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Fetal exposure to ethanol: relationship between ethyl glucuronide in maternal hair during pregnancy and ethyl glucuronide in neonatal meconium

Xavier Joya, Emilia Marchei, Judith Salat-Batlle, Oscar García-Algar, Valeria Calvaresi, Roberta Pacifici and Simona Pichini


Background: In recent years, fatty acid ethyl esters (FAEEs) and ethyl glucuronide (EtG) in meconium emerged as reliable, direct biological markers for establishing gestational ethanol exposure. We investigated whether EtG in maternal hair measured during the three trimesters of pregnancy correlated with EtG and FAEEs in neonatal meconium.

Methods: In a prospective sample of 80 mother-infant dyads from Barcelona (Spain), we measured EtG and FAEE in maternal hair segments and meconium samples using a validated UHPLC-MS/MS method.

Results: Fifty-eight (72.5%) women had EtG concentrations in the hair shafts >7 pg/mg in one or more pregnancy trimesters, and EtG and FAEEs in meconium samples were documented in 50 and 24 of their neonates, respectively. The best significant correlations (p<0.0001) were found between EtG concentration in the proximal 0–3 and 3–6 hair shaft segments corresponding to the last two pregnancy trimesters and EtG in neonatal meconium (ρ=0.609 and ρ=0.577, respectively). Using the combination of EtG in meconium ≥30 ng/g and a median of EtG >11 pg/mg in maternal hair during the second and third trimesters of pregnancy, prenatal ethanol exposure could be predicted with a sensitivity of 85.7% and specificity of 73.7%.

Conclusions: This study provides evidence of proven fetal exposure to ethanol during the second and third trimesters of pregnancy by linking detection of ethanol biomarkers (EtG) in maternal hair segments and EtG in neonatal meconium.

Corresponding author: Simona Pichini, PhD, Department of Therapeutic Research and Medicines Evaluation, Istituto Superiore di Sanitá, Viale Regina Elena 299, 00161 Rome, Italy, Phone: +39 064 9906545, Fax: +39 064 9902016, E-mail:


We thank the women who participated in this study. The authors are grateful to Marta Pulido, MD, PhD, for editing the manuscript and editorial assistance and to Dr. Manuela Pellegrini, Dr. Maria Concetta Rotolo and Dr. Silvia Graziano for technical help.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: This study was supported by grants from Fondo de Investigaciones Sanitarias (FIS) FEDER: Fondo Europeo de Desarrollo Regional (PI13/01135) and Red de Salud Materno-Infantil y del Desarrollo (SAMID) (RD12/0026/0003) from the Instituto Carlos III (Spain), and partially supported by Generalitat de Catalunya (Spain) AGAUR (2014SGR584), RecerCaixa (OG085818) and Fundación Mutua Madrileña (AP150572014) grants.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


1. Lemoine P, Harousseau H, Borteyru JP, Menuet JC. Children of alcoholic parents – observed anomalies: discussion of 127 cases. Ther Drug Monit 2003;25:132–6. Search in Google Scholar

2. Koren G, Nulman I, Chudley AE, Loocke C. Fetal alcohol spectrum disorder. Can Med Assoc J 2003;169:1181–5. Search in Google Scholar

3. Morini L, Marchei E, Vagnarelli F, Garcia Algar O, Groppi A, Mastrobattista L, et al. Ethyl glucuronide and ethyl sulfate in meconium and hair-potential biomarkers of intrauterine exposure to ethanol. Forensic Sci Int 2010;196:74–7. Search in Google Scholar

4. Pichini S, Marchei E, Vagnarelli F, Tarani L, Raimondi F, Maffucci R, et al. Assessment of prenatal exposure to ethanol by meconium analysis: results of an Italian multicenter study. Alcohol Clin Exp Res 2012;36:417–24. Search in Google Scholar

5. Chan D, Bar-Oz B, Pellerin B, Paciorek C, Klein J, Kapur B, et al. Population baseline of meconium fatty acid ethyl esters among infants of nondrinking women in Jerusalem and Toronto. Ther Drug Monit 2003;25:271–8. Search in Google Scholar

6. Bearer CF, Jacobson JL, Jacobson SW, Barr D, Croxford J, Molteno CD, et al. Validation of a new biomarker of fetal exposure to alcohol. J Pediatr 2003;143:463–9. Search in Google Scholar

7. Bakdash A, Burger P, Goecke TW, Fasching PA, Reulbach U, Bleich S, et al. Quantification of fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) in meconium from newborns for detection of alcohol abuse in a maternal health evaluation study. Anal Bioanal Chem 2010;396:2469–77. Search in Google Scholar

8. Garcia-Algar O, Kulaga V, Gareri J, Koren G, Vall O, Zuccaro P, et al. Alarming prevalence of fetal alcohol exposure in a Mediterranean city. Ther Drug Monit 2008;30:249–54. Search in Google Scholar

9. Himes SK, Dukes KA, Tripp T, Petersen JM, Raffo C, Burd L, et al. Clinical sensitivity and specificity of meconium fatty acid ethyl ester, ethyl glucuronide, and ethyl sulfate for detecting maternal drinking during pregnancy. Clin Chem 2015;61: 523–32. Search in Google Scholar

10. Morini L, Marchei E, Tarani L, Trivelli M, Rapisardi G, Elicio MR, et al. Testing ethylglucuronide in maternal hair and nails for the assessment of fetal exposure to alcohol: comparison with meconium testing. Ther Drug Monit 2013;35:402–7. Search in Google Scholar

11. Pichini S, Morini L, Pacifici R, Tuyay J, Rodrigues W, Solimini R, et al. Development of a new immunoassay for the detection of ethyl glucuronide (EtG) in meconium: validation with authentic specimens analyzed using LC-MS/MS. Preliminary results. Clin Chem Lab Med 2014;52:1179–85. Search in Google Scholar

12. Morini L, Groppi A, Marchei E, Vagnarelli F, Garcia Algar O, Zuccaro P, et al. Population baseline of meconium ethyl glucuronide and ethyl sulfate concentrations in newborns of nondrinking women in 2 Mediterranean cohorts. Ther Drug Monit 2010;32:359–36. Search in Google Scholar

13. Wurst FM, Kelso E, Weinmann W, Pragst F, Yegles M, Sundstrom Poromaa I. Measurement of direct ethanol metabolites suggests higher rate of alcohol use among pregnant women than found with the AUDIT – a pilot study in a population-based sample of Swedish women. Am J Obstet Gynecol 2008;198:e1–5. Search in Google Scholar

14. Manich A, Velasco M, Joya X, García-Lara NR, Pichini S, Vall O, et al. [Validity of a maternal alcohol consumption questionnaire in detecting prenatal exposure]. An Pediatr (Barc) 2012;76:324–8. Search in Google Scholar

15. Lange S, Shield K, Koren G, Rehm J, Popova S. A comparison of the prevalence of prenatal alcohol exposure obtained via maternal self-reports versus meconium testing: a systematic literature review and meta-analysis. BMC Pregnancy Childbirth 2014;14:127. Search in Google Scholar

16. Goh YI, Hutson JR, Lum L, Roukema H, Gareri J, Lynn H, et al. Rates of fetal alcohol exposure among newborns in a high-risk obstetric unit. Alcohol 2010;44:629–34. Search in Google Scholar

17. Hutson JR, Magri R, Gareri JN, Koren G. The incidence of prenatal alcohol exposure in Montevideo Uruguay as determined by meconium analysis. Ther Drug Monit 2010;32:311–7. Search in Google Scholar

18. Pichini S, Pellegrini M, Gareri J, Koren G, Garcia-Algar O, Vall O, et al. Liquid chromatography-tandem mass spectrometry for fatty acid ethyl esters in meconium: assessment of prenatal exposure to alcohol in two European cohorts. J Pharm Biomed Anal 2008;48:927–33. Search in Google Scholar

19. Pichini S, Morini L, Marchei E, Palmi I, Rotolo MC, Vagnarelli F, et al. Ethylglucuronide and ethylsulfate in meconium to assess gestational ethanol exposure: preliminary results in two Mediterranean cohorts. Can J Clin Pharmacol 2009;16:e370–5. Search in Google Scholar

20. Pichini S, Puig C, Zuccaro P, Marchei E, Pellegrini M, Murillo J, et al. Assessment of exposure to opiates and cocaine during pregnancy in a Mediterranean city: preliminary results of the “Meconium Project”. Forensic Sci Int 2005;153:59–65. Search in Google Scholar

21. Falcon M, Valero F, Pellegrini M, Rotolo MC, Scaravelli G, Joya J, et al. Exposure to psychoactive substances in women who request voluntary termination of pregnancy assessed by serum and hair testing. Forensic Sci Int 2010;196:22–6. Search in Google Scholar

22. Albermann ME, Musshoff F, Madea B. Comparison of ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs) concentrations in hair for testing abstinence. Anal Bioanal Chem 2011;400:175–81. Search in Google Scholar

23. Kintz P, Nicholson D. Testing for ethanol markers in hair: discrepancies after simultaneous quantification of ethyl glucuronide and fatty acid ethyl esters. Forensic Sci Int 2014;243:44–6. Search in Google Scholar

24. Pragst F, Rothe M, Moench B, Hastedt M, Herre S, Simmert D. Combined use of fatty acid ethyl esters and ethyl glucuronide in hair for diagnosis of alcohol abuse: interpretation and advantages. Forensic Sci Int 2010;196:101–10. Search in Google Scholar

25. Pragst F, Yegles M. Determination of fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) in hair: a promising way for retrospective detection of alcohol abuse during pregnancy? Ther Drug Monit 2008;30:255–63. Search in Google Scholar

26. Kintz P. Consensus for the use of alcohol markers in hair for assessment of both abstinence and chronic excessive alcohol consumption. Forensic Sci Int 2015;249:A1–2. Search in Google Scholar

27. Pichini S, Gottardi M, Marchei E, Svaizer F, Pellegrini M, Rotolo MC, et al. Rapid extraction, identification and quantification of drugs of abuse in hair by immunoassay and ultra-performance liquid chromatography tandem mass spectrometry. Clin Chem Lab Med 2014;52:679–86. Search in Google Scholar

28. Joya X, Papaseit E, Civit E, Pellegrini M, Vall O, Garcia-Algar O, et al. Unsuspected exposure to cocaine in preschool children from a Mediterranean city detected by hair analysis. Ther Drug Monit 2009;31:391–5. Search in Google Scholar

29. Pichini S, Garcia-Algar O, Alvarez A, Gottardi M, Marchei E, Svaizer F, et al. Assessment of unsuspected exposure to drugs of abuse in children from a Mediterranean city by hair testing. Int J Environ Res Public Health 2014;11:2288–98. Search in Google Scholar

30. Vagnarelli F, Palmi I, García-Algar O, Falcon M, Memo L, Tarani L, et al. A survey of Italian and Spanish neonatologists and paediatricians regarding awareness of the diagnosis of FAS and FASD and maternal ethanol use during pregnancy. BMC Pediatr 2011;11:51. Search in Google Scholar

31. Himes SK, Concheiro M, Scheidweiler KB, Huestis MA. Validation of a novel method to identify in utero ethanol exposure: simultaneous meconium extraction of fatty acid ethyl esters, ethyl glucuronide, and ethyl sulfate followed by LC-MS/MS quantification. Anal Bioanal Chem 2014;406:1945–55. Search in Google Scholar

32. Zelner I, Hutson JR, Kapur BM, Feig DS, Koren G. False-positive meconium test results for fatty acid ethyl esters secondary to delayed sample collection. Alcohol Clin Exp Res 2012;36: 1497–506. Search in Google Scholar

33. Jansson-Nettelbladt E, Meurling S, Petrini B, Sjölin J. Endogenous ethanol fermentation in a child with short bowel syndrome. Acta Paediatr 2006;95:502–4. Search in Google Scholar

34. Cabarcos P, Tabernero MJ, Otero JL, Míguez M, Bermejo AM, Martello S, et al. Quantification of fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) in meconium for detection of alcohol abuse during pregnancy: correlation study between both biomarkers. J Pharm Biomed Anal 2014;100:74–8. Search in Google Scholar

35. McDonald SW, Hicks M, Rasmussen C, Nagulesapillai T, Cook J, Tough SC. Characteristics of women who consume alcohol before and after pregnancy recognition in a Canadian sample: a prospective cohort study. Alcohol Clin Exp Res 2014;38:3008–16. Search in Google Scholar

36. Gareri J, Brien J, Reynolds J, Koren G. Potential role of the placenta in fetal alcohol spectrum disorder. Paediatr Drugs 2009;11:26–9. Search in Google Scholar

37. Suesse S, Pragst F, Mieczkowski T, Selavka CM, Elian A, Sachs H, et al. Practical experiences in application of hair fatty acid ethyl esters and ethyl glucuronide for detection of chronic alcohol abuse in forensic cases. Forensic Sci Int 2012;218:82–91. Search in Google Scholar

38. León-Muñoz LM, Galán I, Valencia-Martín JL, López-García E, Guallar-Castillón P, Rodríguez-Artalejo F. Is a specific drinking pattern a consistent feature of the Mediterranean diet in Spain in the XXI century? Nutr Metab Cardiovasc Dis 2014;24:1074–81. Search in Google Scholar

39. Soler-Vila H, Galán I, Donado-Campos J, Sánchez-Alfonso F, Valencia-Martín JL, Morilla F, et al. Binge drinking in Spain, 2008–2010. Alcohol Clin Exp Res 2014;38:810–9. Search in Google Scholar

40. Scafato, E. [Epidemiology of alcohol consumption among women]. Ann Ist Super Sanita 2004;40:25–33. Search in Google Scholar

Received: 2015-6-4
Accepted: 2015-7-18
Published Online: 2015-8-13
Published in Print: 2016-3-1

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