When a hair or body hair is recovered without its root from a crime scene, no conventional genetic analysis can be performed. Lacking nuclear DNA, this biological material has long offered only limited evidentiary value and could neither support the formal identification of an individual nor allow comparison with national DNA databases. Since few years, however, a major shift has occurred: hair proteomics, which exploits the proteins of the hair shaft to reveal individualizing markers. Thanks to advances in mass spectrometry, this approach now provides a new pathway for identification, particularly useful in cold cases or in situations where DNA is absent or unusable.
A biological evidence long underused
Hairs recovered from crime scenes are frequently rootless, preventing any STR (Short Tandem Repeat) analysis. Traditional alternatives (morphological examination or mitochondrial DNA analysis), offer only limited discriminating power [1][9]. In many cases, these items were classified as “weak traces,” with insufficient probative value. Yet a hair is biologically rich. It is composed mainly of keratins and other structural proteins that exhibit remarkable stability and resistance to heat, aging, and environmental degradation [1]. This robustness has led several research teams to explore another avenue: instead of seeking nuclear DNA where it is absent or degraded, why not rely directly on proteins, some of which vary between individuals?

Figure 1: Structure of a hair shaft. Source: cosmeticsdesign.com
From DNA to proteomics
This technological shift relies on high-resolution mass spectrometry (HRMS), combined with bioinformatic analysis of protein polymorphisms. Recent work has confirmed that hundreds of proteins can be identified in a single hair shaft. Among them, certain markers, SAPs (Single Amino acid Polymorphisms), directly reflect individual genetic variation [2]. A major study demonstrated that a single individual presents, on average, more than 600 detectable protein groups and more than 160 polymorphic markers, yielding Random Match Probabilities (RMP) on the order of 10⁻¹⁴ [2]. This protein signature therefore offers strong discriminating power, in some cases comparable to the informational value of mitochondrial DNA, while avoiding several well-known limitations of the latter [10].
Technical obstacles related to protein extraction, made difficult by the highly cross-linked structure of keratin, have also been partially overcome. Protocols combining controlled heat and reducing agents now allow more efficient and reproducible extraction [3]. These advances make the approach more mature and more suitable for forensic practice.

Figure 2: Hair proteomic analysis workflow. Proteins extracted from the hair shaft are fragmented and then analyzed by mass spectrometry to identify individual peptide variations. Source: [2] Parker, G. et al., Deep Coverage Proteome Analysis of Human Hair Shafts, Journal of Proteome Research, 2022.
Concrete opportunities for investigations
Hair-shaft proteomics significantly enhances the usefulness of hair traces in investigations. In cold cases, hairs preserved for decades can now provide individualizing information, even when nuclear DNA was unusable at the time of the original analysis [5]. In extreme contexts (fire scenes, carbonized remains, or highly degraded traces), proteins often persist where DNA has degraded, making them particularly valuable [5][6].
In recent investigations (sexual assaults, abductions, violent incidents, close-contact events), head or body hairs without roots recovered from clothing, vehicles, or victims can now contribute to establishing associations or excluding individuals. Even when it does not yield a formal identification, the protein signature may narrow the suspect pool, confirm or refute an investigative hypothesis, and support evidential assessments presented to judicial authorities [4]. From a legal standpoint, this method must be understood as a probabilistic approach, similar in principle to mitochondrial DNA analysis but based on more stable markers [7]. When integrated carefully, it may become decisive in investigative orientations, the re-examination of older cases, or situations previously left unresolved due to lack of nuclear DNA or usable fingerprints.
Technical limits and challenges
Despite its potential, hair-shaft proteomics remains a technique still in maturation. The first limitation lies in the protocols themselves: protein extraction remains challenging due to the resistant structure of the hair shaft, and full standardization has not yet been achieved [3]. A second challenge is the creation of sufficiently large population databases to compute robust Random Match Probabilities [4]. Inter-laboratory validation, essential before any use in judicial contexts, requires testing on hairs from individuals of different populations, ages, environments, and storage conditions [4][6].
Legal integration also presents challenges. Judges and attorneys will need clear explanations of this emerging probabilistic evidence. Classical admissibility requirements (reliability, reproducibility, methodological transparency, statistical robustness), apply fully [7]. To date, no international standard formally regulates the procedure, although preliminary work is underway [8].
Towards standardization and operational integration?
The outlook for the coming years is particularly promising. Several centres, notably Murdoch University and ChemCentre near Perth, Australia, are working on protocol standardization and the development of diverse reference databases [5][6]. Advances in mass spectrometry and bioinformatic tools now make possible a partial automation of analyses and more seamless integration into routine forensic laboratory workflows. For investigators, police officers, magistrates and forensic experts, this evolution requires adapting collection and preservation practices. From now on, any rootless hair should be systematically collected and retained. Even very small or very old samples may contain an exploitable protein signature. This shift in perspective could transform the re-evaluation of cold cases, fire-scene examinations, and the most complex investigations.
Conclusion
Hair-shaft proteomics represents one of the most promising advances of the coming years in forensic identification. By restoring value to traces long considered underexploited, it offers a reliable and robust alternative when DNA is absent, degraded or otherwise unusable. Although judicial integration still requires validation, standardization and appropriate communication, early results clearly indicate that this approach could play a decisive role in complex investigations, degraded scenes and unresolved cases.
References :
[1] Adav, S.S., Human Hair Proteomics: An Overview, Science & Justice, 2021.
[2] Parker, G. et al., Deep Coverage Proteome Analysis of Human Hair Shafts, Journal of Proteome Research, 2022.
[3] Liu, Y. et al., Individual-specific proteomic markers from protein amino acid polymorphisms, Proteome Science, 2024.
[4] Smith, R.N. et al., Forensic Proteomics: Potential and Challenges, Proteomics, 2023.
[5] Murdoch University – Western Australia, Hair Protein Identification Project (2024–2025).
[6] ChemCentre (Western Australia Government), World-first Forensic Proteomics Research Program, 2024.
[7] Henry, R. & Stoyan, N., The Admissibility of Proteomic Evidence in Court, SSRN, 2020.
[8] ISO / ASTM – Guidelines on Forensic Biology & Novel Analytical Methods, 2022–2024.
[9] Anslinger, K., Hair Evidence in Forensic Science, Wiley, 2019.
[10] Budowle, B., Mitochondrial DNA in Forensic Identification, Elsevier, 2018.
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