Abstracts From Fall 1996
1. TITLE: Evaluation of a Method That Isolates and Quantitates Ecgonine and Other Cocaine Metabolites in Whole Blood
AUTHORS: Eugene W. Schwilke*, Barry R. Logan, Kari E. Blaho and Timothy D. Mandrell, Washington State Toxicology Laboratory, Seattle, WA. 98134
ABSTRACT: This study was designed to evaluate a method of analysis for cocaine, benzoylecgonine, ecgoninemethylester, ecgonine, norcocaine, and cocaethylene in whole blood. This method is of particular interest for its ability to isolate ecognine. Blood proteins were precipitated with acetonitrile. This was followed by propylation of carboxyl and tertiary amine groups, p-fluorobenzoylation of alcohols, and butyl chloride extraction at alkaline pH. Samples and standards spiked with cocaine metabolites were precipitated, derivatized, extracted and analyzed by GC/MS with selected ion monitoring. Calibration curves were linear over the range 0.10 ug/ml to 1.00 ug/ml. Deuterated internal standards were used for quantitation where available. The limit of detectability was 0.010 ug/ml with CV’s from 3-10%. The method was used for testing samples from cocaine overdose patients and samples from juvenile swine from a chronic cocaine study. Ecgonine was found in nearly all patient samples (range 0.061-2.943 ug/ml) and in some pig samples (range 0.043-1.67 ug/mL) where cocaine was also present, indicating that the above described method adequately identifies ecgonine as a significant metabolite of cocaine.
2. TITLE: Ion-trap MS/MS Determination of LSD in Urine
AUTHOR: John D. Laycock*, Wei Huang, Steven T. Fannin, and Rodger L. Foltz, Center for Human Toxicology, Salt Lake City, UT.
ABSTRACT: Lysergic acid diethylamine (LSD) has effective doses in the low microgram range, making it one of the most potent drugs known. LSD gain notoriety in the 1960’s due to its potent hallucinatory effects and its widespread use. There is evidence that LSD continues to be widely abused albeit at considerably lower doses (20-80ug) than were previously used. Detection of LSD use is particularly challenging for several reasons. Extensive and rapid metabolism of LSD mean that only about 1% of the parent drug are excreted in human urine. LSD concentrations in urine drop below detectable concentrations in about 24 hours. Major metabolic products have not yet been identified so methods of detection have focused on the parent drug. Also, LSD can exhibit considerable absorptive losses during chromatography. In order to be effective, methods must be able to selectively detect and quantitate LSD in the low pg/mL concentration range.
3. TITLE: Electron Impact Versus Positive Chemical Ionization : GC/MS for the Forensic Laboratory
AUTHOR: Allan Spanbauer, Center for Human Toxicology, Salt Lake City, UT.
ABSTRACT: Electron impact (EI) is the most common method of ionization in use today. Electrons are obtained from a heated filament under vacuum. Ions are accelerated through a given voltage and detected across an ion chamber. The electrons have energy and an electronic charge. Chemical ionization (CI) is the process by which characteristic ionization of the sample is produced by ionic reactions. The technique requires high reagent gas pressure in the source. EI of the reagent gas produces ions that readily ionize the sample. There are several advantages of CI: molecular weight ions obtained, increased sensitivity, less fragmentation, stereo-isomer sensitivity, decreased time for analysis, and spectra complementary to EI spectra.
1.TITLE: Further Validation of Widmark Calculations Based On Breath Alcohol Concentrations
AUTHORS: Patrick Friel*, Daniel O’Malley, Barry R. Logan, and John S. Baer, Washington State Toxicology Laboratory, Seattle, WA. 98134
ABSTRACT: Objective: To assess the accuracy of estimates of alcohol doses calculated by Widmark’s formula.
Methods: 20 subjects (regular drinkers, age 21-30, 10 females, 10 males) were given bolus doses of ethanol on two occasions: “low dose” (females 0.4875 g/kg, males 0.5325 g/kg; target peak BrAC 0.07 g/210ml) and “high dose” (females 0.8125 g/kg, males 0.8875 g/kg; target peak BrAc 0.10 g/210L). BrAC’s were measured in duplicate using a BAC Verifier. BrAC’s two hours post dosing were used to estimate vodka dose, using Widmark’s formula (female r=0.55, male r=0.68, B=0.015 g/210L/hr).
Results: Calculated vodka doses underestimated actual vodka doses by a mean of 12 ml of 100 proof vodka (p<0.001). For the 40 studies evaluated, Widmark’s formula overestimated dose in 3 cases (maximum overestimate 13 ml), correctly estimated dose (within +/- 5 ml) in 12 cases, and underestimated dose in 25 cases (maximum underestimate 48 ml). Calculated and actual doses were highly correlated. Conclusion: Under the conditions of this study, Widmark calculations using BrAC’s collected two hours post-drinking gave reliable minimum estimates of the amount of ethanol consumed. (Supported by a grant from the University of Washington Alcohol and Drug Abuse Institute).
1. TITLE: Shooting Scene Reconstruction Using Traditional Forensic Methods and Emerging Technologies
AUTHOR: Scott D. Kashuba, B.Sc., M.A., M.B.A., Firearms Section, RCMP Forensic Laboratory, Edmonton, Alberta
ABSTRACT: Traditional forensic firearms methods, including single and multiple gunshot residue patterns, bullet paths, ejection patterns, terminal impact ballistics, and mechanical assessments were combined with the emerging technology of computer animation simulation to determine if a death was an attempted murder/suicide or a premeditated murder. Often in a shooting situation involving only two people, firearms evidence can be the only way of determining the validity of the participants accounts of the events. This is especially true if one of the participants is killed, and cannot give their side of the story. In the case to be presented, the sequence of the two shots fired would confirm or dispute the statement of the survivor of the shooting incident. Forensic examinations of the type mentioned above determined that the version of events described by the surviving participant in the incident could not have occurred. This conclusion was supported by the computer animation simulation that used information from the shooting scene to predict the final location of the fired bullets to within 1/4″ of where they were actually found.
1. Demonstrate the need for a firearms examiner to be aware of, and use the complete crime scene information in reconstructing the events of a shooting.
2. Illustrate how a firearms examiner can utilize the new high technology of computer animation simulation both to confirm the findings achieved through traditional forensic methods, as well as a visual aid in presenting these findings to the court.
3. Reinforce the value of the firearms discipline in the investigation of a criminal offense.
2. TITLE: Integrated Ballistic Imaging System (IBIS) Performance Studies; Test Fired Bullets from Consecutively Rifled Barrels and Fired Cartridge Casing Correlations
AUTHOR: Robert Thompson, Bureau of Alcohol, Tobacco and Firearms, San Francisco, CA
ABSTRACT: The ATF San Francisco Laboratory Center has conducted a performance study of the Integrated Ballistic Identification System (IBIS) hardware and software used in correlating breech face and firing pin impressions on expended cartridge casings. This component, called “BRASSCATCHER” compared pairs of casings from over 200 pistols representing 9mm, .380, .25 and .45 calibers. There was no microscopic pre-screening of the casing pairs prior to selection for testing. Using an updated version of correlation software, the correct “twin” casing was found in the first position of ranked scores between 73 to 87 per cent of the time, and 90 to 93 per cent of the time in the top five positions. The correct “twin” casing was found in the first position 100 per cent of the time for the Glock 9mm caliber pistol database. Visual subjective estimations of cartridge casing “matchability” show that images judged “good” and “fair” in quality constituted the bulk of “first position” matches selected by the computer. However, a sizable number of “poor” quality images of matching casings were still found in the top scoring position. The “BULLETPROOF” component of IBIS was used in comparing ten pairs of expended 9 mm Luger caliber copper jacketed bullets fired from ten consecutively rifled Ruger pistol barrels. The acquired images of the bullet land impressions were compared to a database comprised of 325 additional bullets. The database compared in this testing was not restricted to caliber, number of land bullet impressions, land impression width, or direction of rifling twist. The correct “twin” bullet was selected in the first position ranked score 17 times in the 20 correlations performed. A technical overview of the IBIS image acquisition hardware, image storage, case data input, “ballistic signature” analysis, and correlation scoring to an image database will be presented. A description of upcoming upgrades in computer hardware, database management software, and rimfire firing pin and ejector impression correlations will be discussed.
1. TITLE: Screening Controlled Substances Using Near-Infrared Fourier Transform Raman Spectroscopy
AUTHOR: Roel Ferwerda, Ph.D., Raman Applications Manager, Nicolet Instruments, Madison, WI.
ABSTRACT: The correct identification and analysis of controlled substances is essential to law enforcement departments worldwide. Typical on-site screening methods rely on wet-chemical field test kits. The most common alternative to field test methods is full laboratory analysis using a gas chromatograph coupled with a mass spectrometer. Fourier Transform Infrared (micro) spectroscopy has also been extensively used to analyze drug samples. Although it is a relatively inexpensive technique, FT-IR has some problems associated with sample preparation. Since the development of FT-Raman spectroscopy, the Raman technique has resurfaced. The use of Near-infrared lasers overcomes the fluorescence problems, which are so common in conventional Raman and in many cases mask the Raman spectrum. This paper will describe some of the advantages of this relatively new technique for Forensic research. The Raman method allows materials to be quickly analyzed, without removing them from the sealed evidence bags in which they are collected. The sample can be easily identified by comparing the spectrum with a database of known samples. The identification is largely insensitive to user, sample size or container. Furthermore, the technique is non-destructive. This is essential in a screening technique, since the sample can be saved for later use in court or when a full laboratory analysis is required.
1.TITLE: Household Products as Possible Contaminants at Crime Scenes: An Evaluation Using Two Light Sources and Luminol
AUTHOR: Tilton Davis* and Sandy Mays, Wyoming State Crime Laboratory, Cheyenne, WY. 82002
ABSTRACT: Laser and alternate light sources as well as luminol have been used to evaluate crime scenes and evidence for the presence of biological substances. This paper presents preliminary information about 100 common household products which are possible contaminants at crime scenes. These products were tested under two different conditions: (1) right-out-of-the-bottle and (2) smeared onto white butcher paper. Their reactivity to a laser light (450-540 nm), an alternate light source (450 nm), and luminol are discussed. Most substances did not react with the laser, alternate light source, and luminol. These data provide information about possible contamination reactions when using these tools for evaluating potential evidence. A list of the substances examined will be provided as well as general comments concerning the use of light sources and luminol at crime scenes processed by the Wyoming State Crime Laboratory.
2. TITLE: Automated FTIR Microscopy in Forensic Science
AUTHOR: Steven P. Bouffard, Ph.D., Product Specialist, Perkin-Elmer Corporation, San Jose, CA. 95134
ABSTRACT: The advantages of automated FTIR Microscopy will be discussed as it applies to a forensic scientist. The ability to examine trace amounts of evidence is crucial to the forensic community. The FTIR microscope has become one of the essential components to a trace evidence laboratory. Automated FTIR microscopy provides the means for the most efficient and thorough analysis of trace evidence possible. Automated FTIR microscopy provides automatic focus in the infrared and visible light regions, automatic sample illumination, as well as aperturing of samples. The ability to collect FTIR data from single fibers, paint chips, inks on paper, and trace amounts of drug crystals will be shown.
1.TITLE: Trapping Accelerant Vapors at the Fire Scene Simplifies Laboratory Processing and Increases Efficiency
AUTHOR: William K. Johnston, Ph.D., Portable Arson Samplers, Tooele, UT.
ABSTRACT: The use of a Portable Arson Sampler is described for isolating and trapping flammable fluids at a fire scene. The sampler uses a heated, Teflon line vacuum chamber and commercially available, low cost glass charcoal adsorbent tubes. Debris is placed in the chamber in a card board tub or a paper bag. A vacuum pump controlled by a timer draws vapors from the chamber through an activated charcoal trap available from SKC or Supelco. Sampling times can be controlled by setting the timer. The chamber temperature is controlled with a solid state PID temperature control. After sampling, the trap is removed from the chamber and sealed in a glass vial for shipment to a laboratory. The Portable Arson Sampler can also be used for vapor recovery from surfaces. In this mode the stainless steel lid is removed and the chamber is inverted directly over the area being sampled. The sampler is packaged in a tool box sized, high impact, plastic case and includes vials and adsorbent tubes for fifteen samples. The laboratory analysis of samples is greatly simplified. A glass wool plug is removed from one end of the charcoal trap. The adsorbent material is emptied directly into an auto sampler vial for analysis by high resolution capillary gas chromatography. Using standard 8 X 32mm vials, 1 mL of methanol (for die-hards who will die young, carbon disulfide) is added to each sample. Glass inserts are also available reducing the amount of solvent to as little as 100uL. The greatest level of efficiency is achieved by using an auto sampler and by analyzing samples over night. The auto sampler vial is retained as evidence pending adjudication. In the event that the solvent evaporates, accelerant is readsorbed on the charcoal in the vial further preserving the evidence. Even greater sensitivity can be achieved by using thermal desorption tubes rather than the standard charcoal trap. A thermal desorption system is require to desorb all of the trapped accelerant and introduce the sample on to a capillary column. While the cost of this instrumentation is high, detection limits are increased by a factor of 10-1000. Auto samplers are also available from either Dynatherm or Perkin-Elmer.
No wildlife papers at this meeting
1. TITLE: Validation of a chemiluminescent detection system for RFLP-based DNA typing in the State of Utah Crime Laboratory
AUTHORS: Jay W. Henry* and Todd Van Buren, Utah Department of Public Safety Crime Laboratory, Salt Lake City, UT. 84119
ABSTRACT: The objective of this research was to validate a new method of detection for RFLP-based DNA typing. The current method of detection requires the use of a radioisotope (32P) and is time-consuming (3-5 days per locus). Thus, it would be desirable to eliminate the 32P and its associated safety concerns and decrease the exposure time for each probing. Chemiluminescent detection, based on an alkaline phosphatase labeled oligonucleotide probe combined with Lumi-Phos Plus detection reagent, accomplishes these goals. DNA samples were typed for six VNTR loci using both the 32P and the chemiluminescent detection systems. These samples were sized and evaluated for consistency. Both detection methods were comparable. Sensitivity studies demonstrated that the chemiluminescent system could detect high molecular weight DNA in a range of 3ng-30ng depending on the specific probe used. DNA sample data, technical artifacts, minor procedural refinements and the development of DNA probe quality control guidelines will also be addressed.
2. TITLE: Validation of PCR-based markers PM-DQalpha in the State of Utah Crime Laboratory
AUTHORS: Pilar Shortsleeve* and Lance Newlin, Utah Department of Public of Safety Crime Laboratory, Salt Lake City, UT. 84119
ABSTRACT: The objective of these studies was to determine the reliability and reproducibility of results using the Perkin-Elmer PM + DQA1 Amplitype kit at the Utah State Crime Laboratory. These markers have been evaluated to determine their reproducibility, analytical sensitivity, detectability of mixtures and reaction with exogenous materials. Extraction studies comparing Chelex extraction to organic extraction methods showed a slightly greater yield with the organic method; however, the organic procedure was found to be more time consuming and slightly more cumbersome when compared to the Chelex procedure. Both procedures gave the expected genotype of the donor when applied to a variety of body fluids and tissues. All five polymarker and DQA1 genotypings could be determined when body fluids of different individuals were mixed. Sensitivity studies supported the claims of the manufacturer. Reproducibility studies confirmed that identical profiles were obtained from the same individual with multiple typings and different tissues. A study in which blood was deposited on specific household materials (chosen because they appeared to contain dyes that might cause inhibition of amplification) indicated that inhibition of amplification occurred in some substrates that contained a black indigo dye. All amplified samples yielded the expected genotype of the blood donor
3. TITLE: Insight Into Your PCR-based DNA Laboratory
AUTHORS: Theresa Spear*, Sharyl Barney and Alison Fitzpatrick, California Department of Justice, CCI, Sacramento, CA. 95820
ABSTRACT: This study examined the conditions required for a “contaminating” type to appear in polymerase chain reaction (PCR) typing tests. In addition, the efficacy of various methods to either remove or modify DNA so it could no longer act as viable templates was investigated. Biological samples (unextracted DNA) were added to either DQ Alpha (DQA) or Polymarker (PM) PCR cocktails to determine if they could be successfully amplified and typed. In addition, biological samples, extracted DNA and amplified DNA were tested with a series of “treatments” [UV light/”Stratalinker”, bleach, ethanol, “DNA Away” and autoclaved] to determine if these samples could still act as viable templates. A 20% solution of bleach was found to be effective in inactivating PCR product, extracted and unextracted DNA as templates for PCR. UV light (overnight), autoclaving solutions, and “DNA Away” were found to be effective on biological samples and unextracted DNA.
4. TITLE: DNA Variation and Human Evolution
AUTHOR: Lynn Jorde*, M.J. Bamshad, W.S. Watkins, A.E. Fraley, P. Krakowiak, and S. Sung, Department of Human Genetics, University of Utah Health Sciences Center, Salt Lake City, UT. 84112
ABSTRACT: We present results from an ongoing analysis of nuclear and mitochondrial genetic variation in human populations (75 Africans, 78 Asians, and 90 Europeans). In a recent study (Jorde et al., 1995, Am. J. Hum. Genet. 57: 523-538), we reported results based on 30 nuclear restriction site polymorphisms (RSPs), 30 tetranucleotide short tandem repeats (STRs), 5 trinucleotide STRs, and 200 bp of mtDNA sequence from IIVS2 in the mitochondrial D loop. We now add 430 bp of mtDNA sequence from HVS1 and 27 additional nuclear tetranucleotide STRs. A genetic distance matrix based on the 27 new STRs is highly correlated with a distance matrix based on the original 30 STRs (Mantel matrix correlation = 0.84, p<0.0001 for thirteen subpopulations). The heterozygosity estimates are similar to those obtained in the previous study (0.73 +/- 0.03, 0.68 +/- 0.03, and 0.71 +/- 0.03 for Africans, Asians, and Europeans, respectively). The GST estimate is also similar in the two data sets: 0.046 for the new STRs 0.034 for the original set. As with the original 30 STRs, the new set of STRs yields a neighbor-joining tree in which subpopulations form three well-defined clusters corresponding to the major continental populations (African, Asian, and European). When all 57 tetranucleotide STRs were used to form a genetic distance matrix, the Mantel matrix correlations between this matrix and distance matrices for other sets of polymorphisms (RSPs, trinucleotides, and mtDNA) increased slightly. These comparisons indicate that the STR results are highly reproducible and that both data sets provide an accurate depiction of genetic affinities in human populations. The additional HVS1 sequence yields results similar to those obtained with HVS2 alone. As before, the mtDNA tree demonstrates much more African genetic diversity than do the trees based on nuclear polymorphisms. The Mantel matrix correlations between mtDNA and nuclear data increase with the addition of the HVS1 sequence, but they remain lower than the correlations among the various sets of nuclear polymorphisms. Application of Tajima’s neutrality test shows a statistically significant departure from selective neutrality for the combined HVS1 and HVS2 sequence. Thus, the differences observed between mtDNA and nuclear trees could represent the effects of natural selection on the mitochondrial genome. Supported by NSF (DBS-9310105) and NIH (RR-00064).
5. TITLE: Recovery and Analysis of Ancient DNA
AUTHOR: Scott R. Woodward, Ph.D., Department of Microbiology, Brigham Young University, Provo, UT. 84602
ABSTRACT: Over the last decade it has been possible to isolate, amplify and sequence DNA from a wide variety of ancient sources. Despite the apparent methological problems, aDNA has been recovered from organisms hundreds, thousands and even million of years old. However the consistency with which aDNA is recovered has been highly variable. This seems to be primarily due to the extreme fragmentary nature of the aDNA and the presence of PCR inhibitors that frequently co-purify with the aDNA in many extraction procedures. The difficulty involved in isolating and amplifying aDNA has therefore resulted in many studies that have focused on only one particular locus from a single individual or animal. Even though these reports have proven to be valuable in addressing many phylogenetic issues, and individual cases of identification, the need to perform more informative population-sized studies of ancient groups and in individual cases of extreme importance such as often found in forensic cases, it is necessary to consistently recover reproducible DNA sequences of multiple loci from a large number of individuals. Our laboratory has been developing three approaches to mitigate these problems. 1) The comparative use of resins, chaotropic agents, columns and silica beads have been investigated for the initial isolation of the DNA and separation from inhibitory substances. 2) Reconstruction of the fragmentary pieces of DNA by a pre-PCR repair procedure. 3) Random amplification of any existing DNA sequence by totally degenerate primers followed by locus specific PCR. Using these techniques we have substantially increased our aDNA recovery from Egyptian mummies, ancient parchment, and extinct animals. We have also been able to recover information on nuclear loci that compliments the mitochondrial loci used in most ancient DNA studies.
6. TITLE: Program Updates: CODIS, LABNET and DRUGFIRE
AUTHOR: Jay W. Miller, FBI Laboratory, Washington, D.C.
ABSTRACT: The following topics will be covered: DNA Grants, NIJ Grants: Forensic DNA Improvement Program
-FY 1996 $8,750,000 awarded
-FY 1997 $3,000,000 awarded (Applications now available; closes 11/22/96). Byrne Grants: Forensic DNA testing specifically authorized in DNA Identification Act of 1994.
System Grants: New formula grant program authorized in Anti-Terrorism Act (April 1996) for equipment needed to participate in the FBI’s IAFIS Project, NCIC 2000, and CODIS. Funds distributed only to states having DNA database laws.1 At least $9.5 million will be appropriated in FY 1997 (ie., minimum of $226,000 for 42 states). Split among authorized purposes is at each state’s discretion.
Other topics: LABNET: Status Report, STR Standardization Project: Update, Differential Extraction Robot: New Initiative
National DNA Databank/CODIS Conference — Sponsored by AFIP and FBI
(Tallahassee, Florida – December 9-12, 1996 – hosted by FDLE)
National DNA Index System: Status Report
DNA Laboratory Survey: Preliminary Results
CODIS Program Plans and Priorities
1 Eight states have not passed DNA database laws making them ineligible to receive State Identification System grants: Idaho, Louisiana, Massachusetts, Nebraska, New Mexico, Rhode Island, Vermont and Wyoming.
1. TITLE: Effects of Interviewer Questions on Children’s Statements of Sexual Abuse
AUTHOR: Ron Craig*, Rick Sheibe, John Kircher, David C. Raskin and David Dodd, University of Utah, Salt Lake City, UT.
ABSTRACT: Effects of forensic interview techniques on the production of free narrative and Criteria-Based Content Analysis (CBCA) criteria were assessed in police interviews with 48 children who alleged they had been sexually abused. Allegations of abuse were later confirmed as valid (n = 35) or invalid (n = 13) based on information developed subsequent to the interview. Two raters independently coded interviewer questions and children’s responses, and a separate set of four raters evaluated the transcripts for the presence of CBCA criteria. As predicted, open questions yielded more free narrative and CBCA criteria than did any other type of question. Valid statements of abuse contained more CBCA criteria than invalid statements and statements by older children contained more CBCA criteria than those made by younger children. The results support the use of open questions for eliciting free narrative and the use of CBCA to assess the validity of children’s accusations of sexual abuse.
2. TITLE: Strategies for Grant Seekers
AUTHOR: Laura Lewis, Program Grant Manager, State of Utah Commission on Criminal and Juvenile Justice, Salt Lake City, UT.
ABSTRACT: There has probably never been a better time for criminal justice/law enforcement agencies to request grant funds. This presentation will focus on providing the participants with tools to make them more comfortable when writing grant applications. The Grantsmanship Center in Los Angeles provides a three-to-five day grant writing course somewhere along the Wasatch Front every six months. This is excellent training for those individuals who have not written a grant application. The presenter also recommends that each agency collect samples of well-written grant applications and grant-writing/grant management materials for guidance when writing grant applications. Resource materials may be ordered from the Grantsmanship Center (1-800-421-9512 or (213) 482-9860). Several grant terms are defined, including “grant”;”RFP”; and “NOI”. The presenter also will discuss cash match vs. In-kind match and make suggestions on how to increase an applicant’s chance in obtaining funds. One of the most important recommendations is to follow the grant guidance meticulously. Another is to proof the narrative for typos and the budget for math errors. The presenter will close the presentation by distributing grant program contacts in each state and guidance on how to request funds from private foundations as well as through fundraising.