Simultaneous determination of ethanol's four types of non-oxidative metabolites in human whole blood by liquid chromatography tandem mass spectrometry.
The importance of ethanol non-oxidative metabolites as the specific biomarkers of alcohol consumption in clinical and forensic settings is increasingly acknowledged. Simultaneous determination of these metabolites can provide a wealth of information like drinking habit and history, but it was difficult to achieve because of their wide range of polarity. This work describes development and validation of a simple liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for 4 types of ethanol non-oxidative metabolites (ethyl glucuronide, ethyl sulfate, fatty acid ethyl esters and phosphatidylethanols) in 50 μL of human whole blood. Pretreatment method, column and MS conditions were optimized. For the first time, the four types of ethanol non-oxidative metabolites with enormous discrepancies of property were simultaneously extracted and analyzed in one run within 40 min. The limits of detections (LODs) were among 0.1-10 ng/mL, and good linearity was obtained. Deviations in precision and accuracy were all lower than 15% at three QC levels. This method was then applied to two forensic samples, resulting in information on drinking habits and drinking time which were very useful for the interpretation of the blood alcohol results.
Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, PR China; Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, PR China.
Xinyu
X
Zhang
Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, PR China.
Feng
F
Zheng
Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, PR China.
Zebin
Z
Lin
Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Frederik V's Vej 11, 2100, Copenhagen, Denmark.
Sys Stybe
SS
Johansen
Department of Clinical Medicine, Shanghai Medical College, Fudan University, Shanghai, 200032, PR China.
Tianfang
T
Yu
Department of Clinical Medicine, Shanghai Medical College, Fudan University, Shanghai, 200032, PR China.
Yuming
Y
Liu
Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, PR China.
Zhibin
Z
Huang
Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, PR China.
Jiaolun
J
Li
Department of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha, 410013, Hunan, PR China.
Jie
J
Yan
Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, PR China. Electronic address: yulan_rao@fudan.edu.cn.
Yulan
Y
Rao
D000328:Adult; D000428:Alcohol Drinking / Q000097:blood; D001774:Blood Chemical Analysis / Q000379:methods*; D002853:Chromatography, Liquid; D000431:Ethanol / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism*; D044707:Forensic Sciences; D006184:Habits; D006801:Humans; D057230:Limit of Detection; D008297:Male; D053719:Tandem Mass Spectrometry; D013997:Time Factors
D000431:Ethanol
Ethanol; Human whole blood; LC-MS/MS; Non-oxidative metabolites