Stability of PEth 16:0/18:1, 16:0/18:2, 16:0/20:4, 18:0/18:1, 18:0/18:2, and 18:1/18:1 in authentic whole blood samples (at room temperature)

This article traces the historical development of various biomarkers of acute and/or chronic alcohol consumption. Much of the research in this domain of clinical and laboratory medicine arose from clinics and laboratories in Sweden, as exemplified by carbohydrate deficient transferrin (CDT) and phosphatidylethanol (PEth). Extensive studies of other alcohol biomarkers, such as ethyl glucuronide (EtG), ethyl sulfate (EtS), and 5‐hydroxytryptophol (5‐HTOL), also derive from Sweden. The most obvious test of recent drinking is identification of ethanol in a sample of the person's blood, breath, or urine. However, because of continuous metabolism in the liver, ethanol is eliminated from the blood at a rate of 0.15 g/L/h (range 0.1–0.3 g/L/h), so obtaining positive results is not always possible. The widow of detection is increased by analysis of ethanol's non‐oxidative metabolites (EtG and EtS), which are more slowly eliminated from the bloodstream. Likewise, an elevated ratio of serotonin metabolites in urine (5‐HTOL/5‐HIAA) can help to disclose recent drinking after ethanol is no longer measurable in body fluids. A highly specific biomarker of hazardous drinking is CDT, a serum glycoprotein (transferrin), with a deficiency in its N‐linked glycosylation. Another widely acclaimed biomarker is PEth, an abnormal phospholipid synthesized in cell membranes when people drink excessively, having a long elimination half‐life (median ~6 days) during abstinence. Research on the subject of alcohol biomarkers has increased appreciably and is now an important area of drug testing and analysis.

Brief history of the alcohol biomarkers CDT, EtG, EtS, 5‐HTOL, and PEth

Jones

2023

Storage stability of phosphatidylethanol homologues in whole blood and dried blood spots of nonalcoholics at different temperatures over 60 days

Herzog,

Skopp,

Musshoff

et al. 2023

Phosphatidylethanol (PEth) has recently become a popular direct alcohol marker for evaluating drinking behavior. This study aimed at gaining further information on the long‐term stability of five PEth homologues (16:0/18:1, 16:0/18:2, 16:0/20:4, 18:0/18:1, 18:0/18:2) in whole blood (WB) and dried blood spots (DBS) stored at −80°C, 4°C, and room temperature (18°C) over a period of 60 days. Venous blood was taken from 10 volunteers (five females and five males, aged 21–40 years) with a moderate drinking behavior and a negative breath alcohol test at the time of collection. 100 μL aliquots of WB were prepared in addition to 20 μL DBS samples. The initial PEth concentrations were determined on the day of the blood collection. On days 1, 3, 5, 7, 11, 17, 40, and 60, DBS were analyzed in triplicate by means of LC–MS/MS. On these days, WB aliquots having been stored until that time were used to create further DBS in triplicate, which were subsequently stored at 18°C and analyzed in a single batch after day 60. All homologues, except PEth 16:0/20:4, were stable at −80°C in DBS and WB for 60 days. The initial PEth 16:0/18:1 concentrations remained stable in both DBS and WB in all but one volunteer's specimen at 4 and 18°C. Apart from this exception, simultaneously detected PEth homologues 16:0/18:2, 18:0/18:1, and 18:0/18:2 remained stable over at least 40 days in DBS. Nevertheless, the storage time between sample collection and analysis should be kept as short as possible if an ethanol‐free sample cannot be ensured.

Interlaboratory comparison of phosphatidylethanol in dried blood spots using different sampling devices

Bantle,

Stöth,

Weinmann

et al. 2024

Phosphatidylethanol (PEth) has become an important marker to assess drinking behaviour and monitor abstinence. Despite its increasing use, knowledge on robustness and standardization and comparability of methods and results are still limited. In 2022, the first international consensus for the use of PEth and its interpretation was published. To establish an experience‐based foundation for further harmonization, three rounds of interlaboratory comparison using microsamples were conducted. Participating laboratories sent their sampling devices to the laboratory of Forensic Toxicology at the University of Bern, where for each round, four different authentic blood samples were applied to the devices and sent back. The PEth 16:0/18:1 target concentrations covered a range between 16 and 474 ng/mL (0.023 and 0.676 μmol/L, respectively) and included sample concentrations close to the decision limits of 20 and 200 ng/mL (0.025 and 0.28 μmol/L, respectively). Evaluation of the results based on guidelines by Horwitz and the Society of Toxicological and Forensic Chemistry (GTFCh) showed that 73% of all participating laboratories quantified and reported all samples (N = 4 for each round) within the acceptable limits. More than 90% quantified and reported at least one sample within the acceptable limits.