Determination of blood phosphatidylethanol by liquid chromatography tandem mass spectrometry / 中华检验医学杂志

ABSTRACT

Objective To establish a method for measuring blood phosphatidylethanol by liquid chromatography coupled with tandem mass spectrometry(LC-MS/MS), which can be applied for objective and quantitative of alcohol intake.Methods Whole blood samples were treated with isopropanol to precipitate protein,and phosphopropanol(160/160)was used as the standard.After centrifugation, the supernatants were transferred and evaporated under a stream of nitrogen until dryness.Then the residuals were analyzed by LC-MS/MS.Various methodological parameters, including linearity, limit of detection (LOD), limit of quantitation(LOQ), recovery, and precision, were investigated.Finally, blood samples from 40 Chinese individuals with more than one year of regular drinking habits were analyzed, and distributions of phosphatidylethanol were evaluated.Results The correlation coefficients were higher than 0.9992.The LOD and LOQ were lower than 0.74 and 2.48 ng/ml, respectively.The inter-and total assay coefficient of variations were 0.77%-3.18% and 2.30%-6.95%, respectively, with recoveries ranged from 96.88% to 102.99%.The relationship between phosphatidylethanol level and self-reported alcohol consumption was significantly and positively correlated(r =0.769, P <0.001).Furthermore, Kruskal-Wallis analysis showed a significant difference in total phosphatidylethanol levels among individuals with different levels of alcohol intake(χ2=18.850,P<0.001).Conclusions An LC-MS/MS method for whole blood phosphatidylethanol detection has been developed.This method is simple,sensitive and accurate and can effectively reflect light,moderate and heavy alcohol intake.The method will be applied to the assessment of alcohol consumption and its association with the risks of drinking related diseases.