Analytical and Bioanalytical Chemistry

, Volume 396, Issue 7, pp 2469–2477 | Cite as

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

  • Abdulsallam Bakdash
  • Pascal Burger
  • Tamme W. Goecke
  • Peter A. Fasching
  • Udo Reulbach
  • Stefan Bleich
  • Martin Hastedt
  • Michael Rothe
  • Matthias W. Beckmann
  • Fritz Pragst
  • Johannes Kornhuber
Original Paper

Abstract

Fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) were determined in 602 meconium samples in a maternal health evaluation study for detection of gestational alcohol consumption. A validated headspace solid phase microextraction method in combination with GC-MS was used for FAEE and the cumulative concentration of ethyl palmitate, ethyl linoleate, ethyl oleate, and ethyl stearate with a cut-off of 500 ng/g was applied for interpretation. A new and simple method was developed and validated for quantification of EtG from 10–20 mg meconium with D5-EtG as internal standard consisting of 30 min. extraction with methanol/water (1:1, v/v), evaporation of methanol, filtration of the aqueous solution through a cellulose filter and injection into LC-MS-MS. The limits of detection and quantification for EtG were 10 and 30 ng/g, the recovery 86.6 to 106.4% and the standard deviation of the concentrations ranged from 13% at 37 ng/g to 5% at 46,700 ng/g (N = 6). FAEE above the cut-off were found in 43 cases (7.1%) with cumulative concentrations between 507 and 22,580 ng/g and with one outlier of about 150,000 ng/g (EtG not detected). EtG was detected in 97 cases (16.3%) and concentrations between LOD and 10,200 ng/g with another outlier of 82,000 ng/g (FAEE 10,500 ng/g). Optimal agreement between the two markers was obtained with a cut-off for EtG of 274 ng/g and 547 cases with both FAEE- and EtG-negative, 33 cases with both FAEE- and EtG-positive, nine cases with FAEE-positive and EtG-negative, and seven cases with FAEE-negative and EtG-positive. Differences in physical, chemical, and biochemical properties and in the pharmacokinetic behavior are discussed as reasons for the deviating cases. In none of the 602 cases, serious alcohol consumption was reported by the mothers and no evidence for gestational ethanol exposure was observed in the medical investigation of the newborns. It is concluded that the combined use of FAEE and EtG in meconium as markers for fetal alcohol exposure essentially increases the accuracy of the interpretation and helps to avoid false positive and false-negative results.

Keywords

Ethyl glucuronide Fatty acid ethyl esters Fetal alcohol exposure Headspace solid phase microextraction Liquid chromatography–mass spectrometry Meconium 

References

  1. 1.
    Mukherjee RAS, Hollins S, Turk J (2006) J Royal Soc Med 99:298–302CrossRefGoogle Scholar
  2. 2.
    Manning MA, Hoyme HE (2007) Neurosci Behav Rev 31:230–238Google Scholar
  3. 3.
    Bearer CF, Lee S, Salvator AE, Minnes S, Swick A, Yamashita T, Singer LT (1999) Alcohol Clin Exp Res 23:487–493Google Scholar
  4. 4.
    Bearer CF, Jacobson JL, Jacobson SW, Barr D, Croxford J, Molteno CD, Viljoen DL, Marais AS, Chiodo LM, Cwik AS (2003) J Pediatr 143:463–469CrossRefGoogle Scholar
  5. 5.
    Bearer CF, Santiago LM, O'Riordan MA, Buck K, Lee SC, Singer LT (2005) J Pediatr 146:824–830CrossRefGoogle Scholar
  6. 6.
    Peterson J, Kirchner HL, Xue W, Minnes S, Singer LT, Bearer CF (2008) J Pediatr 152:788–792CrossRefGoogle Scholar
  7. 7.
    Klein J, Karaskow T, Koren G (1999) Ther Drug Monit 21:644–646CrossRefGoogle Scholar
  8. 8.
    Chan D, Bar-Oz B, Pellerin B, Paciorek C, Klein J, Kapur B, Farine D, Koren G (2003) Ther Drug Monit 25:271–278CrossRefGoogle Scholar
  9. 9.
    Chan D, Caprara D, Blanchette P, Klein J, Koren G (2004) Clin Biochem 37:429–438CrossRefGoogle Scholar
  10. 10.
    Chan D, Klein J, Karaskov T, Koren G (2004) Ther Drug Monit 26:474–481CrossRefGoogle Scholar
  11. 11.
    Caprara DL, Nash K, Greenbaum R, Rovet J, Koren G (2007) Neurosci Behav Rev 31:354–360Google Scholar
  12. 12.
    Gareri J, Lynn H, Handley M, Rao C, Koren G (2008) Ther Drug Monit 30:239–245CrossRefGoogle Scholar
  13. 13.
    Hutson JR, Aleksa K, Pragst F, Koren G (2009) J Chromatogr B Analyt Technol Biomed Life Sci 877:8–12CrossRefGoogle Scholar
  14. 14.
    Zelner I, Koren G (2009) Ther Drug Monit 31:170–172CrossRefGoogle Scholar
  15. 15.
    Moore CM, Lewis D (2001) Clin Chim Acta 312:235–237CrossRefGoogle Scholar
  16. 16.
    Moore C, Jones J, Lewis D, Buchi K (2003) Clin Chem 49:133–136CrossRefGoogle Scholar
  17. 17.
    Derauf C, Katz AR, Easa D (2003) Am J Epidemiol 158:705–709CrossRefGoogle Scholar
  18. 18.
    Ostrea EM Jr, Hernandez JD, Bielawski DM, Kan JM, Leonardo GM, Abela MB, Church MW, Hannigan JH, Janisse JJ, Ager JW, Sokol RJ (2006) Alcohol Clin Exp Res 30:1152–1159CrossRefGoogle Scholar
  19. 19.
    Burd L, Hofer R (2008) Birth Defects Res A Clin Mol Teratol 82:487–493CrossRefGoogle Scholar
  20. 20.
    Garcia-Algar O, Kulaga V, Gareri J, Koren G, Vall O, Zuccaro P, Pacifici R, Pichini S (2008) Ther Drug Monit 30:249–254CrossRefGoogle Scholar
  21. 21.
    Pichini S, Pellegrini M, Gareri J, Koren G, Garcia-Algar O, Vall O, Vagnarelli F, Zuccaro P, Marchei E (2008) Pharm Biomed Anal 48:927–933CrossRefGoogle Scholar
  22. 22.
    Chan D, Knie B, Koren G (2003) Placenta 24:A41CrossRefGoogle Scholar
  23. 23.
    Morini L, Marchei E, Pellegrini M, Groppi A, Stramesi C, Vagnarelli F, Garcia-Algar O, Pacifici R, Pichini S (2008) Ther Drug Monit 30:725–732CrossRefGoogle Scholar
  24. 24.
    Pichini S, Morini L, Marchei E, Palmi I, Rotolo MC, Vagnarelli F, Garcia-Algar O, Vall O, Zuccaro P (2009) Can J Clin Pharmacol 16:e370–e375Google Scholar
  25. 25.
    Reulbach U, Bleich S, Knörr J, Burger P, Fasching PA, Kornhuber J, Beckmann MW, Goecke TW (2009) Fortschr Neurol Psychiatr 77:708–713Google Scholar
  26. 26.
    Pragst F, Auwaerter V, Sporkert F, Spiegel K (2001) Forensic Sci Int 121:76–88CrossRefGoogle Scholar
  27. 27.
    Pragst F, Rothe M, Moench B, Hastedt M, Herre S, Simmert D (2010) Forensic Sci Int. doi:10.1016/j.forsciint.2009.12.028
  28. 28.
    Mierswa I, Wurst M, Klinkenberg R, Scholz M, Euler T (2006) Proceedings of the 12th ACM SIGKDD International Conference on Knowledge Discovery and Data Mining (KDD-06)Google Scholar
  29. 29.
    Droenner P, Schmitt G, Aderjan R, Zimmer H (2002) Forensic Sci Int 126:24–29CrossRefGoogle Scholar
  30. 30.
    Høiseth G, Bernard JP, Karinen R, Johnsen L, Helander A, Christophersen AS, Mørland J (2007) Forensic Sci Int 172:119–124CrossRefGoogle Scholar
  31. 31.
    Høiseth G, Morini L, Polettini A, Christophersen A, Mørland J (2009) Forensic Sci Int 188:52–56CrossRefGoogle Scholar
  32. 32.
    Halter CC, Dresen S, Auwaerter V, Wurst FM, Weinmann W (2008) Int J Legal Med 122:123–128CrossRefGoogle Scholar
  33. 33.
    Bergström J, Helander A, Jones AW (2003) Forensic Sci Int 133:86–94CrossRefGoogle Scholar
  34. 34.
    Gerdin E, Gabrielsson J, Lindberg B, Rane A (1990) Pharmacol Toxicol 66:815–819CrossRefGoogle Scholar
  35. 35.
    Gerdin E, Rane A, Lindberg B (1990) J Perinat Med 18:305–312CrossRefGoogle Scholar
  36. 36.
    Garland M, Abildskov KM, Taylor S, Benzeroual K, Caspersen CS, Arroyo SE, Kiu TW, Reznik B, Weldy P, Daniel SS, Stark RI (2006) Drug Metabol Dispos 34:636–646CrossRefGoogle Scholar
  37. 37.
    De Giovanni N, Donadio G, Chiarotti M (2008) J Anal Toxicol 32:156–159Google Scholar
  38. 38.
    Kitchens JM (1994) J Am Med Assoc 272:1782–1787CrossRefGoogle Scholar
  39. 39.
    Pragst F, Yegles M (2008) Ther Drug Monit 30:255–263CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Abdulsallam Bakdash
    • 1
  • Pascal Burger
    • 2
  • Tamme W. Goecke
    • 3
  • Peter A. Fasching
    • 3
  • Udo Reulbach
    • 2
  • Stefan Bleich
    • 2
  • Martin Hastedt
    • 1
  • Michael Rothe
    • 4
  • Matthias W. Beckmann
    • 3
  • Fritz Pragst
    • 1
  • Johannes Kornhuber
    • 2
  1. 1.Institute of Legal MedicineUniversity Hospital CharitéBerlinGermany
  2. 2.Department of Psychiatry and PsychotherapyFriedrich-Alexander-University of Erlangen-NurembergErlangenGermany
  3. 3.Department of Obstetrics and GynecologyUniversity of Erlangen-Nuremberg, ErlangenErlangenGermany
  4. 4.Lipidomix GmbHBerlinGermany

Personalised recommendations