Nonaqueous capillary electrophoresis for analysis of the ethanol consumption biomarker phosphatidylethanol.
Nonaqueous CE (NACE) methodology was developed for the separation and determination of phosphatidylethanol (Peth), a new biomarker of ethanol intake. Peth is an abnormal phospholipid formed in cell membranes only in the presence of ethanol, via the transphosphatidylation reaction of phospholipase D. The NACE separation medium consisted of 80 mM ammonium acetate in 50% ACN, 33% 2-propanol, 12% hexane and 5% methanol. A stacking effect was obtained by reducing the concentration of ammonium acetate in the separation medium for all injected samples. The LOD was estimated to 1 microM (5.6 fmol) of Peth with conventional UV detection, equalling 0.4 micromol/L blood. Peth was successfully determined in extracts of human blood samples. Separation of Peth from other blood lipids in the lipid extract sample was performed in 5 min. The method facilitates smaller sample volumes and performs about ten times faster compared to earlier chromatographical methods.
Varga
Arthur
A
Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, Sweden. arthur.varga@tbiokem.lth.se
Nilsson
Staffan
S
D000428:Alcohol Drinking; D015415:Biomarkers / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D020404:Glycerophospholipids / Q000032:analysis*; D006801:Humans; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D015415:Biomarkers; D020404:Glycerophospholipids; C051521:phosphatidylethanol