Comprehensive Approach for Monitoring Human Tissue Degradation

  • Lena M. DuboisEmail author
  • Pierre-Hugues Stefanuto
  • Katelynn A. Perrault
  • Geraldine Delporte
  • Philippe Delvenne
  • Jean-François Focant


In recent years, comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC × GC–TOFMS) has been reported as a suitable tool for the determination of volatile organic compounds (VOCs) emitted during the process of cadaveric decomposition. The main aim of the present study was to investigate temporal changes in VOC patterns during the decomposition process of various human tissues. The focus of previous research was mainly on the analysis of VOCs produced by whole cadavers. However, this study aimed to identify whether the VOCs produced during decomposition differ between specific organs, and further, to determine the extent of the variation between cadavers. The sampling process developed for this project allowed inter- and intra-cadaveric comparison. The headspace of heart, lung, liver, kidney and blood was monitored during the decomposition process. Tissue samples from five different cadavers were sampled regularly by dynamic pumping onto sorbent tubes that were further thermally desorbed onto a GC × GC–TOFMS system. A large amount of data (n = 774) was obtained, leading to challenges in the integration, interpretation and representation of the results. Eventually, multivariate statistical methods, such as principal components analysis (PCA) and hierarchical cluster analysis (HCA) were applied to the dataset to evaluate trends and differences in subgroups. It was demonstrated that there were subtle differences between the sets of compounds produced from each organ due to the different functions they carry out within the body. However, VOC profiles were more similar among organs from the same cadaver than when comparing samples from different cadavers. Various reasons may cause the differences between the analyzed cadavers, ranging from the individual diet and lifestyle to the time since death.


Human decomposition Forensic science Forensic chemistry GC × GC–TOFMS Volatile organic compounds Organs Death odor 



We wish to thank Restek® Corp. and Trajan Scientific and Medical® for providing us GC phases and various GC consumables. We would also like to thank LECO® for their contribution and their technical support.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The research was approved by the Ethics Committee of Liège University Hospital (CHU), Belgium (Nr. B707201422509, ref: 2014/272).

Supplementary material

10337_2019_3710_MOESM1_ESM.pdf (2.6 mb)
Supplementary material 1 (PDF 2650 KB)


  1. 1.
    Senn DR, Stimson PG (eds) (2010) Forensic dentistry, 2nd edn. CRC Press, Boca RatonGoogle Scholar
  2. 2.
    Haglund WD, Sorg MH (1996) Forensic taphonomy: the postmortem fate of human remains. CRC Press, Boca RatonCrossRefGoogle Scholar
  3. 3.
    Vass AA (2001) Beyond the grave – understanding human decomposition. Microbiol Today 28:190–192Google Scholar
  4. 4.
    Stadler S, Stefanuto P-H, Brokl M et al (2013) Characterization of volatile organic compounds from human analogue decomposition using thermal desorption coupled to comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry. Anal Chem 85:998–1005. CrossRefGoogle Scholar
  5. 5.
    Stefanuto P-H, Perrault K, Stadler S et al (2014) Reading cadaveric decomposition chemistry with a new pair of glasses. ChemPlusChem 79:786–789. CrossRefGoogle Scholar
  6. 6.
    Dekeirsschieter J, Stefanuto P-H, Brasseur C et al (2012) Enhanced characterization of the smell of death by comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC–TOFMS). PLoS One 7:e39005. CrossRefGoogle Scholar
  7. 7.
    Perrault KA, Stefanuto P-H, Stuart BH et al (2015) Reducing variation in decomposition odour profiling using comprehensive two-dimensional gas chromatography. J Sep Sci 38:73–80. CrossRefGoogle Scholar
  8. 8.
    Focant JF, Stefanuto P, Brasseur C et al (2013) Forensic cadaveric decomposition profiling by GC × GC–TOFMS analysis of VOCs. Chem Bull Kazakh Natl Univ 4:177–186. CrossRefGoogle Scholar
  9. 9.
    Stefanuto P-H, Perrault KA, Lloyd RM et al (2015) Exploring new dimensions in cadaveric decomposition odour analysis. Anal Methods 7:2287–2294. CrossRefGoogle Scholar
  10. 10.
    Stefanuto P-H, Perrault KA, Stadler S et al (2015) GC × GC–TOFMS and supervised multivariate approaches to study human cadaveric decomposition olfactive signatures. Anal Bioanal Chem 407:4767–4778. CrossRefGoogle Scholar
  11. 11.
    Perrault KA, Stefanuto P-H, Stuart BH et al (2015) Detection of decomposition volatile organic compounds in soil following removal of remains from a surface deposition site. Forensic Sci Med Pathol 11:376–387. CrossRefGoogle Scholar
  12. 12.
    Perrault KA, Stefanuto P-H, Dubois LM et al (2017) A minimally-invasive method for profiling volatile organic compounds within postmortem internal gas reservoirs. Int J Legal Med 131:1271–1281. CrossRefGoogle Scholar
  13. 13.
    Stefanuto P-H, Perrault K, Grabherr S et al (2016) Postmortem internal gas reservoir monitoring using GC × GC-HRTOF-MS. Separations 3:24–37. CrossRefGoogle Scholar
  14. 14.
    Brasseur C, Dekeirsschieter J, Schotsmans EMJ et al (2012) Comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry for the forensic study of cadaveric volatile organic compounds released in soil by buried decaying pig carcasses. J Chromatogr A 1255:163–170. CrossRefGoogle Scholar
  15. 15.
    Paczkowski S, Schütz S (2011) Post-mortem volatiles of vertebrate tissue. Appl Microbiol Biotechnol 91:917–935. CrossRefGoogle Scholar
  16. 16.
    Verheggen F, Perrault KA, Megido RC et al (2017) The odor of death: an overview of current knowledge on characterization and applications. Bioscience 67:600–613. CrossRefGoogle Scholar
  17. 17.
    Gruber B, Weggler BA, Jaramillo R et al (2018) Comprehensive two-dimensional gas chromatography in forensic science: a critical review of recent trends. TrAC Trends Anal Chem 105:292–301. CrossRefGoogle Scholar
  18. 18.
    Perrault KA, Nizio KD, Forbes SL (2015) A comparison of one-dimensional and comprehensive two-dimensional gas chromatography for decomposition odour profiling using inter-year replicate field trials. Chromatographia 78:1057–1070. CrossRefGoogle Scholar
  19. 19.
    Paczkowski S, Nicke S, Ziegenhagen H, Schütz S (2015) Volatile emission of decomposing pig carcasses (Sus scrofa domesticus L.) as an indicator for the postmortem interval. J Forensic Sci 60(Suppl 1):S130–S137. CrossRefGoogle Scholar
  20. 20.
    Dekeirsschieter J, Verheggen FJ, Gohy M et al (2009) Cadaveric volatile organic compounds released by decaying pig carcasses (Sus domesticus L.) in different biotopes. Forensic Sci Int 189:46–53. CrossRefGoogle Scholar
  21. 21.
    Armstrong P, Nizio KD, Perrault KA, Forbes SL (2016) Establishing the volatile profile of pig carcasses as analogues for human decomposition during the early postmortem period. Heliyon 2:e00070. CrossRefGoogle Scholar
  22. 22.
    Perrault KA, Forbes SL (2016) Elemental analysis of soil and vegetation surrounding decomposing human analogues. Can Soc Forensic Sci J. Google Scholar
  23. 23.
    Forbes SL, Perrault KA, Stefanuto P-H et al (2014) Comparison of the decomposition VOC profile during winter and summer in a moist, mid-latitude (Cfb) climate. PLoS One 9:e113681. CrossRefGoogle Scholar
  24. 24.
    Nizio KD, Ueland M, Stuart BH, Forbes SL (2017) The analysis of textiles associated with decomposing remains as a natural training aid for cadaver-detection dogs. Forensic Chem 5:33–45. CrossRefGoogle Scholar
  25. 25.
    Knobel Z, Ueland M, Nizio KD et al (2018) A comparison of human and pig decomposition rates and odour profiles in an Australian environment. Aust J Forensic Sci 618:1–16. CrossRefGoogle Scholar
  26. 26.
    Dubois LM, Stefanuto P-H, Heudt L et al (2018) Characterizing decomposition odor from soil and adipocere samples at a death scene using HS–SPME–GC × GC–HRTOFMS. Forensic Chem 8:11–20. CrossRefGoogle Scholar
  27. 27.
    Rosier E, Cuypers E, Dekens M et al (2014) Development and validation of a new TD–GC/MS method and its applicability in the search for human and animal decomposition products. Anal Bioanal Chem 406:3611–3619. CrossRefGoogle Scholar
  28. 28.
    Forbes SL, Perrault KA (2014) Decomposition odour profiling in the air and soil surrounding vertebrate carrion. PLoS One 9:e95107. CrossRefGoogle Scholar
  29. 29.
    Vass AA, Smith RR, Thompson CV et al (2008) Odor analysis of decomposing buried human remains. J Forensic Sci 53:384–391. CrossRefGoogle Scholar
  30. 30.
    Perrault K, Stuart B, Forbes S (2014) A longitudinal study of decomposition odour in soil using sorbent tubes and solid phase microextraction. Chromatography 1:120–140. CrossRefGoogle Scholar
  31. 31.
    Dimandja J-MD, Clouden GC, Colón I et al (2003) Standardized test mixture for the characterization of comprehensive two-dimensional gas chromatography columns: the Phillips mix. J Chromatogr A 1019:261–272. CrossRefGoogle Scholar
  32. 32.
    Dubois LM, Perrault KA, Stefanuto P-H et al (2017) Thermal desorption comprehensive two-dimensional gas chromatography coupled to variable-energy electron ionization time-of-flight mass spectrometry for monitoring subtle changes in volatile organic compound profiles of human blood. J Chromatogr A 1501:117–127. CrossRefGoogle Scholar
  33. 33.
    Stefanuto P-H, Perrault KA, Dubois LM et al (2017) Advanced method optimization for volatile aroma profiling of beer using two-dimensional gas chromatography time-of-flight mass spectrometry. J Chromatogr A 1507:45–52. CrossRefGoogle Scholar
  34. 34.
    Hoffman EM, Curran AM, Dulgerian N et al (2009) Characterization of the volatile organic compounds present in the headspace of decomposing human remains. Forensic Sci Int 186:6–13. CrossRefGoogle Scholar
  35. 35.
    Rust L, Nizio KD, Forbes SL (2016) The influence of ageing and surface type on the odour profile of blood-detection dog training aids. Anal Bioanal Chem 408:6349–6360. CrossRefGoogle Scholar
  36. 36.
    Brokl M, Bishop L, Wright CG et al (2014) Multivariate analysis of mainstream tobacco smoke particulate phase by headspace solid-phase micro extraction coupled with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry. J Chromatogr A 1370:216–229. CrossRefGoogle Scholar
  37. 37.
    DeGreeff LE, Furton KG (2011) Collection and identification of human remains volatiles by non-contact, dynamic airflow sampling and SPME-GC/MS using various sorbent materials. Anal Bioanal Chem 401:1295–1307. CrossRefGoogle Scholar
  38. 38.
    DeGreeff LE, Curran AM, Furton KG (2011) Evaluation of selected sorbent materials for the collection of volatile organic compounds related to human scent using non-contact sampling mode. Forensic Sci Int 209:133–142. CrossRefGoogle Scholar
  39. 39.
    Cablk ME, Szelagowski EE, Sagebiel JC (2012) Characterization of the volatile organic compounds present in the headspace of decomposing animal remains, and compared with human remains. Forensic Sci Int 220:118–125. CrossRefGoogle Scholar
  40. 40.
    Fiedler S, Berns AE, Schwark L et al (2015) The chemistry of death—adipocere degradation in modern graveyards. Forensic Sci Int 257:320–328. CrossRefGoogle Scholar
  41. 41.
    Statheropoulos M, Agapiou A, Spiliopoulou C et al (2007) Environmental aspects of VOCs evolved in the early stages of human decomposition. Sci Total Environ 385:221–227. CrossRefGoogle Scholar
  42. 42.
    Kalinová B, Podskalská H, Růzicka J, Hoskovec M (2009) Irresistible bouquet of death: how are burying beetles (Coleoptera: Silphidae: Nicrophorus) attracted by carcasses. Naturwissenschaften 96:889–899. CrossRefGoogle Scholar
  43. 43.
    Vass AA, Smith RR, Thompson CV et al (2004) Decompositional odor analysis database. J Forensic Sci 49:1–10. CrossRefGoogle Scholar
  44. 44.
    Statheropoulos M, Agapiou A, Zorba E et al (2011) Combined chemical and optical methods for monitoring the early decay stages of surrogate human models. Forensic Sci Int 210:154–163. CrossRefGoogle Scholar
  45. 45.
    Can I, Javan GT, Pozhitkov AE, Noble PA (2014) Distinctive thanatomicrobiome signatures found in the blood and internal organs of humans. J Microbiol Methods 106:1–7. CrossRefGoogle Scholar
  46. 46.
    Rosier E, Loix S, Develter W et al (2016) Time-dependent VOC-profile of decomposed human and animal remains in laboratory environment. Forensic Sci Int 266:164–169. CrossRefGoogle Scholar
  47. 47.
    Li W, Chang Y, Han L et al (2018) Trimethylamine in postmortem tissues as a predictor of postmortem interval estimation using the GC method. Leg Med 35:80–85. CrossRefGoogle Scholar
  48. 48.
    Velasquez M, Ramezani A, Manal A, Raj D (2016) Trimethylamine N-oxide: the good, the bad and the unknown. Toxins (Basel) 8:326. CrossRefGoogle Scholar
  49. 49.
    Rendine M, Fiore C, Bertozzi G et al (2018) Decomposing human blood: canine detection odor signature and volatile organic compounds. J Forensic Sci. Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.CART, Organic and Biological Analytical Chemistry GroupUniversity of LiègeLiègeBelgium
  2. 2.Laboratory of Forensic and Bioanalytical Chemistry, Forensic Sciences UnitChaminade University of HonoluluHonoluluUSA
  3. 3.Pathological Anatomy and Cytology LaboratoryUniversity of LiègeLiègeBelgium

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