Skip to main content

Advertisement

SpringerLink
Log in

Quantification of ethyl glucuronide, ethyl sulfate, nicotine, and its metabolites in human fetal liver and placenta

  • Original Article
  • Published:
Forensic Toxicology Aims and scope Submit manuscript

Abstract

Purpose

Tobacco and alcohol use during pregnancy is serious public health concerns and result in adverse developmental outcomes. Identifying in utero exposure is often achieved through meconium analysis or via maternal self-report. In this study, we analyzed fetal liver and placenta to examine second trimester alcohol and smoking exposure.

Methods

A validated liquid chromatography–tandem mass spectrometry method for simultaneous analysis of nicotine and its metabolites and alcohol markers (ethyl glucuronide: EtG and ethyl sulfate: EtS) was employed to analyze 193 fetal liver and 48 placenta (n = 47 paired) samples from electively terminated pregnancies.

Results

EtG, EtS, and nicotine markers’ limits of detection were 0.7–20 ng/g in fetal samples. Ninety-eight fetal liver and 23 placenta samples were EtG/EtS-positive, while 137 liver and 25 placenta samples were positive for tobacco exposure. When both alcohol markers were present in samples, EtG/EtS ratios were 1.6–11.1 in 17 livers and 2.5–31.1 in 10 placentas. Median (range) summed tobacco marker concentrations were 422 (1.0–2776) and 154 (1.6–1621) ng/g in livers and placentas, respectively. Median EtG and nicotine marker concentrations were higher in liver than placenta in paired samples. Strong evidence of exposure occurred in 11 and 22 pairs, respectively, when both samples were positive for alcohol and/or tobacco markers.

Conclusions

These paired fetal liver and placenta alcohol and tobacco data provided a unique means for examining the effects of in utero exposure, a critical first step in selecting fetal samples for proteomic and RNA sequencing studies that could provide mechanisms for adverse developmental outcomes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. DiFranza JR, Lew RA (1995) Effect of maternal cigarette smoking on pregnancy complications and sudden infant death syndrome. J Fam Pract 40:385–394

    CAS  PubMed  Google Scholar 

  2. Odendaal HJ, Steyn DW, Elliott A, Burd L (2009) Combined effects of cigarette smoking and alcohol consumption on perinatal outcome. Gynecol Obstet Invest 67:1–8

    Article  CAS  PubMed  Google Scholar 

  3. Gray TR, Eiden RD, Leonard KE, Connors G, Shisler S, Huestis MA (2010) Nicotine and metabolites in meconium as evidence of maternal cigarette smoking during pregnancy and predictors of neonatal growth deficits. Nicotine Tob Res 12:658–664

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Bailey BA, Sokol RJ (2011) Prenatal alcohol exposure and miscarriage, stillbirth, preterm delivery, and sudden infant death syndrome. Alcohol Res Health 34:86–91

    PubMed  PubMed Central  Google Scholar 

  5. Coriale G, Fiorentino D, Di Lauro F, Marchitelli R, Scalese B, Fiore M, Maviglia M, Ceccanti M (2013) Fetal alcohol spectrum disorder (FASD): neurobehavioral profile, indications for diagnosis and treatment. Riv Psichiatr 48:359–369

    PubMed  Google Scholar 

  6. Mattson SN, Roesch SC, Glass L, Deweese BN, Coles CD, Kable JA, May PA, Kalberg WO, Sowell ER, Adnams CM, Jones KL, Riley EP, CIFASD (2013) Further development of a neurobehavioral profile of fetal alcohol spectrum disorders. Alcohol Clin Exp Res 37:517–528

    Article  PubMed  Google Scholar 

  7. Bublitz MH, Stroud LR (2012) Maternal smoking during pregnancy and offspring brain structure and function: review and agenda for future research. Nicotine Tob Res 14:388–397

    Article  PubMed  Google Scholar 

  8. Day NL, Richardson GA, Goldschmidt L, Cornelius MD (2000) Effects of prenatal tobacco exposure on preschoolers’ behavior. J Dev Behav Pediatr 21:180–188

    CAS  PubMed  Google Scholar 

  9. Substance Abuse and Mental Health Services Administration (SAMHSA) (2016) Results from the 2015 national survey on drug use and health: summary of national findings. NSDUH Series H-51, HHS Publication no. SMA 16-4984, SAMHSA, Rockville

  10. Benowitz NL, Hukkanen J, Jacob P III (2009) Nicotine chemistry, metabolism, kinetics and biomarkers. Handb Exp Pharmacol 192:29–60 (Springer, Heidelberg, PMC2953858)

    Article  CAS  Google Scholar 

  11. Foti RS, Fisher MB (2005) Assessment of UDP-glucuronosyltransferase catalyzed formation of ethyl glucuronide in human liver microsomes and recombinant UGTs. Forensic Sci Int 153:109–116

    Article  CAS  PubMed  Google Scholar 

  12. Schneider H, Glatt H (2004) Sulpho-conjugation of ethanol in humans in vivo and by individual sulphotransferase forms in vitro. Biochem J 383:543–549

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Coughtrie MW, Burchell B, Leakey JE, Hume R (1988) The inadequacy of perinatal glucuronidation: immunoblot analysis of the developmental expression of individual UDP-glucuronosyltransferase isoenzymes in rat and human liver microsomes. Mol Pharmacol 34:729–735

    CAS  PubMed  Google Scholar 

  14. Matlow JN, Lubetsky A, Aleksa K, Berger H, Koren G (2013) The transfer of ethyl glucuronide across the dually perfused human placenta. Placenta 34:369–373

    Article  CAS  PubMed  Google Scholar 

  15. Adjei AA, Gaedigk A, Simon SD, Weinshilboum RM, Leeder JS (2008) Interindividual variability in acetaminophen sulfation by human fetal liver: implications for pharmacogenetic investigations of drug-induced birth defects. Birth Defects Res A Clin Mol Teratol 82:155–165

    Article  CAS  PubMed  Google Scholar 

  16. Coughtrie MW, Bamforth KJ, Sharp S, Jones AL, Borthwick EB, Barker EV, Roberts RC, Hume R, Burchell A (1994) Sulfation of endogenous compounds and xenobiotics-interactions and function in health and disease. Chem Biol Interact 92:247–256

    Article  CAS  PubMed  Google Scholar 

  17. Morini L, Falcón M, Pichini S, Garcia-Algar O, Danesino P, Groppi A, Luna A (2011) Ethyl-glucuronide and ethyl-sulfate in placental and fetal tissues by liquid chromatography coupled with tandem mass spectrometry. Anal Biochem 418:30–36

    Article  CAS  PubMed  Google Scholar 

  18. Mauvais-Jarvis F (2014) Developmental androgenization programs metabolic dysfunction in adult mice: clinical implications. Adipocyte 3:151–154

    Article  PubMed  PubMed Central  Google Scholar 

  19. Bolton JL, Auten RL, Bilbo SD (2014) Prenatal air pollution exposure induces sexually dimorphic fetal programming of metabolic and neuroinflammatory outcomes in adult offspring. Brain Behav Immun 37:30–44

    Article  CAS  PubMed  Google Scholar 

  20. Rkhzay-Jaf J, O’Dowd JF, Stocker CJ (2012) Maternal obesity and the fetal origins of the metabolic syndrome. Curr Cardiovasc Risk Rep 6:487–495

    Article  PubMed  PubMed Central  Google Scholar 

  21. Fall CHD (2006) Chapter 2: Developmental origins of cardiovascular disease, type 2 diabetes and obesity in humans. In: Wintour EM, Marelyn JO (eds) Early life origins of human health and disease advances in experimental medicine and biology, vol 573. Springer, New York, pp 8–28

    Chapter  Google Scholar 

  22. O’Shaughnessy PJ, Monteiro A, Bhattacharya S, Fowler PA (2011) Maternal smoking and fetal sex significantly affect metabolic enzyme expression in the human fetal liver. J Clin Endocrinol Metab 96:2851–2860

    Article  PubMed  Google Scholar 

  23. O’Shaughnessy PJ, Monteiro A, Bhattacharya S, Fraser MJ, Fowler PA (2013) Steroidogenic enzyme expression in the human fetal liver and potential role in the endocrinology of pregnancy. Mol Hum Reprod 19:177–187

    Article  PubMed  Google Scholar 

  24. O’Shaughnessy PJ, Baker PJ, Monteiro A, Cassie S, Bhattacharya S, Fowler PA (2007) Developmental changes in human fetal testicular cell numbers and messenger ribonucleic acid levels during the second trimester. J Clin Endocrinol Metab 92:4792–4801

    Article  PubMed  Google Scholar 

  25. Himes SK, Concheiro M, Scheidweiler KB, Huestis MA (2014) 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 406:1945–1955

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Piller M, Gilch G, Scherer G, Scherer M (2014) Simple, fast and sensitive LC-MS/MS analysis for the simultaneous quantification of nicotine and 10 of its major metabolites. J Chromatogr B 951-952:7–15

    Article  CAS  Google Scholar 

  27. Scientific Working Group for Forensic Toxicology (2013) Scientific Working Group for Forensic Toxicology (SWGTOX) standard practices for method validation in forensic toxicology. J Anal Toxicol 37:452–474

    Article  Google Scholar 

  28. Høiseth G, Bernard JP, Stephanson N, Normann PT, Christophersen AS, Mørland J, Helander A (2008) Comparison between the urinary alcohol markers EtG, EtS, and GTOL/5-HIAA in a controlled drinking experiment. Alcohol Alcohol 43:187–191

    Article  PubMed  Google Scholar 

  29. Høiseth G, Karinen R, Christophersen A, Mørland J (2010) Practical use of ethyl glucuronide and ethyl sulfate in postmortem cases as markers of antemortem alcohol ingestion. Int J Legal Med 124:143–148

    Article  PubMed  Google Scholar 

  30. Schloegl H, Rost T, Schmidt W, Wurst FM, Weinmann W (2006) Distribution of ethyl glucuronide in rib bone marrow, other tissues and body liquids as proof of alcohol consumption before death. Forensic Sci Int 156:213–218

    Article  CAS  PubMed  Google Scholar 

  31. Gray TR, Magri R, Shakleya DM, Huestis MA (2008) Meconium nicotine and metabolites by liquid chromatography-tandem mass spectrometry: differentiation of passive and nonexposure and correlation with neonatal outcome measures. Clin Chem 54:2018–2027

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Braun JM, Daniels JL, Poole C, Olshan AF, Hornung R, Bernert JT, Xia Y, Bearer C, Barr DB, Lanphear BP (2010) A prospective cohort study of biomarkers of prenatal tobacco smoke exposure: the correlation between serum and meconium and their association with infant birth weight. Environ Health 9:53–64

    Article  PubMed  PubMed Central  Google Scholar 

  33. Pastrakuljic A, Schwartz R, Simone C, Derewlany LO, Knie B, Koren G (1998) Transplacental transfer and biotransformation studies of nicotine in the human placental cotyledon perfused in vitro. Life Sci 63:2333–2342

    Article  CAS  PubMed  Google Scholar 

  34. Joya X, Pujadas M, Falcón M, Civit E, Garcia-Algar O, Vall O, Pichini S, Luna A, de la Torre R (2010) Gas chromatography-mass spectrometry assay for the simultaneous quantification of drugs of abuse in human placenta at 12th week of gestation. Forensic Sci Int 196:38–42

    Article  CAS  PubMed  Google Scholar 

  35. Shakleya DM, Huestis MA (2009) Simultaneous quantification of nicotine, opioids, cocaine, and metabolites in human fetal postmortem brain by liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 393:1957–1965

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Urakawa N, Nagata T, Kudo K, Kimura K, Imamura T (1994) Simultaneous determination of nicotine and cotinine in various human tissues using capillary gas chromatography/mass spectrometry. Int J Legal Med 106:232–236

    Article  CAS  PubMed  Google Scholar 

  37. Helander A, Böttcher M, Fehr C, Dahmen N, Beck O (2009) Detection times for urinary ethyl glucuronide and ethyl sulfate in heavy drinkers during alcohol detoxification. Alcohol Alcohol 44:55–61

    Article  CAS  PubMed  Google Scholar 

  38. Wurst FM, Seidl S, Ladewig D, Müller-Spahn F, Alt A (2002) Ethyl glucuronide: on the time course of excretion in urine during detoxification. Addict Biol 7:427–434

    Article  CAS  PubMed  Google Scholar 

  39. Musshoff F, Albermann E, Madea B (2010) Ethyl glucuronide and ethyl sulfate in urine after consumption of various beverages and foods—misleading results? Int J Legal Med 124:623–630

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

This research was supported by the Medical Research Council (UK) grant MR/L01 0011/1 and the Intramural Research Program at the National Institute on Drug Abuse of the National Institutes of Health. Paired fetal liver and placenta samples were graciously provided by the Joint Medical Research Council/Wellcome Trust (grant number 099175/Z/12/Z) Human Developmental Biology Resource (www.hdbr.org).

Author information

Authors and Affiliations

  1. Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD, USA

    Madeleine J. Swortwood, Sarah H. Bartock, Karl B. Scheidweiler & Marilyn A. Huestis

  2. Department of Forensic Science, College of Criminal Justice, Sam Houston State University, Huntsville, TX, USA

    Madeleine J. Swortwood

  3. Institute of Medical Sciences, School of Medicine, Medical Sciences and Nutrition, University of Aberdeen, Foresterhill, Aberdeen, Scotland, UK

    Sophie Shaw, Panagiotis Filis, Alex Douglas & Paul A. Fowler

  4. College of Medical, Veterinary and Life Sciences, Institute of Biodiversity, Animal Health and Comparative Medicine, University of Glasgow, Glasgow, Scotland, G61 1QH, UK

    Peter J. O’Shaughnessy & Ugo Soffientini

  5. MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, Scotland, UK

    Baltasar Lucendo-Villarin & David C. Hay

  6. University of Bristol, Senate House, Tyndall Avenue, Bristol, England, BS8 1TH, UK

    John P. Iredale

  7. University of Maryland School of Medicine, 655 W Baltimore Street, Baltimore, MD, 21201, USA

    Marilyn A. Huestis

  8. 683 Shore Road, Severna Park, MD, 21146, USA

    Marilyn A. Huestis

Authors
  1. Madeleine J. Swortwood
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sarah H. Bartock
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Karl B. Scheidweiler
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sophie Shaw
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Panagiotis Filis
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Alex Douglas
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Peter J. O’Shaughnessy
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ugo Soffientini
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Baltasar Lucendo-Villarin
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. John P. Iredale
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. David C. Hay
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Paul A. Fowler
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Marilyn A. Huestis
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Marilyn A. Huestis.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of all involved institutions and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Collection of fetal liver material was approved by the National Health Service Grampian Research Ethics Committees (REC04/S0802/21). This article does not contain any studies with animals performed by any of the authors. Informed consent was obtained from all individual participants included in the study. No identify information is included in this article.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 44 kb)

Supplementary material 2 (PDF 8 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Swortwood, M.J., Bartock, S.H., Scheidweiler, K.B. et al. Quantification of ethyl glucuronide, ethyl sulfate, nicotine, and its metabolites in human fetal liver and placenta. Forensic Toxicol 36, 102–112 (2018). https://doi.org/10.1007/s11419-017-0389-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11419-017-0389-2

Keywords

Search

Navigation