Analytical and Bioanalytical Chemistry

, Volume 410, Issue 24, pp 6165–6175 | Cite as

A complementary forensic ‘proteo-genomic’ approach for the direct identification of biological fluid traces under fingernails

  • Sathisha Kamanna
  • Julianne Henry
  • Nico Voelcker
  • Adrian Linacre
  • K. Paul KirkbrideEmail author
Research Paper


Violent contact between individuals during a crime can result in body fluids becoming trapped under the fingernails of the individuals involved. The traces under fingernails represent valuable forensic evidence because DNA profiling can indicate from whom the trace originated and proteomic methods can be used to determine the type of fluid in the trace, thus providing evidence as to the circumstances surrounding the crime. Here, we present an initial study of an analytical strategy that involves two complementary techniques, direct PCR DNA profiling and direct mass spectrometry-based protein biomarker detection, for the comprehensive examination of traces of biological fluids gathered from underneath fingernails. With regard to protein biomarker detection, direct MALDI-ToF MS/MS is very sensitive, allowing results to be obtained from biological material present on only a few fibres plucked from a microswab used to collect the traces. Human cornulin, a protein biomarker for vaginal fluid, could be detected up to 5 h after it had been deposited under fingernails whereas haemoglobin, a biomarker for blood, is somewhat more persistent under fingernails and could be detected up to 18 h post-deposition. Bottom-up tandem mass spectrometry techniques were used to provide a high level of confidence in assigning the identity of protein biomarkers. nLC-ESI-qToF MS/MS offered higher levels of confidence and the ability to detect traces that had been present under fingernails for longer periods of time, but this performance came with the cost of longer analysis time and a more laborious sampling approach.

Graphical abstract


Forensic science Haemoglobin Vaginal fluid MALDI-TOF MS 



The authors would like to acknowledge Flinders Analytical and the Proteomics Centre at Flinders University. We also thank the volunteers who provided human body fluid samples and kind thanks to Dr. Jennifer Templeton and Ms. Alice Stephenson (Forensic Science SA) for conducting the DNA-profiling experiments.

Funding information

This project was supported by the Ross Vining Research Fund, which is administered by Forensic Science SA (South Australia).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1223_MOESM1_ESM.pdf (4.1 mb)
ESM 1 (PDF 4226 kb)


  1. 1.
    Kamanna S, Henry J, Voelcker NH, Linacre A, Kirkbride KP. A mass spectrometry based forensic toolbox for imaging and detecting biological fluid evidence in finger marks and fingernail scrapings. Int J Legal Med. 2017;131(5):1413–22.CrossRefPubMedGoogle Scholar
  2. 2.
    Yang H, Zhou B, Prinz M, Siegel D. Proteomic analysis of menstrual blood. Mol Cell Proteomics. 2012;11:1024–35.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Patel E, Cicatiello P, Deininge L, Clench MR, et al. A proteomic approach for the rapid, multi informative and reliable identification of blood. Analyst. 2016;141:191–8.CrossRefPubMedGoogle Scholar
  4. 4.
    Yang H, Zhou B, Deng H, Prinz M, Siegel D. Body fluid identification by mass spectrometry. Int J Legal Med. 2013;127(6):1065–77.CrossRefPubMedGoogle Scholar
  5. 5.
    Van Steendam K, De Ceuleneer M, Dhaenens M, Van Hoofstat D, et al. Mass spectrometry-based proteomics as a tool to identify biological matrices in forensic science. Int J Legal Med. 2013;127(2):287–98.CrossRefPubMedGoogle Scholar
  6. 6.
    Ottens R, Taylor D, Linacre A. DNA profiles from fingernails using direct PCR. Forensic Sci Med Pathol. 2015;11(1):99–103.CrossRefPubMedGoogle Scholar
  7. 7.
    Templeton JEL, Taylor D, Handt O, Skuza P, et al. Direct PCR improves the recovery of DNA from various substrates. J Forensic Sci. 2015;60(6):1558–62.CrossRefPubMedGoogle Scholar
  8. 8.
    Ottens R, Templeton J, Paradiso V, Taylor D, et al. Application of direct PCR in forensic casework. Forensic Sci Int Genet. 2013;4(1):e47–e8.CrossRefGoogle Scholar
  9. 9.
    Kamanna S, Henry J, Voelcker NH, Linacre A, Kirkbride KP. Direct identification of forensic body fluids using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Int J Mass Spectrom. 2016;397:18–26.CrossRefGoogle Scholar
  10. 10.
    Igoh A, Doi Y, Sakurada K. Identification and evaluation of potential forensic marker proteins in vaginal fluid by liquid chromatography/mass spectrometry. Anal Bioanal Chem. 2015;407(23):7135–44.CrossRefPubMedGoogle Scholar
  11. 11.
    Legg KM, Powell R, Reisdorph N, Reisdorph R, Danielson PB. Verification of protein biomarker specificity for the identification of biological stains by quadrupole time-of-flight mass spectrometry. Electrophoresis. 2017;38(6):833–45.CrossRefPubMedGoogle Scholar
  12. 12.
    Legg KM, Powell R, Reisdorph N, Reisdorph R, Danielson PB. Discovery of highly specific protein markers for the identification of biological stains. Electrophoresis. 2014;35(21–22):3069–78.CrossRefPubMedGoogle Scholar
  13. 13.
    Dasari S, Pereira RAP, Michaels JE, et al. Comprehensive proteomic analysis of human cervical-vaginal fluid. J Proteome Res. 2007;6(4):1258–68.CrossRefPubMedGoogle Scholar
  14. 14.
    Gravett MG, Thomas A, Schneider KA, Reddy AP, et al. Proteomic analysis of cervical−vaginal fluid: identification of novel biomarkers for detection of intra-amniotic infection. J Proteome Res. 2007;6(1):89–96.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Sathisha Kamanna
    • 1
  • Julianne Henry
    • 2
  • Nico Voelcker
    • 3
  • Adrian Linacre
    • 1
  • K. Paul Kirkbride
    • 1
    Email author
  1. 1.College of Science and EngineeringFlinders UniversityAdelaideAustralia
  2. 2.Forensic Science SAAdelaideAustralia
  3. 3.Melbourne Centre for NanofabricationMonash UniversityClaytonAustralia

Personalised recommendations