Concentration units used to report blood- and breath-alcohol concentration for legal purposes differ between countries which is important to consider when blood/breath ratios of alcohol are compared and contrasted.

This technical note reviews the plethora of concentration units used to report blood-alcohol concentration (BAC) and breath-alcohol concentrations (BrAC) for legal purposes in different countries. The choice of units sometimes causes confusion when scientific papers originating from a certain country might be introduced into evidence via expert testimony, such as when alcohol-related crimes are prosecuted. The concentration units are also important to consider when blood/breath ratios (BBRs) of alcohol are calculated and compared between countries. Statutory BAC limits for driving in most nations are reported in mass/volume (m/v) units, such as g/100 mL (g%) in the United States, mg/100 mL (mg%) in the United Kingdom and Republic of Ireland, or g/L (mg/mL) in many EU nations. By contrast, Germany and the Nordic countries report BAC as mass/mass (m/m) units, hence g/kg or mg/g, which are ~5.5% lower than m/v units, because whole blood has an average density of 1.055 g/mL. There are historical reasons for reporting BAC in mass/mass units because the aliquots of blood analyzed were measured by weight rather than volume. The difference between m/m and m/v is also important in postmortem toxicology, such as when distribution ratios of ethanol between blood and other biological specimens, such as urine, vitreous humor, and cerebrospinal fluid, are reported.

Dubowski's stages of alcohol influence and clinical signs and symptoms of drunkenness in relation to a person's blood-alcohol concentration-Historical background.

This article traces the origin of various charts and tables delineating the stages of alcohol influence in relation to the clinical signs and symptoms of drunkenness and a person's blood-alcohol concentration (BAC). In forensic science and legal medicine, the most widely used such table was created by Professor Kurt M. Dubowski (University of Oklahoma). The first version of the Dubowski alcohol table was published in 1957, and minor modifications appeared in various articles and book chapters until the final version was published in 2012. Seven stages of alcohol influence were identified including subclinical (sobriety), euphoria, excitement, confusion, stupor, alcoholic coma and death. The BAC causing death was initially reported as 0.45+ g%, although the latest version cited a mean and median BAC of 0.36 g% with a 90% range from 0.21 g% to 0.50 g%. An important feature of the Dubowski alcohol table was the overlapping ranges of BAC for each of the stages of alcohol influence. This was done to reflect variations in the physiological effects of ethanol on the nervous system between different individuals. Information gleaned from the Dubowski table is not intended to apply to any specific individual but more generally for a population of social drinkers, not regular heavy drinkers or alcoholics. Under real-world conditions, much will depend on a person's age, race, gender, pattern of drinking, habituation to alcohol and the development of central nervous tolerance. The impairment effects of ethanol also depend to some extent on whether observations are made on the rising or declining phase of the blood-alcohol curve (Mellanby effect). There will always be some individuals who do not exhibit the expected behavioral impairment effects of ethanol, such as regular heavy drinkers and those suffering from an alcohol use disorder.

Bibliometric evaluation of Journal of Analytical Toxicology as a scholarly publication according to the Web-of-Science citation database.

Soon approaching its 50th anniversary, Journal of Analytical Toxicology (JAT) is an international scholarly publication specializing in analytical and forensic aspects of toxicology. Science Citation Index (SCI) and Journal Citation Reports (JCR), both of which are part of the Web-of-Science (WOS) database, were used to make a bibliometric evaluation of JAT articles. Between 1977 (volume 1) and 2023 (volume 47), a total of n = 4,785 items were published in JAT; the top-ten most highly cited articles and the most prolific authors were identified. Changes in the journal impact factor (JIF) were studied between 1997 and 2022, and this metric varied from a low of 1.24 (2006) to a high of 3.36 (2020).The most recent JIF (2022) dropped to 2.5 and the corresponding 5 year JIF was 2.6. JAT's most highly cited article (590 cites) was a working group (SWGTOX) report dealing with standard practices for the validation of analytical methods in forensic toxicology laboratories. JAT published 62 articles each of which were cited over 100 times and the H-index for JAT was 89. The most prolific author of JAT articles was credited with 119 items, the first in 1980 (volume 4) and the latest in 2023 (volume 47). JAT articles were cited 4,537 times in 2022 by all journals in the JCR database, although 520 of these were self-citations (11.5%). Bibliometric methods are increasingly used to evaluate the published work of individual scientists, university departments, entire universities and whole countries. Highly cited articles are considered more influential and authoritative compared with papers that are seldom or never cited.

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

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.

Bibliometric evaluation of Forensic Science International as a scholarly journal within the subject category legal medicine.

This article presents a bibliometric evaluation of Forensic Science International (FSI) as a scholarly journal within the "legal medicine" subject category. Citation data were retrieved from Science Citation Index (SCI) and Journal Citation Reports (JCR), both of which are part of the Web-of-Science (WOS) database. The most cited articles in FSI were identified along with the most prolific authors. The current journal impact factor (JIF) of FSI is 2.2, which was in good agreement with the 5-year JIF of 2.3. FSI was ranked fourth among 17 journals within the legal medicine subject category. Since 1979, a total of 209 FSI articles were cited over 100 times and the H-index for times cited was 125. Although widely used in academia, bibliometric methods might also prove useful in jurisprudence, such as when evaluating the research and publications of people proposed as expert witnesses.

Who are the most highly cited forensic scientists in the United States?

The most highly cited forensic practitioners in the United States were identified using a publicly available citation database that used six different citation metrics to calculate each person's composite citation score. The publication and citation data were gleaned from Elsevier's SCOPUS database, which contained information about ~7 million scientist each of whom had at least five entries in the database. Each individual was categorized into 22 scientific fields and 176 subfields, one of which was legal and forensic medicine (LFM). The database contained citation records for 13,388 individuals having LFM as their primary research discipline and 282 of these (2%) were classified as being highly cited. Another 99 individuals in the database had LFM as their secondary discipline, making a total of 381 highly cited forensic practitioners from 35 different countries. The career-long publication records of each individual were compared using their composite citation scores. Of the 381 highly cited scientists, 93 (24%) had an address somewhere in the United States. The various branches of forensics they specialized in were anthropology, criminalistics, DNA/genetics, odontology, pathology, statistics/epidemiology, and toxicology. The two most highly cited scientists, according to their composite citation score, were both specialists in DNA/genetics. Bibliometric methods are widely used for evaluating research performance in academia and a similar approach might be useful in jurisprudence, such as when an expert witness is instructed to testify in court and explain the meaning of scientific evidence.

Population Pharmacokinetics as a Tool to Reevaluate the Complex Disposition of Ethanol in the Fed and Fasted States.

The pharmacokinetics (PK) of ethanol are important in pharmacology and therapeutics because of potential drug-alcohol interactions as well as in forensic science when alcohol-related crimes are investigated. The PK of ethanol have been extensively studied since the 1930s, although some issues remain unresolved, such as the significance of first-pass metabolism, whether zero-order kinetics apply, and the effects of food on bioavailability. We took advantage of nonlinear mixed-effects modeling to describe blood-alcohol concentration (BAC) profiles derived from 3 published clinical studies involving oral, intraduodenal, and intravenous administration of ethanol with and without food. The overall data set included 1510 BACs derived from 72 healthy subjects (60 men, 12 women) aged between 20 and 60 years. Two-compartment models with first-order absorption and Michaelis-Menten elimination kinetics adequately described the BAC profiles. Food intake had 2 separate effects: It reduced the absorption rate constant and accelerated the maximum elimination rate. Estimates of the maximum elimination rate (fasted) and the food effect (as a factor) were 6.31 g/h (95%CI, 6.04-6.59 g/h) and 1.39-fold (95%CI, 1.33-1.46-fold), respectively. Simulations showed that the area under the BAC-time curve (AUC) was smaller with lower input rate of ethanol, irrespective of any first-pass metabolism. The AUC from time 0 to 10 hours for a 75-kg subject was 2.34 g â€¢ h/L (fed) and 3.83 g â€¢ h/L (fasted) after an oral dose of 45 g ethanol. This difference was mainly attributable to the food effect on ethanol elimination and depended less on the absorption rate. Our new approach to explain the complex human PK of ethanol may help when BAC predictions are made in clinical pharmacology and forensic medicine.

Insulin murder and the case of Colin Norris.

Although insulin is an essential medicine and a life-saving drug, it has also been incriminated in many poisoning deaths; accidental, suicidal and some with malicious intent. Overdosing with insulin precipitates a life-threatening state of hypoglycemia and if untreated leads to coma, irreversible brain damage and death. Normally, the pancreatic ß-cells secrete equimolar amounts of insulin and C-peptide into the portal venous blood, although under physiological conditions the plasma concentration ratio (insulin/C-peptide) is less than unity, because insulin is more susceptible to hepatic first-pass metabolism. A high ratio of insulin/C-peptide in plasma from a poisoned patient is compelling evidence that pharmaceutical insulin was administered, which does not contain C-peptide. The analysis of insulin and C-peptide was traditionally done by immunoassay methods (RIA and/or ELISA), although high resolution LC-MS/MS is more suitable for forensic purposes and permits the identification of insulin analogues. Use of insulin as a murder weapon is exemplified by the case of Colin Norris, a male nurse found guilty of murdering four elderly patients and the attempted murder of a fifth by injecting them with insulin. However, the prosecution evidence against Norris was mainly circumstantial and hearsay. Toxicological evidence against Norris consisted of a high insulin/C-peptide concentration ratio in plasma from one of the victims. This analysis was done by an immunoassay method at a clinical laboratory and not a forensic laboratory. Analytical procedures, including chain-of-custody routines, are more stringent at forensic laboratories. Since his conviction, some of the medical evidence against Norris has been called into question, especially the prevalence of spontaneous attacks of hypoglycemia in elderly and frail patients with co-morbidities.

Distribution ratios of ethanol and water between whole blood, plasma, serum, and erythrocytes: Recommendations for interpreting clinical laboratory results in a legal context.

This article reviews the scientific literature dealing with the distribution of ethanol and water between whole blood (WB), plasma, serum, and erythrocytes (red-blood cells). Knowledge of the ethanol distribution ratio is important when analytical results derived from hospital clinical laboratories are interpreted in a forensic context, such as during the prosecution of traffic offenders. Statutory blood-alcohol concentration (BAC) limits for driving are defined as the concentration of ethanol in WB and not in plasma, serum or red-blood cells. These bio-fluids differ in their water content and thereby the concentrations of ethanol. Plasma and serum contain ~90%-92% w/w water, WB ~78%-80% w/w and erythrocytes ~64%-66% w/w. The mean plasma/WB and serum/WB distribution ratios of ethanol are therefore expected to be ~1.15:1 (91/79 = 1.15), which is in good agreement with values determined empirically. However, in individual cases, the actual distribution ratio will depend on the person's age, gender, and biochemical and hematological properties of the blood specimen, such as its hematocrit. For legal purposes, we recommend that the concentration of ethanol in plasma or serum determined at hospital laboratories is divided by a factor of 1.2, which would provide a conservative estimate of the co-existing BAC and the chance of overestimating the true value is only 1 in 2000 (0.05%).

The advantages of standardizing exhaled breath-alcohol concentration to a reference respiratory gas-water vapor.

Measuring the concentration of alcohol (ethanol) in exhaled breath (BrAC) provides a rapid and non-invasive way to determine the co-existing concentration in arterial blood (A-BAC). The results of breath-alcohol testing are used worldwide as evidence of excessive drinking, such as when traffic offenders are prosecuted. Two types of breath-alcohol analyzer are in common use; hand-held instruments used as preliminary screening tests of sobriety and more sophisticated evidential instruments, the results of which are accepted as evidence for prosecution of drunken drivers. Most evidential breath-alcohol analyzers are designed to capture the last portion of a prolonged exhalation, which is thought to reflect the alcohol concentration in substantially alveolar air. The basic premise of breath-alcohol analysis is that there is a physiological relationship between A-BAC and BrAC and close agreement between the two analytical methods. This article reviews the principles and practice of breath-alcohol analysis and introduces the concept of standardizing the results to a secondary physiological gas (water vapor), which therefore serves as an internal standard. The measured BrAC is thus adjusted to an alveolar air water content of 43.95 mg l-1at 37 °C. This has several advantages, and means that a sample of breath can be captured without the person having to blow directly into the instrument. Adjusting the breath-alcohol concentration to water vapor concentration also compensates for variations in temperature of the expired air. The contact-free method of sampling breath means that a mouthpiece is unnecessary and the test subject does not need to make a continuous end exhalation.

[Improved access to test results for toxic alcohols can save lives]. / Bättre tillgång till provsvar för toxiska alkoholer kan rädda liv.

Acute poisoning involving toxic alcohols other than ethanol is not uncommon. Poisonings from drinking isopropanol are rarely life threatening, whereas methanol and ethylene glycol without prompt treatment cause severe metabolic acidosis, organ damage, and death, mainly due to toxic metabolites. Rapid identification of the type of alcohol responsible for the poisoning requires access to 24/7 toxicological service. The analysis of alcohols is usually done with gas chromatographic (GC) methods, which are not always available at smaller or medium-sized hospitals. As a complement to GC methods, reliable enzymatic oxidation procedures are now available for the analysis of ethanol, methanol, and ethylene glycol. The present study showed good agreement (r2 = 0.996) between the results of methanol analysis with a new enzymatic method (Catachem Inc.) and with GC over the clinically relevant concentration range (1-50 mmol/l). Moreover, high concentrations of ethanol (up to 80 mmol/l), ethylene glycol (to 40 mmol/l), isopropanol (to 100 mmol/l) or acetone (to 20 mmol/l) did not interfere with the analytical results for methanol. Toxicological analysis of the two most dangerous alcohols (methanol and ethylene glycol) can now be done with rapid and specific enzymatic methods, which makes it possible to diagnose and treat poisoned patients at smaller regional hospitals.

Highly cited forensic practitioners in the discipline legal and forensic medicine and the importance of peer-review and publication for admission of expert testimony.

Peer-review of manuscripts submitted to scholarly journals for publication dates back ~ 350 years and this process represents the foundation of scientific publishing. After a manuscript has undergone and survived a rigorous peer-review, this conveys a stamp of approval, because it signifies the work has been checked by independent experts in the scientific discipline concerned. The publication and citation track records of people instructed to appear as expert witness in civil and criminal litigation are important considerations. Using a publically available database, the most highly cited scientists in the discipline legal and forensic medicine were identified. For each scientist, a composite score was calculated based on six different citation metrics; (i) Total number of citations, (ii) H-index, (iii) Hm-index, which modifies the H-index for multi-authored papers, (iv) Citations to single-author papers, (v) Citations to single and first author papers and (vi) citations to single, first and last author papers. The top 100,000 most highly cited scientists from all disciplines were identified along with the top 2% of the most highly cited in each of 176 sub-fields. The latest version of the citation databases, up to the end of 2020, classified 14.163 people as having legal and forensic medicine as their primary research discipline. Of these, there were 29 names listed among the top 100,000 most highly cited in all disciplines and 299 were among the top cited 2% in their particular sub-field. More than 50% of the highly cited forensic practitioners resided in four countries (USA, Germany, UK and Australia). The top-ten most highly cited individuals were the same in all four versions of the database (2017, 2018, 2019 and 2020) and represented the sub-disciplines of toxicology (n = 3), genetics/DNA/heredity (n = 3), whereas two specialized in pathology/toxicology and two in pathology/genetics.

Scientometric evaluation of highly cited scientists in the field of forensic science and legal medicine.

A publically available database of the most highly cited scientists in all disciplines was used to identify people that belonged to the subject category "forensic science and legal medicine." This bibliometric information was derived from Elsevier's SCOPUS database containing eight million scientists with at least five articles as author or co-author. The top 100,000 most highly cited scientists were identified and ranked according to six citation metrics; total number of citations, H-index, H-index adjusted for co-authorship, citations to single-authored papers, citations to single or first author papers and, citations to single, first, or last-authored papers. The eight million entries in the SCOPUS database were sub-divided into 22 main subject categories and 176 sub-categories, one of which was legal and forensic medicine. The citation databases were provided as supplementary material in two articles published in PLoS Biology in 2019 and 2020. Among the top 100,000 most highly cited scientists, there were only 30 allocated to the legal and forensic medicine category, according to the 2019 PLoS Biology article. The updated database from 2020 also included the names of people within the top-cited 2% of their scientific discipline. This increased the number of forensic practitioners to 215 from a total of 10,158 individuals in this subject category. This article takes a closer look at these highly cited forensic scientists, the countries where they work, the particular research field in which they publish, and their composite citation scores with and without self-citations. The top ten most cited individuals in both databases (2019 and 2020) were the same and these should therefore be considered an elite group among all forensic practitioners.

Inferences and Legal Considerations Following a Blood Collection Tube Recall.

In mid-2019, medical, forensic and legal communities were notified that a certain shipment of evacuated blood sampling tubes were recalled by the manufacturer. This recall order described that the preservative sodium fluoride (100 mg) and anticoagulant potassium oxalate (20 mg) were missing from a small batch of 10-mL evacuated tubes. This gave cause for concern for possible implications in criminal justice (e.g., in drink-driving offenses) when blood-alcohol concentrations are interpreted. In reality, the lack of an anticoagulant would have been immediately obvious during sample preparation, owing to the formation of a large clot in the tube when received. Certain impairing drugs (e.g., cocaine and 6-acetylmorphine) are unstable in blood and tend to degrade without an enzyme inhibitor, such as sodium fluoride, present. In reviewing available literature related to current practices and the stability of ethanol in stored blood samples, there does not appear to be a clear consensus regarding the amount of sodium fluoride preservative necessary, if any at all, when blood is taken from living subjects under sterile conditions for typical forensic ethanol analysis.

Evaluation and review of ways to differentiate sources of ethanol in postmortem blood.

Accurate determination of a person's blood alcohol concentration (BAC) is an important task in forensic toxicology laboratories because of the existence of statutory limits for driving a motor vehicle and workplace alcohol testing regulations. However, making a correct interpretation of the BAC determined in postmortem (PM) specimens is complicated, owing to the possibility that ethanol was produced in the body after death by the action of various micro-organisms (e.g., Candida species) and fermentation processes. This article reviews various ways to establish the source of ethanol in PM blood, including collection and analysis of alternative specimens (e.g., bile, vitreous humor (VH), and bladder urine), the identification of non-oxidative metabolites of ethanol, ethyl glucuronide (EtG) and ethyl sulfate (EtS), the urinary metabolites of serotonin (5-HTOL/5-HIAA), and identification of n-propanol and n-butanol in blood, which are known putrefaction products. Practical utility of the various biomarkers including specificity and stability is discussed.

Rate of elimination of γ-hydroxybutyrate from blood determined by analysis of two consecutive samples from apprehended drivers in Norway.

AIM: Gamma-hydroxybutyrate (GHB) is a common drug of abuse with an elimination half-life of 20-45 min. However, there is some evidence that GHB might exhibit saturation kinetics after ingesting high recreational doses. The aim of this study was to investigate the elimination kinetics of GHB from blood in people apprehended by the police for impaired driving and secondary to describe concentrations in all GHB-positive drivers. METHODS: Two consecutive blood samples were taken about 30-40 min apart from N = 16 apprehended drivers in Norway. GHB was determined in blood by an Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS) method. The changes in GHB between the two consecutive blood samples allowed estimating GHB's elimination half-life, assuming first-order and zero-order elimination kinetics. GHB concentrations are also reported for N = 1276 apprehended drivers with GHB in blood. RESULTS: The median time interval between collecting the two blood samples was 36 min (range 20-56 min). The median concentration of GHB in the first blood sample was 56.5 mg/L (range 14.1-142 mg/L) compared with 47.8 mg/L in the second sample (range 9.75-113 mg/L). The median elimination half-life was 103 min (range 21-187 min), and GHB's median zero-order elimination rate constant was 21.0 mg/L/h (range 6.71-45.4 mg/L/h). Back-calculation to the time of driving resulted in GHB concentrations up to 820 mg/L assuming first-order kinetics and up to 242 mg/L assuming zero-order kinetics. In all drivers (N = 1276), the median GHB concentration was 73.7 mg/L and highest was 484 mg/L. CONCLUSION: The elimination half-life of GHB in blood samples from apprehended drivers was longer than expected compared with results of controlled dosing studies. Zero-order kinetics seems a more appropriate model for GHB when concentrations are back-calculated, and the median elimination rate was 21 mg/L/h.

Reflections on variability in the blood-breath ratio of ethanol and its importance when evidential breath-alcohol instruments are used in law enforcement.

Variability in the blood-breath ratio (BBR) of alcohol is important, because it relates a measurement of the blood-alcohol concentration (BAC) with the co-existing breath-alcohol concentration (BrAC). The BBR is also used to establish the statutory BrAC limit for driving from the existing statutory BAC limits in different countries. The in-vivo BBR depends on a host of analytical, sampling and physiological factors, including subject demographics, time after end of drinking (rising or falling BAC), the nature of the blood draw (whether venous or arterial) and the subject's breathing pattern prior to exhalation into the breath analyzer. The results from a controlled drinking study involving healthy volunteers (85 men and 15 women) from three ethnic groups (Caucasians, Hispanics and African Americans) were used to evaluate various factors influencing the BBR. Ethanol in breath was determined with a quantitative infrared analyzer (Intoxilyzer 8000) and BAC was determined by headspace gas chromatography (HS-GC). The BAC and BrAC were highly correlated (r = 0.948) and the BBR in the post-absorptive state was 2 382 ± 119 (mean ± SD). The BBR did not depend on gender (female: 2 396 ± 101 and male: 2 380 ± 123, P > 0.05) nor on racial group (Caucasians 2 398 ± 124, African Americans 2 344 ± 119 and Hispanics 2 364 ± 104, P > 0.05). The BBR was lower in subjects with higher breath- and body-temperatures (P < 0.05) and it also decreased with longer exhalation times into the breath-analyzer (P < 0.001). In the post-absorptive state, none of the 100 subjects had a BBR of less than 2 100:1.

Forensic Drug Profile: Cocaethylene.

This article is intended as a brief review or primer about cocaethylene (CE), a pharmacologically active substance formed in the body when a person co-ingests ethanol and cocaine. Reference books widely used in forensic toxicology contain scant information about CE, even though this cocaine metabolite is commonly encountered in routine casework. CE and cocaine are equi-effective at blocking the reuptake of dopamine at receptor sites, thus reinforcing the stimulant effects of the neurotransmitter. In some animal species, the LD50 of CE was lower than for cocaine. CE is also considered more toxic to the heart and liver compared with the parent drug cocaine. The plasma elimination half-life of CE is ~2 h compared with ~1 h for cocaine. The concentrations of CE in blood after drinking alcohol and taking cocaine are difficult to predict and will depend on the timing of administration and the amounts of the two precursor drugs ingested. After an acute single dose of cocaine and ethanol, the concentration-time profile of CE runs on a lower level to that of cocaine, although CE is detectable in blood for several hours longer. A strong case can be made for adding together the concentrations of cocaine and CE in forensic blood samples when toxicological results are interpreted in relation to acute intoxication and the risk of an overdose death.

Inter-laboratory proficiency results of blood alcohol determinations at clinical and forensic laboratories in Italy.

BACKGROUND: Inter-laboratory proficiency schemes are widely used to control the performance of clinical and forensic toxicology laboratories. In 2016 the Laboratory of Environmental Hygiene and Forensic Toxicology - Venice (Italy) initiated an inter-laboratory proficiency test of blood-alcohol analysis. The number of participating laboratories gradually increased from 26 to 36. Furthermore, a few clinical laboratories were included if gas chromatographic (GC) methods were used for blood alcohol analysis. PROCEDURE: Whole blood was obtained from the Blood Transfusion Centre of the Venice Hospital and a mixture of sodium fluoride and potassium oxalate was added as a preservative and anticoagulant, respectively. Aliquots of the blood were spiked with certified pure ethanol to obtain target blood-alcohol concentrations (BACs) ranging from 0 to 5.0g/L. Two blood samples (4mL each) were included in each shipment to the participating laboratories. The laboratories were asked to provide information about number of replicate BAC determinations they made, the types of ethanol reference standards used, and inherent measurement uncertainty. The aim of the testing was to obtain a mean consensus value for the target BAC and to assess inter-laboratory imprecision. All procedures for registration and submission of results were done on-line. A confidential report and statistical evaluations were returned to the participants one week later. ANALYTICAL METHODS: All participants used head-space GC (HS-GC) for the analysis of ethanol in blood. More than 85% of participants used HS-GC with flame-ionization detection, whereas the others used mass spectrometry (MS) as a detector. More than 40% of the participating laboratories kept the blood samples frozen (-20°C) prior to analysis, whereas the others used refrigeration (+4°C). The preliminary validation tests showed that there were no statistically significant differences between BAC in frozen or refrigerated samples for a period of 20 days. RESULTS AND CONCLUSION: The statistical evaluation of results was done using an iterative procedure known as Algorithm A (ISO 13528:2015, C.3.1). This provides robust estimates for mean and standard deviation between laboratories and these were used as consensus values. More than 85% of participants provided satisfactory results (z-score <1) and 94% of laboratories were within z-score <2, based on five control samples. When a blood sample without any alcohol (blank) was sent for analysis, laboratories reported this as zero, 0.00g/L, below limit of detection (LOD) or not detected. Some type of consensus should be reached for reporting blank samples.