A rapid, accurate and specific proton nuclear magnetic resonance ((1)H NMR) spectroscopic method is developed to determine ethanol in blood, known as the blood alcohol concentration (BAC). The limits of detection and quantification are 0.02g/L and 0.07g/L, respectively. The (1)H NMR spectra show linearity for whole blood and serum samples of a concentration range of 0.00-3.00g/L (R(2)>0.9995). The (1)H NMR method is applied and validated for whole blood as the sample media. Real driving under influence case samples are analyzed with the reference enzyme-based alcohol dehydrogenase and headspace gas chromatography techniques by the Forensic Medicine in Bonn. The reference results are compared with the (1)H NMR spectroscopic results. The validation and comparison indicate that (1)H NMR is suitable for the quantification of BAC in whole blood. This technique has the advantages of automated analysis with good measurement precision and fast sample throughput. A drop of blood (V=20μL) is adequate for an analysis leading to a possible simplification of the sample collection. Due to the non-destructive method, follow-up examinations by (1)H NMR spectroscopy or DNA determinations by different techniques (PCR, in situ hybridization) are possible in resolving legal disputes.
Keywords: BAC; Blood alcohol concentration; Deuterium oxide (PubChem CID: 24602); Dimethylsulfone (PubChem CID: 6213); Ethanol (PubChem CID: 702); Forensic analysis; Nuclear magnetic resonance; Quantitative (1)H NMR; Validation.
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