The method successfully detected and measured the CPF as well as the antidotes concentrations along the different times tested. Samples were collected in different periods after the antidote treatment (15 min, 30 min, 1 h, 2 h, 3 h, 4 h and 24 h). The validated method was applied to blood samples from rats acutely intoxicated with an OP, chlorpyrifos (CPF) and treated 1 h after poisoning with the antidotes, ATR and 2-PAM, separately or in combination.
The intra-assay accuracy results were within 82.8% (pirimiphos-methyl) to 111.8% (malathion) and inter-assay accuracy ranged from 84.9% (pirimiphos-methyl) to 111.6% (malathion). Intra-assay precision ranged from 1.2% (atropine) to 14.6% (chlorpyrifos), while inter-assay precision ranged from 4.4% (pyrazophos) to 17.2% (chlorpyrifos). Linearity results as well as limits of detection and quantification were within the desired range for all analytes tested. Method validation was achieved using blank blood samples from rats, followed by deproteinization with cold acetonitrile. This study aimed to develop a fully validated LC-MS/MS method for combined analysis of multiple OP compounds and antidotes in blood samples. Despite the profusion of methods described for analysis of either OP compounds or antidotes, to our knowledge, there is no method available that allows simultaneous identification and monitoring of these compounds.
In cases of OP poisoning, identifying the toxic agent, as well as monitoring antidotes and toxic agent's concentration along time may be of great importance for therapeutic success and survival, particularly in view of the marked hemodynamic changes present in the poisoning state, which could interfere with OPs and antidotes levels achieved. Standard treatment after acute OP poisoning employ the antidotes, atropine (ATR), to prevent cholinergic effects and oximes, such as the pralidoxime (2-PAM), to reactivate the inhibited AChE.
These compounds promote inhibition of the acetylcholinesterase (AChE) leading to the accumulation of acetylcholine in the central and peripheral cholinergic synapses. Organophosphorus (OP) compounds stand out as one of the most extensively used pesticides' class worldwide, being associated with a high prevalence of voluntary and accidental poisoning in humans.
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