Exploring Microscopic and Spectroscopic Techniques in Forensic Hair Analysis: A Comprehensive Review
DOI:
https://doi.org/10.37506/xqs1ex03Keywords:
Hair, UV-VIS, Vibrational, Mass,Atomic, NMR, ESR SpectroscopyAbstract
Background: Hair assists as crucial evidence in any forensic investigations that has strong ability to link suspects to crime scenes or victims. One of the most traditional methods of forensic hair examination is microscopy which aids in morphological examination. However, with advent of more advanced techniques such as spectroscopy, deciphering the chemical and structural information. In view to this, the present review aims to provide a comprehensive overview of both microscopic and spectroscopic methods employed in forensic hair analysis.
Methodology: A systematic search was performed in the ScienceDirect and R-Discovery databases. Research articles published between time period 2014-2025 were included in the review. The identification phase yielded 2,399 records from ScienceDirect and R-Discovery databases. After removing 4 duplicates, 2,335 records remained. Following title and abstract review, 2,189 were excluded, leaving 146 for full-text retrieval. After assessment, 39 articles were included in the review.
Conclusion: This review provides a concise overview of microscopic and spectroscopic techniques used for human hair analysis, along with a discussion of the advantages and limitations of each approach.
References
Boll MS, Doty KC, Wickenheiser R, Lednev IK. Differentiation of hair using ATR FT-IR spectroscopy: A statistical classification of dyed and non-dyed hairs. Forensic Chemistry. 2017 Dec 1;6:1-9.
Ha BJ, Lee GY, Cho IH, Park S. Age-and sex-dependence of five major elements in the development of human scalp hair. Biomaterials Research. 2019 Dec 21;23(1):29. https://doi.org/10.1186/s40824-019-0179-5
Airlie M, Robertson J, Brooks E. An evaluation of novice, expert and supervised machine learning model classifications for forensic hair analysis. Australian Journal of Forensic Sciences. 2024 Sep 2;56(5):551-65. https://doi.org/10.1080/00450618.2023.2254337 .
Horiguchi M, Kunimura S. Surface-enhanced Raman scattering and X-ray fluorescence analyses of a single hair colored with a hair dye product. Analytical Sciences. 2020 Nov 10;36(11):1311-5. https://doi.org/10.2116/analsci.20P14
Bhatia D, Sharma CP, Sharma S, Singh R. Species discrimination from hair using ATR-FTIR spectroscopy: Application in wildlife forensics. Science & Justice. 2024 May 1;64(3):314-21.https://doi.org/10.1016/j.scijus.2024.04.002
Houck MM, Siegel JA Forensic Hair Examinations. Fundamentals of Forensic Science. 2015: 291–312. doi:10.1016/b978-0-12-800037-3.00012-1 https://doi.org/10.1016/B978-0-12-800037-3.00012-1
Manheim J, Doty KC, McLaughlin G, Lednev IK. Forensic hair differentiation using attenuated total reflection Fourier transform infrared (ATR FT-IR) spectroscopy. Applied spectroscopy. 2016 Jul;70(7):1109-17.https://doi.org/10.1177/0003702816652321
Houck M. Forensic Sciences. Reference Module in Chemistry, Molecular Sciences and Chemical Engineering. 2015. https://doi.org/10.1016/B978-0-12-409547-2.14397-5
Linacre A, Ottens R. DNA: Hair Analysis. Encyclopedia of Forensic and Legal Medicine. 2016: 337–342. https://doi.org/10.1016/b978-0-12-800034-2.00157-9
Ohira H, Sakamoto Y, Yamamoto I, Yamada Y. DNA analysis of hairs in a suspected case of child abuse. Legal Medicine. 2018 Nov 1;35:77-9. https://doi.org/10.1016/j.legalmed.2018.09.010
Goncalves R, Castaing N, Richeval C, Ducint D, Titier K, Morvan E, Grelard A, Loquet A, Molimard M. Methoxpropamine (MXPr) in powder, urine and hair samples: analytical characterization and metabolite identification of a new threat. Forensic Science International. 2022 Apr 1;333:111215.https://doi.org/10.1016/j.forsciint.2022.111215
Johansen SS. Detection of the antipsychotic drug quetiapine in the blood, urine and hair samples of the victim of a drug-facilitated sexual assault. Forensic science international. 2017 Jan 1;270:e12-5.https://doi.org/10.1016/j.forsciint.2016.12.010
Jang M, Yang W, Jeong S, Park S, Kim J. A fatal case of paramethoxyamphetamine poisoning and its detection in hair. Forensic science international. 2016 Sep 1;266:e27-31.https://doi.org/10.1016/j.forsciint.2016.06.030
Arbouche N, Farrugia A, Walch A, Raul JS, Kintz P. Specific interpretation of hair concentrations in 2 fatal metformin intoxication cases. Legal Medicine. 2021 Feb 1;48:101803.https://doi.org/10.1016/j.legalmed.2020.101803Get rights and content
Epain M, Cartiser N, Bévalot F, Fanton L. Toxicological detection of pholcodine in blood, urine and hair in three cases of fatal intoxication. Forensic science international. 2021 Oct 1;327:110975.https://doi.org/10.1016/j.forsciint.2021.110975
(n.d.). https://www.sciencedirect.com/. https://www.sciencedirect.com(2023, May 7).
Academic Research Reading: Personalised Research Discovery - R Discovery. https://discovery.researcher.life
(n.d.). CADIMA. https://www.cadima.info/
Boyala G, Dehgahi S, Zubair M, Ullah A, Waghmare P, Qureshi AJ. 3D printed human hair-polymer continuous fiber reinforced composites through Vat Photopolymerization process. Materials Today Communications. 2023 Jun 1;35:106096. https://doi.org/10.1016/j.mtcomm.2023.106096
Cloete KJ, Šmit Ž, Gianoncelli A. Multidimensional profiling of human body hairs using qualitative and semi-quantitative approaches with SR-Xrf, ATR-Ftir, Dsc, and SEM-EDX. International Journal of Molecular Sciences. 2023 Feb 19;24(4):4166. https://doi.org/10.3390/ijms24044166
Mills M, Bonetti J, Brettell T, Quarino L. Differentiation of human hair by colour and diameter using light microscopy, digital imaging and statistical analysis. Journal of Microscopy. 2018 Apr;270(1):27-40. https://doi.org/10.1111/jmi.12646
Richena M, Silveira M, Rezende CA, Joekes I. Yellowing and bleaching of grey hair caused by photo and thermal degradation. Journal of Photochemistry and Photobiology B: Biology. 2014 Sep 5;138:172-81.https://doi.org/10.1016/j.jphotobiol.2014.05.017
de Oblitas RL, de Camargo Junior FB, Magalhaes WV, de Sá Teixeira F, Salvadori MC. Characterization of the mechanical properties of the cortex region of human hair fibers by multiparametric atomic force microscopy mapping. Ultramicroscopy. 2024 May 1;259:113925. https://doi.org/10.1016/j.ultramic.2024.113925
Dos Santos JD, Edwards HG, de Oliveira LF. Raman spectroscopy and electronic microscopy structural studies of Caucasian and Afro human hair. Heliyon. 2019 May 1;5(5). https://doi.org/10.1016/j.heliyon.2019.e01582
Fedorkova MV, Brandt NN, Chikishev AY, Smolina NV, Balabushevich NG, Gusev SA, Lipatova VA, Botchey VM, Dobretsov GE, Mikhalchik EV. Photoinduced formation of thiols in human hair. Journal of Photochemistry and Photobiology B: Biology. 2016 Nov 1;164:43-8.https://doi.org/10.1016/j.jphotobiol.2016.09.021
Richena M, Rezende CA. Effect of photodamage on the outermost cuticle layer of human hair. Journal of Photochemistry and Photobiology B: Biology. 2015 Dec 1;153:296-304.https://doi.org/10.1016/j.jphotobiol.2015.10.008
Sharma S, Gupta S, Yadav PK. Sex and blood group determination from hair using ATR-FTIR spectroscopy and chemometrics. International Journal of Legal Medicine. 2024 May;138(3):801-14.https://doi.org/10.1007/s00414-023-03123-w
Kurouski D, Van Duyne RP. In situ detection and identification of hair dyes using surface-enhanced Raman spectroscopy (SERS). Analytical chemistry. 2015 Mar 3;87(5):2901-6. https://doi.org/10.1021/ac504405u .
Holman AP, Peterson M, Linhart E, Kurouski D. Using surface-enhanced Raman spectroscopy to probe artificial dye degradation on hair buried in multiple soils for up to eight weeks. Scientific Reports. 2024 Mar 18;14(1):6469. https://doi.org/10.1038/s41598-024-57147-2
Santos JD, Pinto PF, Edwards HG, de Oliveira LF. Characterization by Raman and infrared spectroscopy and fluorescence microscopy of human hair treated with cosmetic products. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2022 Nov 5;280:121577.https://doi.org/10.1016/j.saa.2022.121577
Juarez I, Kurouski D. Surface-enhanced Raman spectroscopy hair analysis after household contamination. Analytical Methods. 2023;15(38):4996-5001.https://doi.org/10.1039/D3AY01219K .
Rashaid AH, Harrington PB, Jackson GP. Amino acid composition of human scalp hair as a biometric classifier and investigative lead. Analytical Methods. 2015;7(5):1707-18.https://doi.org/10.1039/C4AY02588A .
Szynkowska MI, Marcinek M, Pawlaczyk A, Albińska J. Human hair analysis in relation to similar environmental and occupational exposure. Environmental toxicology and pharmacology. 2015 Sep 1;40(2):402-8. https://doi.org/10.1016/j.etap.2015.07.005 .
Adav SS, Leung CY, Ng KW. Profiling of hair proteome revealed individual demographics. Forensic Science International: Genetics. 2023 Sep 1;66:102914.https://doi.org/10.1016/j.fsigen.2023.102914
Mason KE, Paul PH, Chu F, Anex DS, Hart BR. Development of a protein‐based human identification capability from a single hair. Journal of forensic sciences. 2019 Jul;64(4):1152-9. https://doi.org/10.1111/1556-4029.13995
Wu X, Zhang T, Mao M, Zhang Y, Zhang Z, Xu P. A methodological exploration of distinguishing hair quality based on hair proteomics. Proteome Science. 2024 May 2;22(1):5https://doi.org/10.1186/s12953-024-00229-w
Carelli C, Fais P, Freni F, Moretti M, Vignali C, Morini L. Importance of segmental hair analysis in a suspected case of attempted homicide by flocoumafen and difenacoum. Forensic science international. 2020 Nov 1;316:110466. https://doi.org/10.1016/j.forsciint.2020.110466
Poetzsch M, Steuer AE, Roemmelt AT, Baumgartner MR, Kraemer T. Single hair analysis of small molecules using MALDI-triple quadrupole MS imaging and LC-MS/MS: investigations on opportunities and pitfalls. Analytical chemistry. 2014 Dec 2;86(23):11758-65. https://doi.org/10.1021/ac503193w
Rocha AV, Cardoso BR, Bueno RB, de Bortoli Dr MC, Farias LA. Selenium status and hair mercury levels in riverine children from Rondônia, Amazonia. Nutrition. 2014 Nov 1;30(11-12):1318-23.https://doi.org/10.1016/j.nut.2014.03.013
Zhang S, Hu Z, Zhao Z, Chen F, Tang Y, Sheng Z, Zhang D, Zhang Z, Jin H, Pu H, Guo L. Quantitative analysis of mineral elements in hair and nails using calibration-free laser-induced breakdown spectroscopy. Optik. 2021 Sep 1;242:167067. https://doi.org/10.1016/j.ijleo.2021.167067
Zhang S, Chu Y, Ma S, Chen F, Zhang D, Hu Z, Zhang Z, Jin H, Guo L. Highly accurate determination of Zn and Cu in human hair by ultrasound-assisted alkali dissolution combined with laser-induced breakdown spectroscopy. Microchemical Journal. 2020 Sep 1;157:105018. https://doi.org/10.1016/j.microc.2020.105018
Rafiee A, Delgado-Saborit JM, Sly PD, Quemerais B, Hashemi F, Akbari S, Hoseini M. Environmental chronic exposure to metals and effects on attention and executive function in the general population. Science of The Total Environment. 2020 Feb 25;705:135911. https://doi.org/10.1016/j.scitotenv.2019.135911
Kumakli H, A'ja VD, McDaniel K, Mehari TF, Stephenson J, Maple L, Crawford Z, Macemore CL, Babyak CM, Fakayode SO. Environmental biomonitoring of essential and toxic elements in human scalp hair using accelerated microwave-assisted sample digestion and inductively coupled plasma optical emission spectroscopy. Chemosphere. 2017 May 1;174:708-15. https://doi.org/10.1016/j.chemosphere.2017.02.032.
Cherni I, Nouir R, Daoud F, Hamzaoui S, Ghalila H. Fast diagnostic of osteoporosis based on hair analysis using LIBS technique. Medical Engineering & Physics. 2022 May 1;103:103798. https://doi.org/10.1016/j.medengphy.2022.103798 .
Çam ST, Polat M, Seyhan N. The use of human hair as biodosimeter. Applied Radiation and Isotopes. 2014 Dec 1;94:272-81. https://doi.org/10.1016/j.apradiso.2014.08.021 .
Tintner J, Rennhofer H, Kennedy CJ, Revie WA, Weber H, Pavlik C, Lanszki J. Recalcitrance of hair in historical plasters. Polymer Degradation and Stability. 2020 Nov 1;181:109333.https://doi.org/10.1016/j.polymdegradstab.2020.109333
Pienpinijtham P, Thammacharoen C, Naranitad S, Ekgasit S. Analysis of cosmetic residues on a single human hair by ATR FT-IR microspectroscopy. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2018 May 15;197:230-6.https://doi.org/10.1016/j.saa.2018.01.084
Samanta A, Bhattacharya M, Dalui S, Acharya M, Das PS, Chanda DK, Acharya SD, Sivaraman SK, Nath S, Mandal AK, Ghosh J. Nanomechanical responses of human hair. journal of the mechanical behavior of biomedical materials. 2016 Mar 1;56:229-48. https://doi.org/10.1016/j.jmbbm.2015.10.010
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