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Analytical and Bioanalytical Chemistry

, Volume 406, Issue 1, pp 193–200 | Cite as

NIR Raman spectra of whole human blood: effects of laser-induced and in vitro hemoglobin denaturation

  • P. Lemler
  • W. R. Premasiri
  • A. DelMonaco
  • L. D. Ziegler
Research Paper

Abstract

Care must be exercised in the use of Raman spectroscopy for the identification of blood in forensic applications. The Raman spectra of dried whole human blood excited at 785 nm are shown to be exclusively due to oxyhemoglobin or related hemoglobin denaturation products. Raman spectra of whole blood are reported as a function of the incident 785-nm-laser power, and features attributable to heme aggregates are observed for fluences on the order of 104 W/cm2 and signal collection times of 20 s. In particular, the formation of this local-heating-induced heme aggregate product is indicated by a redshifting of several heme porphyrin ring vibrational bands, the appearance of a large broad band at 1,248 cm-1, the disappearance of the Fe–O2 stretching and bending bands, and the observation of a large overlapping fluorescence band. This denaturation product is also observed in the low-power-excitation Raman spectrum of older ambient-air-exposed bloodstains (2 weeks or more). The Raman spectrum of methemoglobin whole blood excited at 785 nm is reported, and increasing amounts of this natural denaturation product can also be identified in Raman spectra of dried whole blood particularly when the blood has been stored prior to drying. These results indicate that to use 785-nm-excitation Raman spectra as an identification method for forensic applications to maximum effect, incident laser powers need to be kept low to eliminate variable amounts of heme aggregate spectral components contributing to the signal and the natural aging process of hemoglobin denaturation needs to be accounted for. This also suggests that there is a potential opportunity for 785-nm-excitation Raman spectra to be a sensitive indicator of the age of dried bloodstains at crime scenes.

Keywords

Raman spectroscopy Hemoglobin Human blood Forensics 

Notes

Acknowledgments

The support of the National Institutes of Health (grant 1R01AI090815-01) and the Undergraduate Research Opportunities Program of Boston University (support for P.L) is gratefully acknowledged.

References

  1. 1.
    Mazzella WD, Buzzini P (2005) Raman spectroscopy of blue gel pen inks. Forensic Sci Int 152:241–247CrossRefGoogle Scholar
  2. 2.
    Suzuki EM, Carrabba M (2001) In situ identification and analysis of automotive paint pigments using line segment excitation Raman spectroscopy: I. Inorganic topcoat pigments. J Forensic Sci 46:1053–1069Google Scholar
  3. 3.
    Thomas J, Buzzini P, Massonnet G, Reedy B, Roux C (2005) Raman spectroscopy and the forensics analysis of back/grey and blue cotton fibers part 1: investigation of the effects of varying laser wavelength. Forensic Sci Int 152:189–197CrossRefGoogle Scholar
  4. 4.
    West MJ, Went MJ (2011) Detection of drugs of abuse by Raman spectroscopy. Drug Test Anal 3:532–538CrossRefGoogle Scholar
  5. 5.
    Hargreaves MD, Page K, Munshi T, Tomsett R, Lynch G, Edwards HGM (2008) Analysis of seized drugs using portable Raman spectroscopy in an airport environment—a proof of principle study. J Raman Spectrosc 39:873–880CrossRefGoogle Scholar
  6. 6.
    Moore DS, Scharff RJ (2009) Portable Raman explosives detection. Anal Bioanal Chem 393:1571–1578CrossRefGoogle Scholar
  7. 7.
    Bueno J, Sikirzhytski V, Lednev IK (2012) Raman spectroscopic analysis of gunshot residue offering great potential for caliber differentiation. Anal Chem 84:4334–4339CrossRefGoogle Scholar
  8. 8.
    De Wael K, Lepot L, Gason F, Gilbert B (2008) In search of blood—detection of minute particles using spectroscopic methods. Forensic Sci Int 180:37–42CrossRefGoogle Scholar
  9. 9.
    Virkler K, Lednev IK (2008) Raman spectroscopy offers great potential for the nondestructive confirmatory identification of body fluids. Forensic Sci Int 18:e1–e5CrossRefGoogle Scholar
  10. 10.
    Virkler K, Lednev IK (2009) Blood species identification for forensic purposes using Raman spectroscopy combined with advanced statistical analysis. Anal Chem 81:7773–7777CrossRefGoogle Scholar
  11. 11.
    Sikirzhytski V, Virkler K, Lednev IK (2010) Discriminant analysis of Raman spectra for body fluid identification for forensic purposes. Sensors 10:2869–2884CrossRefGoogle Scholar
  12. 12.
    Virkler K, Lednev IK (2010) Raman spectroscopic signature of blood and its potential application to forensic body fluid identification. Anal Bioanal Chem 396:525–534CrossRefGoogle Scholar
  13. 13.
    Virkler K, Lednev IK (2010) Forensic body fluid identification: the Raman spectroscopic signature of saliva. Analyst 135:512–517CrossRefGoogle Scholar
  14. 14.
    Virkler K, Lednev IK (2009) Analysis of body fluids for forensic purposes: from laboratory testing to non-destructive rapid confirmatory identification at a crime scene. Forensic Sci Int 188:1–17CrossRefGoogle Scholar
  15. 15.
    Sikirzhytski V, Sikirzhytskaya A, Lednev IK (2011) Multidimensional Raman spectroscopic signatures as a tool for forensic identification of body fluid traces: a review. Appl Spectrosc 65:1223–1232CrossRefGoogle Scholar
  16. 16.
    Sikirzhytskaya A, Sikirzhytski V, Lednev IK (2012) Raman spectroscopic signature of vaginal fluid and its potential application in forensic body fluid identification. Forensic Sci Int 216:44–48CrossRefGoogle Scholar
  17. 17.
    Boyd S, Bertino MF, Seashols SJ (2011) Raman spectroscopy of blood samples for forensic applications. Forensic Sci Int 208:124–128CrossRefGoogle Scholar
  18. 18.
    Premasiri WR, Lee JC, Ziegler LD (2012) Surface-enhanced Raman scattering of whole human blood, blood plasma, and red blood cells: cellular processes and bioanalytical sensing. J Phys Chem B 116:9376–9386CrossRefGoogle Scholar
  19. 19.
    Bremmer RH, de Bruin DM, de Joode M, Buma WJ, van Leeuwen TG, Aalders MCG (2011) Biphasic oxidation of oxy-hemoglobin in bloodstains. PLoS ONE 6:e21845CrossRefGoogle Scholar
  20. 20.
    Wood BR, Tait B, McNaughton D (2001) Micro-Raman characterisation of the R to T state transition of haemoglobin within a single living erythrocyte. Biochim Biophys Acta 1539:58–70CrossRefGoogle Scholar
  21. 21.
    Wood BR, Caspers P, Puppels GJ, Pandiancherri S, McNaughton D (2007) Resonance Raman spectroscopy of red blood cells using near-infrared laser excitation. Anal Bioanal Chem 387:1691–1703CrossRefGoogle Scholar
  22. 22.
    Hu S, Smith KM, Spiro TG (1996) Assignment of protoheme resonance Raman spectrum by heme labeling in myoglobin. J Am Chem Soc 118:12638–12646CrossRefGoogle Scholar
  23. 23.
    Abe M, Kitagawa T, Kyogoku Y (1978) Resonance Raman spectra of octaethylporphyrinato-Ni(II) and meso-deuterated and 15N substituted derivatives. II. A normal coordinate analysis. J Chem Phys 69:4526–4534CrossRefGoogle Scholar
  24. 24.
    Sato H, Chiba H, Tashiro H, Ozaki Y (2001) Excitation wavelength-dependent changes in Raman spectra of whole blood and hemoglobin: comparison of the spectra with 514.5-, 720-, and 1064-nm excitation. J Biomed Opt 6:366–370CrossRefGoogle Scholar
  25. 25.
    Wood BR, Hammer L, Davis L, McNaughton D (2005) Raman microspectroscopy and imaging provides insights into heme aggregation and denaturation within human erythrocytes. J Biomed Opt 10:014005CrossRefGoogle Scholar
  26. 26.
    Shikama K (1998) The molecular mechanism of autoxidation for myoglobin and hemoglobin: a venerable puzzle. Chem Rev 98:1357–1374CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • P. Lemler
    • 1
  • W. R. Premasiri
    • 1
  • A. DelMonaco
    • 2
  • L. D. Ziegler
    • 1
  1. 1.Department of Chemistry and The Photonics CenterBoston UniversityBostonUSA
  2. 2.Department of Biomedical EngineeringBoston UniversityBostonUSA

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