ATR FT-IR Spectroscopy: A Novel and Non-Destructive Approach for the Detection of Blood on a Single Fiber
Article content
Abstract
Abstract
Blood is often encountered on different types of substrates in criminal investigations. Among such substrates, blood stained fabrics are one of the most commonly encountered evidence. Analysis of bloodstains on fabrics is restricted due to interference of fabric dyes, color of fabric which hampers visual identification, and limited sample quantity. Over the years, a plethora of research has been conducted to analyze recovered blood traces, however, the methods employed are limited by factors such as sample destructibility, chronophagus processing, and vulnerability to false positive results. To circumvent the aforementioned limitations, use of ATR FT-IR spectroscopy has surged as a non-destructive and reliable tool for the detection of traces of blood and other body fluids on fibers. In the present study, an attempt has been made to detect traces of blood in fibers of different types and colors using ATR FT-IR spectroscopy. The obtained results unequivocally concluded that by using ATR FT-IR spectroscopy blood can be successfully detected even on a single fiber through characteristic peaks positioned at 1650 cm-1 and 1543 cm-1 corresponding to amide I and amide II. On the basis of the IR marked protein region (1700-1500 cm-1) bloodstains could easily be discriminated from the blank fibers (except human hair, which overlapped with the peaks of bloodstained fibers) by using PCA chemometric tool. Although, further investigations are needed to be conducted, this proof-of-concept study establishes the potential use of ATR FT-IR spectroscopy for the non-destructive, rapid, eco-friendly.
Keywords
Forensic Science, Forensic Serology, Fibers, Fourier Transform Infrared Spectroscopy
Downloads
References
Virkler K, Lednev IK. Analysis of body fluids for
forensic purposes: From laboratory testing to non-
destructive rapid confirmatory identification at
a crime scene. Forensic Sci Int. 2009;188:1–17.
doi:10.1016/j.forsciint.2009.02.013.
Orphanou CM. The detection and discrimination of
human body fluids using ATR FT-IR spectroscopy.
Forensic Sci Int. 2015;252:e10–6. doi:10.1016/j.
forsciint.2015.04.020.
An JH, Shin KJ, Yang WI, Lee HY. Body fluid
identification in forensics, BMB Rep. 2012; 45: 545–
doi:10.5483/bmbrep.2012.45.10.206.
Zapata F, Fernández de la Ossa MÁ, García-Ruiz C.
Emerging spectrometric techniques for the forensic
analysis of body fluids. TrAC - Trends Anal Chem
;64:53–63. doi:10.1016/j.trac.2014.08.011.
Zapata F, Silva Gregório Martins M. Body Fluids
and Spectroscopic Techniques in Forensics: A Perfect
Match? Journal of forensic medicine. 2016; 1: 1-7.
doi:10.4172/2472-1026.1000101.
Bai P, Deng W, Wang L, Long B, Liu K, Liang W,
et al. Micro RNA profiling for the detection and
differentiation of body fluids in forensic stain analysis.
Forensic Sci Int Genet Suppl Ser. 2013;4:e216–7.
doi:10.1016/j.fsigss.2013.10.111.
Li Z, Bai P, Peng D, Long B, Zhang L, Liang W.
Influences of different RT-qPCR methods on forensic
body fluid identification by microRNA. Forensic Sci
Int Genet Suppl Ser. 2015;5:295–7.doi:10.1016/j.
fsigss.2015.09.117.
Courts C, Madea B. Specific Micro-RNA Signatures
for the Detection of Saliva and Blood in Forensic Body-
fluid Identification. J Forensic Sci. 2011;56:1464–70.
doi:10.1111/j.1556-4029.2011.01894.x.
Hanson EK, Mirza M, Rekab K, Ballantyne J. The
identification of menstrual blood in forensic samples
by logistic regression modeling of miRNA expression.
Electrophoresis 2014;35:3087–95. doi:10.1002/
elps.201400171.
Hanson E, Lubenow H, Ballantyne J. Identification
of forensically relevant body fluids using a panel of
differentially expressed microRNAs. Forensic Sci
Int Genet Suppl Ser. 2009;2:503–4. doi:10.1016/j.
fsigss.2009.08.184.
Zubakov D, Boersma AWM, Choi Y, Kayser
M. MicroRNA markers for forensic body fluid
identification obtained from microarray screening
and quantitative RT-PCR confirmation. Int J Legal
Med. 2010:217–26. doi:10.1007/s00414-009-0402-3.
Haas C, Klesser B, Maake C, Bär W, Kratzer A.
mRNA profiling for body fluid identification by
reverse transcription endpoint PCR and realtime PCR.
Forensic Sci Int Genet. 2009;3:80–8. doi:10.1016/j.
fsigen.2008.11.003.
Juusola J, Ballantyne J. Multiplex mRNA profiling
for the identification of body fluids. Forensic Sci Int.
ASFSFM 2020; Volume 2 Issue (1)Sharma et al.
;152:1–12. doi:10.1016/j.forsciint.2005.02.020.
Fleming R, Harbison S, Lin M. New RNA methods
for the identification of body fluids and cell types.
Forensic Sci Int Genet Suppl Ser. 2013;4:e87–8.
doi:10.1016/j.fsigss.2013.10.044.
Wang S, Wang Z, Tao R, He G, Liu J, Li C. The
potential use of Piwi-interacting RNA biomarkers
in forensic body fl uid identi fi cation : A proof-of-
principle study. Forensic Sci Int Genet. 2019;39:129–
doi:10.1016/j.fsigen.2019.01.002.
Liu B, Song F, Yang Q, Zhou Y, Shao C, Shen Y, et al.
Characterization of tissue-specific biomarkers with
the expression of circRNAs in forensically relevant
body fluids. Int J Legal Med. 2019:1–11.doi: 10.1007/
s00414-019-02027-y
An JH, Choi A, Shin K. DNA methylation-specific
multiplex assays for body fluid identification. Int J
Leg Med. 2013;127:35–43. doi:10.1007/s00414-012-
-1.
Antunes J, Silva DSBS, Balamurugan K, Duncan G,
Alho CS, Mccord B. High-resolution melt analysis of
DNA methylation to discriminate semen in biological
stains. Anal Biochem. 2016;494:40–5. doi:10.1016/j.
ab.2015.10.002.
Choi A, Shin KJ, Yang WI, Lee HY. Body fluid
identification by integrated analysis of DNA
methylation and body fluid-specific microbial DNA.
Int J Legal Med. 2014;128:33–41. doi:10.1007/
s00414-013-0918-4.
Gomaa R, Salehi J, Behl S. DNA Methylation as
a Biomarker for Body Fluid Identification. Arab
J Forensic Sci Forensic Med. 2017;1:681–94.
doi:10.26735/16586794.2017.001.
Fachet C, Quarino L. High resolution melt curve
analysis based on methylation status for human semen
identification. Forensic Sci Med Pathol. 2017:86–91.
doi:10.1007/s12024-016-9825-6.
Forat S, Huettel B, Reinhardt R, Fimmers R, Haidl
G, Denschlag D, et al. Methylation Markers for
the Identification of Body Fluids and Tissues from
Forensic Trace Evidence. PLoS ONE. 2016:1–19.
doi:10.1371/journal.pone.0147973.
Frumkin D, Wasserstrom A, Budowle B, Davidson A.
DNA methylation-based forensic tissue identification.
Forensic Sci Int Genet. 2011;5:517–24. doi:10.1016/j.
fsigen.2010.12.001.
Fu XD, Wu J, Wang J, Huang Y, Hou YP, Yan J.
Identification of body fluid using tissue-specific DNA
methylation markers. Forensic Sci Int Genet Suppl
Ser. 2015;5:151–3. doi: 10.1016/j.fsigen.2014.07.011.
Gomes I, Kohlmeier F, Schneider PM. Genetic
markers for body fluid and tissue identification
ASFSFM 2020; Volume 2 Issue (1)
in forensics. Forensic Sci Int Genet Suppl Ser.
;3:e469–70. doi:10.1016/j.fsigss.2011.09.096.
Zrnec D, Vickovic S, Grs B, Popovic M. DNA
methylation : the future of crime scene investigation ?
Mol Biol Rep. 2013:4349–60. doi:10.1007/s11033-
-2525-3.
Auvdel MJ. Comparison of laser and ultraviolet
techniques used in the detection of body secretions.
J Forensic Sci. 1987;32:326–45.doi: 10.1520/
JFS11136J.
Vandenberg N, van Oorschot RAH. The Use of
Polilight® in the Detection of Seminal Fluid,
Saliva, and Bloodstains and Comparison with
Conventional Chemical-Based Screening Tests. J
Forensic Sci. 2006;51:361–70. doi:10.1111/j.1556-
2006.00065.x.
Fiedler A, Rehdorf J, Hilbers F, Johrdan L, Stribl C,
Benecke M. Detection of Semen(Human and Boar)
and Saliva on Fabrics by a Very High Powered UV-/
VIS-Light Source. Open Forensic Sci J. 2008;1:12–5.
doi:10.2174/1874402800801010012
Zapata F, de la Ossa MÁF, García-Ruiz
C. Differentiation of Body Fluid Stains on
Fabrics Using External Reflection Fourier
Transform Infrared Spectroscopy (FT-IR) and
Chemometrics. Appl Spectrosc. 2016;70:654–65.
d o i : 1 0 . 11 7 7 / 0 0 0 3 7 0 2 8 1 6 6 3 1 3 0 3 .
Muro CK, Doty KC, Bueno J, Halámková L, Lednev
IK. Vibrational Spectroscopy: Recent Developments
to Revolutionize Forensic Science. Anal Chem.
;87:306–27. doi:10.1021/ac504068a.
Boyd S, Bertino MF, Ye D, White LS, Seashols SJ.
Highly Sensitive Detection of Blood by Surface
Enhanced Raman Scattering. J Forensic Sci.
;58:753–6.
doi:10.1111/1556-4029.12120.
Muro CK, Doty KC, de Souza Fernandes L,
Lednev IK. Forensic body fluid identification and
differentiation by Raman spectroscopy. Forensic
Chem. 2016;1:31–8. doi:10.1016/j.forc.2016.06.003.
Virkler K, Lednev IK. Raman spectroscopic signature
of semen and its potential application to forensic body
fluid identification. Forensic Sci Int. 2009;193:56–62.
doi:10.1016/j.forsciint.2009.09.005.
Sikirzhytski V, Sikirzhytskaya A, Lednev IK.
Multidimensional Raman spectroscopic signature of
sweat and its potential application to forensic body
fluid identification. Anal Chim Acta. 2012;718:78–
doi:10.1016/j.aca.2011.12.059.
Feine I, Gafny R, Pinkas I. Combination of prostate-
specific antigen detection and micro-Raman
spectroscopy for confirmatory semen detection.40
ATR FT-IR Spectroscopy: A Novel and Non-Destructive Approach for the Detection of Blood on a Single Fiber
Forensic Sci Int. 2017;270:241–7. doi:10.1016/j.
forsciint.2016.10.012.
Huang Z, Chen X, Chen Y, Feng S, Chen R, Chen
J, et al. Raman spectroscopic characterization and
differentiation of seminal plasma. J Biomed Opt.
;16:1–4. doi:10.1117/1.3650310
McLaughlin G, Sikirzhytski V, Lednev IK.
Circumventing substrate interference in the Raman
spectroscopic identification of blood stains.
Forensic Sci Int. 2013;231:157–66. doi:10.1016/j.
forsciint.2013.04.033.
McLaughlin G, Lednev IK. In Situ Identification of
Semen Stains on Common Substrates via Raman
Spectroscopy. J Forensic Sci. 2015;60:595–604.
doi:10.1111/1556-4029.12708.
McLaughlin G, Doty KC, Lednev IK. Raman
Spectroscopy of Blood for Species Identification.
Anal Chem. 2014;86:11628–33. doi:10.1021/
ac5026368.
Doty KC, Lednev IK. Differentiation of human
blood from animal blood using Raman spectroscopy:
A survey of forensically relevant species.
Forensic Sci Int. 2018;282:204–10. doi:10.1016/j.
forsciint.2017.11.033.
Fujihara J, Fujita Y, Yamamoto T, Nishimoto
N, Kimura-Kataoka K, Kurata S, et al. Blood
identification and discrimination between human and
nonhuman blood using portable Raman spectroscopy.
Int J Legal Med. 2017;131:319–22. doi:10.1007/
s00414-016-1396-2.
Virkler K, Lednev IK. Blood Species Identification
for Forensic Purposes Using Raman Spectroscopy
Combined with Advanced Statistical Analysis. Anal
Chem. 2009;81:7773–7. doi:10.1021/ac901350a.
Mistek E, Halámková L, Doty KC, Muro CK,
Lednev IK. Race Differentiation by Raman
Spectroscopy of a Bloodstain for Forensic Purposes.
Anal Chem. 2016;88:7453–6. doi:10.1021/acs.
analchem.6b01173.
Muro CK, Lednev IK. Race Differentiation Based on
Raman Spectroscopy of Semen Traces for Forensic
Purposes. Anal Chem. 2017;89:4344–8. doi:10.1021/
acs.analchem.7b00106.
Muro CK, de Souza Fernandes L, Lednev IK. Sex
Determination Based on Raman Spectroscopy
of Saliva Traces for Forensic Purposes. Anal
Chem.
;88:12489–93.
doi:10.1021/acs.
analchem.6b03988.
Sikirzhytskaya A, Sikirzhytski V, K Lednev I. Raman
spectroscopy coupled with advanced statistics for
differentiating menstrual and peripheral blood.
Journal of biophotonics. 2014; 7: 59-67 doi:10.1002/
jbio.201200191.
Zapata F, Fernández de la Ossa MÁ, García-Ruiz C.
Emerging spectrometric techniques for the forensic
analysis of body fluids. TrAC Trends Anal Chem
;64:53–63. doi:10.1016/j.trac.2014.08.011.
Quinn AA, Elkins KM. The Differentiation of
Menstrual from Venous Blood and Other Body Fluids
on Various Substrates Using ATR FT-IR Spectroscopy.
J Forensic Sci 2017;62:197–204. doi:10.1111/1556-
13250.
Gregório I, Zapata F, García-Ruiz C. Analysis of
human bodily fluids on superabsorbent pads by
ATR-FTIR. Talanta 2017;162:634–40. doi:10.1016/j.
talanta.2016.10.061.
Elkins KM. Rapid Presumptive “Fingerprinting”
of Body Fluids and Materials by ATR FT-IR
Spectroscopy. J Forensic Sci. 2011;56:1580–7.
doi:10.1111/j.1556-4029.2011.01870.x.
De Wael K, Lepot L, Gason F, Gilbert B. In search
of blood—Detection of minute particles using
spectroscopic methods. Forensic Sci Int. 2008;180:37–
doi:10.1016/j.forsciint.2008.06.013.
Gregório I, Zapata F, Torre M, García-Ruiz C.
Statistical approach for ATR-FTIR screening of
semen in sexual evidence. Talanta 2017;174:853–7.
doi:10.1016/j.talanta.2017.07.016.
Sharma S, Chophi R, Singh R. Forensic discrimination
of menstrual blood and peripheral blood using
attenuated total reflectance (ATR)-Fourier transform
infrared (FT-IR) spectroscopy and chemometrics.
Int J Legal Med 2019. doi:10.1007/s00414-019-
-w.
Lin H, Zhang Y, Wang Q, Li B, Huang P, Wang Z.
Estimation of the age of human bloodstains under the
simulated indoor and outdoor crime scene conditions
by ATR-FTIR spectroscopy. Sci Rep. 2017;7:13254.
doi:10.1038/s41598-017-13725-1.
Lin H, Zhang Y, Wang Q, Li B, Fan S, Wang Z.
Species identification of bloodstains by ATR-
FTIR spectroscopy: the effects of bloodstain age
and the deposition environment. Int J Legal Med.
;132:667–74. doi:10.1007/s00414-017-1634-2.
Zhang Y, Wang Q, Li B, Wang Z, Li C, Yao Y, et
al. Changes in Attenuated Total Reflection Fourier
Transform Infrared Spectra as Blood Dries Out. J
Forensic Sci. 2017; 62:761–7. doi:10.1111/1556-
13324.
Takamura A, Watanabe K, Akutsu T, Ozawa T.
Soft and Robust Identification of Body Fluid Using
Fourier Transform Infrared Spectroscopy and
Chemometric Strategies for Forensic Analysis. Sci
Rep. 2018;8:8459. doi:10.1038/s41598-018-26873-
ASFSFM 2020; Volume 2 Issue (1)Sharma et al.
Takamura A, Watanabe K, Akutsu T, Ikegaya H,
Ozawa T. Spectral Mining for Discriminating Blood
Origins in the Presence of Substrate Interference
via Attenuated Total Reflection Fourier Transform
Infrared Spectroscopy: Postmortem or Antemortem
Blood? Anal Chem. 2017;89:9797–804. doi:10.1021/
acs.analchem.7b01756.
Takamura A, Halamkova L, Ozawa T, Lednev
IK. Phenotype Profiling for Forensic Purposes:
Determining Donor Sex Based on Fourier
Transform Infrared Spectroscopy of Urine Traces.
Anal Chem. 2019;91:6288–95. doi:10.1021/acs.
analchem.9b01058.
Zhang K, Wang Q, Liu R, Wei X, Li Z, Fan S, et
al. Evaluating the effects of causes of death on
postmortem interval estimation by ATR-FTIR
spectroscopy. Int J Legal Med. 2019. doi:10.1007/
s00414-019-02042-z.
Lu Z, DeJong SA, Cassidy BM, Belliveau RG,
Myrick ML, Morgan SL. Detection Limits for Blood
on Fabrics Using Attenuated Total Reflection Fourier
Transform Infrared (ATR FT-IR) Spectroscopy and
Derivative Processing. Appl Spectrosc. 2016;71:839–
doi:10.1177/0003702816654154.
Cai S, Singh BR. Identification of β-turn and random
coil amide III infrared bands for secondary structure
estimation of proteins. Biophys Chem 1999;80:7–20.
doi:10.1016/S0301-4622(99)00060-5.
Garside P, Wyeth P. Identification of Cellulosic
Fibres by FTIR Spectroscopy I: Thread and Single
Fibre Analysis by Attenuated Total Reflectance.
Studies in Conservation. 2003; 48:269-275.
doi:10.2307/1506916.
Peets P, Leito I, Pelt J, Vahur S. Identification
and classification of textile fibres using ATR-
FT-IR spectroscopy with chemometric methods.
Spectrochim Acta Part A Mol Biomol Spectrosc.
;173:175–81.
doi:10.1016/j.saa.2016.09.007.
Lang PL, Katon JE, O’Keefe JF, Schiering DW.
The identification of fibers by infrared and Raman
microspectroscopy. Microchem J. 1986;34:319–31.
doi:10.1016/0026-265X(86)90127-X.
Chen H, Lin Z, Tan C. Classification of different
animal fibers by near infrared spectroscopy and
chemometric models. Microchem J. 2019;144:489–
doi:10.1016/j.microc.2018.10.011.
Kumar R, Sharma V. Chemometrics in forensic
ASFSFM 2020; Volume 2 Issue (1)
science. TrAC Trends Anal Chem 2018;105:191–
doi:10.1016/j.trac.2018.05.010.
Silva CS, Pimentel MF, Amigo JM, Honorato RS,
Pasquini C. Detecting semen stains on fabrics using
near infrared hyperspectral images and multivariate
models. TrAC Trends Anal Chem. 2017;95:23–35.
doi:10.1016/j.trac.2017.07.026.
Gautam R, Vanga S, Ariese F, Umapathy S. Review
of multidimensional data processing approaches for
Raman and infrared spectroscopy. EPJ Tech Instrum.
;2:8. doi:10.1140/epjti/s40485-015-0018-6.
Lee LC, Liong C-Y, Jemain AA. A contemporary
review on Data Preprocessing (DP) practice
strategy in ATR-FTIR spectrum. Chemom
Intell Lab Syst 2017;163:64–75. doi:10.1016/j.
chemolab.2017.02.008.
Narayanan K, Thirunavukkarasu M, Esther Jeyanthi
C, Shenbagarajan P. FTIR and UV-visible spectral
study on normal blood samples. Int J Pharma. Bio.
Sci. 2011; 1:74-81.
Chung C, Lee M, Choe EK. Characterization of
cotton fabric scouring by FT-IR ATR spectroscopy.
Carbohydr Polym 2004;58:417–20. doi:10.1016/j.
carbpol.2004.08.005.
Li L, Frey M, Browning KJ. Biodegradability Study
on Cotton and Polyester Fabrics. J Eng Fiber Fabr.
; 42-53. doi:10.1177/155892501000500406.
Kong J, Yu S. Fourier Transform Infrared Spectroscopic
Analysis of Protein Secondary Structures. Acta
Biochim Biophys Sin (Shanghai) 2007;39:549–59.
doi:10.1111/j.1745-7270.2007.00320.x.
Dong A, Huang P, Caughey WS. Protein secondary
structures in water from second-derivative amide
I infrared spectra. Biochemistry 1990;29:3303–8.
doi:10.1021/bi00465a022.
Pelton JT, McLean LR. Spectroscopic Methods
for Analysis of Protein Secondary Structure.
Anal Biochem. 2000; 277:167–76. doi:10.1006/
abio.1999.4320.
Haris PI. Probing protein–protein interaction
in biomembranes using Fourier transform
infrared spectroscopy. Biochim Biophys Acta -
Biomembr. 2013;1828:2265–71. doi:10.1016/j.
bbamem.2013.04.008.
Morillas AV, Gooch J, Frascione N. Feasibility of
a handheld near infrared device for the qualitative
analysis of bloodstains. Talanta. 2018;184:1–6.
doi:10.1016/j.talanta.2018.02.110.
Article details
You must read and accept the copyright terms and conditions(click here for copyright terms).