Validation of a new method for the detection of Ethyl glucuronide in larvae of Lucilia sericata as a marker of ante-mortem alcohol consumption

Abstract

Larvae and insects are an important and alternative biological matrix in the development anpost-mortem forensic toxicology. They are very useful when conventional matrices are not available, in particular when the loss of biological fluids, due to the decomposition of corpses or to a traumatic death, occurs. The purpose of this study is to develop and validate an analytical method in Ultra High-Performance Liquid Chromatography at High Resolution Mass Spectrometry (HPLC/HR-MS) for the research and quantification of Ethyl glucuronide (EtG) on larvae. The criteria taken into consideration for the validation are linearity, quantitation limits (LOD and LLOQ), accuracy, precision, carryover, interferences and ionization suppression/enhancement. The method was shown to be linear within the tested range, with a coefficient of determination higher than 0.99. LOD was 2 pg mg−1, while LLOQ was 5 pg mg-1. Accuracy, precision and ionization/suppression enhancement fulfilled the criteria indicated in the guidelines used for the validation. The establishment and validation of this method allowed the identification of Ethyl glucuronide on the larvae of Lucilia sericata (Calliphoridae) of a subject found death in an advanced state of decomposition.

Keywords

• Entomotoxicology
• Ethanol
• Forensic toxicology
• Ethyl glucuronide
• Ultra-high-performance liquid cromatography
• Larvae