Analytical and Bioanalytical Chemistry

, Volume 388, Issue 7, pp 1545–1556 | Cite as

Detection of diazepam in urine, hair and preserved oral fluid samples with LC-MS-MS after single and repeated administration of Myolastan® and Valium®

  • Marleen Laloup
  • Maria del Mar Ramirez Fernandez
  • Michelle Wood
  • Viviane Maes
  • Gert De Boeck
  • Yvan Vanbeckevoort
  • Nele Samyn
Original Paper


Sedative agents are used to facilitate sexual assault due to their ability to render the victim passive, submissive and unable to resist. The primary pharmacological effect of the benzodiazepine tetrazepam is muscle relaxation, whereas the benzodiazepine diazepam acts on the central nervous system (CNS) exerting mainly sedation effects. Therefore, contrary to tetrazepam, diazepam is an often-abused drug, which can potentially be used as a date-rape drug. In this study, we describe the detection of low amounts of diazepam in Myolastan® (Sanofi–Synthelabo S.A., Brussels, Belgium) and Epsipam® (Will-Pharma, Wavre, Belgium) 50mg tablet preparations by LC-MS-MS, GC-FID and HPLC-DAD. Considering the important forensic implication of this finding, a study was conducted with volunteers receiving a single or repeated dosage of Myolastan®. Urine, hair and preserved oral fluid samples were analysed using a previously described sensitive and specific LC-MS-MS detection method allowing for the simultaneous quantification of tetrazepam, diazepam, nordiazepam, oxazepam and temazepam. This study demonstrates that diazepam can be observed in urine samples even after a single dose of Myolastan®. In addition, maintaining therapy for 1 week results in the detection of both diazepam and nordiazepam in urine samples and of diazepam in the first hair segment. Importantly, comparing urine and hair samples after a single intake of diazepam versus the single and 1 week administration of Myolastan® shows that the possible metabolic conversion of tetrazepam to diazepam is a more plausible explanation for the detection of diazepam in biological samples after the intake of Myolastan®. As such, these results reveal that the presence of diazepam and/or nordiazepam in biological samples from alleged drug-facilitated assault cases should be interpreted with care.


LC-MS-MS Tetrazepam Diazepam Urine Hair Oral fluid 



The authors would like to thank Dr. Frank Peters and Dr. Jochen Beyer for their valuable discussions and suggestions.


  1. 1.
    Cheze M, Villain M, Pepin G (2004) Forensic Sci Int 145:123–130CrossRefGoogle Scholar
  2. 2.
    ElSohly MA, Gul W, ElSohly KM, Avula B, Khan IA (2006) J Anal Toxicol 30:524–538Google Scholar
  3. 3.
    Hurley M, Parker H, Wells DL (2006) J Clin Forensic Med 13:181–185CrossRefGoogle Scholar
  4. 4.
    Kintz P, Villain M, Dumestre-Toulet V, Ludes B (2005) J Clin Forensic Med 12:36–41CrossRefGoogle Scholar
  5. 5.
    Ohshima T (2006) J Clin Forensic Med 13:44–45CrossRefGoogle Scholar
  6. 6.
    Scott-Ham M, Burton FC (2006) J Clin Forensic Med 13:107–111CrossRefGoogle Scholar
  7. 7.
    Villain M, Cheze M, Dumestre V, Ludes B, Kintz P (2004) J Anal Toxicol 28:516–519Google Scholar
  8. 8.
    Villain M, Cheze M, Tracqui A, Ludes B, Kintz P (2004) Forensic Sci Int 143:157–161CrossRefGoogle Scholar
  9. 9.
    Negrusz A, Gaensslen RE (2003) Anal Bioanal Chem 376:1192–1197CrossRefGoogle Scholar
  10. 10.
    Scott-Ham M, Burton FC (2005) J Clin Forensic Med 12:175–186CrossRefGoogle Scholar
  11. 11.
    Kintz P, Villain M, Cheze M, Pepin G (2005) Forensic Sci Int 153:222–226CrossRefGoogle Scholar
  12. 12.
    Kintz P, Villain M, Dumestre-Toulet V, Capolaghi B, Cirimele V (2005) Ther Drug Monit 27:741–743CrossRefGoogle Scholar
  13. 13.
    Laloup M, Ramirez Fernandez MM, De Boeck G, Wood M, Maes V, Samyn N (2005) J Anal Toxicol 29:616–626Google Scholar
  14. 14.
    Kintz P, Villain M, Ludes B (2004) Ther Drug Monit 26:211–214CrossRefGoogle Scholar
  15. 15.
    Concheiro M, Villain M, Bouchet S, Ludes B, Lopez-Rivadulla M, Kintz P (2005) Ther Drug Monit 27:565–570CrossRefGoogle Scholar
  16. 16.
    Kintz P, Villain M, Cirimele V, Pepin G, Ludes B (2004) Forensic Sci Int 145:131–135CrossRefGoogle Scholar
  17. 17.
    Kintz P, Villain M, Ludes B (2004) J Chromatogr B Anal Technol Biomed Life Sci 811:59–63CrossRefGoogle Scholar
  18. 18.
    Samyn N, Verstraete A, van Haeren C, Kintz P (1999) Forensic Sci Rev 11:1–19Google Scholar
  19. 19.
    Cone EJ, Presley L, Lehrer M, Seiter W, Smith M, Kardos KW, Fritch D, Salamone S, Niedbala RS (2002) J Anal Toxicol 26:541–546Google Scholar
  20. 20.
    Shah VP, Midha KK, Dighe S, McGilveray IJ, Skelly JP, Yacobi A, Layloff T, Viswanathan CT, Cook CE (1991) Eur J Drug Metab Pharmacokinet 16:249–255CrossRefGoogle Scholar
  21. 21.
    Shah VP, Midha KK, Findlay JW, Hill HM, Hulse JD, McGilveray IJ, McKay G, Miller KJ, Patnaik RN, Powell ML, Tonelli A, Viswanathan CT, Yacobi A (2000) Pharm Res 17:1551–1557CrossRefGoogle Scholar
  22. 22.
    Herzler M, Herre S, Pragst F (2003) J Anal Toxicol 27:233–242Google Scholar
  23. 23.
    Kintz P, Villain M, Concheiro M, Cirimele V (2005) Forensic Sci Int 150:213–220CrossRefGoogle Scholar
  24. 24.
    Quintela O, Cruz A, Castro A, Concheiro M, Lopez-Rivadulla M (2005) J Chromatogr B Anal Technol Biomed Life Sci 825:63–71CrossRefGoogle Scholar
  25. 25.
    Bonfiglio R, King RC, Olah TV, Merkle K (1999) Rapid Commun Mass Spectrom 13:1175–1185CrossRefGoogle Scholar
  26. 26.
    Moffat AC, Osselton MD, Widdop B (2004) (eds) Clarke’s analysis of drugs and poisons. Phramaceutical Press, LondonGoogle Scholar
  27. 27.
    Sharp ME, Wallace SM, Hindmarsh KW, Peel HW (1983) J Anal Toxicol 7:11–14Google Scholar
  28. 28.
    Henderson GL, Harkey MR, Zhou C, Jones RT, Jacob P III (1996) J Anal Toxicol 20:1–12Google Scholar
  29. 29.
    Maurer H, Pfleger K (1987) J Chromatogr 422:85–101CrossRefGoogle Scholar
  30. 30.
    Pavlic M, Libiseller K, Grubwieser P, Schubert H, Rabl W (2006) Int J Legal Med (in press)Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Marleen Laloup
    • 1
  • Maria del Mar Ramirez Fernandez
    • 1
  • Michelle Wood
    • 2
  • Viviane Maes
    • 3
  • Gert De Boeck
    • 1
  • Yvan Vanbeckevoort
    • 1
  • Nele Samyn
    • 1
  1. 1.Federal Public Service Justice, National Institute of Criminalistics and Criminology (NICC)Section ToxicologyBrusselsBelgium
  2. 2.Waters CorporationMS Technologies CentreManchesterUK
  3. 3.Department of Clinical Chemistry-ToxicologyAcademic Hospital, Free University of BrusselsBrusselsBelgium

Personalised recommendations