Impact of using Sarcophaga (Liopygia) argyrostoma (Robineau-Desvoidy, 1830) as a toxicological sample in detecting clonazepam for forensic investigation

Abstract

Abstract Background Sarcophagidae along with Calliphoridae (superfamily Oestroidea) were known as important colonizers of cadavers and could be used to estimate the postmortem interval (PMI), through tracking the flies colonizing pattern and/or monitoring their growth rates on cadavers. Many previous researches discussed the impact of toxins in decomposing corpses, on the developmental stages of insects, which would affect the accuracy of PMI estimation. Clonazepam belongs to the benzodiazepines, and it is one of the most routinely used drugs to control humans’ seizures. The current study aimed to investigate the effect of clonazepam on the developmental stages of Sarcophaga argyrostoma, one of the most widespread Sarcophagidae in Giza Governorate. Also, we investigate the ability of these developmental stages to detect the drug. Results The current study used different concentrations of clonazepam (25, 50, and 100 mg/ml) as an in vitro application of the flesh fly S. argyrostoma. The drug has affected significantly the morphological measurements (weight, length, and width) of the different developmental stages, especially in the highest concentration (100 mg/ml). Quantitative and qualitative analyses were applied, by using high-performance liquid chromatography-mass spectrometry (HPLC–MS) and protein carbonyls amount respectively, to detect clonazepam in the second, early third, and late third larval instars and prepupa of S. argyrostoma. The results showed that the relationship between the concentration of the drug and its detection in the same developing instar is interdependent, using HPLC–MS. However, the drug faded from instar to the following one and transformed to its metabolite form. Measuring the protein carbonyls amount (OD/mg protein/min) revealed an elevation in the macromolecules damage, compared to the control groups, in almost all treated groups. Conclusions The current data suggested that clonazepam has oxidative damage in S. argyrostoma. While HPLC–MS was efficient in measuring the concentration of the drug in the insect, protein carbonyls analysis was a time- and cost-saving method and could be used to detect the drug in insects qualitatively. Graphical abstract

Keywords

• Forensic entomology
• Development
• Entomotoxicology
• Sarcophagidae
• Enzodiazepines