In vitro assessment of isopropanol leakage from antiseptic barrier caps into commonly used needleless connectors.

BACKGROUND: Needleless connectors (NCs) can be disinfected using antiseptic barrier caps (ABCs) to reduce the risk of catheter-related bloodstream infections. However, recent evidence suggests that isopropanol can leak from the ABC into the NC, posing concern about their safe use. We sought to determine in vitro which ABC and NC parameters influence the leakage of isopropanol through the infusion circuit. METHODS: We assessed 13 NCs and 4 ABCs available in the European market. In vitro circuits consisting of an isopropanol cap, a NC, and an 11-cm catheter line were created. The circuits were left in place for 1 to 7 days at room temperature to assess the kinetics of isopropanol leakage. Isopropanol content in ABC and in circuit flushing solutions (5 mL NaCl 0.9%) after exposure to the cap were measured using gas chromatography with a flame ionization detector. RESULTS: The leakage of isopropanol from the cap to the NC was dependent on the NC, but not the cap. The NC mechanism did not predict the leakage of isopropanol. The Q-Syte NC exhibited the most isopropanol leakage (7.01±1.03 mg and 28.32±2.62 mg at 24 hours and 7 days, respectively), whereas the Caresite NC had the lowest isopropanol leakage at 7 days (1.69±0.01 mg). CONCLUSION: The use of isopropanol ABCs can cause isopropanol leakage into the catheter circuit according to NC parameters. Caution should be exercised when using these devices, especially in the pediatric and neonatal population.

Identification, synthesis and quantification of eutylone consumption markers in a chemsex context.

Eutylone is a cathinone-derived synthetic amphetamine scheduled by the World Health Organization and European Monitoring Centre for Drugs and Drug Addiction since 2022 due to its growing consumption. We report here an eutylone intoxication involving a 38-year-old man and a 29-year-old woman in a chemsex context. A bag containing a white crystalline powder labelled as a research product was found in their vehicle. Nuclear magnetic resonance and liquid chromatography-high-resolution mass spectrometry (LC-HRMS) analyses identified the powder as eutylone and confirmed purity superior to 99%. LC-HRMS data analysis using molecular networking allowed to propose new eutylone metabolites in blood samples in a graphical manner. We described 16 phase I (e.g. hydroxylated or demethylated) and phase II metabolites (glucuroconjugates and sulfoconjugates). The same metabolites were found both in male and female blood samples. Toxicological analyses measured eutylone concentration in blood samples at 1374 ng/mL and 1536 ng/mL for the man and the woman, respectively. A keto-reduced metabolite (m/z 238.144) was synthesized to permit its quantification at 67 ng/mL and 54 ng/mL in male and female blood samples, respectively. Overall, the identification of these metabolites will increase the knowledge of potential drug consumption markers and allow to implement mass spectrometry databases to better monitor future drug abuse or consumption.

In silico and in vitro metabolism studies of the new synthetic opiate AP-237 (bucinnazine) using bioinformatics tools.

The recent emergence of new synthetic opioids (NSOs) compounds in the illicit market is increasingly related to fatal cases. Identification and medical care of NSO intoxication cases are challenging, particularly due to high frequency of new products and extensive metabolism. As the study of NSO metabolism is crucial for the identification of these drugs in cases of intoxication, we aimed to investigate the metabolism of the piperazine NSO AP-237 (= bucinnazine). Two complementary approaches (in silico and in vitro) were used to identify putative AP-237 metabolites which could be used as consumption markers. In silico metabolism studies were realized by combining four open access softwares (MetaTrans, SyGMa, Glory X, Biotransformer 3.0). In vitro experiments were performed by incubating AP-237 (20 µM) in differentiated HepaRG cells during 0 h, 8 h, 24 h or 48 h. Cell supernatant were extracted and analyzed by liquid chromatography coupled to high-resolution mass spectrometry and data were reprocessed using three strategies (MetGem, GNPS or Compound Discoverer®). A total of 28 phase I and six phase II metabolites was predicted in silico. Molecular networking identified seven putative phase I metabolites (m/z 203.154, m/z 247.180, m/z 271.180, two m/z 289.191 isomers, m/z 305.186, m/z 329.222), including four previously unknown metabolites. Overall, this cross-disciplinary approach with molecular networking on data acquired in vitro and in silico prediction enabled to propose relevant candidate as AP-237 consumption markers that could be added to mass spectrometry libraries to help diagnose intoxication.

Severe 25E-NBOH intoxication associated with MDPHP intake: a case report, metabolism study, and literature review.

N-Benzylphenethylamine derivatives are 5-HT2A receptor agonists with hallucinogenic properties, including NBOMe (N-(2-methoxybenzyl)-2-(3,4,5-trimethoxyphenyl)ethan-1-amine) and NBOH (2-(((2,5-dimethoxyphenethyl)amino)methyl)phenol). We reported here the case of a 23-year-old man who presented a serotoninergic syndrome and a loss of consciousness following the consumption of a powder labelled as 25I-NBOH. Toxicological analyses of biological samples were carried out using a liquid chromatography high-resolution mass spectrometry. Two new psychoactive substances were identified and confirmed with certified reference materials: 25E-NBOH (2-(((4-ethyl-2,5-dimethoxyphenethyl)amino)methyl)phenol) and MDPHP (1-(benzo[d][1,3]dioxol-5-yl)-2-(pyrrolidin-1-yl)hexan-1-one). Pharmaceuticals administered to the patient during his medical care were found in plasma and urine. 25E-NBOH and MDPHP concentrations were respectively at 2.3 ng/mL and 3.4 ng/mL in plasma, and 25.7 ng/mL and 30.5 ng/mL in urine. 25I-NBOH (2-(((4-iodo-2,5-dimethoxyphenethyl)amino)methyl)phenol) was specifically searched in both samples and was not detected. These results are discussed along with a literature review on human cases of exposure to N-benzylphenethylamine derivatives. Using molecular networking approach, we propose the first 25E-NBOH metabolism study using authentic biological samples (plasma and urine). We described seven metabolites (M1 to M7), including two phase I (m/z 330.172; m/z 288.160) and five phase II metabolites (m/z 464.191, m/z 478.207, m/z 492.223, m/z 508.218; m/z 396.156). The M6 (m/z 492.223) was the most intense ion detected in plasma and urine and could be proposed as a relevant 25E-NBOH consumption marker. Overall, we described an original case of 25E-NBOH poisoning and identified metabolites that could potentially be used as consumption markers to detect 25E-NBOH intoxications with a higher confidence level and probably a longer detection window.

Spice-drug interactions: a case report on the use of turmeric, curry and ginger in a renal transplant patient on tacrolimus.

Tacrolimus is a widely used immunosuppressant for the prevention of rejection after transplantation. In vitro studies suggest that interactions exist between spices and tacrolimus. We present the case of a renal transplant patient aged around 70 years who was treated with prednisone, mycophenolate-mofetil and tacrolimus. The patient had a pre-transplant dietary habit of consuming foods spiced with turmeric, curry and ginger. The following protocol was implemented in parallel with close monitoring of plasma tacrolimus concentrations: administration of 10 g/day of turmeric for 4 days, then 10 g/day of curry for 4 days and then 10 g/day of ginger for 4 days. No change in tacrolimus plasma concentrations during and after the implementation of the protocol was observed. The impact of turmeric, curry and ginger on plasma tacrolimus concentrations seems negligible in vivo although further studies are needed. A shared decision to test the impact of spice consumption in a patient with dietary habits involving these spices seems reasonable.

Assessing health literacy in transplant patients to better tailor the content of their therapeutic education: an observational study.

OBJECTIVES: Evaluate health literacy in transplant patients to better tailor the content of their continuing therapeutic education. METHODS: A 20-item questionnaire divided into five themes (sport/recreation, dietary measures, hygiene measures, recognition of the signs of graft rejection and medication management) was sent to transplant patient associations. Participants' responses (a score out of 20 points), were analysed according to demographic characteristics, transplanted organ (kidney, liver or heart), type of donor (living or deceased), participation in a therapeutic patient education (TPE) programme, management of end-stage renal disease (with or without dialysis) and the date of transplant. RESULTS: 327 individuals completed the questionnaires (mean age 63.3±12.7 years, mean time post-transplant 13.1±12.1 years). From 2 years after transplantation, the patients' score decreases significantly compared with the score obtained at hospital discharge. Patients who received TPE had significantly higher scores than patients who did not receive it, but only in the first 2 years post-transplant. The scores were different depending on the organs transplanted. Patients' knowledge varied according to the theme; the percentage of errors being higher for questions related to hygienic and dietary rules. CONCLUSION: These findings highlight the importance of the role of the clinical pharmacist in maintaining the transplant recipient's health literacy level over time to increase graft life. We show the topics on which pharmacists must acquire solid knowledge to best meet the needs of transplant patients.

A pharmacist-led intervention to improve kidney transplant recipient outcomes and identify patients at risk of highly variable trough tacrolimus levels: a cohort study.

OBJECTIVES: Given the positive impact of appropriate medication management on graft outcome and therefore of patient survival and graft function, the pharmacist's role in the kidney transplantation team has evolved over recent decades. The primary objective of this study was to determine whether pharmacist-led intervention after kidney transplantation is associated with a lower graft rejection rate and intra-patient variation in tacrolimus trough concentrations (Cmin). The study's secondary objective was to develop a questionnaire to identify patients at risk for highly variable Cmin. METHODS: We retrospectively analysed kidney transplant recipients at Rennes University Hospital (France) between January 2013 and December 2020. Patients who received pharmacist-led education (intervention group, n=139) were compared with patients who did not (control group, n=131), according to graft survival at 1 year post-transplant, coefficient of variation (%CV) for the tacrolimus Cmin, age, sex, length of hospital stay post-transplantation, body mass index, and Charlson Comorbidity Index. In the intervention group, a questionnaire assessing patient knowledge was introduced to compare scores with the %CV. RESULTS: In the intervention group, 1 year post-transplant graft survival was higher (95.7% vs 88.5%, p=0.0289) and patients had fewer variabilities in Cmin. The %CV was correlated with questionnaire scores (r=-0.9758, p<0.0001). CONCLUSIONS: Pharmacist-led interventions may have contributed to improved graft survival and patient management of immunosuppressants. Because %CV correlates with the patient questionnaire score, its introduction could be useful in identifying kidney transplant patients who would benefit most from a pharmacist-led patient education.

Long-term lithium therapy and risk of chronic kidney disease, hyperparathyroidism and hypercalcemia: a cohort study.

BACKGROUND: Lithium is well recognized as the first-line maintenance treatment for bipolar disorder (BD). However, besides therapeutic benefits attributed to lithium therapy, the associated side effects including endocrinological and renal disorders constitute important parameters in prescribing patterns and patient adherence. The objectives of this study is to (i) determine whether long-term lithium therapy is associated with a decrease in renal function, hyperparathyroidism and hypercalcemia and (ii) identify risk factors for lithium-induced chronic kidney disease (CKD). METHODS: We conducted a single-centered cohort study of adult patients (≥ 18 years) treated with lithium, who were enrolled at Rennes University Hospital in France between January 1, 2018 and June 1, 2020. Required data were collected from the patient's medical records: demographics characteristics (age, sex, body mass index), biologic parameters (GFR, lithium blood level, PTH and calcium), medical comorbidities (hypertension and diabetes), lithium treatment duration and dosage, and length of hospitalization. RESULTS: A total of 248 patients were included (mean age: 60.2 ± 16.5 years). Duration of lithium treatment correlated with (i) deterioration of renal function estimated at - 2.9 mL/min/year (p < 0.0001) and (ii) the development of hyperparathyroidism (p < 0.01) and hypercalcemia (p < 0.01). We also noted that patients with lithium blood level > 0.8 mEq/mL had significantly lower GFR than patients with lithium blood level < 0.8 mEq/mL (61.8 mL/min versus 77.6 mL/min, respectively, p = 0.0134). Neither diabetes mellitus nor hypertension was associated with more rapid deterioration of renal function. CONCLUSION: This study suggests that the duration of lithium treatment contribute to the deterioration of renal function, raising the question of reducing dosages in patients with a GFR < 60 mL/min. Overdoses has been identified as a risk factor for CKD, emphasizing the importance of regular re-evaluation of the lithium dose regimen. Also, long-term lithium therapy was associated with hyperparathyroidism and hypercalcemia. Particular vigilance is required on these points in order to limit the occurrence of endocrinological and renal lithium adverse effects.

Unexpected overdose of oral cyclosporine in a kidney transplant patient: a case report.

Cyclosporine is a widely used immunosuppressive agent to prevent rejection of solid organ transplant. Here, we describe the case of a 71-year-old man who received the prescribed dose of cyclosporine 10 times 6 days after a kidney transplantation because of a concentration miscalculation involving two galenic forms. The patient presented gastrointestinal and neurological disorders. Therapeutic drug monitoring revealed high cyclosporine blood concentrations (693 ng/mL, therapeutic range 100-300 ng/mL). Symptomatic management of digestive disorders was performed, and haemodialysis was started the day after the cyclosporine overdose in the face of acute renal failure. The patient's disorders were quickly resolved. The dosing regimen was adapted in order to administer the most appropriate galenic form and to avoid another administration error. Long-term follow-up showed no failure of renal transplantation. The purpose of this case report is to warn physicians and clinical pharmacists about the vigilance required on cyclosporine prescription, especially when two galenic forms are administered to obtain the prescribed dose.

Use of innovative, cross-disciplinary in vitro, in silico and in vivo approaches to characterize the metabolism of chloro-alpha-pyrrolidinovalerophenone (4-Cl-PVP).

Synthetic cathinones constitute a family of new psychoactive substances, the consumption of which is increasingly worldwide. A lack of metabolic knowledge limits the detection of these compounds in cases of intoxication. Here, we used an innovative cross-disciplinary approach to study the metabolism of the newly emerging cathinone chloro-alpha-pyrrolidinovalerophenone (4-Cl-PVP). Three complementary approaches (in silico, in vitro, and in vivo) were used to identify putative 4-Cl-PVP metabolites that could be used as additional consumption markers. The in silico approach used predictive software packages. Molecular networking was used as an innovative bioinformatics approach for re-processing high-resolution tandem mass spectrometry data acquired with both in vitro and in vivo samples. In vitro experiments were performed by incubating 4-Cl-PVP (20 µM) for four different durations with a metabolically competent human hepatic cell model (differentiated HepaRG cells). In vivo samples (blood and urine) were obtained from a patient known to have consumed 4-Cl-PVP. The in silico software predicted 17 putative metabolites, and molecular networking identified 10 metabolites in vitro. On admission to the intensive care unit, the patient's plasma and urine 4-Cl-PVP concentrations were, respectively, 34.4 and 1018.6 µg/L. An in vivo analysis identified the presence of five additional glucuronoconjugated 4-Cl-PVP derivatives in the urine. Our combination of a cross-disciplinary approach with molecular networking enabled the detection of 15 4-Cl-PVP metabolites, 10 of them had not previously been reported in the literature. Two metabolites appeared to be particular relevant candidate as 4-Cl-PVP consumption markers in cases of intoxication: hydroxy-4-Cl-PVP (m/z 282.1254) and dihydroxy-4-Cl-PVP (m/z 298.1204).

A Transversal Approach Combining In Silico, In Vitro and In Vivo Models to Describe the Metabolism of the Receptor Interacting Protein 1 Kinase Inhibitor Sibiriline.

Sibiriline is a novel drug inhibiting receptor-interacting protein 1 kinase (RIPK1) and necroptosis, a regulated form of cell death involved in several disease models. In this study, we aimed to investigate the metabolic fate of sibiriline in a cross-sectional manner using an in silico prediction, coupled with in vitro and in vivo experiments. In silico predictions were performed using GLORYx and Biotransformer 3.0 freeware; in vitro incubation was performed on differentiated human HepaRG cells, and in vivo experiments including a pharmacokinetic study were performed on mice treated with sibiriline. HepaRG culture supernatants and mice plasma samples were analyzed with ultra-high-performance liquid chromatography, coupled with tandem mass spectrometry (LC-HRMS/MS). The molecular networking bioinformatics tool applied to LC-HRMS/MS data allowed us to visualize the sibiriline metabolism kinetics. Overall, 14 metabolites, mostly produced by Phase II transformations (glucuronidation and sulfation) were identified. These data provide initial reassurance regarding the toxicology of this new RIPK1 inhibitor, although further studies are required.

Arylcyclohexylamine Derivatives: Pharmacokinetic, Pharmacodynamic, Clinical and Forensic Aspects.

Since the 2000s, an increasing number of new psychoactive substances (NPS) have appeared on the drug market. Arylcyclohexylamine (ACH) compounds such as ketamine, phencyclidine and eticyclidine derivatives are of particular concern, given their rapidly increasing use and the absence of detailed toxicity data. First used mainly for their pharmacological properties in anesthesia, their recreational use is increasing. ACH derivatives have an antagonistic activity against the N-methyl-D-aspartate receptor, which leads to dissociative effects (dissociation of body and mind). Synthetic ketamine derivatives produced in Asia are now arriving in Europe, where most are not listed as narcotics and are, thus, legal. These structural derivatives have pharmacokinetic and pharmacodynamic properties that are sometimes very different from ketamine. Here, we describe the pharmacology, epidemiology, chemistry and metabolism of ACH derivatives, and we review the case reports on intoxication.

Carbofuran self-poisoning: forensic and analytic investigations in twins and literature review.

Carbofuran is a pesticide widely used in agricultural context to kill insects, mites, and flies by ingestion or contact. Along with literature review, we aimed to (i) present the clinical, autopsy, and toxicological findings of carbofuran self-poisonings in two 69-year-old twins, resulting in the death of one of them and (ii) assess carbofuran metabolite distribution using molecular networking. Quantitative analysis of carbofuran and its main metabolites (3-hydroxycarbofuran and 3-ketocarbofuran) was carried out using an original liquid chromatography-tandem mass spectrometry method on biological samples (cardiac or peripheral blood, urine, bile, and gastric contents). Toxicological analysis of post-mortem samples (twin 1) highlighted high concentrations of carbofuran and its metabolites in cardiac blood, bile, and gastric contents. These compounds were also quantified in blood and/or urine samples of the living brother (twin 2), confirming poisoning. Using molecular networking approach to facilitate visualization of mass spectrometry datasets and sample-to-sample comparisons, we detected two more metabolites (7-phenol-carbofuran and 3-hydroxycarbofuran glucuronide) in bile (twin 1) and urine (twin 2). These results highlight the value of (i) these compounds as carbofuran consumption markers and (ii) bile samples in post-mortem analysis to confirm poisoning. From an analytical point of view, molecular networking allowed the detection and interpretation of carbofuran metabolite ammonium adducts which helped to confirm their identification annotations, as well as their structural data. From a clinical point of view, the different outcomes between the two brothers are discussed. Overall, these cases provide novel information regarding the distribution of carbofuran and its metabolites in poisoning context.

Mise en évidence d'intoxications par ingestion de champignons supérieurs : expérience au CHU de Rennes.

L'α- et la ß-amanitine sont de puissantes toxines de champignons supérieurs responsables de cytolyses hépatiques graves pouvant menacer le pronostic vital. En France, les données des centres antipoison rapportent un nombre croissant d'intoxications aux champignons depuis 2016, justifiant le besoin de méthodes de diagnostic biologique robustes. En laboratoire de toxicologie hospitalière, l'objectivation d'une intoxication par les amanitines à partir de prélèvements sanguins ou urinaires constitue ainsi un élément important dans la prise en charge du patient intoxiqué. L'objectif de ce travail consiste à réaliser une mini-revue de la littérature sur le dosage des amanitines dans les fluides biologiques pour le diagnostic des intoxications aux amanitines. Les caractéristiques des amanitines, les méthodes analytiques, les données d'interprétation, les applications pratiques ainsi que les perspectives d'utilisation des techniques de dosage y sont présentées. À travers une comparaison de deux techniques analytiques de chromatographie liquide couplée à de la spectrométrie de masse en tandem utilisées au Centre hospitalier universitaire de Rennes (Waters Xevo TQ XSTM et Thermo Scientific Q ExactiveTM), cet article présente également le retour d'expérience de biologistes médicaux dans l'amélioration continue des méthodes de dosages des amanitines.

Comparison of Illicit Drug Seizures Products of Natural Origin Using a Molecular Networking Approach.

Drug powder composition analysis is of particular interest in forensic investigations to identify illicit substance content, cutting agents and impurities. Powder profiling is difficult to implement due to multiple analytical methods requirement and remains a challenge for forensic toxicology laboratories. Furthermore, visualization tools allowing seizure products identification appear to be under-used to date. The aim of this study is to present the utility of molecular networking for the composition establishment of natural origin drugs. A powder suspected to contain heroin and three powders suspected to contain cocaine obtained from law enforcement agency seizures were analyzed using untargeted screening by liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS/MS). Molecular networking and metabolite annotation applied to suspected heroin sample allowed rapid confirmation of its illicit content (heroin), the identification of structurally related major impurities (6-monoacetylmorphine, 6-monoacetylcodeine, noscapine, and papaverine), as well as cutting agents (acetaminophen and caffeine). The cocaine powder profiling allowed the comparison of its constituents in a semi-quantitative manner (cocaine, benzoylecgonine, trans/cis-cinnamoylcocaine, trimethoxycocaine, hexanoylecgonine methylester, caffeine, hydroxyzine, levamisole, and phenacetin), bringing additional information for their identification, including geographically sourcing of natural product and their putative place in the supply chain. Although this approach does not replace the profiling techniques used by forensic laboratories, the use of molecular networks provides a visual overview of structurally related constituents which aids the comparison and investigation of seizure powders. Molecular networks offers here an ideal way to depict structurally related and unrelated compounds in these often complex mixtures of chemicals.

New psychoactive substance cocktail in an intensive care intoxication case elucidated by molecular networking.

BACKGROUND: The recreational use of new psychoactive substances (NPS) is increasing worldwide. Among them, the arylcyclohexylamine family including phencyclidine (PCP) and ketamine derivatives is increasing. We report a non-fatal intoxication mainly due to arylcyclohexylamine compounds illustrated by molecular networking (MN). CASE DETAILS: A 37-year-old man with a history of drug abuse was discovered with several bags labeled as research chemicals around him and traces of powder on his nose. He was rehydrated, intubated, and admitted to the intensive care unit (ICU). Urine and drug were analyzed by liquid chromatography-mass spectrometry for NPS identification. Several NPS were quantified in urine: 3-OH-PCP at 12,085 mg/L, 3-MeO-PCP at 1100 mg/L, 2F-DCK at 147 mg/L, N-ethylhexedrone at 165 mg/L and CMC at 48 mg/L. Using a bioinformatic approach, a molecular network was built to confirm the consumption of powders contained in the bags by comparison with patient's urine. DISCUSSION: This case illustrates the interest of MN to (i) perform sample-to-sample comparison, (ii) target quantification methods, and (iii) allow proper management to confirm the relevance of the treatment.

Increasing vaccine supply with low dead-volume syringes and needles.

As global vaccine production capacity is limited, every optimization strategy must be explored to rapidly increase the number of people vaccinated. The objective of this study is to determine which medical devices allow the extraction of the maximum number of doses from different vaccine vials (Pfizer-BioNTech, AstraZeneca, Moderna and Johnson & Johnson vaccines) by analyzing all the factors involved in the preparation of the injected doses. By measuring the dead-volume of 32 syringe-needle combinations, we show that fixed-needle syringe with a dead-volume of less than 5 µL can extract up to 7 doses from Pfizer vials, 13 doses from AstraZeneca vials, 12 doses from Moderna vials and 6 doses from Johnson & Johnson vials. We found that the syringe accuracy is important, and can compromise the chances of extracting additional doses when withdrawing too large a volume. For Pfizer vaccine, particular attention must be paid to the choice of dilution syringe, which may compromise the extraction of the 7th dose. The withdrawal of extra doses from vaccine vials was not operator-dependent. In this unprecedented health context, the medical device considerations presented here could help to optimize every COVID-19 vaccine vial.

Alcohol and drug consumption among motor vehicle drivers in the Brittany region of France: A 9-year cross-sectional population study.

The primary objective of the present study was to evaluate the frequency of positive tests for alcohol and drugs during roadside testing or after road accidents among drivers in the Brittany region of France. The study's secondary objective was to describe the blood concentrations of the substances found during these tests, in order to provide a scientific basis for the establishment or modification of legislative threshold values for road injuries prevention. We performed a cross-sectional study of a database compiled by Rennes University Hospital's toxicology laboratory in the Brittany region of France between 2010 and 2018. Driver's age, sex, and test status (positive or negative), and blood levels of ethanol, 9-tetrahydrocannabinol (THC), methylene dioxymethamphetamine (MDMA), amphetamine, benzoylecgonine and 6-monoacetylmorphine (6-MAM) were collected. Twelve thousand four hundred and ninety-seven drivers (males: 86.1%; median (range) age: 29 (15-94)) have provided roadside blood samples, giving a total of 25,998 test results. Among the 10,996 drivers with at least one positive test, the median blood concentrations of ethanol, THC, MDMA, amphetamine, benzoylecgonine, and 6-MAM were respectively 1.82 g/L, 2.41 ng/mL, 138.4 ng/mL, 67.7 ng/mL, 173.3 ng/mL, and 0.97 ng/mL. 1159 (10.54%) of the 10,996 drivers tested positive for two or more substances, and 151 (1.4%) tested positive for three or more substances. With the exception of heroin, the currently recommended threshold values appear to be appropriate for road injuries prevention with regard to the concentrations observed in offenders.