Quantitative determination of phosphatidylethanol in dried blood spots for monitoring alcohol abstinence.

Luginbühl, Marc; Stöth, Frederike; Schröck, Alexandra; Gaugler, Stefan; Weinmann, Wolfgang (2021). Quantitative determination of phosphatidylethanol in dried blood spots for monitoring alcohol abstinence. Nature protocols, 16(1), pp. 283-308. Nature Publishing Group 10.1038/s41596-020-00416-x

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Phosphatidylethanol (PEth), which is formed by enzymatic reaction between ethanol and phosphatidylcholine, is a direct marker for alcohol usage. PEth has a long elimination half-life (~5-10 d) and specimens can be sampled using minimally invasive microsampling strategies. In combination with rapid analysis procedures PEth has proved to be advantageous for the detection of abstinence over other direct (e.g., ethyl glucuronide in blood, urine or hair) and indirect (e.g., carbohydrate-deficient transferrin in serum) alcohol markers. Although PEth determination is widely applied around the world, laboratory protocols are not standardized. Here we provide general guidelines for the analysis of PEth in dried blood spots (DBSs), including reference material evaluation, synthesis of a deuterated internal standard, preparation of calibration samples (reference material in teetotaller blood), and analyte separation and detection. The protocol contains information to extract the DBSs either manually or with a fully automated autosampler. Extraction of the analytes from DBS filter paper cards is performed using an organic extraction, followed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). For accurate and reliable measurement of PEth, the two most abundant analogs, PEth 16:0/18:1 and PEth 16:0/18:2, are quantified. We show data that provide guidelines on how to interpret the results for both demographic studies and forensic applications. The described protocol can be applied by experienced laboratory staff with basic LC-MS/MS knowledge and takes 2 d to perform.

Item Type:

Journal Article (Further Contribution)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute of Legal Medicine
04 Faculty of Medicine > Service Sector > Institute of Legal Medicine > Forensic Chemistry and Toxicology

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Stöth, Frederike Theresa and Weinmann, Wolfgang

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1754-2189

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Antoinette Angehrn

Date Deposited:

28 Jan 2021 11:47

Last Modified:

19 Apr 2021 17:22

Publisher DOI:

10.1038/s41596-020-00416-x

PubMed ID:

33288956

BORIS DOI:

10.48350/150503

URI:

https://boris.unibe.ch/id/eprint/150503

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