Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are specific and sensitive biomarkers for the diagnosis of acute or chronic alcohol abuse. Due to postmortem alcohol production in biological tissues, they have recently been evaluated as potential biomarkers of ethanol ingestion. This in vitro study aimed to evaluate all developmental stages of the fly Calliphora vicina L. (Calliphoridae). Different pig substrates were used during the in vitro experiments to rear C. vicina Experiments were performed to: (i) assess the presence of EtG and EtS in larvae, pupae and insects; (ii) study variability due to the substrates characteristics; (iii) evaluate the possibility of false positives due to external alcohol contamination; and (iv) estimate the potential metabolism of ethanol by the insects. EtS was found in all of the samples where the standard was added to the substrate. Muscle provided the most reliable results. EtS was found in larvae, pupae and puparia. EtG and EtS were found in larvae and pupae collected from the body of an alcoholic found dead in his home. This study showed that maggots, pupae and puparia could be a useful matrix for the evaluation of antemortem alcohol ingestion.
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