

yellow-green fluorescence in an alkaline solution. The fluorescence disappears upon the
reaction. The product displays a pink colour in the daylight.
92
2.15.10
O OH
O
2 (R3)2NCH2CH2SH
HOHg HgOH - 2 H2O
COO
O OH
O
(R3)2NCH2CH2SHg HgSCH2CH2N(R3)2
COO
The potential of the detection of the V-series compounds is markedly increased by their
conversion to the G-series compounds or to organophosphorus fluoridates. This derivatization
not only widens the choice of available detection methods, but also increases the volatility of
the otherwise little volatile V-series compounds. This may be of considerable significance,
e.g. when determining the V-series compounds as air dispersions, using devices detecting
atmosphere contamination (M43/M8).
2.15.11
R1 O R1 O
P AgF P AgSCH2CH2N(R3)2
R2O SCH2CH2N(R3)2 R2O F
This reaction is surprisingly fast and, depending on reaction conditions, even quantitative. It
resembles the reaction of the V-series compounds with palladium(II) chloride (see 2.15.3)
which, however, is used directly for their determination. The arising fluoridate can be
detected by many methods (Chapter 2.12).
The liquid V-series compounds in the form of settling crude air dispersion particles are
detected by detector papers, pencils, powders and dyes. As concerns the solubility and
indication of alkaline character of the compounds, examples of suitable dyes are the green
dyes employed in the detector paper PP-1 (AP-1) or PP-3 (ABC-M8).
2.15.12
Br Br
HO H3C OH
PP-1
Bromkresolová zeleň
Bromcresol Green
Br Br
CH3 CAS 76-60-8
O CH3
O
S
93
2.15.13
O2N NO2
PP-3
O Dye Green EDA
CH C CAS 5833-18-1
O C2H5
O2N NO2
Besides the cholinesterase method (Chapter 2.9), the V-series compounds can be
quantitatively determined by classical iodometric and manganometric methods (according to
2.15.6) for the determination of thiols liberated by nucleophilic substitution reaction. Thiols
can be determined by many other redox methods, including electrochemical methods, such as
polarography. Thiols of the V-series can advantageously be determined by mercurimetric
titration (see 2.15.9), potentiometrically indicated by a silver sulfide membrane ion-selective
electrode [16].
Thanks to the increasing number of methods utilizing the excellent properties of the
ELLMAN reagent, thiols liberated from the V-series compounds are often determined by
spectrophotometric methods using absorbance measurements at 412 nm. To obtain accurate
results, it is necessary to prevent oxidation of thiols with air oxygen or, prior to the
determination itself, to reduce the disulfides back to the thiols, e.g. with hydrogen in statu
nascendi [17].
2.15.14
2 (R3)2NCH2CH2SH 1
/2 O2 (R3)2NCH2CH2S SCH2CH2N(R3)2 H2O
(R3)2NCH2CH2S SCH2CH2N(R3)2 2H 2 (R3)2NCH2CH2SH
The reaction of thiols with the ELLMAN reagent can be described by two mechanisms of the
same stoichiometry leading to a yellow product.
2.15.15
R1 O R1 O
P OR P SCH2CH2N(R3)2
R2O SCH2CH2N(R3)2 R2O OR
94
(R3)2NCH2CH2S O2N S S NO2
HOOC COOH
(R3)2NCH2CH2S S NO2 S NO2
COOH COOH
S N
COOH
(R3)2NCH2CH2S SCH2CH2N(R3)2 2 S NO2
COOH
2 (R3)2NCH2CH2S O2N S S NO2
HOOC COOH
So far, the literature prefers the course via the mixed disulfide, with liberation of yellow
2-nitro-5-sulfanylbenzoic acid.

1. Pat. ČR 285 242. TUŠAROVÁ I., HALÁMEK E., OREL, J.: Detekční trubička inhibitorů cholinesteras ve
vzduchu a ve vodě. 16.6. 1999.
2. Užitný vzor ČR 11 391. TUŠAROVÁ I., ŠEBEK, J., OREL, J., HALÁMEK E.: Kolorimetrický biochemický
detektor inhibitorů cholinesteras ve vzduchu, ve vodě a na povrchu.12.07. 2001
3. Pat. ČR 288 576. TUŠAROVÁ I., HALÁMEK E.: Biosenzor pro detekci a rozlišení inhibitorů cholinesteráz,
způsob přípravy zóny biosenzoru s imobilizovanou cholinesterázou, způsob detekce inhibitorů cholinesteráz a
způsob rozlišení inhibitorů cholinesteráz. 22. 5. 2001.
4. Pat. SR 281 395. TUŠAROVÁ, I., HALÁMEK, E.: Biosenzor, spôsob prípravy zóny s imobilizovanou
cholínesterázou, spôsob detekcie a rozlíšenia inhibítorov cholínesteráz biosenzorom. 12. 3. 2001.
5. Pat. SR 281 547. TUŠAROVÁ, I., HALÁMEK, E., OREL, J.: Detekčná rúrka inhibítorov cholinesteráz vo
vzduchu a vo vode. 9.4. 2001.
6. SOUČEK, J., HALÁMEK, E., KYSILKA, R.: Extrakčně spektrofotometrické stanovení O-ethyl-S-2-diiso-
propylaminoethylmethylthiofofonátu s bromkresolovou zelení a bromthymolovou modří. In: Sborník VVŠ PV,
řada „B“, 1, Vyškov, VVŠ PV, 1985, s.33 - 43.
95
7. KOBLIHA, Z., HALÁMEK, E., SOUČEK, J.: Extrakčně spektrofotometrické stanovení látky VX
azobarvivem kalkon. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1986, s. 44 - 52.
8. Pat. ČSSR 2716. HALÁMEK E., KOBLIHA Z.: Způsob stanovení aminoalkylthiofosfonátů nitrofenoly.
3.11.1988.
9. Pat. ČSSR 2717. HALÁMEK, E., KOBLIHA, Z.: Způsob stanovení aminoalkylthiofosfonátů azobarvivy.
3.11.1988.
10. Pat. ČSSR 2738. HALÁMEK, E., KOBLIHA, Z.: Způsob stanovení aminoalkylthiofosfonátů sulfoftaleiny.
3.11.1988.
11. HALÁMEK, E., KOBLIHA, Z.: Stanovení O-ethyl-S-(2-diisopropylaminoethyl)-methylthiofosfonátu (VX)
s vybranými barvivy metodou extrakční spektrofotometrie. In: Sborník VVŠ PV, řada B, č. 1, Vyškov, VVŠ
PV, 1991, s. 83-95.
12. SOUČEK, J., HALÁMEK, E., PŘIKRYL, F., KYSILKA, R.: Spektrofotometrické stanovení látky VX a
některých doprovodných sloučenin. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1985, s. 23 - 32.
13. SOUČEK, J., HALÁMEK, E., PŘIKRYL, F., KYSILKA, R.: Extrakčně spektrofotometrické stanovení
diisopropylaminoethanthiolu. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1985, s. 2 - 11.
14. SOUČEK, J., HALÁMEK, E., PŘIKRYL, F., SKALIČAN, Z.:Spektrofotometrické stanovení thiolů ve
vzorku látky VX. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1984, s. 1 - 9.
15. HALÁMEK, E., KOBLIHA, Z., SKALIČAN, Z.: Spektrofotometrické stanovení 2-di-
methylaminoethanthiolu s Guinea Green B a Malachite Green Carbinol Hydrochlorid. In: Sborník VVŠ PV, 3,
Vyškov, VVŠ PV, 1998, s.237 - 247.
16. PŘIKRYL, F., TOMEČEK, I., HALÁMEK, E., KELLNER, J., SOUČEK, J.: Potenciometrické stanovení
látek typu V. In: Studijní sborník, 3, Vyškov, VVŠ PV, 1977, s. 87 - 96.
17. SOUČEK, J., HALÁMEK, E., KYSILKA, R.: Stanovení thiolů a disulfidů ve vzorku látky VX
s Ellmanovým činidlem. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1985, s. 44 - 53.
96
2.16 GP Compound
CH3
H3C
N O CH3 N O
H3C
H3C P HOCH2CH2N P CH3
F F CH3 - HF F OCH2CH2N
CH3
The potential binary chemical warfare agent, [2-(dimethylamino)ethyl]

(dimethylphosphoramido)fluoridate, is a strong acetylcholinesterase inhibitor and as such it
belongs to nerve CWAs [1-4]. Its structure and some physico-chemical properties as well as
chemical behaviour correspond to a borderline between the G- and V-series compounds and,
therefore, it is also named GV. The substance is a basic amine compound that, similarly to the
V-series compounds, does not afford reactions characteristic of the G-compounds (e.g. the
SCHÖNEMANN reaction). On the other hand, its molecule contains a fluorine atom bonded
to the central atom of phosphorus like most organophosphorus fluoridates of the G-series.
With the amide grouping it resembles the otherwise structurally exotic tabun. GP compound
is fairly stable in solutions, whereas in the pure state it is rapidly converted into its non-toxic
isomer. The conversion rate depends on the temperature; under usual conditions it is relatively
high (1-2 hours) [5,6].
2.16.1
H3C H3C
N O CH3 N O
H3C P CH3 N H3C P
F OCH2CH2N CH3 F O
CH3
On the basis of LARSSON’s studies of TAMMELIN’s esters, the present authors have
proven that GP compound is converted to the isomeric dimethylaziridinium dimethylamido-
fluorophosphate, a non-toxic solid. Analogously to the original GP compound, its isomer
undergoes many precipitation reactions for tertiary (here quaternary) amines and alkaloids
(see Chapters 2.7 and 2.15) but, unlike GP compound, it does not afford extractable ion pairs
with acidic dyes. For the determination of GP compound, the present authors have suggested
an advantageous procedure with Bromoxylenol Blue in a phosphate buffer, pH 5-6. The
yellow ion associate is then extracted into chloroform [7-12].
2.16.2
CH3 CH3
H3C O OH
N O
H3C P CH3
Br Br
F OCH2CH2NH CH3 CH3
CH3 SO3
97
H3C CH3 CH3
N O O OH
H3C P CH3
F OCH2CH2NH Br Br
CH3 CH3
CH3
SO3
CH3 CH3
HO O
H3C Br Br
CH3 CH3
N O
CH3
H3C P
F OCH2CH2N HO3S
CH3
org
The reaction is not very selective; a similar product is formed with several other nitrogen
compounds such as VX, R33, BZ and alkaloids. The sensitivity is good, 1 µg in a sample
extract. A standard colour chart enables a semiquantitative determination. A
spectrophotometric method for mobile laboratories has also been developed. Upon alkaline
hydrolysis it is possible to detect the presence of fluorides (see Chapter 2.12) and on
saponification the presence of phosphates as well (see Chapter 2.11).
On detector paper PP-3, the liquid GP compound develops spots of the same colour as VX
compound, but, due to higher volatility of this CWA, the spots are surrounded with a green
gradually merging halo.
As a tertiary amine, the organophosphorus GP compound can easily be determined in an acid
medium by titration with a sodium tetraphenylborate solution, the equivalence point being
indicated by a softened PVC membrane ion-selective electrode [13-15].

1. SOUČEK, J., TUŠAROVÁ, I., HALÁMEK, E., KOBLIHA, Z.: Inhibice cholinesteras
2-dimethylaminoethyldimethylamidofluorfosfátem. Civilní obrana, 31, 1989, 2, s. 43 - 45.
2. HALÁMEK, E., TUŠAROVÁ, I.: Jednoduchý detektor nervově paralytických látek. In: Zborník
vojenskovedeckých prác, 4, 2, Liptovský Mikuláš, VVTŠ, 1989, s. 134 - 150.
3. HALÁMEK, E., KOBLIHA, Z., TUŠAROVÁ, I.: The new Tammelin‘s esters: A group of extremely toxic
organophosphates. The ASA Newletter, 48, 95-3, 1995, p. 1, 14-15.
4.TUŠAROVÁ, I., HALÁMEK, E., KOBLIHA, Z.: Study on reactivation of enzyme-inhibitor complexes by
Mikrobiological Technology, 25, 1999, p.400 – 403.
98
5. HALÁMEK, E., TUŠAROVÁ, I., KOBLIHA, Z., SOUČEK, J., FÖLDEŠI, V.: Identifikace produktu
spontánního rozkladu modelové látky se střední těkavostí. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV,
1988, s. 2 - 16.
6. HALÁMEK, E., KOBLIHA, Z., HRABAL, R: Identification of the Isomeric Transformation Product from
2-(Dimethylamino)ethyl-(dimethylphosphoramido)fluoridate. Phosphorus, Sulphur, and Silicon and Related
Elements, Vol. 179( No. 1), 2004, p. 49-53. ISSN 1042-6507.
7. HALÁMEK, E., KOBLIHA, Z., SOUČEK, J.: Extrakčně spektrofotometrické stanovení 2-dimethylamino-
ethyldimethylamidofluorfosfátu. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1987, s. 25 - 34.
8. SOUČEK, J., HALÁMEK, E., KOBLIHA, Z., TUŠAROVÁ, I.: Stanovení aktivní komponenty nové otravné
látky IVA. Civilní obrana, 31, 1989, 6, s. 45 – 48.
9. HALÁMEK, E., KOBLIHA, Z., SOUČEK, J.: Iontové asociáty dialkylaminoethyl-dialkylamidofluorfosfátů a
jejich prekurzorů s vybranými barvivy. In: Zborník vojenskovedeckých prác, 5, 1, Liptovský Mikuláš, VVTŠ,
1990, s.174-196.
10. HALÁMEK, E., KOBLIHA, Z.: The ion associates of methyl-, ethyl- and isopropyl derivatives of
dialkylaminoethyl dialkylamidofluorophosphate with bromophenol blue. Collect. Czech. Chem. Commun.,
57,1992, s. 56-63.
11. Pat. ČSSR 2605. HALÁMEK E., SOUČEK J., KOBLIHA Z.: Způsob analýzy aminofluorfosfátů. 6.5.1987.
12. Pat. ČSSR 2640. HALÁMEK, E., SOUČEK, J., KOBLIHA, Z.: Způsob analýzy aminofluorfosfátů.
6.5.1987.
13. ČAPOUN, T., HALÁMEK, E.: Možnosti použití iontově selektivních elektrod s PVC membránou ke
stanovení některých otravných látek. III. Některé aspekty praktického využití potenciometrických titrací
založených na tvorbě iontových asociátů. Civilní obrana, 3O, 1988, 6, s. 39 - 42.
14. HALÁMEK, E., ČAPOUN, T.: Použití skleněné elektrody k indikaci titrací založených na tvorbě iontových
asociátů. Chemické listy, 83, 1989, s. 431 - 434.
stanovení některých otravných látek. IV. Indikace titrací skleněnou elektrodou s PVC membránou. Civilní
obrana, 31, 1989, 5, s. 43 - 45.
2.17 Novichok (A-230, A-232, A-234)
99
dichlor(fluor)nitrosomethan
dichloro(fluoro)nitrosomethane
Cl
O N C F
Cl
R´ Cl R´ O N C
O O
F
P F P F
Cl
R O R O
Cl O N C Cl
F
chloro(fluoro)formoxime
chlor(fluor)formoxim chloride
chlorid
4,5-dialkyl-2-fluor-1,3,2-dioxafosfolan
4,5-dialkyl-2-fluoro-1,3,2-dioxaphospholane
Cl
R´ R C N O O
F
H H A-230 P
Cl CH CH O F
H CH3 A-232
CH3 CH3 A-234 R´ R
O-chlorfluorkarbiminoyl-O-[(2-chlor-1,2-dialkyl)ethyl]fosforofluoridát
O-chlorofluorocarbiminoyl-O-[(2-chloro-1,2-dialkyl)ethyl] phosphorofluoridate
This group of potential binary nerve warfare agents, developed in Russia probably
during the eighties and nineties, is unofficially regarded as the third and the fourth generation
of nerve CWAs [1-7]. These agents allegedly have been developed as part of the FOLIANT
project and the compounds as well as their precursors are called NOVICHOK (newcomer)
[8]. There are only scarce pieces of information on them and almost no analytical data. They
are strong ACHE inhibitors, allegedly 10x stronger than VX compound [9] and, therefore,
they can be determined by the cholinesterase reaction and by chemical reconnaissance and
check tools based on this reaction [10-12]. Moreover, it is known that the effect on ACHE is
rapid, and thus a modified cholinesterase reaction, utilizing the effect of oxime reactivators on
the enzyme-inhibitor complex, will have no marked effect (see Supplement to Chapters 2.9-
2.17). As evident from the structure of these compounds, it will be easy to liberate fluoride or
chloride ions using a nucleophilic substitution reaction and then to detect them.
From what has been said above it is clear that within the framework of systematic analysis of
a sample of nerve compounds it will not be difficult to differentiate and determine this new
group of CWAs, although some analytical procedures and reactions still remain to be verified.
Because of an order of magnitude higher toxicity of these new and potentially very dangerous
CWAs, the chemical reconnaissance and check tools necessarily should exhibit a
correspondingly higher sensitivity. For majority of the existing automatic warning systems
based on physical detection principles, this fundamental requirement will be met only with
great difficulty. On the other hand, one may expect that devices and methods based on the
cholinesterase reaction will still meet this requirement because the toxicity of these
compounds corresponds with the inhibitory effects on ACHE and thus with the detection limit
of the cholinesterase method and the devices based on them.
Supplement to Chapters 2.9-2.17
100
Table 3 shows detection reactions for current organophosphorus CWAs which may
serve as a basis for frontal or systematic analysis of a sample according to the development
diagram.
Table 3
Outline of detection reactions for current organophosphorus CWAs (a variant)
Reaction GA GB GD GF VX R33 GP A-230/2/4

1. ⊕ ⊕ ⊕ ⊕ ⊕ ⊕ ⊕ ⊕
2. ⊕ ⊕ ⊕ ⊕ - - - ?
3. ⊕ - - - - - - -
4. - ⊕ ⊕ ⊕ - - ⊕ ⊕
5. - ⊕ - - - - - ?
6. - - ⊕ - - - - ?
7. -* - - - ⊕ ⊕ ⊕ -
8. - - - - ⊕ ⊕ - -
9. - - - - - - - ⊕
10. ⊕ ⊕ ⊕ ⊕ ⊕ - - -
Caption: ⊕ / - positive / negative reaction; ? – information missing
* tabun can lose dimethylamine
Detection reactions:
1. Biochemical cholinesterase reaction, the determination of ACHE inhibitors.
2. SCHÖNEMANN aminoperoxide reaction, differentiation between the G- and V-series
compounds according to nucleophilic substitution reactions.
3. KÖNIG-ZINCKE reaction, the detection of cyanides for the identification of tabun.
4. Indirect proof of fluoride ions by zirconium-alizarin lake after the hydrolysis.
5. Reaction with p-dimethylaminobenzaldehyde in dichloroethane and with concentrated
sulfuric acid, the proof of alkoxy (isopropoxy) group.
6. Reaction in dichloroethane with vanillin in sulfuric acid, the proof of alkoxy
(pinacolyloxy) group.
7. DRAGENDORFF reagent and other reactions for tertiary amines and alkaloids.
8. ELLMAN reagent or precipitation with Hg 2+, reaction for thiols after the alkaline
hydrolysis.
9. Detection of chlorides with silver nitrate after the alkaline hydrolysis.
10. Modified cholinesterase reaction with an oxime (N-ethylpyridinium-2-aldoxime).
With regard to point 10, the modified cholinesterase reaction with oximes is based on the
formation of characteristic enzyme-inhibitor complexes between ACHE and the individual
inhibitors (organophosphorus CWAs). These inhibitors are acylating and irreversible, but
immediately after the formation of the complex, they remain reversible for some time. This
time interval is different for particular inhibitors as well as the possibility of cleaving the
complex acetylcholinesterase-organophosphorus CWA by the reaction with a suitable oxime.
A great number of current organophosphorus agents may be selectively determined by the
combination of the enzyme incubation time and the inhibitor, the time of action of the
reactivator on the enzyme-inhibitor complex, and by the reactivator concentration and
structure. The use of specific enzyme, acetylcholinesterase, immobilized on a solid
101
polysaccharide-based support is very advantageous. Such enzyme chimera of high stability
has been used in the construction of the DETEHIT colorimetric biosensor. Currently, the
distinction of VX and R-33 isomeric compounds that are homologues to each other at the
same time is not possible without use of physico-chemical separation and spectral methods
[13-19].
Similarly to the GP compound, A-230, 232 and 234 compounds (or their precursors) denoted
as NOVICHOKs will not give the modified cholinesterase reaction with oximes, even in view
of the fact that they are halophosphorylated oximes.

1. NIFANTJEV, E.J. Chimija fosfororganičeskich sojediněnij. Izdatělstvo Moskovskovo univerzitěta.
Moskva,1971. 352 s.
2. PURDELA, D., VILCEANU, R. Chimija organičeskich sojediněnij fosfora. Izdatělstovo Chimija.
Moskva,1972. 752 s.
3. MARTYNOV, I.V., KRUGLAK, J.L., PRIVEZENCEVA, N.F. Zh. Obshch. Khim, 37, 1967, s. 1125.
4. MARTYNOV, I.V., KRUGLAK, J.L., LEJBOVSKAJA, G.A., CHROMOVA, Z.I., STRUKOV O.G. Zh.
Obshch. Khim, 39, 1969, s. 996.
5. KRUGLAK, J.L., MALEKIN, S.I., MARTYNOV, I.V., STRUKOV, O.G. Zh. Obshch. Khim, 39, 1969,
s.1265.
6. KRUGLAK, J.L., LEJBOVSKAJA, G.A., MARTYNOV, I.V. Zh. Obshch. Khim, 39, 1969, s.1263.
7. KRUGLAK, J.L., MALEKIN, S.I., MARTYNOV, I.V. Zh. Obshch. Khim, 42, 1971, s. 811.
8. FEDOROV, L.A. Chemical weapons in Russia: History, Ecology, Politics. Center of Ecological Policy of
Russia.Moskva,1994. 71 s.
9. SMITHSON, E.A., MIRZAYANOV, V.S., LAJOIE, R., KREPON, M. Chemical Weapons. H.L.S. Center.
Report No. 17. October 1995.
10. HOENIG, S.L. Compedium of Chemical Warfare Agents. Springer. New York, 2007.
222 s.
11. HALÁMEK, E., KOBLIHA,Z.: Poslední generace chemických zbraní. Vojenské rozhledy,17(49), č.4, 2008,
s.137-146. ISSN 1210-3292.
12. ELLISON, D.H.: Handbook of Chemical and Biological Warfare Agents. Taylor and Francis Group. 2008,
Second edition. 762 p.
13. TUŠAROVÁ, I., HALÁMEK, E., KOBLIHA, Z.: Study on reactivation of enzyme-inhibitor complexes by
Mikrobiological Technology, 25, 1999, p.400 – 403.
14. HOSKOVCOVÁ, M., HALÁMEK, E., KOBLIHA, Z.: Porovnání reaktivní účinnosti reaktivátorů
pralidoximového typu na imobilizovanou acetylcholinesterázu inhibovanou sarinem a somanem. In Sborník
UO, řada „B“ 1/2005. Brno: Univerzita obrany, 2005, s.165-178. ISSN 1211-1023.
5. HOSKOVCOVÁ M.: Studium imobilizovaných enzym-inhibitorových komplexů typu acetylcholinesteráza –
organofosforová bojová látka. [Doktorská práce]. Univerzita obrany Brno, Vyškov 2006, 114 s.
16. HOSKOVCOVÁ, M., HALÁMEK, E., KOBLIHA, Z., TUŠAROVÁ, I..: Efficacy of 2-PAM, MMB-4,
TMB-4, toxogonin and HI-6 in reactivation of acetylcholinesterase inhibited by warfare organophosphorus
compounds of the G series. Current Topics in Pharmacology, 10, 2, 2006 p. 71-76.
17. HOSKOVCOVÁ, M., HALÁMEK, E., KOBLIHA, Z., TUŠAROVÁ, I.: Efficacy of 2-PAM, TMB-4, HI-6
and toxogonin in reactivation of immobilized acetylcholinesterase inhibited by compound R-33. Toxicology
Mechanisms and Methods, 17, 2007, s.161-164.
18. HOSKOVCOVÁ, M., HALÁMEK, E., KOBLIHA, Z., TUŠAROVÁ, I.: Reactivation of immobilized
acetylcholinesterase-tabun complex by methoxime and its homologues. Drug and Chemical Toxicology, 30, 2,
2007, s. 1-7.
19. HOSKOVCOVÁ, M., HALÁMEK, E., KOBLIHA, Z., TUŠAROVÁ, I.: Efficacy of metoxime and its
homologues in reaktivation of immobilized acetylcholinesterase inhibited by sarin, cyklosarin and soman.
Current Topics in Toxicology, 4, 2007 p. 25-29.
102
2.18 Bromobenzyl cyanide
Br
CH
CN
Bromobenzyl cyanide is a relatively stable compound, nevertheless in the time of its

military use it was disqualified by its instability in contact with metals. Upon the alkaline
hydrolysis it dimerizes and decomposes into bromides and salts of diphenylmaleic acid.
2.18.1
Br
C CN
2 CH 2 OH 2 Br H2O
CN C CN
O 2 H2O O
C C NH2 C C ONH4
2 H2O
C C NH2 C C ONH4
O O
2 OH +2H - 2 NH4
O - 2 H2O, 2 NH3 O - H2O
C C O C C
2H
O
- H2O
C C O C C
O O
In an acid medium, the salt of diphenylmaleic acid is converted into crystalline
diphenylmaleic anhydride which serves for the identification of bromobenzyl cyanide (m.p.
156 °C).
Ammonium salts or ammonia, arising in hydrolysis, can be proved using the NESSLER
reagent (an alkaline solution of potassium tetraiodomercurate). Ammonium salts and
ammonia form a yellow-brown precipitate of oxodimercuriumimine iodide.
2.18.2
Hg
2 K2[HgI4] 4 OH - NH4 + O N.I 4K+ 7I- 3 H2O
Hg
The older literature also suggests another composition of the precipitate: dimercuriumamine
triiodide.
2.18.3
NH4 +
2 [HgI4] 2- 2 OH -
Hg2I3NH2 5I- 2 H2O
103
The sensitivity of the reaction is 1 µg.ml-1. Its selectivity is good; the same reaction may be
obtained with nitrile-containing CWAs, such as AC, CK or CS.
The bromides, arising by the decomposition of bromobenzyl cyanide, are oxidized with
chromates in acetic acid to give free bromine which can be proved by the reaction
with fluorescein affording red tetrabromofluorescein (eosin).
2.18.4
2 CrO4 2- 16 H +
6 Br - 3 Br2 2 Cr 3+ 8 H2O
2.18.5
Br Br
O OH O OH
O O
4 Br2 Br Br 4 HBr
COO COO
The sensitivity is 10 µg.ml-1 and the selectivity is very high.

On melting with sodium or potassium hydroxide, bromobenzyl cyanide affords, inter alia,
phenylglycolate and cyanide [1,2].
2.18.6
Br O
2 CH 6 OH - 1
/2 O2 CH 2 CN -
CN COO
2 Br - 4 H2O
The cyanide is determined by usual detection reactions (see Chapter 2.4). As an aromatic
compound, bromobenzyl cyanide reacts with concentrated sulfuric acid under formation of a
characteristic red-brown coloration. The reaction can be accelerated by heating. The red
coloration is assigned to arylcarbonium radicals present in concentrated sulfuric acid and
usually disappears on the dilution with water. As a compound with reactive hydrogen,
bromobenzyl cyanide undergoes the ZIMMERMANN reaction with 1,3-dinitrobenzene in a
strongly alkaline medium.
104
2.18.7
O2N
Br
CH NO2 3 OH -
CN
NO2
O O
2 H2O Br - CN - C N
NO2
OH O
C N
The reaction affords a red-brown colour, the intensity of which increases by heating or
standing. It is certainly not surprising that bromobenzyl cyanide reacts with a group of
reagents for alkyl and aryl halides, e.g. with 4-(4'-nitrobenzyl)pyridine in an alkaline medium
with formation of red-violet coloration [3].
2.18.8
Br
CH N CH2 NO2 OH -
CN
O2N CH N C Br - H2O
CN
O
N CH N CH
O
CN
For the quantitative determination of bromobenzyl cyanide, the method of choice is the
argentometric determination of bromides liberated quantitatively by treatment with sodium
sulfide in alcohol.
2.18.9
Br CH CN
2 CH Na2S S 2 NaBr
CN
CH CN
105
Prior to titration using silver nitrate, it is necessary to remove the excess sulfide by boiling
with diluted sulfuric acid and hydrogen peroxide. Bromides can be determined by the
VOLHARD method, i.e. by precipitation with an excess silver nitrate solution and the
titration of the excess silver with ammonium thiocyanate with an iron(III) salt as an indicator
(formation of red iron(III) thiocyanate). A simpler method consists in the direct titration with
the silver salt, potentiometrically indicated by a metallic silver electrode or a silver salt
membrane ion-selective electrode [4].

3. SOBOROVSKIJ, L.Z, EPŠTEJN, G.J.: Chimija i těchnologija bojevych otravljajuščich věščevstv, Oborongiz,
Moskva, 1936.
106
2.19 Chloroacetophenone
CH2 Cl
Similarly to bromobenzyl cyanide, chloroacetophenone contains an active hydrogen

atom of the methylene group and thus readily undergoes the ZIMMERMANN detection
reaction with 1,3-dinitrobenzene in a strongly alkaline medium [1-3].
2.19.1
O O2N
C 2 OH -
NO2
CH2 Cl
NO2
O O
Cl - 2 H2O C CH N
O
or
nebo
NO2
O O
Cl - H2O C CH2 O N
The literature describes the reaction course by two reactions, leading to two different
products, out of which only the former structurally corresponds to deep violet coloration of a
fully conjugated product. The sensitivity of the reaction is very good, less than 1 µg.ml-1 in a
sample. Its selectivity is not very high because many other compounds give a positive
reaction; however, the violet shade is typical for chloroacetophenone, acetophenone and
acetone. Colours, obtained with other compounds, are usually red and red-brown to brown.
The reaction may be significantly accelerated by warming in a water bath. The reaction can be
employed for the construction of a detector tube with ampoule. A silica gel support is
impregnated with 1,3-dinitrobenzene and the ampoule contains 20% potassium hydroxide
solution. In the presence of chloroacetophenone the white layer turns pink to raspberry red.
The sensitivity is high: 0.5 µg.l-1 in air. Bromobenzyl cyanide manifests itself by a light violet
to brown-violet coloration; the detection limit is 1 µg.l-1 in air.
In an alkaline medium, ethanolic solution of chloroacetophenone reacts with 4-(4'-nitro-
benzyl)pyridine affording a violet product. The same reaction is observed also with a detector
tube suggested by the present authors and originally designed for the detection of sulfur and
nitrogen yperites.
107
2.19.2
O
C N CH2 NO2 OH -
CH2 Cl
H2O Cl - C
CH2 N CH NO2
C O
CH2 N CH N
The reaction with 4-(4'-nitrobenzyl)pyridine is relatively sensitive, but little selective. The
reagent reacts with many compounds of acyl and alkyl halide character. An analogous
reaction of chloroacetophenone with pyridine affords yellow phenacylpyridine. It is also used
for the quantitative photometric determination of chloroacetophenone in air: after trapping in
a pyridine-dimethylformamide mixture, the reaction mixture is heated for 15 minutes and then
it is measured. The sensitivity is reported to be 1 µg.ml-1 in a sample or 0.6 µg.l-1 in air.
2.19.3
O O
C N C Cl
CH2 Cl CH2 N
The authors have described some variants of detector tubes for chloroacetophenone (and
o-chlorobenzylidenemalononitrile), employing furazans or thiadiazole as chromogenic
reagents [benzofurazan 1-oxide (I), 5-chlorobenzofurazan 3-oxide (II), 7-chloro-4-
nitrobenzofurazan (III) and 4-nitro-2,1,3-benzothiadiazole (IV)] [4-6].
2.19.4
O O
Cl
N Cl N N N
O O O S
N N N N
NO2 NO2
I II III IV
108
With chloroacetophenone these new reagents afford orange to violet coloration. So far, the
reaction mechanism has not been sufficiently studied. The sensitivity is good, 0.2-1.4 µg.l-1 in
air.
The reaction using the MARQUIS reagent (concentrated sulfuric acid with dissolved
formaldehyde [7]) has not been elucidated so far. However, it gives a yellow colour which is
not usual for aromatic compounds.
The reactions with ethanolic ammonia or ammonium sulfide belong to highly selective
detection reactions. The condensation of chloroacetophenone with ammonia gives indoxyl.
This reacts slowly with air oxygen or faster with oxidation reagents, such as hydrogen
peroxide, to give blue indigo.
2.19.5
O O
C 2 NH3 1
/2 O2 NH4Cl H2O
N
CH2 Cl H
O O
2 2 H2O2 4 H2O
H
N
N N
H H
However, the execution of the reaction is somewhat tedious and the reaction mixture requires
boiling.
Another modification of this method is the condensation of indoxyl with p-dimethylamino-
benzaldehyde in an acid medium, leading to a red product [8].
2.19.6
O CH3
OHC N H+
N CH3
H
O
CH3
N CH NH H2O
H
CH3
O
CH3
N CH N
H
CH3
OH
CH3
N CH N
H
CH3
109
The condensation of chloroacetophenone with thiourea under formation of 4-phenyl-2-
aminothiazole represents another interesting detection reaction of high selectivity and
quantitative yield.
2.19.7
O
NH2
C S C N HCl H2O
NH2
CH2 Cl S NH2
It is possible to determine quantitatively the arising hydrochloric acid or to couple the formed
4-phenyl-2-aminothiazole with a suitable diazonium salt, such as p-nitrophenyldiazonium
salt, to obtain the corresponding azo dye which is spectrophotometrically evaluated.
19.8
H3N
N N N
S
N
N
S NH2
NO2 O2N N
Chloroacetophenone is usually determined quantitatively by stoichiometric reactions with an

ethanolic solution of sodium sulfide or sodium phenoxide. Similarly as in the determination of
bromides in the case of bromobenzyl cyanide (see Chapter 2.18), the excess of sodium sufide
has to be removed by boiling with diluted sulfuric acid and hydrogen peroxide.
2.19.9
O O O
2 C 2 Na2S C C
- 2 NaCl
CH2 Cl CH2 S CH2
2.19.10
O O
C ONa C
- NaCl
CH2 Cl CH2 O
110
In both cases it is necessary to heat the mixture to complete the reaction. The arising chlorides
are determined by the VOLHARD method, i.e. by the titration of excess silver nitrate with
ammonium thiocyanate to an iron(III) salt as a visual indicator. The direct titration, as well as
the back titration, can be indicated potentiometrically by a silver metal electrode or a
membrane electrode of insoluble silver salts (AgCl, AgBr, AgI, Ag2S) [9-11].

1. JANOVSKY, J.V.: Ueber eine Reaction der Dinitrokörper. Ber. 24, 971-972 (1891).
2. NAKADATE, M., MATSUYAMA, C., KIMURA,M.: Janovsky reaction of polynitrodiphenyl compounds.
Chem. Pharm. Bull. 12, 1138-1143 (1964).
3. ZIMMERMANN, W.: Verglech verschiedener Modifikation der m-Dinitrobenzol-Reaktion auf 17-
ketosteroide. Hoppe Seylers Z. Physiol. Chem. 300, 141-146 (1955).
4. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z.: Detekční trubička s deriváty benzofurazanu ke
zjišťování dráždivých látek ve vzduchu. Sborník Univerzity obrany, řada „B“ (Technické a přírodní vědy),
2005, číslo 1, s. 179-186.
5. ORITEST. Detekční trubička ke stanovení dráždivých látek. Původce: Pitschmann, V., Halámek, E.,
Kobliha, Z. Int. G01N31/22. CZ 14444. Zapsáno 14. 6. 2004.
6. ORITEST. Detekční trubička ke stanovení lewisitu a policejních plynů. Původce: Pitschmann, V.,
Halámek, E., Kobliha, Z. Int. G01N31/22. CZ 14443. Zapsáno 14. 6. 2004.
7. MARQUIS, E., ANTHEAUME, A., MOUNEYRAT, A.: Ueber den Nachweis des Morphins. Z. Anal. Chem.
38(7), 466-468 (1899).
8. COX, H.E.: Test available for the identification of small quantities of the war gas. Analyst 64, 807-813 (1939).
obrana, 28, 1986, 2, s . 75-87.
111
2.20 o-Chlorobenzylidenemalononitrile, CS compound
Cl
CN
CH C
CN
A considerable part of detection reactions for CS compound is based on reactions of

malononitrile arising in hydrolysis that occurs readily already in a mildly alkaline medium.
2.20.1
Cl Cl
CN CN
CH C H2O CHO H2C
CN CN
The ZIMMERMANN reaction with 1,3-dinitrobenzene in a strongly alkaline medium

represents an exception; another hydrolysis product, o-chlorobenzaldehyde as a compound
with reactive hydrogen can also provide it [1].
2.20.2
Cl
O2N
CH O OH -
NO2
Cl NO2
OH O
H2O C N
Cl NO2
O O
C N
H O
The reaction gives a red-brown coloration. Its sensitivity is 1 µg.ml-1 in a sample. The
selectivity is not high because a similar reaction occurs also with other compounds.
The reaction with 1,3,5-trinitrobenzene was described as a reaction of malononitrile with
aromatic nitro compounds. In the presence of dimethylformamide it gives an orange colour.
112
2.20.3
CN
O2N CH
CN
O2N NO2 CN NO2
H2C HNO2
CN CN
NO2
O2N C
CN
NO2
CS compound is reported to react with the MARQUIS reagent under formation of a yellow
coloration, the sensitivity being up to 0.5 µg.ml-1 in a sample.
One of the most employed detection reactions of malononitrile is its condensation with
quinones. Thus, e.g. the reaction with perchloro-1,4-benzoquinone (chloranil, tetrachloro-
benzoquinone) or 1,4-naphthoquinone is used even in detector tubes [2].
2.20.4
O
Cl Cl
CN
O Cl CH
CN Cl Cl O CN
H2C
HCl
CN Cl Cl
OH
O
Cl Cl
CN
Cl C
O CN
The reaction gives a blue, blue-green or green coloration, depending on the solvent employed
(water, ethanol, chloroform, tetrachloromethane). The reaction rate increases with heating and
using lipophilic solvents. Its sensitivity with chloranil is 3 µg.ml-1 in a sample. Better results
can be obtained with 1,4-naphthoquinone.
113
2.20.5
O
CN
O CH
CN O CN
H2C 1
/2 O2
H2O
CN OH
O
CN
O CN
The coloration is up to deep blue, the sensitivity up to 0.5 µg.ml-1 in a sample solution and the
reaction course allows even quantitative photometric determination.
An analogous reaction with ninhydrin gives a yellow coloration. The reaction is also utilized
in detector tubes [3].
2.20.6
O O
CN CN
O H2C C H2O
CN CN
O O
Detector tubes for CS compound make use of sodium nitroprusside as a reagent for
malononitrile.
2.20.7
Na2[Fe(NO)(CN)5] H2C(CN)2 Na2[Fe{H 2C(CN)2}(CN)5] NO
Yellow sodium nitroprusside is converted into an orange compound. According to some

authors, it is not a mere replacement of one ligand by another but a true reaction between
these two ligands.
2.20.8
Na2[Fe(NO)(CN)5] H2C(CN)2 Na 2[Fe{N=C(CN)2}(CN)5] H 2O
The sensitivity of the reaction is 2 µg.ml-1 in a sample. Its selectivity is not high; a similar
coloration is obtained with many reducing compounds such as sulfides, thiols or sulfites.
The present authors have described detector tubes for CS compound (and
chloroacetophenone) that use furazans and a thiadiazol as chromogenic agents (see Chapter
2.19). CS compound gives a violet to blue-violet coloration [4]. The sensitivity of these
114
detector tubes is very good: 0.2-0.7 µg.l-1 in air. The reaction mechanism is not hitherto
elucidated.
The present authors have also described another method for the identification of malononitrile
arising by hydrolysis of CS compound. Malononitrile is hydrolyzed relatively rapidly to
ammonium salt of malonic acid. Ammonium salts are detected by the NESSLER reagent (see
Chapters 2.18.2 and 2.18.3).
2.20.9
CN NH2 NH2 ONH4 ONH4
2 H2O + 2 H2O
H2C
C C C C
CN O C O O C O
H2 H2
In an alkaline medium, malononitrile behaves as an acid and forms an anion.
2.20.10
CN CN
H2C OH HC H2O
CN CN
This anion can be utilized as a passive component in the reaction with a suitable diazonium
salt, such as Fast Red TR or Fast Blue B (2.20.11), which is a reactive dye formed by
diazotation of o-dianisidine and stabilized as tetrafluoroborate.
The coupling of the diazonium salt with malononitrile affords an intermediary ion associate
that gradually reacts to give a corresponding azo dye. This is extracted from water into a
water-immiscible solvent, chloroform. The azo dye is extracted mainly in the hydrazone form.
The reagent is dissolved in methanol. For the quantitative or semiquantitative determination, a
mixture of the analyte and the reagent is allowed to react for 10 minutes and then is extracted
with chloroform. A yellow coloration of the organic phase shows the presence of
malononitrile (CS compound) in the sample. The sensitivity is 1 µg in a sample and the
procedure is sufficiently selective. The method is used in many mobile laboratories [5-10].
2.20.11
H3CO OCH3
CN
2 HC N N N N
CN
H3CO OCH3
NC CN
CH N N N N HC
NC CN
w
115
H3CO OCH3
NC CN
CH N N N N CH
NC CN
H3CO OCH3
NC CN
C N NH NH N C
NC CN
org
One of the valued methods for determination of CS compound is that suggested by MAKLES.
It is based on the oxidation of the benzylidene double bond to epoxide and subsequent
rearrangement and alkaline hydrolysis under formation of cyanide ions.
2.20.12
Cl Cl
CN O CN
(KMnO4)
CH C O CH C
CN CN
Cl Cl
CN O CN O
-
+ 2 OH
CH C O CN - H2O CH C CN
The procedure can be used for the detection of CS compound as well as for the quantitative
determination either by titration or by spectrophotometry, with application of some of the
known methods for the detection and determination of cyanides (see Chapter 2.4). The
method is used in field chemical laboratories. The present authors have described a detector
tube [11], containing an indication layer impregnated with a reagent for cyanogen chloride
(4-benzylpyridine, dimedone), chlorination layer, a glass wool filtering layer and an ampoule
with KMnO4.
The present authors have described a very dependable method for the determination of CS
compound, based on the addition of hydrogen cyanide to a multiple bond. An excess of the
volumetric solution of potassium cyanide in ethanol and a solution of potassium hydrogen
carbonate are added to an ethanolic solution of CS compound. After standing for 10 minutes,
the excess cyanide is titrated with a silver nitrate solution. Turbid and coloured solutions can
be advantageously titrated with potentiometric indication by a silver electrode or insoluble
silver salt membrane electrode [12].
116
2.20.13
Cl Cl
CN CN CN
CH C KCN KHCO3 CH CH CN K2CO3
CN
2 CN Ag [Ag(CN)2] Ag 2 AgCN
The method mentioned is very easy to be carried out. It is also advantageous because the
reaction is performed with the whole molecule of the analyte and not only with its
decomposition product. Its drawback is the controlled commercial accessibility of cyanides
[13-15].

1. ISSA, F.M., ISSA, Y.M., YOUSSEF, H.Z., ISSA, R.M.: Spectrophotometric determination of
arylidenemalononitrile derivates using m-dinitrobenzene. Z. Anal. Chem. 277, 125 (1975).
2. WIACEK, K., MAKLES, Z.: Rurka wskaznikowa do wykrywania dwunitrylu o-chloro-
benzylidenomalonowego i nitrilu malonowego. Int. Cl. G01N31/22. PL 83474. (15.11.1976). WOJSKOWY
INSTITUT CHEMII I RADIOMETRII.
3. EICHARDT, A., OLTHOFF, U., FRIEDEL, B.: Verfahren zur Herstellung eines Indikatorrörchens. Int. Cl.
G01N31/22. DD 299 087 A5. (26.3.1993). JENAPHARM.
4. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z.: Detekční trubička s deriváty benzofurazanu ke
zjišťování dráždivých látek ve vzduchu. Sborník Univerzity obrany, řada „B“ (Technické a přírodní vědy),
2005, číslo 1, s. 179-186.
5. HALÁMEK, E., KOBLIHA, Z.: Spektrofotometrické stanovení orthochlorbenzalmalononitrilu a
malononitrilu reaktivními barvivy typu diazoniových solí. In: Zborník vojenskovedeckých prác, 3, 1,
Liptovský Mikuláš, VVTŠ, 1988, s. 136 - 159.
6. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Metody analýzy o-chlorbenzalmalononitrilu (CS) pro aplikaci
v polních chemických laboratořích. In: Sborník VVŠ PV, řada“B“, č. 1, Vyškov, VVŠ PV, 1990, s. 57-68.
7. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Ověření metod stanovení o-chlorbenzalmalononitrilu (CS) na
reálných vzorcích.In: Sborník VVŠ PV, řada „B“, č. 1, Vyškov, VVŠ PV, 1990, s. 69 - 78.
8. HALÁMEK, E., KOBLIHA, Z.: Extraction spectropfotometric [(2-chlorophenyl)methylene]propane dinitrile
with reactive dyes of the stabilized diazonium salt type. Collect. Czech. Chem. Commun., 57,1992, s. 1221-
1229.
9. HALÁMEK, E., KOBLIHA,Z: Analýza bojových otravných látek extrakční spektrofotometrií v přenosné
chemické laboratoři PCHL-90. In: Informační zpravodaj ICO ČR 10, č.1, 1999, s. 10 - 27. (Sborník přednášek
z 13. celostátního semináře o separační chemii, Lázně Bohdaneč, 28.-29. 6. 1999.)
10. Pat. ČSSR 2700. KOBLIHA, Z., HALÁMEK, E.: Způsob spektrofotometrického stanovení o-chlorbenzal-
malononitrilu diazoniovými solemi. 12.2.1988.
11. ORITEST. Detekční trubička pro stanovení látky CS. Původce: Pitschmann, V., Tušarová, I., Halámek, E.,
Kobliha, Z. Int. Cl. G01N31/22. CZ 17644. Zapsáno 2. 7. 2007.
12.PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J., ABERLE, J.: Stanovení
o-chlorbenzalmalonnitrilu membránovými elektrodami. In: Studijní sborník, 3, Vyškov, VVŠ PV, 1977, s. 104
- 112.
13.HALÁMEK, E.: Příspěvek k elektrochemickému stanovení některých bojových otravných látek.
obrana, 28, 1986, 2, s . 75-87.
117
2.21 Dibenzo[b,f]-1,4-oxazepine, CR compound
N CH
CR compound is a relatively stable compound of aromatic character, with fully

conjugated system of bonds which is also contributed by the ether oxygen atom with its two
free electron pairs and the heterocyclic tertiary nitrogen atom with one electron pair. Thanks
to the aromatic conjugation, CR compound behaves as a very weak base that is quaternized
only in a strongly acid medium.
2.21.1
H
N CH N CH
H
O O
With the MARQUIS reagent dibenzo[b,f]-1,4-oxazepine affords a yellow coloration. The

reaction is not selective, but it is quite sensitive, about 1 µg in a sample [1-3].
Similarly to tertiary amines and alkaloids, CR compound is precipitated relatively readily by
many metal complexes as an ion pair, similarly to other CWAs of amine character (see
Chapters 2.7 and 2.15). Precipitation with the DRAGENDORFF reagent that gives an orange
precipitate may serve as one of many examples.
2.21.2
H
N CH HN CH
[BiI4] [BiHI4]
O O
The formation of ion pairs with acidic dyes is much less facile. CR compound reacts in this
way only with some strongly acidic sulfonated dyes that remain in the dissociated form even
at low pH (1-2). Thus, e.g. sodium 1-(2-hydroxynaphthylazo)-4-benzenesulfonate (Orange II)
with protonized CR compound forms an orange associate, extractable from an aqueous
medium with chloroform [4-12].
2.21.3
HN CH
O N N SO3
OH
118
HN CH
N N SO 3
O
OH
w
N H O S N N
O
CH
O HO
org
This method, described by the present authors, is performed in the buffer of pH 1. Its
sensitivity is good, 2 µg in a sample. It is interfered by amines. The procedure is used in
mobile laboratories.
In spite of the aromatic and strongly conjugated character of the molecule, the azomethine
bond in CR compound behaves as an isolated bond that is a potential site of addition and
nucleophilic reactions, its polarization being marked only in a strongly acid medium. The
nitrous acid-induced cleavage of the azomethine bond in CR compound, described by the
present authors, affords, surprisingly at the first glance, a diazonium salt which can be
determined by several methods, e.g. by coupling with a suitable passive component under
formation of the corresponding dye. This unusual reaction does not obey the generally
respected rule of preparative organic chemistry that only primary, preferably aromatic amines
are diazotized, whereas tertiary heterocyclic amines do not react in this way [13,14].
2.21.4
N CH N N
NaNO2 2 HCl
O - H2O, - NaCl, - Cl
O
OHC
119
N N
NH2
NH
O
OHC
NH3
N N NH
OHC
NH3
NH N N
OHC
N-(1-Naphthyl)ethylenediamine (BRATTON-MARSHALL reagent) is used as a passive

component. The formed azo dye, 2-[2-(4-aminoethylamine)-1-naphthylazo)phenoxy]-
benzaldehyde, has a sharp red colour. The name itself - Irritant Red VF (λmax 556 nm) -
suggests its aetiology. The reaction is highly selective and its sensitivity is sufficient: it is
possible to detect 5 µg in a sample. The procedure is performed also as a spot test in mobile
laboratories. The mentioned principle serves as a basis for spectrophotometric quantitative
determinations using a number of passive components, and also for a detector tube [15,16].
Recently, the workers of the Military University for CBR Defense in Moscow have also
described a similar chromogenic reaction for a detection tube. CR compound, after being
changed from a nitrogen heterocyclic compound to primary amine using acid hydrolysis,
reacts readily with 7-chloro-4,6-dinitrobenzofuroxan resulting in a red product (λmax 500 nm)
[17].
120
2.21.5
O
HC
N CH NH3
H , H 2O
O O
O O
NO 2 N
O OH
NO2
HC N N
NH3
N O O
+ H2O
O N N
O 2N O 2N H
N O - HCl O O O
O 2N - H2O O
HN CH NH CH
O
Cl
O O
A detector tube conforming to required selectivity attains the high detection limit of 0.05
mg.m-3.
CR compound can be quantitatively determined by acid-base titration with perchloric acid in
anhydrous acetic acid, the equivalence point being indicated with an acid-base indicator or a
glass membrane electrode. Another procedure consists in the potentiometric determination of
the diazotation cleavage of the azomethine bond in CR compound with an excess of nitrite in
an acid medium and back titration with sulfanilic acid, potentiometrically indicated by a redox
platinum electrode. A relatively simple titration method is the direct titration of CR compound
with sodium tetraphenylborate in an acid medium, indicated by an electrode with liquid
membrane of softened PVC [18,19].

1. FÖLDEŠI, V., HALÁMEK, E., PŘIKRYL, F.: I. Syntéza látky CR cyklizáciou N-(2-fenoxyfenyl)-
formamidu. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1987, s. 14 - 24.
2. FÖLDEŠI, V., HALÁMEK, E., PŘIKRYL, F.: II. Syntéza látky CR z „laktámu“. In: Sborník VVŠ PV, řada
„B“, 1, Vyškov, VVŠ PV, 1987, s. 25 - 40.
3. Pat. ČSSR 2677. FÖLDEDŠI, V., HALÁMEK, E., KUBUŠ, J.: Spôsob prípravy dibenz[b,f]-l,4-oxazepínu.
25.9.1987.
4. KOBLIHA, Z., HALÁMEK, E., ČAPOUN, T., KRYKORKOVÁ, J: CO-3-16 Analýza vysoce toxických látek
v chemických laboratořích Civilní ochrany ČR. Příloha 5: Extrakčně spektrofotometrické metody. MO Praha.
28 s.
5. HALÁMEK, E., KOBLIHA, Z., SOUČEK, J.: Spektrofotometrické stanovení látky CR. In: Zborník
vojenskovedeckých prác, 3, 2, Liptovský Mikuláš, VVTŠ, 1988, s. 156 - 173.
6. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Metody analýzy dibenz[b,f]-1,4-oxazepinu pro aplikaci
v polních chemických laboratořích. In: Sborník VVŠ PV, řada „B“, č. l, Vyškov, VVŠ PV, 1990, s.40 - 47.
7. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Ověření metod stanovení dibenz[b,f]-1,4-oxazepinu na
reálných vzorcích. In: Sborník VVŠ PV, řada „B“, č. 1, Vyškov, VVŠ PV, 1990, s. 48 - 56.
8. HALÁMEK, E., KOBLIHA, Z., SOUČEK J.: Determination of dibenzo[b,f]-1,4-oxazepine by extraction
spectrophotometry. Talanta 40, (2), 1993, s. 287-289.
9. Pat. ČSSR 2641. HALÁMEK, E., FÖLDEŠI, V.: Spôsob analýzy dibenz[b,f]-1,4-oxazepínu. 25.9.1987.
10. Pat. ČSSR 2731. HALÁMEK, E., KOBLIHA, Z., SOUČEK, J.: Způsob extrakčně spektrofotometrického
stanovení dibenz[b,f]-1,4-oxazepinu antrachinonovými barvivy. 16.6.1988.
121
stanovení dibenz[b,f]-1,4-oxazepinu sulfoftaleiny. 16.6.1988.
stanovení dibenz[b,f]-1,4-oxazepinu azobarvivy. 16.6.1988.
13. HALÁMEK, E., KOBLIHA, Z., FÖLDEŠI, V.: Determination of dibenzo[b,f]-1,4-oxazepine by diazotation
cleavage of azomethine bond. Talanta 40, 1993, 1189 – 1192.
z 13. celostátního semináře o separační chemii, Lázně Bohdaneč, 28.-29. 6. 1999).
15. Pat. ČSSR 2702. HALÁMEK, E., FÖLDEŠI, V.: Trubičkový detektor k zisteniu dibenz[b,f]-1,4-oxazepínu
v ovzduší. 25.9.1987.
16. Pat. ČR 2761. HALÁMEK, E., FÖLDEŠI, V.: Trubičkový detektor k zisteniu dibenz[b,f]-1,4-oxazepínu
v ovzduší. 11.5.1993.
17. EVGEN´EV, M.I., GARMONOV, S.Yu., BELOV, P.E., TSEKHMISTER, V.I., DRUZHININ, A.A.: Test
method for the determination of toxic irritants in air. Журн. Аналит. Химии 58, 5, 2003, 542-545.
laboratořích Civilní ochrany ČR. Příloha 6: Potenciometrické titrace. MO Praha. 23 s.
obrana, 31, 1989, 5, s. 43 - 45.
122
2.22 Diphenylchloroarsane
As
Cl
When treated with solutions of alcoholates, diphenylchloroarsane (DA) liberates

chlorides that can be determined in a solution, e.g. as a silver salt.
2.22.1
As OR - As Cl -
Cl OR
Even on heating, diphenylchloroarsane is not completely decomposed by the treatment with

potassium or sodium hydroxide in an aqueous-alcoholic solution. The reaction affords a salt
of diphenylarsonic acid.
2.22.2
As 2 OH -
As Cl - H 2O
Cl O
Similarly to other warfare organoarsenic compounds, diphenylchloroarsane can be detected

by a detector tube for lewisite filled with colourless osmium(VIII) oxide which is reduced to
brown-black osmium(IV) oxide. In order to enhance the indication effect, the detector tube
may contain the redox indicator benzidine.
2.22.3
O
As OsO4 H 2O As OsO2 HCl
Cl OH
Diphenylchloroarsane is completely decomposed on oxidative melting with alkalies (e.g. with

a sodium peroxide-potassium nitrate mixture) leading to an arsenate [1,2].
2.22.4
As 28 NaOH 31 O Na3AsO4 NaCl 12 Na2CO3 19 H 2O
Cl
The arising arsenates afford a number of characteristic reactions. On longer standing with a
“molybdenum solution” (ammonium molybdate in nitric acid), they form a yellow precipitate
of ammonium tetrakis(trimolybdato)arsenate.
123
2.22.5
AsO4 3- 12 (NH4)2MoO4 24 H + (NH4)3[As(Mo3O10)4] x H2O (12-x) H2O 21 NH4 +
The sensitivity of the reaction is 20 µg in a sample. The reaction is interfered by phosphates.

Molybdatoarsenate is reduced with tin(II) chloride to give Molybdenum Blue. Analogously,
arsenates can also be detected with “magnesium solution” (magnesium chloride and
ammonium chloride in ammonium hydroxide) as a white precipitate.
2.22.6
3-
AsO4 MgCl2 NH4Cl 6 H 2O NH4MgAsO4 .6 H2O 3 Cl -
The determination is interfered by phosphates.

Arsenates react with silver, copper(II) and iron (III) ions to give a brown, light blue and white
precipitate.
2.22.7
3 Ag + AsO4 3- Ag3AsO4
2+
Cu HAsO4 2- CuHAsO4
Fe 3+ AsO4 3- FeAsO4
The As(V) atom in arsenates can be converted into the As(III) atom by the reduction with
iodides.
2.22.8
extraction
extrakce
As 5+ 2 I- As 3+ I2 {I2}org
This equilibrium reaction can be shifted markedly to the right by e.g. extraction of iodine into
a non-polar, water-immiscible solvent (red coloured solution).
The formed arsenites afford characteristic precipitation reactions. Silver ions precipitate a
yellow product, whereas copper(II) ions give rise to a green precipitate.
2.22.9
3 Ag + AsO3 3- Ag3AsO3
Cu 2+ HAsO3 2- CuHAsO3
Arsenites are not precipitated by either a “molybdenum solution” or “magnesium solution”.

Yellow dichromates are reduced to green chromites.
2.22.10
3 AsO3 3- Cr2O7 2-
3 AsO4 3- 2 CrO2 -
Aluminium in an alkaline medium reduces only arsenites but not arsenates.
2.22.11
3- 3-
AsO3 2 Al 3 OH - H3As 2 AlO3
Arsane can be proved by the classical MARSH-LIEBIG test [3,4] (decomposition to metallic
arsenic) or by the GUTZEIT test (formation of silver, mercury(II) or lead(II) arsenide; see
Chapter 2.8) [5,6]. WINKLER suggested to use gold(III) chloride on paper which forms a
red-violet coloration; sensitivity up to 0.2 µg in a sample. The colour corresponds to colloidal
gold (see 2.8.10).
124
So far the most sensitive reaction for arsane, employed together with the modified GUTZEIT
test in mobile laboratories and detector tubes, is that with silver diethyldithiocarbamate in
pyridine or dimethyl sulfoxide (see Chapter 2.8). Diphenylchloroarsane can be quantitatively
determined by direct titration with iodine or back titration with sodium thiosulfate.
2.22.12
As I2 2 H2O As 2 HI HCl
Cl OH
Another alternative is the titration of chlorides liberated in the reaction with hydroxide
(2.22.2) or alcoholate ions (2.22.1) with silver nitrate indicated potentiometrically by a silver
or insoluble silver salt membrane electrode [7]. The determination of chlorides can also be
used according to VOLHARD by treatment with an excess of silver nitrate and back titration
with ammonium thiocyanate to iron(III) salt as a visual indicator [8].

Moskva 1990. 272 s.
5. VAŠÁK, V., ŠEDIVEC, V.: Kolorimetrické stanovení arsenu. Chem. Listy 46, 341-344 (1952).
1977. 512/616 s.
7. ČIHALÍK, J.: Potenciometrie. Nakladatelství ČSAV. Praha. 1961. 772 s.
8. TOMÍČEK, O.: Odměrná analýza. Československá chemická společnost pro vědu a průmysl. Praha 1949.
380 s.
125
2.23 Diphenylcyanoarsane
As
CN
As concerns its determination, diphenylcyanoarsane (DC) differs from the preceding

CWA only by losing cyanide ions when treated with nucleophilic reagents, e.g. hydroxide or
alcoholate ions.
2.23.1
As OR - As CN -
CN OR
The cyanides are then detected by a number of formerly described procedures and methods
(see Chapter 2.4). Other reactions and analytical procedures are analogous to, or identical
with, those described for diphenylchloroarsane (Chapter 2.22).

126
2.24 10-Chloro-9,10-dihydrophenarsazine (adamsite)
H
N
As
Cl
Adamsite is sparingly soluble in organic solvents, even on boiling. Therefore, one

usually handles samples of a solid compound or evaporation residues.
The best-known detection reaction for adamsite is its reaction with concentrated sulfuric acid
affording a marked red-violet coloration that arises with only a few aromatic compounds.
2.24.1
H
N NH
H2SO4 SO4 2- HCl
As As
Cl H
The formation of deep-coloured aromatic products in concentrated sulfuric acid is usually

explained by extension of the conjugated bond systems and formation of carbonium ions and
radicals with unpaired electrons. This explanation is supported by the fact that the colour
disappears on dilution of the coloured reaction mixture with water. The reaction is sensitive,
capable of detecting up to 0.4-0.5 µg of analyte in a sample. The same effect is induced also
by hot formic acid; however, the red colour disappears on cooling or shaking the test tube.
When the mixture is heated again, the colour reappears. The procedure with sulfuric acid is
used in field laboratories and also in detector tubes. Equally sensitive (1.0-0.5 µg in a
sample), but selective, is the reaction with concentrated nitric acid or nitration mixture (a
mixture of concentrated sulfuric acid and nitric acid) which affords yellow coloration; on
subsequent treatment with sodium hydroxide the sample turns sharp red due to the formation
of the so-called aci-salt (Na-salt) [1]
2.24.2
H
N NH2
3 HNO3 NO3 - NO2 HCl
As As NO2
O OH
Cl
N + 3 OH -
3 H2O NO3 -
O
As N
O O
O
127
A related reaction, utilized in designing detector tubes, is the appearance of green coloration
on treatment with mercury(I) nitrate in concentrated sulfuric acid. Glass wool is employed as
support and the reagent is sealed in an ampoule. This chromogenic reaction is evidently based
on the formation of a mercury(II) salt of oxidized and nitrated adamsite[2].
2.24.3
H
N NH2
2 Hg2(NO3)2 2 H2SO4 2 HCl SO2
SO4 2-
As As NO2
O OHg
Cl 2
The tube is highly selective and the detection limit is also very good, 0.2 µg.ml-1 in a sample.
The very widespread and specific reaction with silver nitrate in hot acetic acid, affording a
green-yellow coloration, may have a similar course.
2.24.4
H
N NH2
2 AgNO 3 NO 3 - AgCl
As As NO2
O OAg
Cl
The treatment of adamsite with uranyl nitrate also affords a green coloration.
The tube, published by the present authors, is based on the reaction of adamsite with nitrous
acid, affording yellow to orange nitroso or isonitroso (oxime) derivative [3,4]
2.24.5
H NH2
N
NaNO2 2 HCl Cl - NaCl H2O
As As NO
Cl
Cl
NH
Cl -
As N
OH
Cl
The selectivity as well as sensitivity of the reaction is good, the detection limit being 0.5 µg in
air.
As verified by the authors, the substitution reaction with thiocyanate leading to yellow
thiocyanate derivative of adamsite represents another reaction employed in a detector tube
[3,5,6] Its selectivity is good but the tube sensitivity is only about 5 µg in air.
128
2.24.6
H H
N N
NH4SCN NH4Cl
As As
Cl SCN
Besides alkaline oxidative mineralization, adamsite can be decomposed to arsenic salts by

boiling with hydrochloric or hydroiodic acid at 100-160 °C under formation of diphenylamine
and arsenic(III) chloride.
2.24.7
H H
N N
2 HCl AsCl3
As
Cl
Diphenylamine treated with nitrates and concentrated sulfuric acid or with other strong
oxidation agents (chlorine in an H+-medium) affords blue coloration. Another possibility is
the reaction of diphenylamine with p-dimethylaminobenzaldehyde in an acid medium giving
a yellow coloration. Other reactions of arsenic (AsIII and AsV) or arsane are described in
Chapters 2.8 and 2.22.
The reaction of bromine with a solution of adamsite in a glacial acetic acid is a typical
halogenation reaction in which the As-C bonds are cleaved and diphenylamine is brominated
to give bis(2,4-dibromophenyl)amine.
2.24.8
Br Br
H H
N N
4 Br2 AsBr3 HCl HBr
As Br Br
Cl
The quantitative determination of adamsite utilizes with advantage the easy loss of chloride
ion that is then titrated according to VOLHARD or potentiometrically with silver nitrate [7,8].
2.24.9
H H
N N
OR - Cl -
As As
Cl OR
129
Adamsite can also be determined by redox methods, however, its insolubility is a problem.
2.24.10
H H
N N
H2O2 HCl
As As
O OH
Cl
Besides hydrogen peroxide, adamsite can also be oxidized with other reagents, such as
chloroamines, hypochlorites or potassium permanganate [9].
The mechanism of reaction of adamsite with a chromogenic reagent based on chlorodinitro
derivative of benz-2,1,3-oxadiazole (4-chloro-5,7-dinitrofurazan) has been described only
recently.
2.24.11
NO2 Cl Cl
H
N N As + H2O As
O
N As - HCl N
O2N - H2O N
Cl Cl O2N H
O 2N
N N
O O
N N
OH
NO 2 N
O O
The reaction affords a deep blue product ( λmax= 590 nm). The procedure has been verified
using a detector tube of required selectivity and high detection limit (0.05 mg.m-3) [10].

1977. 512/616 s.
3. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z.: Simple detection of adamsite based on substitution-
type reactions. Science & Military 2(1), 63-66 (2007).
4. ORITEST. Trubičkový detektor pro určení dráždivé látky DM. Původce: Pitschmann, V., Halámek, E.,
Kobliha, Z. Int. Cl. G01N21/78. CZ 15059. Zapsáno 25. 1. 2005.
6. ORITEST. Detekční trubička ke stanovení adamsitu. Původce: Pitschmann, V., Halámek, E., Kobliha, Z. Int.
G01N31/22. CZ 14426. Zapsáno 7. 6. 2004.
obrana, 28, 1986, 2, s . 75-87.
Moskva 1990. 272 s.
10. EVGEN´EV, M.I., GARMONOV, S.Yu., BELOV, P.E., TSEKHMISTER, V.I., DRUZHININ, A.A.: Test
method for the determination of toxic irritants in air. Журн. Аналит. Химии 58, 5, 2003, 542-545.
130
2.25 Quinuclidin-3-yl diphenylhydroxyacetate, BZ compound
OH O N
The structure of BZ compound (3-quinuclidinyl benzilate) offers various reaction

possibilities, however, only some of them are suitable for its identification.
As a heterocyclic tertiary amine, BZ compound readily undergoes typical group reactions for
tertiary amines and alkaloids. Thanks to its spatial arrangement, the quinuclidine moiety has
an extraordinary position among amines and alkaloids.
If a solution of BZ compound, preferably in an organic solvent, is evaporated to dryness,
treated with methyl iodide, and then again heated on a water bath to evaporate all the methyl
iodide, the residue consists of methoiodide of BZ compound [3-(benziloyloxy)-N-methyl-
quinuclidinium iodide].
2.25.1
CH3
OH O N OH O N
CH3I I
O O
On addition of an oxidation reagent, e.g. hydrogen peroxide, the iodide liberates iodine which
colours starch blue or is extracted into a nonpolar, water-immiscible solvent forming a red
solution.
2.25.2
OH O N OH O N
H2O2 2H+
2 I I2 2
CH3 - 2 H2O O CH3
O
The formation of protonated quaternary cation of BZ compound also underlies other reactions
based on ion pair formation. First of all, this concerns a number of group precipitation
reactions for tertiary amines and alkaloids. Due to the high basicity of the quinuclidine
moiety, BZ compound readily undergoes such reactions (described sufficiently in Chapters
2.7 and 2.15). In the case of BZ compound, some of them proceed stoichiometrically and
enable a quantitative determination. The method of determination of BZ compound, described
by MAKLES, can serve as an example. A solution of ammonium hexathiocyanatocobaltate is
added to a sample of BZ compound; the mixture is acidified with phosphoric acid and then
extracted with chloroform. The presence of BZ compound manifests itself by a grey-blue
coloration of the chloroform layer [1].
131
2.25.3
OH O N
3 (NH4)3[Co(SCN)6] H3PO4
O
H
OH O N
[Co(SCN)6] (NH4)3PO4
O
3 w
H
OH O N
[Co(SCN)6]
O
3 org
Although the sensitivity is not very high and amounts to only 100 µg in a sample, the reaction
affords reproducible results and, therefore, it has been recommended for quantitative
determination of BZ.
With acidic dyes, BZ compound relatively readily forms ion pairs extractable into organic
water-immiscible solvents. The present authors have described a number of protocols based
on this behaviour. In field laboratories, Bromocresol Green or better Bromopyrogallol Red in
an acid buffer of pH 4.0-5.0 is used as a reagent [2-17].
2.25.4
OH OH
O OH
OH O N
H+
O C
Br Br
COO
132
OH OH
H
O OH
OH O N
O C
Br Br
COO
OH OH
H
O OH
OH O N
O C
Br Br
COO
org
The sensitivity is very good. It is possible to detect visually 1 µg in a sample. The

concentration or the amount of BZ compound can be assessed semiquantitatively by
comparing the red chloroform extract with a standard colour chart. Other amine compounds
interfere.
Besides being a tertiary and heterocyclic amine, BZ is also an aromatic compound and,
therefore, it affords typical reactions with sulfuric acid under formation of red-violet
coloration due to diphenylcarbonium and triphenylcarbonium ions [18].
2.25.5
OH O N
HOS2O
2 H2SO4 C COOH H
N
O
H2O SO4 2-
133
2 C COOH C
CO2 H+
C
H COOH
- CO2
- 2 H2O H2SO4 HOOC C
COOH - SO2
C C C
These triphenylcarbonium ions are stable only in concentrated sulfuric acid or in some other
acid with strong dehydrogenation and dehydration properties, such as concentrated
phosphoric or perchloric acid. The coloration disappears on dilution. The detection limit is
satisfactory: 5 µg in a sample. The selectivity is questionable because an identical coloration
is observed with adamsite, and many aromatic compounds give a red-brown and brown
coloration.
A similar reaction is described for spectrophotometric determination of BZ compound after
hydrolysis [19,20]. Hydrolysis affords free benzilic acid (diphenylhydroxyacetic acid,
diphenylglycolic acid) which in concentrated sulfuric acid undergoes condensation
with 1-naphthol to give a purple dye (naphthofuchsone) whose absorbance is then measured
at 558 nm.
2.25.6
OH
O OH
OH OH
H2SO4
OH
O - H2O
- CO2
H2SO4 - 2 H2O
- SO2
H2SO4
C OH HSO4
O
Esters and salts of benzilic acid can be selectively detected by the MARQUIS reagent based
on reaction with concentrated sulfuric acid. This reagent consists of concentrated sulfuric acid
and formaldehyde in the ratio 40:1. Formaldehyde is a gas and, therefore, the reagent is
prepared from its 40% aqueous solution or from the solid polymer – paraformaldehyde or
from hexamethylenetetramine (urotropine) or other compounds which on reaction with
concentrated sulfuric acid afford formaldehyde (acetate complex of iron, etc.). The sulfuric
134
acid must be chemically pure and its concentration must be higher than 86% because at a
lower concentration it loses the required dehydrogenation and dehydration properties [21].
The MARQUIS reagent acts here again as a reagent for aromatic moieties reacting selectively
with BZ compound and other benzilates with formation of a deep blue condensation and
dehydration product.
2.25.7
OH O N
2 CH2O H2SO4
O
- H2O
H H
N O OH OH O N
CH2 SO4 2-
O O
H2SO4 - H2O
H H
N O OH O N
CH C SO4 2-
O O
Depending on the execution, the sensitivity of the reaction ranges from 0.5 to 5 µg in a
sample, in detector tubes up to 0.1 µg.l-1 in air [22]. Its selectivity is good; out of CWAs only
BZ compound gives a blue coloration.
Due to the basic quinuclidine moiety, BZ compound also reacts with reagents for amines, for
example with perchloro-1,4-benzoquinone (chloranil). Heating a benzene or toluene solution
on a water bath produces a green coloration. The sensitivity is low, 60 µg.ml-1 in a sample,
with insufficient selectivity. The same coloration is obtained with other amines.
2.25.8
O
OH N Cl Cl
O
O
Cl Cl
135
O
Cl Cl
N O OH Cl
Cl
O O
BZ compound can be quantitatively determined by acid-base titration with perchloric acid in

anhydrous acetic acid. More advantageous, however, are potentiometric titrations, described
by the present authors, using ion-selective electrodes. As an example may serve the titration
method based on precipitation of tetraiodocadmate, indicated by a silver iodide membrane
electrode or direct mercurimetric determination of BZ compound in a medium of iodide ions
or titration with mercury(II) chloride under formation of 3-quinuclidinium benzilate
tetraiodomercurate(II) indicated by a silver sulfide membrane electrode or, possibly, titration
with tetraphenyl borate using a plastified PVC membrane electrode [23-36].
2.25.9
OH O N
2
2H+ CdI2 2 KI
O
OH O NH
+
2K [CdI4]
O
OH O N
2
2H+ HgCl2 4 KI
O
OH O NH
2K + 2 KCl [HgI4]
O
136
2.25.10
OH O N
B H+
O
4
H
OH O N
B
O
Another possible method for the determination of BZ compound is also the cholinesterase
reaction because the molecule of quinuclidin-3-yl diphenylhydroxyacetate is structurally
related to the neurotransmitter acetylcholine.
2.25.11
N CH
C
H2 OH
O
Acetylcholin
Acetylcholine / 3-chinuklidinylbenzilát
/ 3-quinuclidine benzilate
C C
As concerns the direct effect on cholinesterase, BZ compound behaves as a medium strong,

nonacylating, reversible inhibitor that can be determined by the cholinesterase reaction, as
used in one of the common biochemical methods [37-42].

2. SOUČEK, J., PŘIKRYL, F., HALÁMEK, E.: Extrakčně spektrofotometrické stanovení 3-chinuklidinyl-
benzilátu. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1983, s. 34 - 40.
3. KOBLIHA, Z., HALÁMEK, E., SOUČEK, J.: Extrakčně spektrofotometrické stanovení 3-chinuklidinyl-
benzilátu azobarvivem kalkon. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1985, s. 12 - 21.
4. SOUČEK, J., HALÁMEK, E., BELICKÝ, L.: Vliv některých rozpouštědel na extrakční rovnováhu iontového
asociátu látky BZ a bromthymolové modři. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1986, s. 53-
72.
5. SOUČEK, J., HALÁMEK, E.: Spektrofotometrické stanovení látky BZ. Civilní obrana, 28, 1986, 1, s. 88-96.
6. SOUČEK, J., HALÁMEK, E., PŘIKRYL, F.: Stanovení látky BZ bromkresolovou zelení. Civilní obrana, 28,
1986, 3, s. 77-85.
137
7. SOUČEK, J., HALÁMEK, E., PŘIKRYL, F., KOBLIHA, Z.: Spektrofotometrické metody analýzy látky BZ.
In: Zborník vojenskovedeckých prác, 1, 1, Liptovský Mikuláš, VVTŠ, 1986, s.162 – 182.
8. SOUČEK, J., HALÁMEK, E., FLORUS, S.: Extrakčně spektrofotometrické stanovení látky BZ. In: Sborník
VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1987, s. 48 - 66.
9. SOUČEK, J., HALÁMEK, E., PŘIKRYL, F.: Možnosti využití extrakční spektrofotometrie pro stanovení
otravných látek. In: Zborník vojenskovedeckých prác, 2, 1, Liptovský Mikuláš,m VVTŠ, 1987, s. 96 -119.
10. HALÁMEK, E., SOUČKOVÁ, L., PŘIKRYL, F.: Extrakčně spektrofotometrická analýza l-(l`-fenylcyklo-
hexyl)piperidinu, 3-chinuklidinylbenzilátu a N,N-diethyllysergamidu. In: Zborník vojenskovedeckých prác,2,
3, Liptovský Mikuláš, VVTŠ, 1987, s.114 - 131.
11. HALÁMEK, E., KOBLIHA, Z.: Extrakčně spektrofotometrické stanovení 3-chinuklidinylbenzilátu (BZ)
vybranými barvivy. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1988, s. 63 – 77.
12. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Metody analýzy 3-chinuklidinylbenzilátu (BZ) pro aplikaci
v polních chemických laboratořích. In: Sborník VÚ 070, č. 1, Brno, VÚ 070, 1991, s. 25-39.
13. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Ověření metod stanovení 3-chinuklidinylbenzilatu (BZ)
v reálných vzorcích. In: Sborník VÚ 070, č. 1, Brno, VÚ 070, 1991, s. 41-51.
14. HALÁMEK, E., KOBLIHA, Z.: Investigation of the conditions of extraction of ion-associates of 3-
quinuclidinyl benzilate with acidic dyes. Collect. Czech. Chem. Commun., 58, 1993, s. 315-319.
15. Pat. ČSSR 1784. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.: Způsob analýzy glykolátů
sulfoftaleiny. 22.12. 1979.
16. Pat. ČSSR 1788. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.,: Způsob analýzy glykolátů
sulfogaleiny a anilinsulfogaleiny. 22.12.1979.
17. Pat. ČSSR 1789. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.: Způsob analýzy glykolátů
azobarvivy. 22.12.1979.
18. PŘIKRYL, F., SOUČEK, J., HALÁMEK, E., KELLNER, J.: Spektrofotometrické stanovení
3-chinuklidinylbenzilátu s kyselinou sírovou. In: Sborník VVŠ PV, 4, Vyškov, VVŠ PV, 1980, s. 50 - 58.
19. PŘIKRYL, F., HALÁMEK, E., KELLNER, J., SOUČEK, J.: Spektrofotometrické stanovení
3-chinuklidinylbenzilátu některými karboxylovými kyselinami. In: Sborník VVŠ PV, 4, Vyškov, VVŠ PV,
1980, s. 32-42.
20. KAMER, D.N., HACKLEY, E.B.: A specific and sensitive method for the determination of benzilic acid.
Anal. Lett. 1(14), 919-927 (1968).
21. KOBLIHA, Z., HALÁMEK, E., SOUČEK, J.: Stanovení 3-chinuklidinylbenzilátu s acetátovým komplexem
železa a Marquisovým činidlem. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1985, s. 138 – 140.
22. ORITEST. Detekční trubička ke zjišťování psychoaktivní bojové otravné látky BZ ve vzduchu. Původce:
Pitschmann, V., Halámek, E. Int. Cl. G01N31/22. CZ 8223. Zapsáno 1.2.1999.
23. PŘIKRYL, F., SOUČEK, J., HALÁMEK, E., KELLNER, J.: Potenciometrické stanovení
3-chinuklidinylesteru kyseliny benzilové (BZ). In: Studijní sborník, 3, Vyškov, VVŠ PV, 1977, s. 97 - 103.
24. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.: Využití potenciometrie při stanovení
3-chinuklidinylbenzilátu a N-methyl-3-piperidinylbenzilátu Dragendorffovým činidlem. In: Sborník VVŠ PV,
4, Vyškov, VVŠ PV, 1980, s. 21 - 25.
25. PŘIKRYL, F., KELLNER, J., HALÁMEK, E., SOUČEK, J.: Stanovení 3-chinuklidinylbenzilátu
heteropolykyselinami. In: Sborník VVŠ PV, 4, Vyškov, VVŠ PV, 1980, s.43 - 49.
s kyselinou pikrovou. In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1984, s. 18 - 25.
s heteropolykyselinami. In: Sborník VVŠ PV, řada „B“, 2, Vyškov, VVŠ PV, 1984, s. 10 - 23.
28. HALÁMEK, E., SOUČEK, J., PŘIKRYL, F., PITSCHMANN, V.: Potenciometrické stanovení
psychoaktivních látek. Civilní obrana, 28, 1986, 6, s. 88-95.
stanovení některých otravných látek. I. Potenciometrické titrace založené na tvorbě iontových asociátů. Civilní
obrana, 29, 1987, 4, s. 50 - 60.
stanovení některých otravných látek. II. Vliv plastifikátoru PVC membrány na odezvu elektrody. Civilní
obrana, 29, 1987, 5, s. 82 - 9l.
stanovení některých otravných látek. III. Některé aspekty praktického využití potenciometrických titrací
založených na tvorbě iontových asociátů. Civilní obrana, 30, 1988, 6, s. 39 - 42.
obrana, 31, 1989, 5, s. 43 - 45.
138
33. Pat. ČSSR 1855. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J., GRMELOVÁ, V.: Způsob
potenciometrického titračního stanovení 3-chinuklidinylbenzilátu. 3.11.1980.
34. Pat. ČSSR 1880. PŘIKRYL, F., SOUČEK, J., HALÁMEK, E., KELLNER, J., GRMELOVÁ, V.: Způsob
potenciometrického titračního stanovení 3-chinuklidinylbenzilátu. 14.3.1980.
35. Pat. ČSSR 1884. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J., GRMELOVÁ, V.: Způsob
potenciometrického titračního stanovení 3-chinuklidinylbenzilátu kyselinou tetrakis(triwolframáto)křemičitou.
3.11.1980.
36. Pat. ČSSR 2013. PŘIKRYL, F., KELLNER, J., HALÁMEK, E., SOUČEK, J., GRMELOVÁ, V.: Způsob
potenciometrického titračního stanovení 3-chinuklidinylbenzilátu kyselinou
testrakis(triwolframáto)křemičitou. 3.8.1981.
37. HALÁMEK, E., PŘIKRYL, F., SOUČEK, J., SOUČKOVÁ, L.: Stanovení látky BZ biochemickou metodou.
In: Sborník VVŠ PV, řada „B“, 1, Vyškov, VVŠ PV, 1983, s. 50 - 57.
38. SOUČEK, J., HALÁMEK, E.: Inhibice butyrylcholinesterasy 3-chinuklidinylbenzilátem. In: Sborník VVŠ
PV, řada „B“, 2,Vyškov, VVŠ PV, 1985, s. 55 - 69.
39. Pat. ČSSR. 1783. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.: Způsob analýzy cholinolytik
cholinesterasovou reakcí s O-alkyl-S-dialkylaminoalkylthiofosfonáty a O,O‘-dialkyl-S-dialkylaminoalkyl-
thiofosfáty. 22.12.1979.
40. Pat. ČSSR 1785. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.: Trubičkový detektor pro
zjištění přítomnosti 3-chinuklidinylbenzilátu a O-ethyl-S-diisopropylaminoethylmethylthiofosfonátu
v ovzduší. 22.12.1979.
41. Pat. ČSSR 1786. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.: Způsob analýzy cholinolytik
cholinesterasovou reakcí s fysostigminem. 22.12.1979
42. Pat. ČSSR 1787. PŘIKRYL, F., HALÁMEK, E., SOUČEK, J., KELLNER, J.: Způsob analýzy cholinolytik
cholinesterasovou reakcí s O-alkylalkylhalogenfosfonáty a O,O‘-dialkylhalogenfosfáty. 22.12.1979.
139
3. Chemical reconnaissance and chemical check devices
3.1 Detector papers
Detector papers (detector strips, detector tapes) are simple technical devices based on
visual evaluation of colour changes due to solubility of organic dyes in liquid CWAs,
decomposition of complexes under the formation of coloured components (such as K-7a
paper, see reaction 2.6.11) or chemical reaction of CWAs with chromogenic reagents.
Organic dyes, complexes, and chromogenic reagents are immobilized on cellulose, fabric or
other supports in the form of paper, strip or tape. According to the construction principle, the
detector papers can be used for the detection of liquid compounds or for the detection of gases
and vapours. Some known detector papers have also a combined use. Special attention is to be
paid to the PP-1 detector tape introduced into the Czech Army and particularly to the PP-3
detector paper.
PP-1 detector tape
The PP-1 detector tape is used for the easy and rapid detection of liquid VX
compound. This compound can be identified in the form of sedimenting liquid particles of an
aerosol cloud or as liquid droplets or a liquid spray on the surface of features, military
equipment and material.
The PP-1 tape is a self-adhesive paper tape impregnated with an acid-base indicator,
Bromocresol Green (see 2.15.12) which on exposure dissolves in VX compound and affords a
blue coloration. The PP-1 kit comprises a block of 20 self-adhesive tapes, instructions for use
and a standard colour scale. The whole block measures 170x80x8 mm and weighs 60 g. The
working temperature ranges from -40 to +50 °C.
PP-3 detector paper
The PP-3 detector paper (CALID-3, ABC-M8 VGH) is used for the easy and rapid
detection of liquid CWAs, such as yperite and compounds of the G and V types. Such
compounds can be detected as liquid particles falling from an aerosol cloud or as liquid
droplets or a liquid spray on the surface of features, military equipment and material.
The PP-3 detector paper is a self-adhesive paper label whose paper material contains a
mixture of organic dyes. On exposure they dissolve relatively selectively in the above-
mentioned liquid CWAs and afford characteristic colorations. Yperite is detected by Dye Red
E (see 2.6.12), the V-type compounds are detected by Dye Green EDA (see 2.15.13), and the
G-type compounds by Disperse Yellow 23 (see 2.13.3). Coloration occurs practically
immediately, in 30 seconds at the latest, being red in the presence of yperite and lewisite,
yellow for the G-type compounds and green for the V-type as well as GP compounds.
The PP-3 detector paper kit contains a block of 12 self-adhesive labels, instructions for use,
and a standard colour scale. The whole block has dimensions of 100x65x4 mm and weighs
22 g. The working temperature ranges between -40 and +60 °C. The PP-3 detector paper is
part of the CHP-71 chemical detector set and of the ORI-217analytical set.
140
3.2 Colorimetric biosensor
The DETEHIT colorimetric biosensor is a simple device for the detection of nerve
compounds of G and V types in air, water, liquid homogenates, food, soil and on the surface
of objects.
The colorimetric biosensor is a detection strip of special hardened PVC, layered with a
detection zone comprising a fabric with immobilized and stabilized enzyme, a yellow
comparison zone, and a paper zone impregnated with the substrate and indicator. The
colorimetric biosensor detection is based on the inhibition of cholinesterase by a nerve
compound, resulting in blocking the normal enzymatic hydrolysis of a substrate, in this case
hydrolysis of acetylthiocholine to thiocholine which is then indicated using the ELLMAN
reagent (see 2.9.11).
The moistened detection zone of the colorimetric biosensor is exposed to air, immersed into
contaminated water or pressed to the tested surface. The paper zone is then pressed to the
exposed detection zone for 2 minutes and then the colour change of the detection zone is
evaluated visually. In the absence of nerve compounds, the originally white detecting zone
changes to yellow; in their presence the detection zone remains white. Detection limits of the
colorimetric biosensor depend on the nature of the analyzed compound and exposure time
(see Table 4). The working temperature ranges from -20 to +40 °C. Working at temperatures
below 0 °C requires warming the detection part of the biosensor, e.g. by hand.
Table 4: Detection limits of colorimetric biosensor for nerve compounds
Compound Detection limit

-1
in air (mg.l ) in water (mg.l-1), on surface (g.m -2)
Exposure time
2 min 20 min 5 min 30 min
-5 -7 -2
Sarin 1.10 4.10 1.10 1.10-3
Soman 2.10-5 2.10-7 5.10-3 4.10-4
-5 -7 -3
VX Substance 2.10 5.10 8.10 5.10-4
GP Substance 5.10-6 2.10-7 5.10-3 2.10-4
Tabun 8.10-5 1.10-6 6.10-2 9.10-3
-6 -7 -3
Cyclosine 1.10 1.10 2.10 1.10-4
R-33 4.10-5 7.10-7 2.10-2 8.10-4
Each DETEHIT colorimetric biosensor (detector strip) is enclosed in a waterproof metallized

foil. The set contains 2x5 pieces of these strips. They are packed either in a PE tube with 10
strips or in a plastic package containing 5, 10 or 20 strips. Each package contains instructions
for use.
The DETEHIT colorimetric biosensor is part of the CHP-71 chemical detector set, PCHL-90
portable chemical laboratory, ORI-217 detection set and SCHP chemical observer set. The
DETEHIT biosensor is also part of the SONDA-CH and PPCHL-AL-2/ch truck-mounted
laboratory equipment as well as of the LR-CH light reconnaissance vehicle of radiation and
chemical reconnaissance squad.
3.3 Chemical detectors
The term “chemical detectors” usually refers to devices based on detector tubes.
Chemical detectors differ mainly in the sampling method for the contaminated air and in the
141
number and parameters of detector tubes. In the Czech Army, the development and use of
chemical detectors and detector tubes has had a long tradition. In the Czech Army the CHP-71
chemical detector and the subsequent CHP-5 chemical detector are used. The ORI-217
analytical set is of the same character as well.
The CHP-71 chemical detector is designed for chemical reconnaissance and the
detection of CWAs in air, soil, terrain and on the surface of various objects. It is intended for
detection under various climatic and weather conditions, by day as well as by night. The
device can be placed in a reconnaissance vehicle or used out of doors as a portable detector. It
is fed from the vehicle’s power network or by single-cell batteries. Simultaneous detection of
up to four different CWAs or CWA groups is possible. The working air flow rate is 3 l.min-1
(maximum 4 l.min-1). In addition to standard detector tubes, the CHP-71 detector may be
equipped with other tube types for the detection of CWAs, industrial harmful substances or
other militarily important toxic compounds. Dimensions of the device itself are
235x200x80 mm, its weight is 2.8 kg.
The CHP-71 chemical detector set consists of the device itself and accessories which
comprise a satchel, a set of PT-44/2, PT-36/4 and PT-145/2 detector tubes (20 pieces each), a
tube and an ampoule opener, a set of inlet filters (25 pieces), two outlet filters, a rubber tube,
an extension piece for the detection of CWAs in soil and on the surface of objects, a set of
plastic caps for solid samples (5 pieces), a small shovel for taking samples, a thermometer, a
heating element (20 pieces), a feeding cable, 4 single-cell 1.5 V batteries, a cleaning cloth, a
cleaning needle, a strap, instructions for use, a notebook and a standard colour chart for
evaluation of detector tubes. The CHP-71 set also comprises the DETEHIT colorimetric
biosensor (10 pieces) and PP-3 detector papers (12 sheets).
The CHP-5 chemical detector is a modern semi-automatic device replacing the CHP-
71 chemical detector. It can work simultaneously with up to five tubes, control the function of
individual tubes and set the suction time or the volume of the air analyzed. The standard flow
rate of air through one tube amounts to 1 l.min-1; in a special mode it can be varied between
0.3 and 5 l.min-1. If required, tubes of other size and construction, produced practically by any
world manufacturer, may also be used. Unlike its predecessor, this device is fed from the Li-
Ion rechargeable large-capacity source (14.4 V, 9200 mAh). The CHP-5 chemical detector is
also equipped with many auxiliary functions affording a high user-comfort, such as automatic
warming of the tubes, temperature measurement, display of the current function, data storing,
etc. Dimensions of the device itself (without the source box) are 230x82x161 mm and its
weight is 2.8 kg. The source box dimensions are 296x80x50 mm and its weight is 1.6 kg.
The CHP-5 chemical detector kit contains the device itself and accessories. The device
consists of a device box, the chamber and a source box. A tube and an ampoule opener are
attached to the device box. The accessories consist of an accessory satchel, a set of detector
tubes, the Universal 86 manual suction pump with spare parts, an adapter with a set (5 pieces)
of anti-smoke filters, a set of plastic cups for the adapter (5 pieces), a small shovel, a cleaning
needle, a set of flexible tubes, the OF-90 outlet filter compatible with the OM-90 protecting
mask, a transport bag for the device itself, a strap, a small emergency bag, an operation,
service and maintenance instruction handbook, a standard colour chart, a notebook and a
repair manual [1,2].
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ORI-217 detection set
The ORI-217 set for the detection of chemical warfare agents is a multipurpose device
for chemical reconnaissance and chemical check to be used for the determination of principal
CWAs in air, on the terrain, military equipment, on the surface of various objects as well as in
water and in solutions. It is included in the equipment of inspecting teams of the Organization
for Prohibition of Chemical Weapons (OPCW) and of national authorities for verifying
compliance with the Chemical Weapons Convention. In the Czech Army, this set is part of
equipment of SONDA-CH and PPCHL-AL-2/ch mobile chemical laboratories.
The scope and potential of ORI-217 result from its composition. It contains the modified
CHP-71 chemical detector with a set of rechargeable NiCd batteries instead of mono-cells and
the PT-44/2 (TT-11), PT-31/1 (TT-13), PT-38/2 (TT-14) and PT-145/2 (TT-12) sets of
detector tubes, 30 pieces each. Further the ORI-217 set contains three blocks of PP-3 detector
papers and four sets (20 pieces each) of the DETEHIT colorimetric biosensor.
An indispensable part of the ORI-217 detector set is a satchel with accessories to CHP-71, the
Universal 86 manual suction pump, a battery charger, instructions for use and an instruction
book.
The ORI-217 set is kept in a portable 470x350x120 mm metal case with a foam lining. The
whole set weighs 9.5 kg.
Detector tubes
Detector tubes (tube detectors) are usually sealed glass tubes filled with a sorbent
(support) and chemical reagents. The most frequent support is silica gel, but also cellulose,
crushed glass, glass wool, porcelain, pumice or diatomaceous earth and various synthetic
materials. Chemical reagents can be immobilized directly on the support or they can be kept
in a sealed ampoule, usually as solutions. The presence of a toxic substance in air is indicated
by the colour change of the carrier and its concentration is usually assessed by the length of
the coloured zone, intensity of coloration or degree of decoloration [3]. Detector tubes of
national origin that can be used in the established CWA chemical detectors are listed in Table
5.
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Table 5: Reaction principles and detection limits for selected detector tubes
Detector tube Substance Reaction principle, reagents Detection Reference to the

limit, µg.l-1 reaction principle
PT-43 G Aminoperoxide reaction 0.5 2.10.3
PT-44/1 G,V Biochemical reaction 0.005 2.9.3
(Phenol Red)
PT-44/2 G,V Biochemical reaction (Ellman) 0.005 2.9.1, 2.9.11
PT-145/2 CG,DP,CK,AC 4-(4'-nitrobenzyl)pyridine 5 2.1.5
4-benzylpyridine, dimedone, 2.5.5
picric acid 2.4.11
PT-145/3 CG,DP,AC,CK 4-(4'-nitrobenzyl)pyridine 5 2.1.5
chlorinating agent chloramine T
4-benzylpyridine, dimedone 2.5.5
PT-31/1 HD,HN 4-(4'-nitrobenzyl)pyridine, NaOH 1 2.6.16
PT-35 HD, SA gold(III) chloride, chloramine B 1 2.6.7
PT-38/2 L Ilosvay reagent 1 2.8.14
PT-38/3 L,DA,DC zinc, HCl, Ag salt 1 2.8.7, 2.8.8
PT-38/4 SA Ag salt 0.5 2.8.8
PT-36/3 HD HgBr2, ethyl-Michler's ketone 0.5 2.6.10
PT-36/4 HD MgClO4 , ethyl-Michler's ketone 3 Chapter 2.6
PT-37 HN Dragendorff reagent 1 2.7.2
PT-147 AC,CK chlorination agent chloramine T 0.5
4-benzylpyridine, dimedone 2.5.5
PT-147/1 AC 4-nitrobenzaldehyde 0.5 2.4.8
PT-147/2 AC,CK 4-nitrobenzaldehyde, pyridine 3 2.4.8
pyridine, dimedone 2.5.5
PT-148 CG,DP 4-(4'-nitrobenzyl)pyridine 0.5 2.1.5
PT-361 HD-CG,DP MgClO4 , ethyl-Michler ketone 3 Chapter 2.6
4-(4'-nitrobenzyl)pyridine 5 2.1.5
PT-51 BZ Marquis reagent 1 2.25.7
PT-25 CN m-dinitrobenzene 0,5 2.19.1
PT-26 CS chloranil, NH4OH 1 2.20.4
PT-26/X CS benzofurazane, NaOH 1 2.19.4
PT-146 CK dimedone, 4-benzylpyridine 0,5 2.5.5
PT-147/3 AC picric acid 10 2.4.11
PT-381 DM mercury(I) nitrate, sulfuric acid 3 2.24.3
PT-27 CR Bratton-Marshall reagent, NaNO2 0,1 2.21.4
PT-44/2 detector tube for nerve agents
The PT-44/2 detector tube for nerve agents labelled with three red stripes makes use of a
biochemical reaction based on the inhibition of acetylcholinesterase, an enzyme catalyzing
acetylthiocholine hydrolysis (see 2.9.8). The tube contains two layers: an indication layer and
a comparison one, and two ampoules with solutions. The indication layer contains white
granulated cellulose with immobilized and stabilized acetylcholinesterase. The comparison
layer contains yellow crushed glass impregnated with acetylthiocholine substrate and a
chromogenic reagent (ELLMAN reagent, see 2.9.11). Both ampoules are filled with a buffer
solution of pH 8 with ethanol added.
Working with the tube requires strict adherence to the written protocol. The upper ampoule is
crushed and its content is carefully shaken down onto the cellulose. One litre of contaminated
air is taken (using CHP-71 or CHP-5, sampling time 1 min at a rate of 1 l.min-1, with manual
pumps usually 10 strokes per 100 ml) and after 2 minutes, the second ampoule is crushed and
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its content vigorously shaken through the crushed glass up to the cellulose. The colour change
of the indication layer is then evaluated. If the layer turns yellow during the given time, the
atmosphere does not contain CWA; in the opposite case, the indication layer remains
unchanged or the yellow colour appears much later, at least twice as late (1-2 min) as without
air suction (2-4 min).
The tube can detect all types of nerve agents (G, V series) and possibly other volatile
acetylcholinesterase inhibitors (some organophosphorus or carbamate pesticides) as well.
Detection limits for sarin and soman vapours amount to 0.005 µg.l-1. The tube sensitivity is
lowered by high concentration of oxidation, reduction, acid or basic gases and vapours.
The detector tube may also be used for water analysis by dipping its tip into the sample to
moisten the cellulose layer; the next procedure is identical with that described for air samples.
PT-36/4 detector tube for sulfur yperite
The PT-36/4 detector tube for sulfur yperite, labelled with two yellow stripes, is based on the
reaction with an ethyl analogue of MICHLER’s ketone (see 2.6.10). The tube comprises one
indication layer, consisting of silica gel impregnated with chromogenic reagents
4,4'-bis-(diethylamino)benzophenone and magnesium perchlorate.
For the detection of yperite in air, a sample of about 3 litres is taken (sampling time 3 min at
flow rate of 1 l.min-1 for CHP-71 or CHP-5, or 30 to 50 strokes for manual pumps) and the
layer coloration is compared with the standard. The coloration deepens with time and,
therefore, it is useful to perform the evaluation 1 to 2 min after sampling.
The tube can determine all types of sulfur yperites (technical H, distilled HD, sesqui-yperite
Q, oxygen yperite T), as well as other chlorinated alkyl sulfides, the colour always being
orange to red. The detection limit is 3 µg.l-1.
If phosgene, diphosgene or other acylating agents, such as acetyl chloride or benzoyl chloride,
are present at concentrations exceeding 20 µg.l-1, a green coloration can be observed. High
concentrations of hydrogen chloride and ammonia as well as acid and alkaline gases result in
the decoloration of the indication layer.
At lower temperatures of the ambient air (below at least 10 °C), it is recommended to warm
the tube. Additional warming can result in a significantly higher sensitivity and faster onset of
the colour effect.
PT-145/2 detector tube for phosgene (diphosgene), cyanogen chloride and hydrogen cyanide
The PT-145/2 detector tube for phosgene (diphosgene), cyanogen chloride and hydrogen
cyanide, labelled with two green stripes, makes use of colour reactions of these compounds
with analytical agents. Phosgene (diphosgene) is detected by the reaction
with 4-(4'-nitrobenzyl)pyridine (see 2.1.5), cyanogen chloride by the KÖNIG-ZINCKE
reaction (see 2.5.5) and hydrogen cyanide by its reaction with sodium picrate (see 2.4.11).
The tube contains three indication layers consisting of carrier with immobilized chromogenic
agents. The upper layer contains silica gel impregnated with 4-(4'-nitrobenzyl)pyridine and
N-phenylbenzylamine in the presence of sodium carbonate and 3-phenyl-1-propanol as a
stabilizer. The middle layer consists of silica gel impregnated with 4-benzylpyridine and
dimedone, and the bottom layer contains silica gel impregnated with sodium picrate in
dimethyl sulfoxide.
To detect choking and generally poisonous substances in air, a sample of about 3 litres is
taken (sampling time 3 min at flow rate 1 l.min-1 for CHP-71 or CHP-5, or 30 strokes for
manual pumps) and the coloration of the individual layers is compared with the standard. In
the presence of phosgene (diphosgene) the upper layer turns red and the length of the
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coloured zone is proportional to the concentration of the harmful substance in air. If cyanogen
chloride is present, the middle layer turns pink to red-violet, and the presence of hydrogen
cyanide manifests itself by an orange to red-brown coloration of the bottom layer. In the case
of cyanogen chloride and hydrogen cyanide the intensity of coloration is proportional to the
concentration of the substance in air. The detection limit is 5 µg.l-1.
The detection of phosgene (diphosgene) is affected by the presence of other acylating
substances, such as chloromethyl formate, acetyl chloride or benzoyl chloride; these
compounds cause similar, usually pink coloration. However, high concentrations of hydrogen
chloride and other strongly acid gases and vapours interfere with the determination. Nitrogen
dioxide causes yellowing of the upper indication layer. The cyanogen chloride-indicating
layer also responds to cyanogen bromide or to phosgene oxime and turns pink by action of
nitric oxide. Other reducing agents may also react in a similar way as hydrogen cyanide.
PT-31/1 detector tube for sulfur and nitrogen yperite
The PT-31/1 detector tube for sulfur and nitrogen yperite, labelled with one yellow stripe,
makes use of the reaction with 4-(4'-nitrobenzyl)pyridine in the presence of a strong base (see
2.6.16). The tube contains an indication layer and an ampoule with the detection solution. The
indication layer consists of silica gel saturated with 4-(4'-nitrobenzyl)pyridine and the
detection solution contains sodium perchlorate and sodium hydroxide.
To detect yperites in air, a sample of about 3 litres is taken (sampling time 3 min at flow rate
1 l.min-1 for CHP-71 or CHP-5, or 30 strokes for manual pumps), then the ampoule with the
detection solution is broken and its content is vigorously shaken down to the indication layer
whose coloration is compared with the respective standard.
Using this tube can detect all types of sulfur yperites (technical, H, distilled, HD, sesquic
yperite, Q, and oxygen yperite, T) and nitrogen yperites (HN-1, HN-2, and HN-3) and other
chlorinated alkyl sulfides or amines. Detection limit is 1 µg.l-1. When warming the tube after
the sampling of the contaminated air (prior to breaking the ampoule with a detection solution)
higher sensitivity can be achieved.
Phosgene and diphosgene afford a red coloration; coloured products are also obtained with
CN and CS compounds. Some other compounds also react in this way: thus, e.g. dimethyl
sulfate affords a blue reaction product, benzoyl chloride an orange product and benzyl
chloride a violet product. At higher concentrations, the G-series compounds give a blue
coloration.
PT-38/2 detector tube for lewisite
The PT-38/2 detector tube for lewisite, labelled with one yellow stripe and a yellow dot,
makes use of the decomposition of lewisite with a base under formation of acetylene which is
detected by a modified ILOSVAY reagent (see 2.8.14). The tube contains an indication layer
and an ampoule with a detection solution. The indication layer comprises silica gel,
impregnated with copper(I) chloride, hydroxylamine and ammonium chloride. The detection
solution contains 20% sodium hydroxide.
To detect lewisite in air, first a sample of about 3 litres is taken (sampling time 3 min at flow
rate 1 l.min-1 for CHP-71 or CHP-5, or 30 strokes for manual pumps), then the ampoule with
the detection solution is broken and its content shaken down to the indication layer. The
arising coloration is compared with the standard. The test is selective for lewisite which
affords a brick-red colour. The detection limit is 1 µg.l-1.
L-2 and L-3 lewisite analogues do not react in this way and no positive response is observed
with other arsenic-based CWAs. The presence of sulfane interferes, which gives brown
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coloration with copper as well as carbon disulfide in high concentrations. A possible bluish
coloration of the layer does not hamper the determination due to ammoniacal products of
divalent copper
3.4 Mobile laboratories
Mobile laboratories represent the basic device of chemical check. They provide more
accurate and complete information obtained by chemical reconnaissance. They may vary from
small pocket-size laboratories, portable case-type laboratories to large laboratories mounted
on truck carriages or, possibly, housed in one or more containers for transport by rail, ship or
aircraft. The Czech Army is equipped with the PCHL-90 portable chemical laboratory, the
AL-1 truck-mounted chemical laboratory, the SONDA-CH truck-mounted laboratory and the
PPCHL-AL-2/ch container laboratory. The DETEGAS-1 kit introduced as part of the
SONDA-CH laboratory is also of a portable chemical laboratory character.
PCHL-90
The PCHL-90 portable chemical laboratory is a device of the Czech Army chemical
units designed to obtain the information not available by standard chemical reconnaissance. It
is included in the equipment of the Land Rover/rch vehicle for radiation and chemical
reconnaissance, the SONDA/ch mobile laboratory and the AL-2/ch container laboratory.
The PCHL-90 is intended for qualitative and semiquantitative analysis of water, terrain,
military material, and other structures contaminated with nerve agents (VX, GB, GP), blister
agents (HD), irritating agents (CS, CR), psychoactive substances (BZ), alkaloids,
phenoxyacetic acid-based herbicides and heavy metals. In addition, it checks air contaminated
with nerve agents and the content of active chlorine in decontamination mixtures. Potentially,
due to the methods applied, the laboratory can also determine other CWAs, at least all other
organophosphorus CWAs, such as GD, GF, R-33, GA, A-230/2/4 and nitrogen yperites HN-
1/2/3.
The underlying analytical principles are based on colour reactions in a test tube with the
extraction of the arising products into chloroform. These methods are complemented with the
DETEHIT colorimetric biosensor and some identification spot colour reactions on a special
glass fibre support. An overview of detection and semiquantitative methods is given in Table
6.
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Table 6: Overview of detection and semiquantitative methods of PCHL-90
Compound Method Coloration Detection Reaction

limit, µg principle
VX Amido Black 10B Blue 5 2.15.4
GB Aminoperoxide reaction Yellow-orange 1 2.10.3
GP Bromoxylenol Blue Yellow 1 2.16.2
HD o-Cresolphthalein Yellow 5 2.6.22
BZ Bromopyrogallol Red Red 10 2.25.4
Identification: Marquis reagent Blue-green 10 2.25.7
CS Diazonium salt Fast Blue B Yellow 1 2.20.11
CR Orange II Orange 2 2.21.3
HNO2 + Violet 20 2.21.4.
1-naphthylethylenediamine
Alkaloids Bromoxylenol Blue Yellow 1
Herbicides Fuchsin Red-violet 5
Heavy Cupral White to 1-30
metals brown
The PCHL-90 is contained in a tough plastic box measuring 590x420x300 mm. Its total
weight is 21 kg. It comprises a set of powder reagents (20 capsules each), a set of liquid
reagents (10 ampoules each), bottles with chloroform (5x300 ml), the DETEHIT colorimetric
biosensor set (200 strips) and laboratory equipment necessary for taking samples, their
processing and analysis. The laboratory capacity is 200 complete analyses made by one set of
reagents. A trained crew is able to analyze an unknown sample in 30 minutes. At
temperatures below 5 °C it is recommended to move the device to a heated place. The
laboratory is also equipped with means for the decontamination of samples, laboratory
equipment and aids [4,5].
DETEGAS-1
The DETEGAS-1 kit is designed for mobile analytical check of at least 12 industrial
harmful agents including phosgene (diphosgene), hydrogen cyanide and cyanogen chloride. It
can perform simple tests with universal tube detectors (detector tubes with a sorbent layer)
that prior to use are activated with a set of chromogenic agents. The methods are based on the
colorimetric principle with visual evaluation by comparison with standard colour charts.
The detection of phosgene (diphosgene) is based on the colour reaction with
4-(4'-nitrobenzyl)pyridine (see 2.1.5). The liquid reagent in a bottle contains an ethanolic
solution of 4-(4'-nitrobenzyl)pyridine, N-phenylbenzylamine and triethylene glycol. The tube
detector is activated by dipping its tip into the reagent and then 1 litre of contaminated air is
taken using a manual pump (10 strokes). The presence of phosgene (diphosgene) manifests
itself by a red coloration of the support. The detection limit is 0.5 µg.l-1. An alternative test is
based on using the Methyl Orange acid-base indicator that on reaction with phosgene changes
from yellow to red. The detection limit is 5 µg.l-1.
The detection of hydrogen cyanide is based on the colour reaction with the ELLMAN reagent
(see 2.4.10). The content of the reagent capsule is poured in a test tube, the amount of 2 ml of
a dimethyl sulfoxide-dimethylformamide mixture is added and the tube detector is activated.
One litre of contaminated air is collected using a manual pump (10 strokes). The presence of
hydrogen cyanide is revealed by an orange to red coloration of the support. The detection
limit is 2 µg.l-1; the reaction is interfered by sulfane, sulfur dioxide and thiols.
148
The detection of cyanogen chloride is based on the KÖNIG-ZINCKE colour reaction (see
2.5.3). A mixture of 4-(4'-nitrobenzyl)pyridine and 1,3-dimethylbarbituric acid from a capsule
is placed in a test tube and 2 ml of aqueous ethanol is added. The tube detector is activated by
dipping its tip into the reagent and 1 litre of contaminated air is taken using a manual pump
(10 strokes). In the presence of cyanogen chloride the support turns violet, the coloration
slowly changing to blue. This method can also detect hydrogen cyanide. A chlorination agent
solution (sodium dichloroisocyanurate or monochloramine B) is dropped onto an activated
and exposed tube detector. The detection limit is 3 µg.l-1.
The DETEGAS-1 kit is kept in a portable metal case. It contains 150 universal tube detectors,
four 70 ml solvent bottles, 9 tubes with 15 powder reagent capsules, 10 test tubes with
stoppers, syringes, a manual suction device and other necessary laboratory instruments. The
DETEGAS-1 kit has been introduced into the Czech Army as part of the SONDA-CH and
PPCHL-AL-2/ch laboratories.
AL-1 truck-mounted laboratory
The truck-mounted laboratory first appeared in the eighties as a modernised version of

the already physically as well as morally outdated ACHL-1 chemical laboratory. Basically, it
represents a combined chemical and radiometrical laboratory. The chemical part is located in
the body of the P-V3S terrain truck, the radiometric part in the A-3 truck trailer. The
laboratory is designed for the analysis of samples by classical methods of chemical analysis
and comprises a UV/VIS spectrophotometer (SPEKOL-11). The laboratory is well equipped
with a wide choice of laboratory glass and material, however, the laboratory instruction book
concerns only basic analytical procedures for main CWAs and decontamination reagents. In
addition to common analytical equipment, the laboratory is equipped with the CHP-
71chemical detector.
¨
SONDA-CH truck- mounted chemical laboratory
The laboratory was constructed relatively rapidly reflecting the changes in the Armed
Forces. Its construction was significantly influenced by the knowledge and experience gained
during the first Gulf War in 1991 where the Czechoslovak chemical unit was deployed with
great success. The laboratory is housed in a van body of the T 815 terrain truck. Besides
others, it is equipped with a GC/MS (EM 640) mobile set. Other devices of chemical
reconnaissance and check in the laboratory include the PCHL-90 portable chemical laboratory
with the DETEHIT colorimetric biosensor to detect nerve CWAs, the ORI-217 detection kit
with the CHP-71 modified chemical detector, the DETEHIT detector for nerve CWAs and the
PP-3 detector paper for the detection of liquid CWAs of the V-, G-, and H-types and the
DETEGAS-1 set for the analysis of important industrial toxic substances. The laboratory also
contains the GSA-12 automatic nerve agent detector based on the photometric evaluation of
the cholinesterase reaction (2.9.10).
PPCHL-AL-2/ch mobile field chemical laboratory
This modern device, at present being introduced into the equipment of the Czech
Army Chemical Corps, reflects the current demands and potentials for mobile chemical
analysis. The laboratory itself is housed in the ISO-1C standard army container. When the
laboratory is packed, it is transported on the T 815 vehicle equipped with a side loader
149
(KLAUS-STEELBRO SB 30). The rich and varied equipment of the air-tight and air-
conditioned laboratory includes physico-chemical instruments and chemical material. The
main instrument is a mass spectrometer coupled with the GC/MS gas-liquid chromatograph
(EM 640, capillary quartz column), a gas-liquid chromatograph (SRI), a high-performance
liquid chromatography apparatus (KNAUER), a thin-layer chromatography (TLC) kit
(CAMAG), and the UV/VIS spectrophotometer (HELIOS-β). All the physico-chemical
instruments are connected to a computer network. Simultaneously, the network functions as a
laboratory information system that enables not only running the instruments, evaluating the
analytical results and their saving, backup and transferring data, but also the network’s
functioning as a library and teaching system (LIMS). The laboratory equipment also
comprises a manual detector of CWAs (RAID-1M), the PCHL-90 mobile field chemical
laboratory and the ORI-217 CWA detection kit. The DETEHIT colorimetric biosensor for
nerve agents and the PP-3 detector paper for liquid CWAs are also included. Besides a spare 4
kW power source, housed in the laboratory container, the main 16 kW source is mounted on
the carriage of the accompanying LR Defender light terrain vehicle.
1. Normativ MO ČR/Chem-22-4/: Jednoduché prostředky chemického průzkumu a chemické průkazníky CHP-71
a CHP-5. Praha.2008. V tisku.
2. Průmyslový vzor ČR 33315. PITSCHMANN V., ČERNÝ, M., HÁJEK, J., HALÁMEK, E., OREL, J.,
POSEJPAL, J., SCHIMER, T., HEJKAL, T.: Chemický průkazník. 8.3. 2006.
3. Užitný vzor ČR 10473. PITSCHMANN V.,HALÁMEK E., TUŠAROVÁ I.., OREL, J., KOBLIHA, Z.,
VEVERKA, V.: Souprava simulačních trubiček ST-1 pro imitaci bojových otravných látek. 29.09.2000.
z 13. celostátního semináře o separační chemii, Lázně Bohdaneč, 28.-29. 6. 1999.)
5. Užitný vzor ČR 9036 HALÁMEK E., KOBLIHA, Z., OREL, J.,TUŠAROVÁ I..: Souprava indikačních
prostředků DO-2 pro přenosnou chemickou laboratoř PCHL-90. 6.9.1999.
150
4. Trends in development within the system of technical devices used in chemical
reconnaissance and check
It can be stated that the detection and analytical check of chemical warfare agents and
other militarily important compounds are being studied and developed as a system at all levels
of technical solution. As far as simple devices are concerned, several trends can be observed.
First of all, it is the research on detection systems for nerve agents, based on biochemical
reactions with stabilized and possibly even immobilized enzymes, particularly
cholinesterases of various etiologies. The employment of immunochemical procedures
especially for the detection of toxins of antigen character may also be an important
contribution. The investigation into potentialities offered by chromogenic systems, such as
e.g. thin layers of cholesterol derivatives (liquid crystals), still remains a classical direction of
development.
Small size and weight, easy training and use, independence of power sources, low
price and long-term storability are the reasons why this category of CWA detectors is
promising from the long-term perspective. This category can easily be introduced into
an individual's equipment and thus become fundamental for his/her survival in extreme
combat conditions under a possible chemical attack, in case of separation from the unit
or in other unexpected extreme situations.
There is still a steady interest in chemical detectors that make use of detector tubes
(tube detectors, indication tubes) as a most widespread device for the detection and
monitoring of known CWAs. The development aims at extending the choice of available
detectors to detect other current CWAs, to provide tube detectors for long-term suction of the
monitored atmosphere, tubes for the simultaneous detection of several substances and linear
tubes for semi-quantitative determination.
Chemical detectors using detector tubes represent an economically as well as

methodically accessible, traditional and easy way how to determine the presence and
kind of the warfare agent in air and on the surface of military equipment and material.
The choice of tube detectors based on chromogenic reactions can be extended easily and
rapidly according to the current situation. Besides tube detectors for CWAs, a wide
variety of tubes for the detection of industrial toxic substances is also available.
According to the current opinion, the system for the detection and monitoring of
CWAs is based on a rapid automatic detector attached to the data collection and evaluation
network. The devices based on the separation of clusters which are formed by the ionization
of a vapour-air mixture (IMS method) are also promising. Methods and procedures for
designing sensors, particularly biosensors, intended for direct monitoring of the atmosphere
are also being developed. Great attention is also paid to multidetection systems which are the
basis of the so-called electronic nose. Undoubtedly, nanomaterials and nanotechnologies are
an important contribution to the research in this field. Because of their slow response,
automatic detectors based on chromogenic reactions are not considered to be promising;
however, this opinion seems to be rather premature. As concerns long-range detection
research, systems utilizing infrared and differential spectroscopy are preferred. Devices useful
for the military, not only for stationary monitoring, have already appeared.
Rapid automatic detectors based on instrumental physico-chemical methods are

limited in practice, which results from insufficient sensitivity and partially from
151
selectivity as well. Most of the contemporary chemical reconnaissance instruments based
on ionization and using air as carrier gas face fundamental problems in the detection of
small molecules and, on the other hand, the problems with air moisture and pollution,
and thus with false alarms. It is, therefore, still necessary to have a choice of several
CWAs detection devices that use various reaction principles.
As for mobile laboratories, there is a marked shift from often complicated and very
time-consuming procedures of classical chemical analysis to physico-chemical
instrumentation, particularly to separation methods and general methods of organic structural
analysis, namely because of the existing legislation on international check of chemical
weapons. Small portable chemical sets based on classical analytical methods are still of great
importance for specific tasks of chemical check.
Chemical check in mobile as well as stationary laboratories supplements and

extends the information obtained by chemical reconnaissance devices. An unequivocal
identification of a toxic substance, its precursors and decomposition products is
required. Now, a dramatically greater stress is being laid on the exact and qualified
sampling, preservation, and transport of analytical samples of toxic substances.
Only a device or a method based on a characteristic (bio)chemical reaction, consistent with

the nature of the specific toxic effect, can guarantee permanent and satisfactory sensitivity
necessary for the protection of manpower. Only such device remains operational even if a
new, more toxic (bio)chemical warfare agent of a given CWA group appears.
It is the cholinesterase reaction specifically for nerve agents - at present the most important
group of CWAs. If we admit that a warfare agent with anticholinesterase activity which is
much more toxic than that of VX has potentially existed, then no other known device used in
chemical reconnaissance and check, with the exception of those devices which take advantage
of the cholinesterase biochemical reaction, has the required sensitivity and provides required
level of protection of the military and civilians.
1. HALÁMEK, E.: Trendy vývoje prostředků chemického průzkumu a kontroly. In: Vojenské rozhledy, 9 (41),
2000, zvláštní číslo, s. 89-94.
2. HALÁMEK, E., KOBLIHA,Z.: Poslední generace chemických zbraní. Vojenské rozhledy, 17 (49), č. 4, 2008,
s. 137-146.
152
5. Recommended reviews
1. MEYER, J.: Der Gaskampf und die chemischen Kampfstoffe. 2.ed. Verlag von S.
Hirzel. Leipzig 1926. 470 p.
2. SARTORI, M.: Die Chemie der Kampfstoffe. Friedr. Viewg & Sohn. Braunschweig
1935. 507 p.
3. ETTEL, V.: Chemická válka. Vědecký vojenský ústav. Praha 1932. 415 p.
4. PRENTISS, A.M.: Chemicals in War. Washington 1937. 740 p.
5. LOHS, K.: Synthetische Gifte. Verlag des Ministerium für Nationale Verteidigung.
Berlin 1958. 212 p.
6. TOMEČEK, I., MATOUŠEK, J.: Analýza bojových otravných látek. SPN. Praha 1961.
235 p.
7. TOMEČEK, I.: Chemie a analýza bojových otravných látek. VA Brno 1966. 166 p.
8. PŘIKRYL, F., HALÁMEK, E.: Bojové otravné látky. VVŠ PV. Vyškov 1975. 170 p.
9. FRANKE, S.: Lehrbuch der Militärchemie. 2. ed. Vol. 1 and 2. Deutscher
Militärverlag der DDR. Berlin 1977. 512/616 p.
10. STÖHR, R.: Chemische Kampfstoffe und Schutz vor chemischen Kampfstoffen.
Militärverlag. Berlin 1977. 447 p.
11. KARAKČIJEV, N.I.: Toksikologija otravljajuščich věščestv i zasčita ot jadernogo
oružija. 3. ed. Medicina. Taškent 1978. 440 p.
13. ALEXANDROV, V.N., JEMELJANOV, V.I.: Otravljajuščije věščestva. 2. ed.
Vojennoje izdatělstvo. Moskva 1990. 272 p.
14. SOMANI, S. M.: Chemical Warfare Agents. Academic Press, Inc. New York 1992.
444 p.
15. FRANKE, S., KOEHLER, K.F., KOSTYANOVSKY, R.G., KUNTSEVICH, A.D.:
Chemie der Kampfstoffe. vol I. Gesellschaft für Kampfmittelbeseitigung, Dr. Koehler
GmbH. Monster/Burg 1994. 494 p.
16. FEDOROV, L.A.: Chimičeskoje oružije v Rossii. Centr ekologičeskoj politiky Rossii.
Moskva 1994. 120 p.
17. MARRS, T.C., MAYNARD, R.L., SIDELL, F.R.: Chemical Warfare Agents,
Toxicology and Treatment. Wiley. New York 1996. 244 p.
18. CLARK, K.: The Chemical Weapons Convention: Chemical and Toxin Warfare
Agents and Disarmament. Royal Military College of Science. Shrivenham 1997.112 p.
19. PITSCHMANN, V.: Historie chemické války. Military System. Kounice 1999. 172 p.
20. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Úvod do problematiky bojových
chemických látek. AZIN CZ. Praha 2000. 61 p.
21. ELLISON, D.H.: Handbook of Chemical and Biological Warfare Agents. CRC Press.
Boca Raton 2000. 507 p.
22. PITSCHMANN, V., HALÁMEK, E., KOBLIHA, Z.: Boj ohněm, dýmem a jedy.
Military System. Kounice 2001. 178 p.
23. SOMANI, S. M., ROMANO, J.A.: Chemical Warfare Agents: Toxicity at Low Levels.
CRC Press. New York 2001. 448 p.
24. HALÁMEK, E., KOBLIHA, Z.: Přehled bojových chemických látek. VVŠ PV.
Vyškov 2002. 112 p.
25. HOENIG, S.L.: Handbook of Chemical Warfare and Terrorism. Greenwood Press.
London 2002. 178 p.
26. LEDGARD, J.B.: The Preparatory Manual of Chemical Warfare Agents. The
Paranoid Publisher Group. Colombus 2003. 265 p.
153
27. STŘEDA, L., HALÁMEK, E., KOBLIHA, Z.: Bojové chemické látky. AZIN CZ.
Praha 2004.120 p.
28. PITSCHMANN, V.: Vojenská chemie kyanovodíku. Ekoline 2004. 110 p.
29. LANGFORD, R.E.: Introduction to weapons of mass destruction. Wiley. New Jersey
2004. 394 p.
30. PITSCHMANN, V.: Analýza toxických látek detekčními trubičkami. 2.ed. Econt
Consulting. Drahelčice 2005. 194 p.
31. PATOČKA, J. a kolektiv: Vojenská toxikologie. 1.ed. Grada Publishing. Praha 2004.
178 p.
32. MATOUŠEK, J., LINHART, P.: CRBN, Chemické zbraně. Sdružení požárního a
bezpečnostního inženýrství. Ostrava 2005. 151 p.
33. Field Manual FM 3-11.9: Potential Military Chemical/Biological Agents and
Compounds. Eximdyne. Wentzeville 2005.
34. HOENIG, S.L.: Compendium of Chemical Warfare Agents. Springer. New York 2007.
222 p.
35. MARRS, T.C., MAYNERD, R.L., SIDELL, F.R.: Chemical warfare agents,
toxikology and treatment. Wiley. Chichester 2007. 738 p.
36. RICHARDT, A., BLUM, M-M.: Decontamination of warfare agents. Wiley-VCH.
Weinheim 2008, 331 p.
154
Název: Analysis of chemical warfare agents,
Autoři: Prof. Ing. Emil HALÁMEK, CSc., prof. Ing. Zbyněk KOBLIHA, CSc.,
doc. Ing. Vladimír PITSCHMANN, CSc.
Recenzenti: Ing. Ladislav Středa, CSc., Ing. Ivana TUŠAROVÁ, CSc.
Ředitel ústavu: plk. prof. Ing. Dušan VIČAR, CSc.
Rok vydání: 2009
Náklad: 50
Počet stran: 156
Vydavatel: Univerzita obrany
Tisk: Vydavatelská skupina UO
Číslo zakázky: ??/2009
Číslo EP: 54/2009
Cena pro vnitřní potřebu:
ISBN: 978-80-7231-269-6
Obsah skript byl projednán na metodickém shromáždění Ústavu ochrany proti zbraním
hromadného ničení dne . června 2009.
Skripta prošla jazykovou úpravou.
155
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Analýza bojových chemických látek /

Emil Halámek Zbynek Kobliha

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Analysis of chemical warfare agents View project

Carriers for the enzymatic detection of cholinesterase inhibitors View project

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Prof. Ing. Emil HALÁMEK, CSc.

The discovery of poison has undoubtedly been one of the first

© Halámek, E., Kobliha, Z., Pitschmann, V. Brno 2009

Ing. Ladislav Středa, CSc.

Ing. Ivana Tušarová, CSc.

PhDr. Alena Langerová

A-230 O-chlorofluoroformimino-O-(2-chloroethyl) phosphoro-

Advances in physico-chemical instrumentation used for analytical checking has long

Vyškov, April 4, 2007

Preface to the amended and revised second edition

Vyškov, April 24, 2009

1.1 Objectives and duties of chemical reconnaissance and chemical check

Table 1: System of technical devices used in chemical reconnaissance and check

Type B – Detector of liquid CWAs

Type C – Chemical detector

Type D – Fast detector of CWAs

Type E – Portable field chemical laboratory

Type F – Mobile chemical laboratory

Type G – Long-range detector of CWAs

Phosgene is a typical acylating and halogenating compound easily undergoing

(H3C)2N C N(CH3)2 2 NH(CH3)2 Cl

(H3C)2N CH N(CH3)2 N(CH3)2

(H3C)2N (H3C)2N CH N(CH3)2 Cl

(H3C)2N C N(CH3)2 Cl H2O

COCl2 2 O2N CH2 N

O2N CH2 N C N CH2 NO2 2 Cl

The formed condensation product of substituted urea character is converted by

3 H2O CO2 2 H2O CO2

COCl2 4 NH NH2 O C 2 NH NH3 Cl

COCl2 2 NaOH CO2 2 NaCl H2O

Analogously, the argentometric determination of formed chlorides is possible by titration with

Diphosgene as well as its chemical properties is related to phosgene. From the

References to chapter 2.1-2.2

Chloropicrin (trichloronitromethane) reacts readily with aqueous-alcoholic solutions

Cl3C NO2 4 KI I C I 3 KCl KNO2

NO2 HCl HNO2 Cl

2 Cl3C NO2 KCN O2N C C NO2 Cl CN KCl

References to chapter 2.3

SCN - [FeSCN] 2+ [Fe(SCN)2] +

SCN - [Fe(SCN)2] + Fe(SCN)3

Its sensitivity is 10 µg.ml−1 in a sample or up to 5 µg.l−1in air.

2 Cu(CN)2 2 CuCN (CN)2

2 (H3C)2N CH2 N(CH3)2 (CN)2

The arising products of mesoquinoid character are stabilized in charge-transfer complexes.

HCN C NO2 NC C NO2

HCN O2N CH C NO2

red colour pH < 10

O2N S S NO2 HCN H2O HOCN 2 HS NO2

HOOC COOH COOH

O2N NO2 O2N NH OH O

Another possibility is the titration with a nickel(II) salt solution to dimethylglyoxime as a

The nickel salt solution is usually standardized chelatometrically [34].

References to chapter 2.4

Although cyanogen chloride is genetically related to hydrogen cyanide, formally it is

ClCN 2 NaOH NaCl NaOCN H2O

Cyanates do not undergo reactions characteristic of cyanides. Similarly to other halides,