Abstract
In this work a novel bioassay for mustard agent detection was proposed. The bioassay is based on the capability of these compounds to inhibit the enzyme choline oxidase. The enzymatic activity, which is correlated to the mustard agents, was electrochemically monitored measuring the enzymatic product, hydrogen peroxide, by means of a screen-printed electrode modified with Prussian Blue nanoparticles. Prussian Blue nanoparticles are able to electrocatalyse the hydrogen peroxide concentration reduction at low applied potential (−50 mV vs. Ag/AgCl), thus allowing the detection of the mustard agents with no electrochemical interferences. The suitability of this novel bioassay was tested with the nitrogen mustard simulant bis(2-chloroethyl)amine and the sulfur mustard simulants 2-chloroethyl ethyl sulfide and 2-chloroethyl phenyl sulfide. The bioassay proposed in this work allowed the detection of mustard agent simulants with good sensitivity and fast response, which are excellent premises for the development of a miniaturised sensor well suited for an alarm system in case of terrorist attacks.
Highlights
Chemical warfare agents (CWAs) have been deliberately produced and employed in the battlefields during the 20th century with the purpose of killing or debilitating living organisms
From an analytical point of view, these mechanisms should be clarified since the reversible inhibition does not require incubation time and the measurement is characterised by a short time of analysis
The obtained results have demonstrated the possibility of determining nitrogen and sulfur mustard agents using a portable and easy-to-use system based on the inhibition of choline oxidase, extending the few biochemical studies present in the literature on the inhibitory effect of mustards on choline oxidase only focused on nitrogen mustards [18,28,29]
Summary
Chemical warfare agents (CWAs) have been deliberately produced and employed in the battlefields during the 20th century with the purpose of killing or debilitating living organisms. Several studies on battlefield victims have demonstrated that exposure to mustards is a traumatic event having long-lasting effects on mental health [5] For these reasons, MAs remain some of the agents of highest internationally concern and are receiving increasing attention regarding their decontamination and degradation, but in particular for their detection in water and food or in human biological fluids. We developed an amperometric bioassay to reveal sulfur and nitrogen mustards based on their capability to inhibit the enzyme choline oxidase This inhibitory effect was demonstrated in the work of Barron et al in 1948, focused on the effects of mustards on the structural properties and chemical activity of different enzymes involved in metabolic processes. Choline oxidase from Alcaligenes sp. was chosen as biological recognition element for the development of the bioassay, and the simulants nitrogen mustard bis(2-chloroethyl)amine and sulfur mustards 2-chloroethyl ethyl sulfide and 2-chloroethyl phenyl sulfide (Scheme 1) were selected as target analytes and amperometrically detected by following the enzymatic product hydrogen peroxide
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