Detection of benzodiazepines in decomposing rabbit tissues and certain necrophagic dipteran species of forensic importance

The determination of benzodiazepines (carbamazepine and clobazam) in rabbit carcass tissues and larvae of three Calliphorid flies is described. After feeding the rabbits with lethal and toxic doses, samples of larvae and carcass tissues were studied. Residual drugs were determined using Ultra-high-performance liquid chromatography – quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS). Benzodiazepines and its main active metabolites have been detected in the rabbit tissues at different retention times depending on the dosage used (lethal or toxic). A total of 1150 insects were collected and 800 larvae of the flies Chrysomya albiceps, Lucilia sericata and L. silvarum were used in the analysis. The presence of benzodiazepines in the rabbit tissues has been shown to typically affect the larval development cycle of the three necrophagous flies. Chrysomya albiceps larvae feed on drugs developed faster, while the development of L. sericata and L. silvarum larvae slowed. These results indicate that drugs have an impact on the life cycles of insects, which suggests that the presence of these substances is a factor that needs to be taken into account when estimating the post-mortem interval (PMI).