Development and validation of headspace gas chromatography with a flame ionization detector method for the determination of ethanol in the vitreous humor.

Introduction: Qualitative and quantitative testing of ethanol in post-mortem samples is an important analytical procedure that provides accurate, precise, and reliable results. Given the complexity of the issue, obtaining a realistic picture of lifelong alcoholemia requires supporting blood ethanol findings with analyses of alternative samples, primarily vitreous humor (VH).

Objective: The objective of this study was to develop and validate a headspace gas chromatography with flame ionization detection (HS/GC-FID) method for determining ethanol concentration in VH.

Materials and methods: Conditions for the HS/GC-FID method were established and the method was validated according to the guidelines of the European Medicines Agency. Validation parameters such as precision, accuracy, specificity, sensitivity, and linearity over a wide concentration range were evaluated through statistical analysis.

Results: The method demonstrated precision, accuracy, specificity, and sensitivity. Additionally, it proved to be linear across a wide concentration range and relatively fast, making it suitable for rapid and routine determination of ethanol concentration in VH, particularly for forensic applications.

Conclusion: Results from validation and application of the method to VH samples indicate that ethanol concentration in VH can be reliably determined using the presented HS/GC-FID method, making it a valuable tool in forensic investigations.