False Positives in Luminal Testing
Year 2024,
, 129 - 138, 15.01.2024
Yakup Gülekçi
,
Fatma Cavus Yonar
Abstract
The blood sample is one of the most essential pieces of evidence that helps criminal experts in the elucidation of the crime. However, cleaning the blood found at the crime scene after the crime is committed makes it difficult to detect the crime. Therefore, experts have attached great importance to research on blood samples left at the crime scene. Although many test kits are used, especially in detecting erased blood, the luminol kit is widely used at crime scenes. In the luminol kit, the reaction takes place with the addition of hydrogen peroxide (H2O2). Since it is known that hydrogen peroxide can react with other substances containing Fe (II) ions that may be present in the environment during the reaction, it was aimed to carry out research to determine which substances luminol interferes with, especially those that may be found in a domestic crime scene in this study. In domestic crime scenes, there are limited substances that can replicate the distinct, enduring luminescence characteristic of a reaction between luminol and genuine bloodstains, while the uncharacteristic glow produced from many other surfaces is likely to be recognised with the naked eye by a good expert, experienced and knowledgeable in the field. In this regard, the prepared samples were treated with luminol in a dark environment, and the reactions occurring in the first seconds were recorded. The results obtained with this reagent are recommended to be supported by other blood test reagents or confirmatory tests, as there are substances that give false positive results with luminol reagents in the present study.
Ethical Statement
The authors declared that the article was one of the studies that did not require ethics committee approval.
Supporting Institution
The authors declare no conflict of interest. The funders had no role in the design of the study, in the collection, analyses, or interpretation of data, in the writing of the manuscript, or in the decision to publish the results.
Thanks
The authors would like to thank Istanbul University-Cerrahpasa, Institute of Forensic Medicine and Forensic Sciences, Forensic Sciences Laboratory and Kutahya Health Sciences University, Application and Research Centre Laboratory for providing laboratory facilities and a convenient working environment for this study.
References
- Adair TW, Gabel R, Shimamoto S, Tewes R. 2008. A comparison of the luminol and Blue star blood reagents in detecting blood in soil nearly four years after deposition. IABPA News, 24(4): 5-8.
- An JH, Shin KJ, Yang WI, Lee HY. 2012. Body fluid identification in forensics. BMB Rep, 45(10): 545–53. https://doi.org/10.5483/BMBRep.2012.45.10.206.
- Arnhold J, Mueller S, Arnold K, Grimm E. 1991. Chemiluminescence intensities and spectra of luminol oxidation by sodium hypochlorite in the presence of hydrogen peroxide. J Biolumin Chemilumin, 6(3): 189-192.
- Arnhold J, Mueller S, Arnold K, Sonntag K. 1993. Mechanisms of inhibition of chemiluminescence in the oxidation of luminol by sodium hypochlorite. J Biolumin Chemilumin, 8(6): 307-313. https://doi.org/10.1002/bio.1170080604.
- Barni F, Lewis SW, Berti A, Miskelly GM, Lago G. 2007. Forensic application of the luminol reaction as a presumptive test for latent blood detection. Talanta, 72(3): 896–913. https://doi.org/10.1016/j.talanta.2006.12.045.
- Castello A, Alvarez M, Verdu F. 2002. Accuracy, reliability, and safety of luminol in bloodstain investigation. Canadian Soc Foren Sci J, 35(3): 113-121.
- Cheyne M. 2011. Illuminating latent blood. Application methods, fixatives, alternatives and new formulas for luminol. Corpus, 1-222.
- Chourasiya L, Mahakalkar A. 2017. To compare and study differential detection of human and animal blood using phenolphthalein, tetramethylbenzidine and luminol assays. Int J Recent Trends Eng Res, 3(11): 82-102.
- Cox M. 1991. A study of the sensitivity and specificity of four presumptive tests for blood. J Foren Sci, 36(5): 1503-1511. https://doi.org/10.1520/JFS13170J.
- Creamer JI, Quickenden TI, Apanah MV, Kerr KA, Robertson P. 2003. A comprehensive experimental study of industrial domestic and environmental interferences with the forensic Luminol test for blood. Lumin, 18: 193-198.
- Creamer JI, Quickenden TI, Leah BC. 2005. At tempted cleaning of bloodstains and its effect on the forensic luminol test. Lumin, 20: 411– 413.
- Da Silva RR, Agustini BC, da Silva ALL, Frigeri HR. 2012. Luminol in the forensic science. J Biotech Biodiv, 3(4):172-177.
- Dilbeck L. 2006. Use of BlueStar forensic in lieu of luminol at crime scenes. J Forensic Identif, 56(5): 706–20.
- Divortex, 2023. Kimyasal dayanıklı sıvı püskürtme (Sprey) şişesi 1 Lt. URL: https://www.divortex.com.tr/aksesuarlar/kimyasal-dayanikli-sivi-puskurtme-sprey-sisesi-1-lt. (accessed date: Sep 23, 2023).
- Fatoki TH. 2020. In-Silico investigation of luminol, its analogues and improved mechanism of chemiluminescence for blood identification beyond forensics. bioRxiv, 01.
- Fereja TH, Kitte SA, Gao W, Yuan F, Snizhko D, Qi L, Xu G. 2019. Artesunate-luminol chemiluminescence system for the detection of hemin. Talanta, 204: 379-385.
- Finnis J, Lewis J, Davidson A. 2013. Comparison of methods for visualizing blood on dark surfaces. Sci Justice, 53(2): 178-186.
- Grodsky M, Wright K, Kirk PL. 1951. Simplified preliminary blood testing--An improved technique and a comparative study of methods. J Crim L Criminol Police Sci, 42: 95.
- James SH, Kish PE, Sutton TP. 2005. Principles of bloodstain pattern analysis: theory and practice, CRC Press, Boca Raton, 1st ed., New York, USA, pp: 533.
- James SH, Nordby JJ, Forensic Science: An introduction to scientific and investigative techniques, CRC Press, New York, USA, 2nd ed., pp: 667.
- Johnston S, Newman J, Frappier R. 2003. Validation study of the Abacus Diagnostics ABAcard® HemaTrace® membrane test for the forensic identification of human blood. Can Soc Forensic Sci J, 36(3): 173–83. https://doi.org/10.1080/00085030.2003.10757560.
- Karabchevsky A, Mosayyebi A, Kavokin AV. 2016. Tuning the chemiluminescence of a luminol flow using plasmonic nanoparticles. Light Sci Applicat, 5(11): 16164-16164.
- King R, Miskelly G. 2005. The inhibition by amines and amino acids of bleach-induced luminol chemiluminescence during forensic screening for blood. Talanta, 67(2):345–353. https://doi.org/10.1016/j.talanta.2005.01.034.
- Klein A, Feudel E, Türk E, Püschel K, Gehl A. 2007. Luminescence after the use of luminol: Positive or false positive? Rechtsmedizin, 17: 146-152.
- Laux L. 2005. The detection of Blood Using Luminol. in: James SH, Kish PE, Sutton TP. (Eds.), Principles of bloodstain pattern analysis: theory and practice. CRC Press, Boca Raton, 1st ed., New York, USA, pp: 369–389.
- Li R. 2015. Forensic Biology. CRC Press, New York, USA, 2nd ed., pp: 533.
- Lytle LT, Hedgecock DG. 1978. Chemiluminescence in the visualization of forensic bloodstains. J Foren Sci, 23(3): 550-562.
- Manna AD, Montpetit S. 2000. A novel approach to obtaining reliable PCR results from luminol treated bloodstains. J Foren Sci, 45(4): 886-890.
- Marengo-Rowe AJ. 2006. Structure-function relations of human hemoglobins. Proc Bayl Univ Med Cent, 19(3): 239-245.
- Ming Lin J, Shan X, Hanaoka S, Yamada M. 2001. Luminol chemiluminescence in unbuffered solutions with a cobalt(II) – ethanolamine complex immobilized on resin as catalyst and its application to analysis. Anal Chem, 73: 5043–5051. https://doi.org/10.1021/ac010573+.
- Misencik A, Laux DL. 2007. Validation study of the Seratec HemDirect hemoglobin assay for the forensic identification of Human Blood. MAFS Newsl. URL: https://pdfs.semanticscholar.org/2c56/c17d14e06a7472f6246596d10f76164844bb.pdf. (accessed date: Sep 25, 2023).
- Novelli BC. 2020. A review of substances reported to cause false positives and negatives in forensic blood identification tests. MSc thesis, Boston University, Boston, USA, pp: 66.
- Patzelt D, 2004. History of forensic serology and molecular genetics in the sphere of activity of the German Society for Forensic Medicine. Forensic Sci Int, 144: 185-191.
- Petersen D, Kovacs F. 2014. Phenolphthalein false-positive reactions from legume root nodules. J Forensic Sci, 59(2): 481–484. https://doi.org/10.1111/1556-4029.12352.
- Quickenden TI, Cooper PD. 2001a. Increasing the specificity of the forensic luminol test for blood. Lumin, 16: 251–253.
- Quickenden TI, Creamer JI. 2001b. A study of common interferences with the forensic luminol test for blood. Lumin 16: 295–298. https://doi.org/10.1002/bio.657
- Quickenden TI, Ennis CP, Creamer JI. 2004. The forensic use of Luminol chemiluminescence to detect traces of blood inside motor vehicles. Lumin, 19: 271-277.
- Rogiski da Silva R, Agustini B, Lopes da Silva AL, Frigeri HR. 2012. Luminol in the forensic science. J Biotech Biodiv, 3(4): 172–177.
- Seitz WR, Hercules DM. 1972. Determination of trace amounts of iron(II) using chemiluminescence analysis. Anal Chem, 44: 2143–2149. https://doi.org/10.1021/AC60321A020.
- Shivangi G, Apoorva G, Suresh B, Yadav DS, Mahanta P. 2021. Comparative study of presumptive and confirmatory tests for detection of blood on serial dilutions and washed stains. Int J Health Res Medico Leg Prae, 7(1): 59-64.
- Sirchie. 2011. Luminol. URL: https://www.sirchie.com/media/resourcecenter/item/l/u/luminol_ti02-48eng-rev10e.pdf. (accessed date: Sep 23, 2023).
- Stoica BA, Bunescu S, Neamtu A, Bulgaru‐Iliescu D, Foia L, Botnariu EG. 2016. Improving luminol blood detection in forensics. J Foren Sci, 61(5): 1331-1336.
- Stott RAW, Kricka LJ. 1987. Biolumin chemilumin symp. John Wiley & Sons Ltd., Chichester, New York, USA, pp: 237–240.
- Tajani, A. 2014. Photonics for forensic applications. Photon Safet Secur, 1987: 368-397.
- Tobe SS, Watson N, Daéid NN. 2007. Evaluation of six presumptive tests for blood, their specificity, sensitivity, and effect on high molecular-weight DNA. J Forensic Sci, 52(1): 102–109. https://doi.org/10.1111/j.1556-4029.2006.00324.x.
- 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. Foren Sci Inter, 188(1-3): 1-17.
- Weber K. 1966. Die Anwendung der chemiluminescenz des luminols in der gerichtlichen medizin und toxikologie: i. der nachweis von blutspuren. Deutsche Zeitschrift Medizin, 57: 410-423.
- Wells PG, Winn LÇM. 1996. Biochemical toxicology of chemical teratogenesis. Critical Review in Biochem. Mol. Biol. 31 (1): 1-40.
- Yeh HC, Lin WY. 2003. Enhancement of Chemiluminescence from the Oxidation of Luminol with Hydrogen Peroxide Catalyzed by Mn (III)‐Microperoxidase 8. J Chinese Chem Soc, 50(1): 81-88.
False Positives in Luminal Testing
Year 2024,
, 129 - 138, 15.01.2024
Yakup Gülekçi
,
Fatma Cavus Yonar
Abstract
The blood sample is one of the most essential pieces of evidence that helps criminal experts in the elucidation of the crime. However, cleaning the blood found at the crime scene after the crime is committed makes it difficult to detect the crime. Therefore, experts have attached great importance to research on blood samples left at the crime scene. Although many test kits are used, especially in detecting erased blood, the luminol kit is widely used at crime scenes. In the luminol kit, the reaction takes place with the addition of hydrogen peroxide (H2O2). Since it is known that hydrogen peroxide can react with other substances containing Fe (II) ions that may be present in the environment during the reaction, it was aimed to carry out research to determine which substances luminol interferes with, especially those that may be found in a domestic crime scene in this study. In domestic crime scenes, there are limited substances that can replicate the distinct, enduring luminescence characteristic of a reaction between luminol and genuine bloodstains, while the uncharacteristic glow produced from many other surfaces is likely to be recognised with the naked eye by a good expert, experienced and knowledgeable in the field. In this regard, the prepared samples were treated with luminol in a dark environment, and the reactions occurring in the first seconds were recorded. The results obtained with this reagent are recommended to be supported by other blood test reagents or confirmatory tests, as there are substances that give false positive results with luminol reagents in the present study.
Ethical Statement
Yazarlar makalenin etik kurul izni gerektirmeyen araştırmalardan biri olduğunu beyan etmiştir.
Supporting Institution
Bu çalışma sırasında, yapılan araştırma konusu ile ilgili doğrudan bağlantısı bulunan herhangi bir kurum veya firmadan, çalışmanın değerlendirme sürecinde, çalışma ile ilgili verilecek kararı olumsuz etkileyebilecek maddi ve/veya manevi herhangi bir destek alınmamıştır.
Thanks
Bu çalışmanın yapılmasında; laboratuvar olanak ve elverişli çalışma ortamı sunan, İstanbul Üniversitesi-Cerrahpaşa, Adli Tıp ve Adli Bilimler Enstitüsü Adli Bilimler Laboratuvarı’na ve Kütahya Sağlık Bilimleri Üniversitesi, Uygulama ve Araştırma Merkezi Laboratuvarı’na teşekkürlerimizi sunarız.
References
- Adair TW, Gabel R, Shimamoto S, Tewes R. 2008. A comparison of the luminol and Blue star blood reagents in detecting blood in soil nearly four years after deposition. IABPA News, 24(4): 5-8.
- An JH, Shin KJ, Yang WI, Lee HY. 2012. Body fluid identification in forensics. BMB Rep, 45(10): 545–53. https://doi.org/10.5483/BMBRep.2012.45.10.206.
- Arnhold J, Mueller S, Arnold K, Grimm E. 1991. Chemiluminescence intensities and spectra of luminol oxidation by sodium hypochlorite in the presence of hydrogen peroxide. J Biolumin Chemilumin, 6(3): 189-192.
- Arnhold J, Mueller S, Arnold K, Sonntag K. 1993. Mechanisms of inhibition of chemiluminescence in the oxidation of luminol by sodium hypochlorite. J Biolumin Chemilumin, 8(6): 307-313. https://doi.org/10.1002/bio.1170080604.
- Barni F, Lewis SW, Berti A, Miskelly GM, Lago G. 2007. Forensic application of the luminol reaction as a presumptive test for latent blood detection. Talanta, 72(3): 896–913. https://doi.org/10.1016/j.talanta.2006.12.045.
- Castello A, Alvarez M, Verdu F. 2002. Accuracy, reliability, and safety of luminol in bloodstain investigation. Canadian Soc Foren Sci J, 35(3): 113-121.
- Cheyne M. 2011. Illuminating latent blood. Application methods, fixatives, alternatives and new formulas for luminol. Corpus, 1-222.
- Chourasiya L, Mahakalkar A. 2017. To compare and study differential detection of human and animal blood using phenolphthalein, tetramethylbenzidine and luminol assays. Int J Recent Trends Eng Res, 3(11): 82-102.
- Cox M. 1991. A study of the sensitivity and specificity of four presumptive tests for blood. J Foren Sci, 36(5): 1503-1511. https://doi.org/10.1520/JFS13170J.
- Creamer JI, Quickenden TI, Apanah MV, Kerr KA, Robertson P. 2003. A comprehensive experimental study of industrial domestic and environmental interferences with the forensic Luminol test for blood. Lumin, 18: 193-198.
- Creamer JI, Quickenden TI, Leah BC. 2005. At tempted cleaning of bloodstains and its effect on the forensic luminol test. Lumin, 20: 411– 413.
- Da Silva RR, Agustini BC, da Silva ALL, Frigeri HR. 2012. Luminol in the forensic science. J Biotech Biodiv, 3(4):172-177.
- Dilbeck L. 2006. Use of BlueStar forensic in lieu of luminol at crime scenes. J Forensic Identif, 56(5): 706–20.
- Divortex, 2023. Kimyasal dayanıklı sıvı püskürtme (Sprey) şişesi 1 Lt. URL: https://www.divortex.com.tr/aksesuarlar/kimyasal-dayanikli-sivi-puskurtme-sprey-sisesi-1-lt. (accessed date: Sep 23, 2023).
- Fatoki TH. 2020. In-Silico investigation of luminol, its analogues and improved mechanism of chemiluminescence for blood identification beyond forensics. bioRxiv, 01.
- Fereja TH, Kitte SA, Gao W, Yuan F, Snizhko D, Qi L, Xu G. 2019. Artesunate-luminol chemiluminescence system for the detection of hemin. Talanta, 204: 379-385.
- Finnis J, Lewis J, Davidson A. 2013. Comparison of methods for visualizing blood on dark surfaces. Sci Justice, 53(2): 178-186.
- Grodsky M, Wright K, Kirk PL. 1951. Simplified preliminary blood testing--An improved technique and a comparative study of methods. J Crim L Criminol Police Sci, 42: 95.
- James SH, Kish PE, Sutton TP. 2005. Principles of bloodstain pattern analysis: theory and practice, CRC Press, Boca Raton, 1st ed., New York, USA, pp: 533.
- James SH, Nordby JJ, Forensic Science: An introduction to scientific and investigative techniques, CRC Press, New York, USA, 2nd ed., pp: 667.
- Johnston S, Newman J, Frappier R. 2003. Validation study of the Abacus Diagnostics ABAcard® HemaTrace® membrane test for the forensic identification of human blood. Can Soc Forensic Sci J, 36(3): 173–83. https://doi.org/10.1080/00085030.2003.10757560.
- Karabchevsky A, Mosayyebi A, Kavokin AV. 2016. Tuning the chemiluminescence of a luminol flow using plasmonic nanoparticles. Light Sci Applicat, 5(11): 16164-16164.
- King R, Miskelly G. 2005. The inhibition by amines and amino acids of bleach-induced luminol chemiluminescence during forensic screening for blood. Talanta, 67(2):345–353. https://doi.org/10.1016/j.talanta.2005.01.034.
- Klein A, Feudel E, Türk E, Püschel K, Gehl A. 2007. Luminescence after the use of luminol: Positive or false positive? Rechtsmedizin, 17: 146-152.
- Laux L. 2005. The detection of Blood Using Luminol. in: James SH, Kish PE, Sutton TP. (Eds.), Principles of bloodstain pattern analysis: theory and practice. CRC Press, Boca Raton, 1st ed., New York, USA, pp: 369–389.
- Li R. 2015. Forensic Biology. CRC Press, New York, USA, 2nd ed., pp: 533.
- Lytle LT, Hedgecock DG. 1978. Chemiluminescence in the visualization of forensic bloodstains. J Foren Sci, 23(3): 550-562.
- Manna AD, Montpetit S. 2000. A novel approach to obtaining reliable PCR results from luminol treated bloodstains. J Foren Sci, 45(4): 886-890.
- Marengo-Rowe AJ. 2006. Structure-function relations of human hemoglobins. Proc Bayl Univ Med Cent, 19(3): 239-245.
- Ming Lin J, Shan X, Hanaoka S, Yamada M. 2001. Luminol chemiluminescence in unbuffered solutions with a cobalt(II) – ethanolamine complex immobilized on resin as catalyst and its application to analysis. Anal Chem, 73: 5043–5051. https://doi.org/10.1021/ac010573+.
- Misencik A, Laux DL. 2007. Validation study of the Seratec HemDirect hemoglobin assay for the forensic identification of Human Blood. MAFS Newsl. URL: https://pdfs.semanticscholar.org/2c56/c17d14e06a7472f6246596d10f76164844bb.pdf. (accessed date: Sep 25, 2023).
- Novelli BC. 2020. A review of substances reported to cause false positives and negatives in forensic blood identification tests. MSc thesis, Boston University, Boston, USA, pp: 66.
- Patzelt D, 2004. History of forensic serology and molecular genetics in the sphere of activity of the German Society for Forensic Medicine. Forensic Sci Int, 144: 185-191.
- Petersen D, Kovacs F. 2014. Phenolphthalein false-positive reactions from legume root nodules. J Forensic Sci, 59(2): 481–484. https://doi.org/10.1111/1556-4029.12352.
- Quickenden TI, Cooper PD. 2001a. Increasing the specificity of the forensic luminol test for blood. Lumin, 16: 251–253.
- Quickenden TI, Creamer JI. 2001b. A study of common interferences with the forensic luminol test for blood. Lumin 16: 295–298. https://doi.org/10.1002/bio.657
- Quickenden TI, Ennis CP, Creamer JI. 2004. The forensic use of Luminol chemiluminescence to detect traces of blood inside motor vehicles. Lumin, 19: 271-277.
- Rogiski da Silva R, Agustini B, Lopes da Silva AL, Frigeri HR. 2012. Luminol in the forensic science. J Biotech Biodiv, 3(4): 172–177.
- Seitz WR, Hercules DM. 1972. Determination of trace amounts of iron(II) using chemiluminescence analysis. Anal Chem, 44: 2143–2149. https://doi.org/10.1021/AC60321A020.
- Shivangi G, Apoorva G, Suresh B, Yadav DS, Mahanta P. 2021. Comparative study of presumptive and confirmatory tests for detection of blood on serial dilutions and washed stains. Int J Health Res Medico Leg Prae, 7(1): 59-64.
- Sirchie. 2011. Luminol. URL: https://www.sirchie.com/media/resourcecenter/item/l/u/luminol_ti02-48eng-rev10e.pdf. (accessed date: Sep 23, 2023).
- Stoica BA, Bunescu S, Neamtu A, Bulgaru‐Iliescu D, Foia L, Botnariu EG. 2016. Improving luminol blood detection in forensics. J Foren Sci, 61(5): 1331-1336.
- Stott RAW, Kricka LJ. 1987. Biolumin chemilumin symp. John Wiley & Sons Ltd., Chichester, New York, USA, pp: 237–240.
- Tajani, A. 2014. Photonics for forensic applications. Photon Safet Secur, 1987: 368-397.
- Tobe SS, Watson N, Daéid NN. 2007. Evaluation of six presumptive tests for blood, their specificity, sensitivity, and effect on high molecular-weight DNA. J Forensic Sci, 52(1): 102–109. https://doi.org/10.1111/j.1556-4029.2006.00324.x.
- 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. Foren Sci Inter, 188(1-3): 1-17.
- Weber K. 1966. Die Anwendung der chemiluminescenz des luminols in der gerichtlichen medizin und toxikologie: i. der nachweis von blutspuren. Deutsche Zeitschrift Medizin, 57: 410-423.
- Wells PG, Winn LÇM. 1996. Biochemical toxicology of chemical teratogenesis. Critical Review in Biochem. Mol. Biol. 31 (1): 1-40.
- Yeh HC, Lin WY. 2003. Enhancement of Chemiluminescence from the Oxidation of Luminol with Hydrogen Peroxide Catalyzed by Mn (III)‐Microperoxidase 8. J Chinese Chem Soc, 50(1): 81-88.