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Determination of alcohol in human biological fluids An alternative alcotest Panagiota Papaspyridakou, Pinelopi Moutzouri, Christos Kontoyannis and Malvina Orkoula* Department of Pharmacy, University of Patras ICE/HT-FORTH, Patras, Greece Ethanol is contained in beverages and affects the central neural system (CNS) with many consequences to health. The determination of ethanol in biological fluids is considered an important issue in toxicology and forensic sciences. Quantitative analysis of ethanol is mainly conducted with gas chromatography (GC-MS), which is accurate but also time consuming, expensive and requires expert personnel. A lowcost and simple way to measure alcohol is by using breathalyzers, applied on-site for intoxication testing though is often disputed by drivers. In the current project, the detection and quantitative analysis of ethanol in human biological fluids (saliva, urine and blood serum) is attempted employing Raman spectroscopy. It is an easy to handle, fast but reliable technique and does not require sample preparation. Furthermore, it can be applied on-site using portable systems. A special cell was designed for the prevention of evaporation of volatile ethanol during spectral acquisition. It comprised of a highly reflective glass slide carrying a cavity in the centre where the sample is placed and covered by a transparent membrane. Biological fluids (urine, blood serum and saliva) were collected from healthy volunteers and mixtures with ethanol were prepared in concentrations ranging from zero to the lethal alcohol dosage (0-5μl/ml). Accurately performed quantitative analysis showed that the Limit of Detection was 0,29μl/ml in blood serum, which is lower than the threshold for intoxication (1,27μl/ml) and close to the limits for conviviality and dizziness (0,63μl/ml). Finally, biological fluids were collected from healthy volunteers after beverage consumption and analyzed. The results were in accordance with the measured alcohol concentration with a breathalyzer. Key words: Biological fluids, ethanol, alcohol testing, Raman spectroscopy