Development and Validation of a Method for Quantitative Determination of Phosphatidylethanol in Whole Blood

Introduction. Phosphatidylethanol (PEth) is a phospholipid slowly forming in cell membranes as a result of the phospholipase D enzymatic reaction between phosphatidylcholine and ethanol. As a result of the analysis of published scientific papers, high performance liquid chromatography with mass spectrometric detection is used to determine phosphatidylethanol as a method with high specificity, accuracy, precision and the ability to determine substances at minimum concentrations, which is fundamental for clinical laboratory diagnostics when monitoring alcohol abuse in the population. Aim. The aim of the study was to develop and validate a method for the quantitative determination of a direct marker of alcohol abuse, phosphatidylethanol, in human blood. Materials and methods. Phospholipid was quantified by high performance liquid chromatography with tandem mass spectrometry. Results of the study and their discussion. The developed method was validated for the following parameters: selectivity, lower limit of quantitation, linearity, sample transfer, accuracy, precision, matrix effect, stability. The analytical range was 0.0025 µmol/L – 3.0 µmol/L; LPCO level PEth 16:0/18:1 0.0025 was equal to µmol/l. Conclusions. The developed method for the quantitative determination of phosphatidylethanol is suitable for use in laboratory diagnostics in order to monitor the population in the analysis of alcohol abuse.