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The Effect of Luminol on the Serological Analysis of Dried Human Blood~Stains

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The Effect of Luminol on the Serological Analysis of Dried Human Blood~Stains If you have issues viewing or accessing this file contact us at NCJRS.gov. January 1990 U.S. Department of Justice Volume 17, No.1 Federal Bureau of Investigation Crime Laboratory Digest Review Article In This Issue Use of HPLC with Diode Array Spectrophotometric Detection in Forensic Drug Analysis $.:&'...., 1 ....... 1 T. w •• C'rr.' 1"""o:J 9.98 mjn Swjvel 45 deg 482 nm Tjlt 35 deg EDITOR ASSOCIATE EDITOR Barry L. Brown David T. Stafford Quantico, Virginia Memphis, Tennessee ASSISTANT EDITOR Denise K. Bennett Quantico, Virginia EDITORIAL BOARD William Anderson Dean Fetterolf Robert Sibert Reno, Nevada Quantico, Virginia Washington, D.C. F. Samuel Baechtel Barry Gaudette Robert Spalding Quantico, Virginia Ottawa, Canada Washington, D.C. Robert Cravey Robert Koons Irving Stone Santa Ana, California Quantico, Virginia Dallas, Texas Harold Deadman Dale Moreau Phillip Wineman Quantico, Virginia Quantico, Virginia Rockville, Maryland Peter DeForest Thomas Munson New York, New York St. Cloud, Minnesota PUBLICATIONS COMMITTEE AMERICAN SOCIETY OF CRIME LABORATORY DIRECTORS David T. Stafford*, Chairman Gerald Smith Rondal Bridgemon Memphis, Tennessee Knoxville, Tennessee Tucson, Arizona Arnold Melnikoff Kenneth Jonmaire John Murdock* Missoula, Montana Lockport, New York Martinez, California *These individuals also serve on the editorial board. PUBLICATION POLICY The Crime Laboratory Digest is published quarterly by the FBI Laboratof'j in cooperation with the American Society of Crime Laboratory Directors (ASCLD). It is intended to serve as a rapid means of communication between crime laboratories, permitting information of in­ terest and value to be disseminated among crime laboratory scientists. Inclusion of an article in the Crime Laboratory Digest in no way represents an endorsement or recommendation of any part of that article by the Federal Government, the Department of Justice or the FBI. Contributing authors assume total responsibility for the contents and accuracy of their submission. Que3tions or requests concerning an article should be directed to the contributing agency. All submissions are subject to editorial review in accordance with the editorial policy established by the FBI Laboratory and ASCLD. The editorial staff of the Crime Laboratory Digest reserves the right to edit all articles for style, grammer and punctuation. Comments and let­ ters to the editor are encouraged and will be published when appropriate and as space permits. These should be forwarded to: Crime Laboratory Digest - Editor FSRTC, FBI Academy Quantico, Virginia 22135 Contents Message from the Assistant Director in Charge 1 of the FBI Laboratory Editor's Column 2 Review Article: The Use of HPLC with Diode Array 5 . Spectrophotometric Detection in Forensic Drug Analysis Barry K. Logan, H. Steve Nichols, G. Scott Fernandez and David T ... Stafford discuss the benefits of combining diode array ultraviolet visible spectrophotometric detection with HPLC for increased efficiency in drug analysis/identification. Feature Article: The Effect of Luminol on the Serological 13 Analysis of Dried Bloodstains 131354 Robert R. J. Grispino examines the effects of luminol applications on the complete serological analysis of dried bloodstains and addresses the advantages and disadvantages of using the luminol test at crime scenes. DNA Technology Seminar Videotape Order Form 24 Meeting Announcements 25 Instructions for Submitting Articles 28 Crime Laboratory Digest Subscriber Form 29 131353- U.s. Department of Justice 131354 Natlonallnstitute of Justice This document has been reproduced exactly as received from the person or organization originating it. Points of view or opinions stated in this document are those of the authors and do not necessarily represent the official position or poliCies of the National Institute of Justice. Permission to reproduce this Q ~ material has been granted by Public IXJI!1ain/El3I .IL-8. Deparb'Jent of !TU8±ic...,e~ __ to the National Criminal Justice Reference Service (NCJRS). Further reproduction outside of the NCJRS system requires permis­ sion of the ~ owner. CRIME LABORATORY DIGEST Vol 17, No.1, January 1990 ISSN NO. 0743-1872 Mont. 24 11 Minn. VT 33 (488) (3,131) I (362) NH 76 (3,294) ~ OI'~ell:l. 20 S. Oak. (334) MA 102 (5,047) Wyo. CT 54 (2,7 45) 12 H13(~80) Ri 105 (12,218) (212) Neb. NJ 170 (9,331 ) 15 DE 139 (15,664) (466) MD 507 (24,642) Col. OC 521 (28,835) 31 Kan. (2,442) , I. 35__ .,..,,. Ariz. I N. Mex. 31 16 (3,408) (1,008) Texas 212 Foreign Governments (15,803) 140~·i\ (1.978) . ~ ~;R Australia 2 (297) Bahamas 6 (87) British West Indies 4 (903) .. ~.- ,Q Canada 4 (420) '(.\ \j '-} Cayman Islands 1 (95) Hawai~ Jamaica 1 (12) 101 (13,269) {> FBIIDOJ IFeature Miele I The Effect of Luminol on the Serological Analysis of Dried Human Blood~stains Robert R. J. Grispino Serology Unit FBI Laboratory Washington, D. C. 20535 To a great degree today, crime laboratories In 1936, Gleu and Pfannstiel confirmed are being provided with crime scene evidence Albrecht's observations regarding luminol and which has been treated with luminol reagent blood. They noted that luminol would not mixtures by law enforcement personnel at the chemiluminesce in the presence of boiled veg­ scene. Laboratory examiners are then requested etable peroxidases and observed that luminol to perform a complete serological analysis of mixed with pure hematin produced the most the questioned samples. However, uninformed brilliant blue luminescence. crime scene personnel sometimes opt to use the The first proposed forensic use of luminol as luminol test as a preferred field blood test at a preliminary blood test was reported by Specht all crime scenes. Since the reagents are com­ in 1937. He sprayed blood on bushes, stone mercially available in packaged crime scene kits, walls, rusty iron fences, furniture, stone steps the luminol test is susceptible to abuse and mis­ and a garden. After allowing the blood to use by untrained officers. This article discusses remain exposed to the elements for 14 days, the effects of luminol sprays on the complete Specht sprayed a luminol reagent mixture onto serological analysis of neat dried bloodstains. In the blood and photographed the results. 'fhe addition, the advantages and disadvantages of mixture was 0.1% luminal in 5% aqueous sodium the luminol test are addressed, and suggestions carbonate (Na2C03) with a 15% solution of HP2 are offered concerning the proper application added immediately before spraying. All blood­ and protocol of the luminol test at crime scenes. stained areas glowed with blue light for 10 to In summary, the results of this study demon­ 15 minutes. Blood was also detected in water, strate that the use of luminol denatures most soapy water and sewage. The luminol test blood enzymes after a short exposure, thus worked well with both fresh and old blood­ causing further serological comparisons to be stains; in fact, the older the bloodstain, the questionable. more pronounced the positive reaction. Proescher and Moody confirmed Specht's findings in 1939 using Specht's spray mixtures. REVIEW They detected bloodstains on paper, fabrics and iron pipes exposed to the elements for 3 years, with 3-year-old putrefied blood exhibiting bril­ The compound 3 aminophthalhydrazide liant luminescence. In addition, they observed (5- amino 2, 3- dihydrophthalazine 1, 4- dione) that dried and decomposed blood elicited a was first synthesized by Schmitz in 1902. At the stronger and longer lasting luminol reaction than time of synthesis, he noticed that the compound fresh blood. When the luminescence disap­ exhibited a strong blue fluorescence in aciti so­ peared, it could be reproduced by application lutions. In 1927, Lommel first observed a blue of fresh luminol spray. Dried bloodstains were chemiluminescence after oxidation of the com­ made luminescent many times. Fresh dried pound in alkaline solution. His work was never bloodstains were made more luminescent by published, but Albrecht, one of his coworkers, spraying the blood with 1 to 2% hydrochloric confirmed and published Lommel's original acid solution before luminol application. The findings in 1928. Albrecht also pointed out that luminol reaction was elicited with both animal blood and fresh potato juice caused the chemical and human blood. Hematin was detected in a 8 to exhibit strong luminescence in the presence of dilution of 1:10 • Most importantly, Proescher hydrogen peroxide (HPJ. In 1934, Huntress et and Moody (1939) made the following emphatic al. termed this compound luminol as a producer supportive statement on luminol's application to of light. forensic serology: 13 CRIME LABORATORY DIGEST Vol 17, No.1, January 1990 "Luminol does not interfere with the The c:hemical basis of the luminol test is an spectroscopic, chemical or precipitation oxidatjon reaction. An alkaline solution of lu­ tests for the definite identification of minol will oxidize in the presence of HP2 and blood. Hematin and hemochromogen a hematin-catalyzed peroxidase system. Hydro­ crystals of dried blood were obtained gen peroxide is the oxidizing agent. Blood is a after rep~atedly treating blood with hematin-catalyzed peroxidase system. The re­ luminol. The precipitin test can be sulting oxidation reaction is visualized by a blue applied if the blood is not decomposed." chemiluminescence. In 1985, Thornton and Maloney reviewed the Lytle and Hedgecock (1978) experimented chemistry of the luminol reaction'. Figure 1 is with the effects of alkaline luminol/sodium an amalgamation presented by Thornton of the perborate (NaB OJ mixtures on blood. They work of a number of researchers on a proposed concluded: oxidation pathway for luminol. Not all of the steps of the oxidation pathway have been ".... luminol is relatively nondestructive elucidated to date. The important points are: to the surroundings (it is noncorrosive and nonstaining) and to the blood (it 1) Oxygen is required. does not prevent subsequent identi- 2) The anion of luminol is the reactant. fication tests or ABO blood grouping 3) Free radicals are involved which provide the analysis although it does interfere with 40 to 70 kcal./mol of chemical energy needed electrophoretic analysis of.
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