{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T15:29:01Z","timestamp":1784734141417,"version":"3.55.0"},"reference-count":42,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,2,5]],"date-time":"2025-02-05T00:00:00Z","timestamp":1738713600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Alcohol is responsible for an ever-increasing number of deaths worldwide, and many road accidents are caused by irresponsible drinking and driving. The use of biomarkers that can support a diagnosis of alcohol abuse is a very important tool that can improve the prevention of many alcohol-related diseases and serious traffic accidents. The main aim of our study was the full validation of a rapid and simple method by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS\/MS) to detect ethyl glucuronide in hair (hEtG). The method was successfully applied to n = 171 real hair samples collected from drivers convicted of driving while impaired by alcohol or drugs. A comparison of hEtG and serum Carbohydrate-Deficient Transferrin percentages (% CDT) was also performed to carefully evaluate the data in relation to the specific detection windows of the two different biomarkers. Most of the drivers with hEtG &gt; 30 pg\/mg were males in their thirties. None of the hEtG-positives had a serum % CDT above the cutoff (\u22652%). Although some researchers suggest caution until solid data are available on the possible effects of interindividual variability that may influence EtG incorporation and metabolism, hEtG is a very useful biomarker of long-term alcohol exposure that shows greater reliability than traditional blood markers.<\/jats:p>","DOI":"10.3390\/ijms26031344","type":"journal-article","created":{"date-parts":[[2025,2,5]],"date-time":"2025-02-05T10:09:52Z","timestamp":1738750192000},"page":"1344","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development and Validation of a Fast and Sensitive UPLC-MS\/MS Method for Ethyl Glucuronide (EtG) in Hair, Application to Real Cases and Comparison with Carbohydrate-Deficient Transferrin (CDT) in Serum"],"prefix":"10.3390","volume":"26","author":[{"given":"Leonardo","family":"Romani","sequence":"first","affiliation":[{"name":"Laboratory of Forensic Toxicology, Section of Legal Medicine, Social Security and Forensic Toxicology, Department of Biomedicine and Prevention, Faculty of Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"},{"name":"PhD School in Medical-Surgical Applied Sciences, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3157-0968","authenticated-orcid":false,"given":"Giulio","family":"Mannocchi","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Toxicology, Section of Legal Medicine, Social Security and Forensic Toxicology, Department of Biomedicine and Prevention, Faculty of Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7098-7830","authenticated-orcid":false,"given":"Federico","family":"Mineo","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Toxicology, Section of Legal Medicine, Social Security and Forensic Toxicology, Department of Biomedicine and Prevention, Faculty of Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2514-5471","authenticated-orcid":false,"given":"Francesca","family":"Vernich","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Toxicology, Section of Legal Medicine, Social Security and Forensic Toxicology, Department of Biomedicine and Prevention, Faculty of Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucrezia","family":"Stefani","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Toxicology, Section of Legal Medicine, Social Security and Forensic Toxicology, Department of Biomedicine and Prevention, Faculty of Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"},{"name":"PhD School in Medical-Surgical Applied Sciences, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luigi Tonino","family":"Marsella","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Toxicology, Section of Legal Medicine, Social Security and Forensic Toxicology, Department of Biomedicine and Prevention, Faculty of Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0561-3442","authenticated-orcid":false,"given":"Roberta","family":"Tittarelli","sequence":"additional","affiliation":[{"name":"Laboratory of Forensic Toxicology, Section of Legal Medicine, Social Security and Forensic Toxicology, Department of Biomedicine and Prevention, Faculty of Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Via Montpellier 1, 00133 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,5]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (WHO) (2024, December 13). Alcohol. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/alcohol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Trius-Soler, M., Pratic\u00f2, G., G\u00fcrdeniz, G., Garcia-Aloy, M., Canali, R., Fausta, N., Brouwer-Brolsma, E.M., Andr\u00e9s-Lacueva, C., and Dragsted, L.O. (2023). Biomarkers of Moderate Alcohol Intake and Alcoholic Beverages: A Systematic Literature Review. Genes Nutr., 18.","DOI":"10.1186\/s12263-023-00726-1"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1080\/15504260802628684","article-title":"Novel Objective Biomarkers of Alcohol Use: Potential Diagnostic and Treatment Management Tools in Dual Diagnosis Care","volume":"5","author":"Kalapatapu","year":"2009","journal-title":"J. Dual Diagn."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1007\/s40429-021-00402-7","article-title":"Blood Biomarkers of Alcohol Use: A Scoping Review","volume":"8","author":"Harris","year":"2021","journal-title":"Curr. Addict. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Fiorelli, D., Romani, L., Treglia, M., Pallocci, M., Passalacqua, P., Coppeta, L., Marsella, L.T., and Tittarelli, R. (2023). Carbohydrate-Deficient Transferrin (CDT) as a Biomarker of Alcohol Abuse: A Retrospective Study of the Italian Drinking Trend among Drivers from 2016 to 2022. Toxics, 11.","DOI":"10.3390\/toxics11110914"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Triolo, V., Span\u00f2, M., Buscemi, R., Gio\u00e8, S., Malta, G., \u010caplinskiene, M., Vaiano, F., Bertol, E., Zerbo, S., and Albano, G.D. (2022). EtG Quantification in Hair and Different Reference Cut-Offs in Relation to Various Pathologies: A Scoping Review. Toxics, 10.","DOI":"10.3390\/toxics10110682"},{"key":"ref_7","unstructured":"Andraus, M., Appenzeller, B., Binz, T., Crunelle, C., Favretto, D., Kintz, P., and Kronstrand, R. (2024, December 13). Consensus for the Use of Alcohol Markers in Hair for Supporting the Assessment of Abstinence and Chronic Alcohol Consumption. Available online: https:\/\/www.soht.org\/images\/pdf\/Alcohol%20consensus_final_aug19.pdf."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.forsciint.2007.01.025","article-title":"Segmental Determination of Ethyl Glucuronide in Hair: A Pilot Study","volume":"173","author":"Appenzeller","year":"2007","journal-title":"Forensic Sci. Int."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1520\/JFS14267J","article-title":"Ethyl Glucuronide Concentration in Serum of Human Volunteers, Teetotalers, and Suspected Drinking Drivers","volume":"42","author":"Schmitt","year":"1997","journal-title":"J. Forensic Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1093\/alcalc\/34.1.71","article-title":"Ethyl Glucuronide\u2014A Marker of Alcohol Consumption and a Relapse Marker with Clinical and Forensic Implications","volume":"34","author":"Wurst","year":"1999","journal-title":"Alcohol Alcohol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1111\/j.1530-0277.2010.01171.x","article-title":"Clinical (Nonforensic) Application of Ethyl Glucuronide Measurement: Are We Ready?","volume":"34","author":"Jatlow","year":"2010","journal-title":"Alcohol. Clin. Exp. Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Shin, D.W., Kim, S.Y., Suh, S.I., and Kim, J.Y. (2024). Simultaneous Determination of Ethyl Glucuronide, Cocaine, Cocaethylene, and Benzoylecgonine in Hair by Using LC\u2013MS\/MS. J. Anal. Sci. Technol., 15.","DOI":"10.1186\/s40543-023-00412-2"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3161","DOI":"10.1016\/j.jchromb.2010.09.034","article-title":"Determination of Ethyl Glucuronide in Hair Samples of Chinese People by Protein Precipitation (PPT) and Large Volume Injection\u2013Gas Chromatography\u2013Tandem Mass Spectrometry (LVI\u2013GC\/MS\/MS)","volume":"878","author":"Shi","year":"2010","journal-title":"J. Chromatogr. B"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.forsciint.2011.01.044","article-title":"Ethyl Glucuronide in Human Hair after Daily Consumption of 16 or 32 g of Ethanol for 3 Months","volume":"215","author":"Kronstrand","year":"2012","journal-title":"Forensic Sci. Int."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.forsciint.2016.08.012","article-title":"Effects of Various Sample Pretreatment Procedures on Ethyl Glucuronide Quantification in Hair Samples: Comparison of Positivity Rates and Appraisal of Cut-off Values","volume":"267","author":"Salomone","year":"2016","journal-title":"Forensic Sci. Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1093\/jat\/bkaa141","article-title":"Accelerated Extraction and Analysis of Ethyl Glucuronide in Hair by Means of Pressurized Liquid Extraction Followed by Liquid Chromatography\u2013Tandem Mass Spectrometry Determination","volume":"45","author":"Vincenti","year":"2021","journal-title":"J. Anal. Toxicol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1002\/dta.3516","article-title":"Comparison of Two Methods for the Extraction of Ethylglucuronide from Hair","volume":"16","author":"Groff","year":"2024","journal-title":"Drug Test. Anal."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1080\/10937400490432400","article-title":"Carbohydrate-Deficient Transferrin (CDT)\u2014A Biomarker for Long-Term Alcohol Consumption","volume":"7","author":"Golka","year":"2004","journal-title":"J. Toxicol. Environ. Health Part B"},{"key":"ref_19","first-page":"30","article-title":"Biomarkers for Alcohol Use and Abuse\u2014A Summary","volume":"28","author":"Peterson","year":"2004","journal-title":"Alcohol Res. Health"},{"key":"ref_20","first-page":"309","article-title":"Alcohol Biomarkers in Clinical and Forensic Contexts","volume":"115","author":"Glahn","year":"2018","journal-title":"Dtsch. \u00c4rztebl. Int."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1093\/alcalc\/agq092","article-title":"Pregnancy and Variations of Carbohydrate-Deficient Transferrin Levels Measured by the Candidate Reference HPLC Method","volume":"46","author":"Bianchi","year":"2011","journal-title":"Alcohol Alcohol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.1111\/j.1530-0277.1997.tb04457.x","article-title":"Is Carbohydrate-Deficient Transferrin a Specific Marker for Alcohol Abuse? A Study in Patients with Chronic Viral Hepatitis","volume":"21","author":"Perret","year":"1997","journal-title":"Alcohol. Clin. Exp. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1114","DOI":"10.1111\/j.1530-0277.1996.tb01955.x","article-title":"Carbohydrate-Deficient Transferrin in Healthy Women: Relation to Estrogens and Iron Status","volume":"20","author":"Stauber","year":"1996","journal-title":"Alcohol. Clin. Exp. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1093\/alcalc\/ags053","article-title":"N-Latex CDT Results in Liver Diseases","volume":"47","author":"Chrostek","year":"2012","journal-title":"Alcohol Alcohol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Fagan, K.J., Irvine, K.M., McWhinney, B.C., Fletcher, L.M., Horsfall, L.U., Johnson, L., O\u2019Rourke, P., Martin, J., Scott, I., and Pretorius, C.J. (2014). Diagnostic Sensitivity of Carbohydrate Deficient Transferrin in Heavy Drinkers. BMC Gastroenterol., 14.","DOI":"10.1186\/1471-230X-14-97"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Cerioni, A., Buratti, E., Mietti, G., Ribeiro, I.L.A., Cippitelli, M., Tassoni, G., Cingolani, M., Froldi, R., and Scendoni, R. (2024). Comparative Analysis between CDT in Serum and Ethyl Glucuronide in Hair to Define the Best Reliable Tool for the Diagnosis of Alcohol Abuse. Drug Alcohol Depend., 257.","DOI":"10.1016\/j.drugalcdep.2024.111128"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Bornhorst, J.A., and Mbughuni, M.M. (2019). Alcohol Biomarkers. Critical Issues in Alcohol and Drugs of Abuse Testing, Elsevier.","DOI":"10.1016\/B978-0-12-815607-0.00003-4"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.drugalcdep.2014.05.014","article-title":"Hair Ethyl Glucuronide as a Biomarker of Alcohol Consumption in Alcohol-Dependent Patients: Role of Gender Differences","volume":"141","author":"Crunelle","year":"2014","journal-title":"Drug Alcohol Depend."},{"key":"ref_29","unstructured":"Official Gazette of the Italian Republic (2024, December 30). D.Lgs. n. 285\/1992\u2014Art. 186-Bis. Available online: https:\/\/www.gazzettaufficiale.it\/atto\/serie_generale\/caricaArticolo?art.versione=6&art.idGruppo=11&art.flagTipoArticolo=0&art.codiceRedazionale=092G0306&art.idArticolo=186&art.idSottoArticolo=2&art.idSottoArticolo1=10&art.dataPubblicazioneGazzetta=1992-05-18&art.progressivo=0."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/j.alcohol.2010.05.001","article-title":"Influence of Ethanol Dose and Pigmentation on the Incorporation of Ethyl Glucuronide into Rat Hair","volume":"44","author":"Kharbouche","year":"2010","journal-title":"Alcohol"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.forsciint.2016.01.009","article-title":"Formation and Inhibition of Ethyl Glucuronide and Ethyl Sulfate","volume":"265","author":"Stachel","year":"2016","journal-title":"Forensic Sci. Int."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1097\/01.ALC.0000134230.21414.11","article-title":"On Sensitivity, Specificity, and the Influence of Various Parameters on Ethyl Glucuronide Levels in Urine\u2014Results from the WHO\/ISBRA Study","volume":"28","author":"Wurst","year":"2004","journal-title":"Alcohol. Clin. Exp. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1002\/dta.3526","article-title":"The Society of Hair Testing Consensus on General Recommendations for Hair Testing and Drugs of Abuse Testing in Hair","volume":"15","author":"Favretto","year":"2023","journal-title":"Drug Test. Anal."},{"key":"ref_34","first-page":"192","article-title":"Linee Guida per La Determinazione Di Sostanze Stupefacenti e Psicotrope Su Campioni Biologici Con Finalit\u00e0 Tossicologico-Forensi e Medico-Legali","volume":"19","author":"Frison","year":"2023","journal-title":"Riv. Ital. Della Med. Lab."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1093\/alcalc\/agt012","article-title":"Ethyl Glucuronide in Scalp and Non-Head Hair: An Intra-Individual Comparison","volume":"48","author":"Pianta","year":"2013","journal-title":"Alcohol Alcohol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1046\/j.1359-6357.2003.00581.x","article-title":"Traditional Markers of Excessive Alcohol Use","volume":"98","author":"Conigrave","year":"2003","journal-title":"Addiction"},{"key":"ref_37","unstructured":"Eureka (2023, September 04). CDT TEST IN SERUM BY UV\/VIS-FAST\u2014MONOREAGENT\u2014Code Z68215. Available online: https:\/\/www.eurekakit.com\/wp-content\/uploads\/2021\/05\/Z68215_CDT-FAST-MONOREAGENT-500-TEST_002.pdf."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.cca.2016.12.022","article-title":"IFCC Approved HPLC Reference Measurement Procedure for the Alcohol Consumption Biomarker Carbohydrate-Deficient Transferrin (CDT): Its Validation and Use","volume":"465","author":"Schellenberg","year":"2017","journal-title":"Clin. Chim. Acta"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Porpiglia, N.M., Tagliaro, F., Micciolo, R., Canal, L., Musile, G., and Bortolotti, F. (2022). New Evidence of High Association between Carbohydrate Deficient Transferrin (CDT) and Alcohol-Related Road Traffic Accidents. A Retrospective Study on 929 Injured Drivers. Forensic Sci. Int., 340.","DOI":"10.1016\/j.forsciint.2022.111438"},{"key":"ref_40","unstructured":"GTFI-SIMLA (2024, December 13). Transferrina Carboidrato Carente (Carbohydrate-Deficient Transferrin, CDT) Strategie Analitiche Ed Interpretative. Available online: https:\/\/www.gtfi.it\/wp-content\/uploads\/2022\/06\/LineeGuida-SIBioC-GTFI-SIMLA_CDT-rev01-2015.pdf."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"5442","DOI":"10.2174\/1381612823666170714154444","article-title":"Update of Standard Practices for New Method Validation in Forensic Toxicology","volume":"23","author":"Wille","year":"2018","journal-title":"Curr. Pharm. Des."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3019","DOI":"10.1021\/ac020361s","article-title":"Strategies for the Assessment of Matrix Effect in Quantitative Bioanalytical Methods Based on HPLC\u2212MS\/MS","volume":"75","author":"Matuszewski","year":"2003","journal-title":"Anal. Chem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/3\/1344\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:27:23Z","timestamp":1760027243000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/3\/1344"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,5]]},"references-count":42,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["ijms26031344"],"URL":"https:\/\/doi.org\/10.3390\/ijms26031344","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,5]]}}}