Current Status and Prospects of Bloodstain Age Estimation Technology

doi:10.12116/j.issn.1004-5619.2023.431208

Abstract

Abstract:

Bloodstains are important biological evidence at violent crime scenes and contain rich information about the crime. The relatively accurate estimation of bloodstain age is of great significance for estimating the time of the crime, confirming or excluding the association of relevant persons （objects） with the case. In recent years, researchers at home and abroad have conducted extensive studies on the bloodstain age estimation from the aspects of bloodstain image and biochemical characteristics. This paper reviews the methods of bloodstain age estimation in the past two decades, discusses the obstacles of these methods in investigative practice, and provides an outlook on the development of bloodstain age estimation technology.

Key words: forensic pathology, chromatography, spectroscopy, bloodstain age, influencing factor, review

CLC Number:

Qi-rui HAN, Wen-ji ZHANG, Hao-yang LI, Ying-chao LUO. Current Status and Prospects of Bloodstain Age Estimation Technology[J]. Journal of Forensic Medicine, 2024, 40(5): 468-475.

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URL: http://www.fyxzz.cn/EN/10.12116/j.issn.1004-5619.2023.431208

http://www.fyxzz.cn/EN/Y2024/V40/I5/468

References 63

1	ZADORA G， MENŻYK A. In the pursuit of the holy grail of forensic science — Spectroscopic studies on the estimation of time since deposition of bloodstains[J]. TrAC Trends Analyt Chem，2018，105：137-165. doi：10.1016/j.trac.2018.04.009 .
2	LEERS O. Die forensische Blutuntersuchung： Ein Leitfaden für Studierende， beamtete und sachverständige Ärzte und für Kriminalisten[M]. Berlin： Julius Springer，1910：31-69.
3	温永启. 立体显微镜下依据滴落状血迹干燥度推断伤亡时间[J].中国人民公安大学学报（自然科学版），2013，19（3）：17-21.
	WEN Y Q. Inferring casualty time according to the dryness of dripping bloodstains under stereomicroscope[J]. Zhongguo Renmin Gongan Daxue Xuebao （Science and technology），2013，19（3）：17-21.
4	钱尊磊，李响，贾振军，等. 基于Image J软件的血迹遗留时间推断研究[J].中国法医学杂志，2023，38（1）：43-47. doi：10.13618/j.issn.1001-5728.2023.01.008 .
	QIAN Z L， LI X， JIA Z J， et al. Research on bloodstain age inference based on Image J software[J]. Zhongguo Fayixue Zazhi，2023，38（1）：43-47.
5	CHOI W， SHIN J， HYUN K A， et al. Highly sensitive and accurate estimation of bloodstain age using smartphone[J]. Biosens Bioelectron，2019，130：414-419. doi：10.1016/j.bios.2018.09.017 .
6	THANAKIATKRAI P， YAODAM A， KITPIPIT T. Age estimation of bloodstains using smartphones and digital image analysis[J]. Forensic Sci Int，2013，233（1/2/3）：288-297. doi：10.1016/j.forsciint.2013.09.027 .
7	SHIN J， CHOI S， YANG J S， et al. Smart Forensic Phone： Colorimetric analysis of a bloodstain for age estimation using a smartphone[J]. Sens Actuators B Chem，2017，243：221-225. doi：10.1016/j.snb.2016.11.142 .
8	郑吉龙，温洪洋，章彪，等. 一体化微量血痕成像及遗留时间分析装置的研发[J].法医学杂志，2019，35（2）：230-233，239. doi：10.12116/j.issn.1004-5619.2019.02.018 .
	ZHENG J L， WEN H Y， ZHANG B， et al. Development of integrated device of trace bloodstains imaging and age analysis[J]. Fayixue Zazhi，2019，35（2）：230-233，239.
9	郑吉龙，倪首涛，章彪，等. 血痕颜色时序性变化的实验性研究[J].中国医科大学学报，2018，47（5）：415-419，425. doi：10.12007/j.issn.0258-4646.2018.05.008 .
	ZHENG J L， NI S T， ZHANG B， et al. Experimental study on the time regularity of color changes of blood stains[J]. Zhongguo Yike Daxue Xuebao，2018，47（5）：415-419，425.
10	LEE Y R， LEE S， KWON S， et al. Bloodstain metabolite markers： Discovery and validation for estimating age of bloodstain within 7 days[J]. Anal Chem，2022，94（39）：13377-13384. doi：10.1021/acs.analchem.2c01903 .
11	LEE S， MUN S， LEE Y R， et al. Validation of the metabolite ergothioneine as a forensic marker in bloodstains[J]. Molecules，2022，27（24）：8885. doi：10.3390/molecules27248885 .
12	SEOK A E， LEE J， LEE Y R， et al. Estimation of age of bloodstains by mass-spectrometry： A metabolomic approach[J]. Anal Chem，2018，90（21）：12431-12441. doi：10.1021/acs.analchem.8b01367 .
13	HEO T M， GWON S Y， YANG J H， et al. Hemoglobin subunit beta protein as a novel marker for time since deposition of bloodstains at crime scenes[J]. Forensic Sci Int，2022，336：111348. doi：10.1016/j.forsciint.2022.111348 .
14	SCHNEIDER T D， ROSCHITZKI B， GROSSMANN J， et al. Determination of the time since deposition of blood traces utilizing a liquid chromatography-mass spectrometry-based proteomics approach[J]. Anal Chem，2022，94（30）：10695-10704. doi：10.1021/acs.analchem.2c01009 .
15	LEE Y R， LEE S， KWON S， et al. Effect of environmental conditions on bloodstain metabolite analysis[J]. Environ Res，2023，216（Pt 3）：114743. doi：10.1016/j.envres.2022.114743 .
16	JAMES S H， KISH P E， SUTTON T P. Principles of bloodstain pattern analysis：Theory and practice[M]. Boca Raton： CRC Press，2005：41-58.
17	MEINKE M， MÜLLER G， HELFMANN J， et al. Optical properties of platelets and blood plasma and their influence on the optical behavior of whole blood in the visible to near infrared wavelength range[J]. J Biomed Opt，2007，12（1）：014024. doi：10.1117/1.2435177 .
18	GOSCH A， BHARDWAJ A， COURTS C. TrACES of time： Transcriptomic analyses for the contextualization of evidential stains — Identification of RNA markers for estimating time-of-day of bloodstain deposition[J]. Forensic Sci Int Genet，2023，67：102915. doi：10.1016/j.fsigen.2023.102915 .
19	GIULIETTI N， DISCEPOLO S， CASTELLINI P， et al. Neural network based hyperspectral imaging for substrate independent bloodstain age estimation[J]. Forensic Sci Int，2023，349：111742. doi：10.1016/j.forsciint.2023.111742 .
20	LI B， BEVERIDGE P， O’ HARE W T， et al. The age estimation of blood stains up to 30 days old using visible wavelength hyperspectral image analysis and linear discriminant analysis[J]. Sci Justice，2013，53（3）：270-277. doi：10.1016/j.scijus.2013.04.004 .
21	GUO K， ACHILEFU S， BEREZIN M Y. Dating bloodstains with fluorescence lifetime measurements[J]. Chemistry，2012，18（5）：1303-1305. doi：10.1002/chem.201102935 .
22	SHINE S M， SUHLING K， BEAVIL A， et al. The applicability of fluorescence lifetime to determine the time since the deposition of biological stains[J]. Anal Methods，2017，9（13）：2007-2013. doi：10.1039/C6AY03099H .
23	HANSON E K， BALLANTYNE J. A blue spectral shift of the hemoglobin soret band correlates with the age （time since deposition） of dried bloodstains[J]. PLoS One，2010，5（9）：e12830. doi：10.1371/journal.pone.0012830 .
24	COSSETTE M L， STOTESBURY T， SHAFER A B A. Quantifying visible absorbance changes and DNA degradation in aging bloodstains under extreme temperatures[J]. Forensic Sci Int，2021，318：110627. doi：10.1016/j.forsciint.2020.110627 .
25	BERGMANN T， LEBERECHT C， LABUDDE D. Analysis of the influence of EDTA-treated reference samples on forensic bloodstain age estimation[J]. Forensic Sci Int，2021，325：110876. doi：10.1016/j.forsciint.2021.110876 .
26	吴雪梅，王琪，陈卫平，等. 基于ATR-FTIR光谱技术研究血痕形成的时序性变化[J].中国法医学杂志，2020，35（5）：475-478，483. doi：10.13618/j.issn.1001-5728.2020.05.005 .
	WU X M， WANG Q， CHEN W P， et al. Study on the time-dependent changes of blood stain formation using ATR-FTIR spectroscopy[J]. Zhongguo Fayixue Zazhi，2020，35（5）：475-478，483.
27	EDELMAN G， MANTI V， VAN RUTH S M， et al. Identification and age estimation of blood stains on colored backgrounds by near infrared spectroscopy[J]. Forensic Sci Int，2012，220（1/2/3）：239-244. doi：10.1016/j.forsciint.2012.03.009 .
28	KUMAR R， SHARMA K， SHARMA V. Bloodstain age estimation through infrared spectroscopy and Chemometric models[J]. Sci Justice，2020，60（6）：538-546. doi：10.1016/j.scijus.2020.07.004 .
29	LIN H， ZHANG Y， WANG Q， et al. Estimation of the age of human bloodstains under the simulated indoor and outdoor crime scene conditions by ATR-FTIR spectroscopy[J]. Sci Rep，2017，7（1）：13254. doi：10.1038/s41598-017-13725-1 .
30	STOTESBURY T， COSSETTE M L， NEWELL-BELL T， et al. An exploratory time since deposition analysis of whole blood using metrics of DNA degradation and visible absorbance spectroscopy[J]. Pure Appl Geophys，2020，178（3）：735-743. doi：10.1007/s00024-020-02494-0 .
31	DOTY K C， MURO C K， LEDNEV I K. Predicting the time of the crime： Bloodstain aging estimation for up to two years[J]. Forensic Chem，2017，5：1-7. doi：10.1016/j.forc.2017.05.002 .
32	DOTY K C， MCLAUGHLIN G， LEDNEV I K. A Raman “spectroscopic clock” for bloodstain age determination： The first week after deposition[J]. Anal Bioanal Chem，2016，408（15）：3993-4001. doi：10.1007/s00216-016-9486-z .
33	GAUTAM R， PEOPLES D， JANSEN K， et al. Feature selection and rapid characterization of bloodstains on different substrates[J]. Appl Spectrosc，2020，74（10）：1238-1251. doi：10.1177/00037028209 37776 .
34	ZHANG R， WANG P， CHEN J， et al. Age estimation of bloodstains based on Raman spectroscopy and chemometrics[J]. Spectrochim Acta Part A Mol Biomol Spectrosc，2023，290：122284. doi：10.1016/j.saa.2022.122284 .
35	LI B， BEVERIDGE P， O’HARE W T， et al. The estimation of the age of a blood stain using reflectance spectroscopy with a microspectrophotometer， spectral pre-processing and linear discriminant analysis[J]. Forensic Sci Int，2011，212（1/2/3）：198-204. doi：10.1016/j.forsciint.2011.05.031 .
36	SUN H， DONG Y， ZHANG P， et al. Accurate age estimation of bloodstains based on visible reflectance spectroscopy and chemometrics methods[J]. IEEE Photonics J，2017，9（1）：6500614. doi：10.1109/jphot.2017.2651580 .
37	BEREZIN M Y， ACHILEFU S. Fluorescence lifetime measurements and biological imaging[J]. Chem Rev，2010，110（5）：2641-2684. doi：10.1021/cr900343z .
38	EDELMAN G J， AALDERS M C G. Blood degradation and bloodstain age estimation[M]// SCHOTSMANS E M J， MÁRQUEZ-GRANT N， FORBES S L. Taphonomy of human remains： Forensic analysis of the dead and the depositional environment. Hoboken： John Wiley & Sons， Ltd.，2017：53-64.
39	GUO K， ZHEGALOVA N， ACHILEFU S， et al. Bloodstain age analysis： Toward solid state fluorescent lifetime measurements[C]// MAHADEVAN-JANSEN A， VO-DINH T， GRUNDFEST W S. Proceedings of SPIE. Advanced biomedical and clinical diagnostic systems Ⅺ. San Francisco， California， United States，2013.
40	DAYER M R， MOOSAVI-MOVAHEDI A A， DAYER M S. Band assignment in hemoglobin porphyrin ring spectrum： Using four-orbital model of Gouterman[J]. Protein Pept Lett，2010，17（4）：473-479. doi：10.2174/092986610790963645 .
41	LEMLER P， PREMASIRI W R， DELMONACO A， et al. NIR Raman spectra of whole human blood： Effects of laser-induced and in vitro hemoglobin denaturation[J]. Anal Bioanal Chem，2014，406（1）：193-200. doi：10.1007/s00216-013-7427-7 .
42	BREMMER R H， NADORT A， VAN LEEUWEN T G， et al. Age estimation of blood stains by hemoglobin derivative determination using reflectance spectroscopy[J]. Forensic Sci Int，2011，206（1/2/3）：166-171. doi：10.1016/j.forsciint.2010.07.034 .
43	KAUR S， SAINI V， DALAL R. UV-Visible spectroscopic effect on haemoglobin & DNA degradation： A forensic approach[J]. Forensic Sci Int，2020，307：110078. doi：10.1016/j.forsciint.2019.110078 .
44	BAUER M， POLZIN S， PATZELT D. Quantification of RNA degradation by semi-quantitative duplex and competitive RT-PCR： A possible indicator of the age of bloodstains?[J]. Forensic Sci Int，2003，138（1/2/3）：94-103. doi：10.1016/j.forsciint.2003.09.008 .
45	ZHAO H， WANG C， YAO L， et al. Identification of aged bloodstains through mRNA profiling： Experiments results on selected markers of 30- and 50-year-old samples[J]. Forensic Sci Int，2017，272：e1-e6. doi：10.1016/j.forsciint.2017.01.006 .
46	ANDERSON S， HOWARD B， HOBBS G R， et al. A method for determining the age of a bloodstain[J]. Forensic Sci Int，2005，148（1）：37-45. doi：10.1016/j.forsciint.2004.04.071 .
47	ANDERSON S E， HOBBS G R， BISHOP C P. Multivariate analysis for estimating the age of a bloodstain[J]. J Forensic Sci，2011，56（1）：186-193. doi：10.1111/j.1556-4029.2010.01551.x .
48	许炎，蒋巍，平原，等. 应用RNA分析技术推断现场血痕形成时间[J].法医学杂志，2010，26（5）：340-342. doi：10.3969/j.issn.1004-5619.2010.05.006 .
	XU Y， JIANG W， PING Y， et al. Determination of bloodstain formation time by RNA analysis[J]. Fayixue Zazhi，2010，26（5）：340-342.
49	QI B， KONG L， LU Y. Gender-related difference in bloodstain RNA ratio stored under uncontrolled room conditions for 28 days[J]. J Forensic Leg Med，2013，20（4）：321-325. doi：10.1016/j.jflm.2012.09.014 .
50	WEI Y， WANG J， WANG Q， et al. The estimation of bloodstain age utilizing circRNAs and mRNAs biomarkers[J]. Forensic Sci Int，2022，338：111408. doi：10.1016/j.forsciint.2022.111408 .
51	HENEGHAN N， FU J， PRITCHARD J， et al. The effect of environmental conditions on the rate of RNA degradation in dried blood stains[J]. Forensic Sci Int Genet，2021，51：102456. doi：10.1016/j.fsigen.2020.102456 .
52	CHENG F， LI W， JI Z， et al. Estimation of bloodstain deposition time within a 24-h day-night cycle with rhythmic mRNA based on a machine learning algorithm[J]. Forensic Sci Int Genet，2023，66：102910. doi：10.1016/j.fsigen.2023.102910 .
53	DISSING J， SØNDERVANG A， LUND S. Exploring the limits for the survival of DNA in blood stains[J]. J Forensic Leg Med，2010，17（7）：392-396. doi：10.1016/j.jflm.2010.08.001 .
54	SEARS D A， UDDEN M M， THOMAS L J. Carboxyhemoglobin levels in patients with sickle-cell anemia： Relationship to hemolytic and vasoocclusive severity[J]. Am J Med Sci，2001，322（6）：345-348. doi：10.1097/00000441-200112000-00007 .
55	WALD N J， IDLE M， BOREHAM J， et al. Carbon monoxide in breath in relation to smoking and carboxyhaemoglobin levels[J]. Thorax，1981，36（5）：366-369. doi：10.1136/thx.36.5.366 .
56	CANENE-ADAMS K， ERDMAN JR J W. Absorption， transport， distribution in tissues and bioavaila-bility[M]// BRITTON G， LIAAEN-JENSEN S， PFANDER H. Carotenoids. Basel， Switzerland： Birkhäuser Verlag Basel，2009：115-148.
57	BREMMER R H， DE BRUIN D M， DE JOODE M， et al. Biphasic oxidation of oxy-hemoglobin in bloodstains[J]. PLoS One，2011，6（7）：e21845. doi：10.1371/journal.pone.0021845 .
58	FUJITA Y， TSUCHIYA K， ABE S， et al. Estimation of the age of human bloodstains by electron paramagnetic resonance spectroscopy： Long-term controlled experiment on the effects of environmental factors[J]. Forensic Sci Int，2005，152（1）：39-43. doi：10.1016/j.forsciint.2005.02.029 .
59	BERGMANN T， HEINKE F， LABUDDE D. Towards substrate-independent age estimation of blood stains based on dimensionality reduction and k-nearest neighbor classification of absorbance spectroscopic data[J]. Forensic Sci Int，2017，278：1-8. doi：10.1016/j.forsciint.2017.05.023 .
60	ARANY S， OHTANI S. Age estimation of bloodstains： A preliminary report based on aspartic acid racemization rate[J]. Forensic Sci Int，2011，212（1/2/3）：e36-e39. doi：10.1016/j.forsciint.2011.05.015 .
61	STRASSER S， ZINK A， KADA G， et al. Age determination of blood spots in forensic medicine by force spectroscopy[J]. Forensic Sci Int，2007，170（1）：8-14. doi：10.1016/j.forsciint.2006.08.023 .
62	WU Y， HU Y， CAI J， et al. Time-dependent surface adhesive force and morphology of RBC measured by AFM[J]. Micron，2009，40（3）：359-364. doi：10.1016/j.micron.2008.10.003 .
63	MARRONE A， LA RUSSA D， MONTESANTO A， et al. Short and long time bloodstains age determination by colorimetric analysis： A pilot study[J]. Molecules，2021，26（20）：6272. doi：10.3390/molecules 26206272 .

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