Midwest Forensics Resource Center - ameslab.gov · biochemistry offers opportunities for developing microscopic analytical devices with unique characteristics. This project develops

Midwest Forensics Resource Center

Research and DevelopmentProgram Summary

October 2008

Acknowledgement

These research projects were funded by the U.S. Department of Justice COPSProgram under contract number 2005 CKWX0466, and by the National Instituteof Justice, through the Midwest Forensics Resource Center at Ames Laboratoryunder interagency agreement number 2008-DN-R-038. The Ames Laboratory isoperated for the US Department of Energy by Iowa State University, undercontract No. DE-AC02-07CH11358.

Midwest Forensics Resource Center (MFRC)Research and Development Program

Introduction

The mission of the MFRC Research and Development Program is to provide technologicaladvances in forensic science for the benefit of our regional partners as well as the forensiccommunity at large. Key areas of forensic science need are identified through our interactionswith our Midwest partners and our R&D advisory group, as well as through our participationin national meetings in forensic science. Under the sponsorship of the National Institute ofJustice and the U.S. Department of Justice COPS Program, the MFRC solicits proposals forthe development of practical and useful technology, instrumentation, and methodology thataddress needs in areas related to forensic science and its application to operational crimelaboratories. The MFRC facilitates proposal development by working to establish partnershipsbetween researchers and our regional partners. The MFRC administers a peer-review ofthe proposals and then funds the selected projects at a cost of approximately $55,000 each,with a 12-month period of performance.

The process for selection of these projects includes the following steps: 1) drafting of a callfor proposals by MFRC staff, 2) review of the draft call by members of the R&D advisorycommittee, 3) review and approval of the call by NIJ, 4) issuance of the call to ISU, AmesLaboratory, regional partners, and research organizations, 5) receipt of proposals, 6) reviewof proposals by R&D advisory committee, 7) ranking and selection by MFRC staff usingadvisory committee reviews, with concurrence by NIJ, 8) notification of proposers, 9) receiptand review of progress reports by MFRC, 10) receipt and review of final reports by MFRC,R&D advisory committee, and NIJ.

The decision to fund any specific project is based upon a peer-reviewed call-for-proposalsystem administered by the MFRC. The reviewers are crime laboratory specialists andscientists who are asked to rate the proposals on four criteria areas including: 1) relevanceto the mission of the MFRC, 2) technical approach and procedures, 3) capabilities, teaming,and leveraging, and 4) dissemination and implementation plan. A successful proposaldemonstrates knowledge of the background for the research and related work in the fieldand includes a research plan with a defined plan to implement the technology to benefit ourpartners at the crime laboratories.

Program Summary Technical Sheets

The following project summaries, while not a complete summary of all research areas, aremeant to demonstrate the range of research funded by the MFRC. The project summariesdescribe the forensic need the projects serve as well as the benefits derived from thetechnology. The summaries provide a brief description of the technology and theaccomplishments to date. In addition, the collaboration with regional partners and the statusof the dissemination of project results and implementation of the product are highlighted.These technical summaries represent the development and implementation of practical anduseful technology for crime laboratories that the MFRC hopes to accomplish.

TABLE of CONTENTSPage

Abstracts of Funded Projects..................................................................................................................3

CHEMISTRY

Analysis of Automotive Clear Coat Paints by Micro-Laser Raman Spectroscopy.........................9

Determination of Heavy Metals in Whole Blood Using Inductively CoupledPlasma - Mass Spectrometry...............................................................................................................11

Application of Chemometric Procedures to Differentiate Ignitable Liquid Residuesfrom Substrate Interferences..............................................................................................................14

Evaluation of a Portable Raman Analyzer for Testing Drugs...........................................................16

Fast Gas Chromatography Capabilities in Arson Debris Analysis..................................................18

Micromechanical Cantilever-Based Sensors for Chemical Species Detection.............................22

Optimization of HeadSpace-Solid Phase Microextraction for OrganicImpurity Profiling of Illicit MDMA Tablets...........................................................................................25

The Temporal Fate of Drugs in Decomposing Tissues....................................................................28

Ultra-Fast Gradient Elution HPLC as a High Throughput, High Information ContentScreening Tool for Drugs of Abuse....................................................................................................31

DIGITAL EVIDENCE

A Steganalyzer Package for Forensic Applications..........................................................................34

PATTERNED EVIDENCE

Application of a Crystal Orientation Method to Forensic Physical Matching of Metal Surfaces along a Fracture Line...........................................................................................................36

High-Speed Digital Video Analysis of Bloodstain Pattern Formation from Common Bloodletting Mechanisms....................................................................................................................41

CD-ROM Based Digital Information Database on Pipe and Tubing Utilized in Improvised Explosive Devices..........................................................................................43

Quantification of the Individual Characteristics of the Human Dentition.......................................46

Spectral Analysis of the 3D Fracture Surfaces for Enhanced Matching.........................................50

A Method for Lifting Bloody Impressions Using a Lifting Strip Containing Titanium Dioxide...............................................................................................................53

Testing for Potential Contextual Bias during the Verification Stage of theACE-V Methodology when Conducting Latent Print Examinations........................................55

BIOLOGY/DNA

Testing DNA Samples for Population of Origin.........................................................................59

Abstracts of Funded Projects

CHEMISTRY

• Analysis of Automotive Clear Coat Paints by Micro-Laser RamanSpectroscopy. Jay Siegel, School of Science, Indiana University � PurdueUniversity, Indianapolis (IUPUI), Indianapolis, Indiana.

This project partially funds the purchase of a micro-laser Raman spectrometer toevaluate the feasibility and discriminating power of Raman spectrometry in thecharacterization and analysis of automotive clear coat paints. This represents onephase of an ongoing project in the analysis of clear coats undertaken by the Forensicand Investigative Sciences Program at IUPUI. The methods used include micro-laserRaman spectroscopy, Fourier transform infrared spectrophotometry and UV-visible-nearinfrared spectrophotometry. The Raman phase of the project is conducted in partnershipwith the Indiana State Police Forensic Science Lab.

• Determination of Heavy Metals in Whole Blood Using Inductively CoupledPlasma � Mass Spectrometry. Joseph Wermeling, Wisconsin State CrimeLaboratory-Madison, Madison, Wisconsin and Charles Cornett, Department ofChemistry and Engineering Physics, University of Wisconsin � Platteville, Platteville,Wisconsin.

The forensic investigation of toxicological cases involving acute intoxication, poisoning,and death is heavily focused on the analysis of organic and pharmaceutical analytes ofinterest. The determination of inorganic analytes, especially heavy metals, is oftenlimited to a few metals. Tools used include relatively insensitive wet chemicaltechniques or single-element methods employing atomic absorption or graphite furnacemethods. This project addresses the need for the development of rapid, sensitive, multi-elemental methods for the analysis of metal toxins in whole blood, thereby expandingthe range of inorganic analysis in toxicological investigations.

• Application of Chemometric Procedures to Differentiate Ignitable LiquidResidues from Substrate Interferences. Ruth Waddell-Smith, School of CriminalJustice, Michigan State University, East Lansing, Michigan.In arson investigations, fire debris is typically extracted and analyzed to determine thepresence of ignitable liquid residues. Yet, identification of these residues can becomplicated by interferences from substrates. This project targets development of anobjective methodology for differentiating ignitable liquids from burned substrates bycreating reference collections. The project also demonstrates the potential of statisticaland chemometric procedures to associate and differentiate classes of liquids and todifferentiate liquids from burned substrate interferences.

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• Evaluation of a Portable Raman Analyzer for Testing Drugs. Susan Gross,Minnesota Department of Public Safety, Bureau of Criminal Apprehension, St. Paul,Minnesota.

Large case backlogs can cause slowdowns and delays in the judicial system. Speedytrial demands by the defendant are difficult to maintain with these backlogs. This projectevaluates a portable drug identification system, using laser-based Raman technology, toease some of the backlog problems. The evaluation of the Raman StreetLab®is performed in the laboratory and in the field. By implementing a plan to analyzecontrolled substances in the field, the laboratory case backlog can be alleviated and theprosecution of controlled substance cases can continue in a timely manner.

• Fast Gas Chromatography Capabilities in Arson Debris Analysis. CharlesCornett, Department of Chemistry and Engineering Physics, University of Wisconsin� Platteville, Platteville, Wisconsin.

This project assesses the potentially large impact that fast gas chromatography (FastGC) may have on the determination of ignitable liquids. Forensic Sciences: Review ofStatus and Needs clearly defines a need for further advances in this field. In addition,current analytical literature appears to support Fast GC as a separation techniquecapable of comparable resolution to conventional GC in less time per sample. Thisresearch compares resolution capabilities of the two techniques in separating a widerange of ignitable liquids in an array of matrices. With sufficient resolution, Fast GC mayhelp clear casework in a more timely manner.

• Micromechanical Cantilever Based Sensors for Chemical Species Detection.Pranav Shrotriya, Department of Mechanical Engineering, Iowa State University andMarit Nilsen-Hamilton, Department of Biochemistry, Biophysics and MolecularBiology, Iowa State University, Ames, Iowa.

The merging of silicon microfabrication techniques with surface functionalizationbiochemistry offers opportunities for developing microscopic analytical devices withunique characteristics. This project develops a proof-of-concept for high specificity andsensitivity micromechanical cantilever-based (MC) sensors. Successful completion ofthe research may help the development of portable sensors capable of detectingchemical species at concentrations of parts-per-billion. Such sensitivity will revolutionizeforensic analysis of controlled substances, explosives, biological molecules, and DNArequired for crime-scene investigation.

• Optimization of HeadSpace - Solid Phase Microextraction for Organic ImpurityProfiling of Illicit MDMA Tablets. Ruth Waddell, Department of Chemistry,Michigan State University, East Lansing, Michigan.

Organic impurity profiling of illicit synthetic drug tablets aims to identify similaritiesamong tablets. Similar impurity profiles indicate a common production method, andsimilar levels of the same impurities potentially indicate common production

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laboratories. In this research project, headspace solid phase micro-extraction (HS-SPME) procedures for the extraction of organic impurities from illicit MDMA (�ecstasy�)tablets are developed and compared with conventional liquid-liquid extractionprocedures, in terms of the number and level of impurities extracted. HS-SPME is rapid,requiring no solvent and yielding selective extraction of impurities. Hence, HS-SPME isa promising extraction procedure that offers attractive advantages over conventionalprocedures.

• The Temporal Fate of Drugs in Decomposing Tissues. John Wyman, FranklinCounty Coroner�s Office, Columbus, Ohio.

Interpreting drug concentrations found in decomposed remains is always difficult.Postmortem tissues that are routinely collected and analyzed (blood, urine, vitreoushumor), and therefore provide the largest comparative database for interpretation, arefrequently lost in the early stages of putrefaction. Consequently, when drugs are foundin weathered tissues, there is currently little or no information available to help guide thetoxicologist, and subsequently the pathologist, in evaluating whether a drug(s) playedany significant role in causing the death. This study follows the fate of sixteen drugs,which commonly cause intoxications, in seven different tissues collected fromdecomposing pigs.

• Ultra-Fast Gradient Elution HPLC as a High Throughput, High InformationContent Screening Tool for Drugs of Abuse. Peter Carr, Department ofChemistry, University of Minnesota, Minneapolis, Minnesota.

This project is a continuation of previously funded work titled �Ultra-Fast GradientElution Reverse-Phased HPLC with Diode-Array Detection.� That activity improved boththe speed and selectivity of older HPLC methods through a combination of preciseretention measurements and chemometric analysis of the spectro-chromatograms. Inthe currently funded two projects, several enhancements are examined. First, toimprove selectivity, three HPLC phases are studied to find the pair that is mostorthogonal so as to dramatically reduce the number of indistinguishable analytes.Second, to improve speed, recently commercialized very small stationary phases thatuse the �core-shell� concept are applied. Finally, the sensitivity and specificity of the newmethodology is studied by evaluating real samples whose compositions have been pre-assessed by accepted confirmatory methods, especially LC-MS.

DIGITAL EVIDENCE

• A Steganalyzer Package for Forensic Applications. Jennifer Davidson,Department of Mathematics / Electrical Engineering, Iowa State University, Ames,Iowa.The use of steganographic software for hiding information in image files for illicitpurposes is becoming more widespread. While steganalysis algorithms are abound inthe academic literature, there are few software programs that address the needs oflocalpolice departments who perform computer forensic functions for steganalysis. This

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project extends the proof-of-concept established on the use of an artificial neuralnetwork for wavelet steganalysis in an earlier study. Specific aims are to enhance thecapabilities of the software and to make it available for practical use in computerforensics.

PATTERNED EVIDENCE• Application of Crystal Orientation Method to Forensic Physical Matching of

Metal Surfaces along a Fracture Line. Barbara Lograsso, School of Technology,Michigan Technological University, Houghton, Michigan; Thomas Lograsso, AmesLaboratory, Iowa State University, Ames, Iowa; and James Kreiser, Springfield,Illinois.

A physical match between two surfaces is routinely viewed as definitive proof that thefracture surfaces were generated from the same part. The practice of fracture matchinghas recently been challenged in court. Therefore, there is a need for a method tomeasure the nature and characteristics of materials with a known error rate that can beapplied to performing physical matches. This study examines the feasibility of usingsurface crystal orientation to associate or differentiate metal fracture fragments found atcrime scenes.

• High Speed Digital Video Analysis of Bloodstain Pattern Formation fromCommon Bloodletting Mechanisms. Terry Laber, Minnesota Department of PublicSafety, Bureau of Criminal Apprehension, St. Paul, Minnesota; Bart Epstein, Edina,Minnesota; and Michael Taylor, Institute of Environmental Science and Research,Christchurch, New Zealand.

The analysis of bloodstain patterns is used by criminal investigators to draw inferencesabout the events that led to the formation of the pattern. An understanding of thedynamics of a blood transfer event is therefore critical to the sound interpretation of theresultant bloodstain pattern. This project studies the formation of some commonbloodstain patterns by using a high-speed digital video camera to record the bloodtransfer as it occurs. The outcome of the research will strengthen the science behindbloodstain pattern analysis (BPA).

• CD-ROM Based Digital Information Database on Pipe and Tubing Utilized inImprovised Explosive Devices. Jamie Crippin, Western Forensic LawEnforcement Training Center, Pueblo, Colorado; David Green, Lake County CrimeLaboratory, Painesville, Ohio; and Will Randle, Missouri State Highway PatrolLaboratory, Jefferson City, Missouri.

The rise in terrorist bombings has made the identification of bomb materials a priority forthe forensic scientist. The volume of evidence to be analyzed and the importance ofsuch testing for preventive and investigative purposes have created a need for reliable,fast, and cost-effective tools that can help identify specific tubing material used forbombs. The objective of this project is to collect data on all known types of pipes andtubing and to store the information in a searchable CD-ROM-based database. Such a

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database greatly reduces the amount of time and effort spent by forensic scientists toprocess bomb samples.

• Quantification of the Individual Characteristics of Human Dentition. ThomasJohnson, School of Dentistry, Marquette University, Milwaukee, Wisconsin.

Although DNA can be associated with a human bite, it is not always recovered. In suchcases, an analysis of bitemark evidence is required. Yet, forensic odontologists areunder attack for lacking a scientific basis for their analysis. This project studies theability to scientifically quantify the occurrence of special dental characteristics and errorrates in the analysis of bitemarks. Using six measurements, a 3D quantificationtechnique and 400 samples, a database is created that may provide the criminal justicesystem with the beginning of a tool and the hard science for objective statement ofprobability, in either exculpating or incriminating a suspect from patterned injuries causeby human teeth.

• Spectral Analysis of the 3D Fracture Surfaces for Enhanced Matching. AshrafBastawros, Department of Aerospace Engineering, Iowa State University, Ames,Iowa; Barbara Lograsso, School of Technology, Michigan Technological University,Houghton, Michigan; and Jeremiah Morris, Johnson County, Sheriff�s OfficeCriminalistics Laboratory, Mission, Kansas.

A fractal surface carries sets of unique signatures dictated by the intrinsic materialmicrostructure and the external loading conditions. This project combines the basicunderstanding of fracture mechanics with the practical applications of forensic scienceto develop testing protocols for improved matching of fractured surfaces. A 3D spectralanalysis of the fracture surface is developed to identify these fracture signatures and toshow their uniqueness for each fractured specimen. Fracture of metal couponspecimens are used to generate proof-of-concept and to establish the feasibility of theapproach. The matching ability is then compared against the techniques currently usedin forensic labs.

• A Method for Lifting Bloody Impressions Using a Lifting Strip ContainingTitanium Dioxide. Jessica Zarate, Northville City Police Department, Northville,Michigan.Bloody impressions are of great importance to the forensic community as they arefrequently encountered at crime scenes. Impression evidence in blood cannot alwaysbe removed from the crime scene for analysis in a laboratory setting. Because of this,many potentially identifiable impressions can only be photographed and not enhanced,thereby complicating the latent print identification process. In an earlier study, a liftingstrip containing titanium dioxide was successful in lifting and enhancing bloodyfingerprints from several non-porous and semi-porous surfaces of contrasting colors.This study improves upon the methods used to lift bloody palm prints, foot prints, andfootwear impressions, in addition to bloody fingerprints from porous, semi-porous, andnon-porous surfaces.

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• Testing for Potential Contextual Bias during the Verification Stage of the ACE-V Methodology when Conducting Latent Print Examinations. GlennLangenburg, Minnesota Department of Public Safety, Bureau of CriminalApprehension, St. Paul, Minnesota.

Recent Daubert challenges to fingerprint evidence and the ACE-V methodology havequestioned the reliability of the methodology as it is currently applied by experts. Thisproject studies the potential effect of contextual bias on expert opinion. Experimentswere conducted at an International Association for Identification conference in Boston,MA, and at a local community college where the principal investigator teaches anintroductory course in forensic science. The results of the experiments provide valuableinsight on the effects of potentially-biasing context information, when present, during theverification of latent print examinations.

BIOLOGY/DNA

• Testing DNA Samples for Population of Origin. Raymond Miller, School ofMedicine, Washington University, St. Louis, Missouri.

This activity funds the development and implementation of a forensic test to help fieldinvestigators with unidentified DNA samples and to provide information about thepopulation of origin of the donor of the DNA sample. The test uses available equipmentsto genotype ancestry informative single nucleotide polymorphisms (SNPs). The resultswill be compared with known population frequencies of self-described ethnic groups tonarrow the possible ancestry of the sample.

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Analysis of Automotive Clear Coat Paints by Micro-Laser RamanSpectroscopy

FORENSIC TECHNOLOGY NEED

Little attention to date has been paid to clearcoats. This may be due, in part, to the lack ofinorganics in clear coats, which are almostalways present in color or base coats. Recentadvances in micro-laser Ramaninstrumentation make it easier to obtain highquality spectra of small materials, like the crosssection of a chip of automotive paint.

TECHNOLOGY DESCRIPTION

Micro-laser Raman spectroscopy is a verypowerful sampling and analysis technique thatacquires spectra of extremely small areas non-destructively and without touching the sample.Although widely used in many different fields,the technique has seen little application byforensic scientists.

The main objective of this research is toevaluate the effectiveness of micro-laserRaman spectroscopy in discriminating amongautomobile clear coat paints.

A secondary objective is to evaluate micro-laserRaman spectroscopy as a part of a scheme toanalyze automotive clear coats. Othertechniques that are used in such a scheme arealso evaluated. They include Fourier TransformInfrared (FTIR) spectrophometry and Ultraviolet(UV)-Visible-Near Infrared (NIR)spectrophotometry.

METHODOLOGY

To achieve these objectives, a micro-laserRaman spectrometer will be purchased toevaluate the feasibility and discriminating powerof Raman spectrometry in thecharacterization and analysis of clear coats.

Specific activities include:• evaluation of micro-laser Raman

spectroscopy and optimization ofsampling and analysis oftechniques.

• partial validation of the techniquesand proposed scheme.

• use of the scheme that is developed

ACCOMPLISHMENTS AND ONGOINGWORK

A laser Raman spectrometer was purcha-ed for the analysis of clear coats. The inst-rument was defective and replaced by aCRAIC, model CTR-1, 2.5 MW laser at785 nm.

Figure 1. 2.5 mw, 785 nm microlaser Raman with 20x objective

More than 200 samples of automobile paintchips with make and model informationwere obtained and a method for sectioningthe chips was developed. Paint chips weretested under several thickness conditions

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(whole chip, 15 µ, 10 µ,and 5 µ). It wasfound that 5 µ cross sections generated theclearest Raman spectra.

Spectra of different clear coats werecollected using both 50x and 20x objectivelenses. Data revealed that spectra obtainedfrom 50x objective were no better thanthose from the 20x objective, yet they tooklonger to collect.

The PI is currently working on optimizingthe integration time and number of aver-ages necessary to obtain the best Ramanspectrum for the clear coats. Based onparameters tested, integration has beennarrowed to between 20 and 60 seconds,and number of averages between 5 and 10.

TECHNOLOGY BENEFITS

Clear coats on automobile paints are verycommon today. Yet, they have not beenanalyzed forensically to any degree. Thisproject shows the value of laser Ramanspectroscopy when applied to the analysisof trace evidence. Use of the instrument willimprove the scientific value of paint evi-dence and increase the association value ofpaint evidence to a suspect source.

COLLABORATION

The research is conducted by the Forensicand Investigative Sciences Program atIndiana University, Purdue University,Indianapolis (IUPUI). The Raman phase ofthe project is conducted in partnership withthe Indiana State Police Forensic ScienceLaboratory.

Using IUPUI blind samples and samplesfrom previous casework, forensic scientistsfrom the Indiana State Police (ISP) use theirown FTIR and UV-Visible-NIR instrument aswell as the Raman instrument to evaluatepaint chips. They will also determine the

effectiveness of the scheme and of Micro-laser Raman, in particular, to characterizeand compare known and unknown paintchips.

The ISP will further prepare a number ofproficiency tests to use for further validationof the Raman technique and of the scheme.

DISSEMINATION

The findings of the research will be pre-sented at the 2009 America Academy ofForensic Science meeting in Denver, CO. Apaper will also be prepared for submittal tothe Journal of Forensic Sciences, while afinal technical report on the project and itsfindings will be posted on the MFRC website.

PUBLICATIONS AND PRESENTATIONS

This is a new project with no publications orpresentations to date.

IMPLEMENTATION

ISP will use the scheme (developed form thisresearch) in the analysis of case samples ofpaint as they come into the lab, and as asupplement to their normal analysis proto-cols. They will have use of the Ramaninstrument and microtome for their casework.

CONTACTS

Principal Investigator

Jay SiegelSchool of ScienceIndiana University, Purdue University,Indianapolis402 North Blackford StreetIndianapolis, IN 46202Phone: [email protected]

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Determination of Heavy Metals in Whole Blood UsingInductively Coupled Plasma - Mass Spectrometry


The forensic investigation of toxicologicalcases involving acute intoxication, poisoning,and death investigations is heavily focusedon the analysis of organic and pharmaceuti-cal analytes of interest.

The determination of inorganic analytes,especially heavy metals, is often limited to afew metals, using relatively insensitive wetchemical techniques or single-elementmethods employing atomic absorption orgraphite furnace methods. Because thesemethods are limited and time-consuming,testing for toxic metals in blood samplesoften goes undone.


The primary objective of this project is toemploy the sensitivity, wide dynamic range,and multi-element capabilities of inductivelycoupled plasma mass spectrometry (ICP-MS) to the elemental analysis of samplesrelated to forensic toxicology.

Specific goals of the project are to:

• develop a rapid screening method forheavy metal toxins using the semi-quantitative feature of ICP-MS

• develop and validate a quantitativemethod for arsenic, cadmium, mercury,thallium, and lead in whole blood

• asses the use of microwave-assisteddigestion of whole blood samples inaddition to currently used dilutionprotocols

METHODOLOGY

This project addresses the need for thedevelopment of rapid, sensitive, multielemental methods for analyzing metaltoxins in whole blood.Two differentapproaches are used in establishing themethods.

First, the diluents for several well-established biomonitoring procedures areevaluated for both semi-quantitative andquantitative analyses.

Second, the use of microwave-assistedacid digestion of whole blood samples isevaluated. Such digestion procedures havebeen shown to remove effects fromdissolved solids and the carbonaceouscomponents of biological samples.Optimization of both dilution and digestionis carried out in an interactive fashion.


Two separate methods for the determina-tion of metals in blood were developed. Thefirst method employs microwave digestionof whole blood samples in HNO3 / H2O2,followed by matrix-matched standardadditions analysis in the ICP-MS. Thesecond method involved direct analysis ofwhole blood following dilution with adiluent, also employing matrix-matchedstandard additions analysis.

Both methods were linear (R²=0.999) overthe ranges calibrated. Arsenic was cali-brated from 10-250 µg/L, cadmium andthallium were calibrated from 1-10 µg/L,

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mercury was calibrated from 1-100 µg/L,and lead was calibrated from 10-250 µg/L.The lower limit of calibration for mercurywas changed to 3 µg/L later in the study.

Table 1. Detection limits for five heavy metals utilizing microwave and dilution methods.

The detection limit (DL) for each elementwas calculated blanks as the mean blanksignal plus three times the background. Insome cases (*) the DL exceeded the limit ofquantification (LOQ) as estimated by thedeviation in the low bench standard. Thegeneral trend was for lower DLs in themicrowave method compared to the dilutionmethod. This was somewhat expectedgiven fewer matrix issues with the micro-wave digestion method.

Table 2. The 95% confidence intervals of the microwave and dilution methods.

Analysis of the medium and high benchquality control standards exhibited verygood accuracy for both the dilution and

microwave digestion methods. However, themicrowave method consistently providedstatistically significant (p < .05) better preci-sion (RSD from 2 to 10%) than the dilutionmethod (RSD from 8 to 45%). The bulk ofthe accuracy data for the microwave methodfalls within 15% of the true value or within20% near the LOQ.

TECHNOLOGY BENEFITS

With this procedure, toxicology samples canbe screened for common heavy metals andfor a suite of toxic elements in a singleanalysis. This ability to routinely test biologi-cal samples for a broad spectrum of elemen-tal toxins adds a valuable dimension toforensic toxicological analysis.

Since this project represents collaborationbetween the toxicology and trace evidenceunits at the crime laboratory, this work canexpand ICP-MS capabilities to other disci-plines in the forensic sciences, particularly intrace evidence.

COLLABORATION

This project is a collaborative effort betweenthe Wisconsin State Crime Laboratory-Madison (WSCL-M) and the University ofWisconsin-Platteville. It also representscollaboration between the Trace Evidenceand Toxicology Units within the WSCL-M.

DISSEMINATION

The project and its findings were presentedas a poster session at the Midwestern Asso-ciation of Forensic Scientists. The work wasalso presented at the annual America Acad-emy of Forensic Science meeting as part ofthe Young Forensic Scientist forum. Furtherdissemination plans include presentations atthe Midwestern Association of ForensicScientists Fall Meeting and at the Annual

DL (ug/L) Microwave Method

Dilution Method

As 1.3 5.2 Cd 0.3 1.9* Hg 1.3 4.6* Tl 0.6* 0.4 Pb 2.1 4.4

111Cd Accepted Value (µg/L)

Microwave Method (µg/L)

Dilution Method (µg/L)

Low Bench QC

2.0 ± N/A # 1.77 ± 0.17 (9)

1.6 ± 0.5 (4)

Medium Bench QC

5.0 ± N/A 4.8 ± 0.2 (9) 4.8 ± 0.6 (4)

High Bench QC

8.0 ± N/A 7.7 ± 0.4 (9) 7.9 ± 0.6 (4)

QMEQASO7B

-06 14.0 ± 1.1 *12.8 ± 0.8 (5) 12.8 ± 2.5 (4)

QMEQASO6B-08

2.0 ± 0.3 2.6 ± 0.7 (2) *1.60 ± 0.17 (1)

QMEQASO7B-03

1.31 ± 0.28 *0.9 ± 0.3 (2) *0.90 ± 0.25 (1)

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Pittsburgh Conference and Exposition onAnalytical Chemistry and Applied Spectros-copy (Pittcon). A final technical report on theproject and its findings will also be posted onthe MFRC web site.


• Schweitzer, L., Wermeling, J., Cornett, C.�Determination of Heavy Metals in WholeBlood Using Inductively Coupled PlasmaMass Spectrometry: A Comparison ofMicrowave and Dilution Techniques.�Presentation at the MidwesternAssociation of Forensic Scientists FallMeeting, October 2007, Traverse City, MI.

• Schweitzer, L., Wermeling, J., Cornett, C.�Determination of Heavy Metals in WholeBlood Using Inductively Coupled PlasmaMass Spectrometry: A Comparison ofMicrowave and Dilution Techniques.�America Academy of Forensic Sciences60th Annual Meeting, February 2008,Washington, D.C.

• Wermeling, J., Cornett, C., Schweitzer, L.�Determination of Heavy Metals in WholeBlood Using Inductively Coupled Plasma -Mass Spectrometry: Dilution andMicrowave Digestion Compared.� Oralpresentation at the 37th Annual MidwesternAssociation of Forensic Scientist Meeting,October 2008, Des Moines, IA.

• Schweitzer, L., Wermeling, J., Cornett,C.�Determination of Heavy Metals inWhole Blood Using Inductively - CoupledPlasma-Mass Spectrometry.� Manuscriptsubmitted to the 60th Annual PittsburghConference and Exposition on AnalyticalChemistry and Applied Spectroscopy(Pittcon), March 2009, Chicago, IL.

IMPLEMENTATION

Following instrument overhaul, this workwill continue on an unfunded basis. Underthe aegis of the WSCL-M Toxicology Unit,acquired data from this project will be usedto develop an inorganic database. Whencompleted, the blood metal analysismethods will be added to the WSCL-MToxicology Unit Procedure Manual.

Also, based on a request from a submittingagency, a protocol for the analysis ofarsenic, selecium, cadmium, thallium, andlead in water and clear beverages wasdeveloped using the basic outline of theblood methods. This request resulted in acase report. The technique will now beconsidered as a routine offering of theWSCL-Trace Unit.

CONTACTS

Principal Investigators

Joseph WermelingWisconsin State Crime Laboratory-Madison4626 University AvenueMadison, WI 53705Phone: [email protected]

Charles CornettDepartment of Chemistry andEngineering Physics1 Plaza DriveUniversity of Wisconsin-PlattevillePlatteville, WI 53818Phone: [email protected]

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Application of Chemometric Procedures to DifferentiateIgnitable Liquid Residues from Substrate Interferences


In arson investigations, fire debris is typicallyextracted and analyzed to determine thepresence of ignitable liquid residues (ILR).Identification of these ILRs can becomplicated by interference from substrates,such as inherent hydrocarbons and pyrolysisproducts, making identification of the ILRsdifficult and potentially subjective.


The primary objective of this research is todevelop an objective methodology fordifferentiating ignitable liquids from burnedsubstrates. This is achieved by creatingreference collections and by demonstratingthe potential of statistical and chemometricprocedures to associate and differentiatenot only classes of liquids but also todifferentiate liquids from burned substrateinterferences.

METHODOLOGY

Specific goals of the research are to:

• create reference collections of neat andevaporated ignitable liquids

• create reference collections of burnedhousehold substrates

• optimize data pre-treatment proceduresfor statistical and chemometric analyses

• assess correlations among neatignitable liquids, evaporated liquids, andburned substrates using Pearsonproduct moment correlation (PPMC)coefficients

• assess association and discrimination ofneat ignitable liquids, evaporated liquidsand burned substrates using principalcomponent analysis (PCA)


This is a new project that has not startedwork yet.

TECHNOLOGY BENEFITS

Reference collections for burned substratesare not currently available. The creation ofa reference collection of burned householditems will improve forensic arson investiga-tions. Also, by developing mathematicalprocedures, subjectivity associated with thecomparison of chromatograms will beimproved and objective distinction ofignitable liquids from substrate interferen-ces increased. This will reduce the risk offalse positive identifications of liquids in firedebris.

COLLABORATION

This is a collaborative effort betweenMichigan State University and the MichiganState Police. Troy Ernst, of the TraceEvidence Unit of Michigan State Police �Grand Rapids Forensic Laboratory, willserve as consultant on the development ofthe methodologies and their implications toforensics crime laboratories.

DISSEMINATION

The results of this study will bedisseminated through presentations atforensic science conferences, including the

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Midwestern Association of ForensicScientists, and through submittal of amanuscript for publication in the Journal ofForensic Sciences. The project and its resultswill also be posted on the MFRC web site inthe form of a technical report.


This is a new research project with nopublications or presentations to date.

IMPLEMENTATION

DVDs of completed reference collections willbe created and made available to forensiccrime laboratories.

Based on the success of this research, thePIs may seek funding from the NIJ tocontinue the research.

CONTACTS


Ruth Waddell-SmithSchool of Criminal Justice560 Baker HallMichigan State UniversityEast Lansing, MI 48824Phone: [email protected]

Victoria McGuffinDepartment of ChemistryMichigan State UniversityEast Lansing, MI 48824Phone: 517-355-9715, ext. [email protected]

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Evaluation of a Portable Raman Analyzer for Testing Drugs


Numerous liquid samples from clandestinelaboratories are submitted for analysis. Theseanalyses are very time consuming andoftentimes result in case backlogs. Such casebacklogs cause slowdowns and delays in thejudicial system. Speedy trial demands aretherefore difficult to maintain.

This research project attempts to ease someof the backlog problems by evaluating aportable drug identification system usinglaser-based Raman technology. The portableRaman system will be evaluated for ease ofuse and for its ability to analyze controlledsubstances.


A screening test (non-confirmatory) is oftenall that is needed to begin the process ofprosecuting individuals with controlled sub-stances. Having a portable Raman unit in thefield should expedite this process. The Ra-man is a portable system that can identifycontrolled substances.

Figure 1. Portable Raman Analyzer Manufact- ured by Ahura Scientific.

METHODOLOGY

This project does not intend to repeat thealready published studies, but to furtherinvestigate the analysis of controlled sub-stances utilizing the portable Raman ana-lyzer. Specific types of samples for thisproject include known standards, liquidsamples from clandestine laboratories,tablets, powders, and other street drugsthat law enforcement personnel encounter.


Ahura Scientific (the manufacturer of thePortable Raman instrument) providedtraining in operation and use of the ana-lyzer. They also provided training in how torun the samples and how to incorporatethem into a searchable Bureau of CriminalApprehension (BCA) library.

So far, 200 samples have been run on theinstrument and incorporated into the library.Approximately 200 new vials will be pur-chased, as cleaning the vials is time con-suming and cross-examination is an issue.These samples will also be incorporatedinto the library.

TECHNOLOGY BENEFITS

By implementing a plan to analyze con-trolled substances in the field, the labora-tory case backlog can be alleviated and theprosecution of controlled substance casescan continue in a timely manner. Lawenforcement personnel trained on theequipment can obtain results within min-utes, thereby making it possible to analyzesamples while the suspect is still in cus-tody. Portable Raman analyzers for testing

16

drugs have the potential to become ascommonly used as the Breath Test Unitsfor determining blood alcohol levels.

COLLABORATION

The laboratory portion of this project tookplace in the Drug Chemistry Section of theMinnesota BCA in St. Paul, Minnesota.Available drug standards were utilized foranalyzing known samples. Some previ-ously in the laboratory analyzed �street�samples were also used.

The field portion of this project will takeplace in the Law Enforcement Center(LEC) in Rochester, Minnesota. The LEChouses various law enforcement agenciesincluding field offices for the BCA and theGang Strike Force. The Southeast Minne-sota Drug Task Force, the RochesterPolice Department, and the OlmstedCounty Sheriff�s Office are also located inthe LEC.

DISSEMINATION

The results of this research project werepresented to BCA agents and were alsoshared with other law enforcement person-nel. The BCA hosts several trainingclasses including a Clandestine Metham-phetamine Lab Certification Course and aDrug Investigation Class.

Research findings will further be sharedwith the manufacturer and be presented atthe fall meeting of the Midwestern Asso-ciation of Forensic Scientists. Uponcompletion of this project, a manuscript willbe submitted to a scientific journal forpublication, while a final technical report onthe project and its findings will be postedon the MFRC web site.


• Gross, S. �First Defender Grant Project.�Presentation to BCA agents, January2008, St. Paul, MN.

• Gross, S. �First Defender Grant Project.�Presentation to Minnesota lawenforcement agents, February 2008, St.Paul, MN.

• Engebretson, A., Patterson, M., Radthke,J. �Validation Study of the First Defender.�Oral presentation at the 37th AnnualMidwestern Association of ForensicScientist Meeting, October 2008, DesMoines, IA.

IMPLEMENTATION

The instrument is currently being used bylocal law enforcement in the Rochester areaof Minnesota. Special agents are testing theinstrument by running unidentified sub-stances and liquids from a methamphetam-ine lab. In the future, the instrument will beoffered to other counties to increase thenumber of scans performed on street drugs.

CONTACTS


Susan GrossMinnesota Department of Public SafetyBureau of Criminal Apprehension1430 Maryland Avenue, EastSt. Paul, MN 55106-2801Phone: [email protected]

Mark PattersonMinnesota Department of Public SafetyBureau of Criminal Apprehension1430 Maryland Avenue, EastSt. Paul, MN 55106-2801Phone: [email protected]

17

Fast Gas Chromatography Capabilities in Arson DebrisAnalysis

TECHNOLOGY NEED

While autosamplers have increased samplethroughput in trace units across the nation�scrime laboratories, many laboratories stillfind themselves needing more throughput.

As defined in Forensic Sciences: Review ofStatus and Needs (NIJ/OLES, 1999). Thereis a need for method development in therecovery of ignitable liquid residues from avariety of matrices.


Current analytical literature identifies FastGas Chromatography (Fast GC) as a sepa-ration technique capable of increasing thesample throughput of trace element groups.

The goal of this project is to assess themerit of Fast GC in the determination ofignitable liquids in a variety of matrices. Thestable, rapid heating cycle of Fast GCcoupled with narrower capillary columnsand high phase ratio potentially results inmore plates per meter. This increase willenable fast separations with potentiallysuperior resolution compared to conven-tional GC if hydrogen replaces helium asthe carrier gas.

METHODOLOGY

This project tests the hypothesis that FastGC is a beneficial technique in the area ofignitable liquids analysis. The experimentaldesign uses both solid phasemicroextraction (SPME) and traditionalactivated charcoal strip (ACS) to pre-con-centrate single compound ignitable liquids(i.e. toluene) and common ignitable liquids

These volatiles are analyzed by both con-ventional GC (helium mobile phase) andFast GC. The results are statistically com-pared to test the hypothesis that Fast GCwith hydrogen mobile phase provides muchfaster separation and higher resolution thanconventional GC.


Data from this study illustrated that Fast GCalone reduces chromatographic retentiontimes by 60% in the analysis of ignitableliquids. The study also demonstrated thatthe gains in resolution by switching fromhelium to hydrogen as a carrier gas (mobilephase) far exceeded the resolution lossesfrom the shorter Fast GC minbore column.

The combination of these approaches (FastGC and hydrogen carrier gas) producedphenomenal improvements in the analysis ofignitable liquids as the 60% reduction inretention times is accompanied by increasesin resolution of up to 500%.

These improvements in separation by FastGC (hydrogen) compared with conventionalGC (helium) were consistently observed in avast variety of ignitable liquid analyses aswell as analysis of ignitable liquids recoveredfrom controlled burn matrices such as as-phalt shingles, carpet, carpet padding, andsoftwood lumber.

Yet, there is a need for serious safety proce-dures if hydrogen is used as a carrier gas ina gas chromatography-mass spectrometer,rather than the gas chromatography-flameionization detector (FID) system in this study.

18

Table 1. Statistical comparison of Fast GC and conventional GC retention times for the separation of 95% evaporated gasoline.

Fast GC analysis of primary standards andsamples in dilute CS2 will be completed inthe near future and final statistical datacomparisons will be conducted. SPME workwill extend the library and comparison as faras possible in the remaining time period.This will be followed by analyzing the impactof Fast GC on a practicing laboratory.

Meritorious results will be used to asses thefeasibility of and seek out funding related toprojects addressing the need for portableinstruments and technologies in detectingcompounds of interest in arson debris.

TECHNOLOGY BENEFITS

The practical significance of this work forcrime laboratories is two-fold: 1) a potentialincrease in sample throughput and 2) animproved detection of potential ignitableliquids through better resolution.

COLLABORATION

This project is a collaborative effort betweenthe University of Wisconsin-Platteville (UWP)and the Wisconsin State Crime Laboratory-Madison. The principal investigator supervisesundergraduate researchers and experimentsperformed at UWP using the FGC-FID/ECD(Electron Capture Detector), while the co-principal investigator supervises studentsusing the GC-MS at WSCL-M and lendsexpertise in the area of arson residue sam-pling.

DISSEMINATION

Initial research data and findings were pre-sented at the Midwestern Association ofForensic Scientists Fall Meeting, September2007, in Traverse City, MI. Final results will bepresented at Pitcon 2009. The project and itsresearch results will also be disseminated ina final technical report posted on the MFRCweb site.


• Cornett, C., Wermeling, J. �Advancesin Ignitable Liquid Identification: AComparison of Fast and ConventionalGas Chromatography.� Presentation atthe Midwestern Association of ForensicScientists Annual Meeting, September2007, Traverse City, MI.

• Halligan, A., Selle, A., McCrary, C.,Jacobs, J., Larsen, L., Vircks, K.,Vyhnanek, A., Wermeling, J., Cornett, C.�Fast Gas Chromatography Capabilities inArson Debris Analyis�. American Academyof Forensic Sciences 60th Annual Meeting,February 2008, Washington, D.C.

• Cornett, C., Wermeling, J. �Fast GasChromatography in Arson DebrisAnalysis.� To be presented at the 60th

Compound Class

Retention Time (min) (Fast GC)

Retention Time (min)

(Conventional)

C4-alkylbenzene 3.93# 10.69# C4-alkylbenzene 4.14 11.28 C4-alkylbenzene 4.16 11.37 C5-alkylbenzene 4.29 11.74 C5-alkylbenzene 4.36 11.95 C5-alkylbenzene 4.44 12.16 C5-alkylbenzene 4.49 12.32

naphthalene 4.57 12.58 C5-alkylbenzene 4.65 12.77 C5-alkylbenzene 4.70 12.93 C5-alkylbenzene 4.76 13.09 C5-alkylbenzene 4.86 13.39 C1-naphthalene

(2-methylnaphthalene)

4.95 13.64

C1-naphthalene (1-

methylnaphthalene)

5.04 13.91

C2-naphthalene 5.21 14.43 C2-naphthalene 5.31 14.70

19

Annual Pittsburgh Conference andExposition on Analytical Chemistry andApplied Spectroscopy (Pittcon), March2009, Chicago, IL.

• Cornett, C., Wermeling, J., Halligan, A.,Selle, A., Vircks, K., Jacobs, J.,Vyhnamek, A., McCrary, C. �Building aBetter Mouse Trap for Ignitable Liquids:Fast GC Meets Hydrogen Carrier Gas.�Oral presentation at the 37th AnnualMidwestern Association of ForensicScientist Meeting, October 2008, DesMoines, IA.

• Halligan, A., Wermeling, J., Jacobs, J.,Selle, A., Vircks, K., Vyhnamek, A.,McCrary, C., Cornett, C. �Putting Fast GCto the Test in Ignitable Liquid Analysis: AnEvaluation of Standards and Matrices�Poster presentation at the 37th AnnualMidwestern Association of ForensicScientist Meeting, October 2008, DesMoines, IA.

IMPLEMENTATION

Switching to Fast GC from a conventionalunit is relatively simple. It involves upgradingthe power source to either a 220 V or 240 V(preferred) service, and installing a high-heatoven shroud, power cord, and narrowercapillary column. The instrumentation upgrademay cost as little as $1,000 - $2,000.

CONTACTS


Charles CornettDepartment of Chemistry andEngineering Physics1 Plaza DriveUniversity of Wisconsin-PlattevillePlatteville, WI 53818Phone: [email protected]

Joseph WermelingState of Wisconsin Crime Laboratory-Madison4626 University AvenueMadison, WI 53705-2174Phone: [email protected]

20

(Conventional GC)

(Fast GC)

Figure 1. A Comparison of Fast GC (Hydrogen Carrier Gas) and Conventional GC (HeliumCarrier Gas) Separations of 95% Evaporated Gasoline

min3.5 4 4.5 5 5.5 6 6.5

pA

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FID1 B, (C:\CHEM32\1\DATA\GASOLINE STANDARDS 2\10-4-2007 95% EVAP BP 2.D)

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747

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21

Micromechanical Cantilever Based Sensors forChemical Species Detection


Current sensor systems require extensivesample preparation or specialized instru-mentation to identify molecules of controlledsubstances such as cocaine. High specific-ity and sensitivity Micromechanical Cantile-ver (MC)-based sensors provide an invalu-able tool for forensic science because oftheir portability, capability for detection, andcapability for identification with high sensi-tivity and specificity. Successful completionof the project will buttress research effortstowards developing portable sensors forchemical species detection by integration ofreceptor layer coated micro-cantilevers andhigh resolution interferometry into a singlemicrofabricated chip.


Specific aims of this project are to:

• develop robust miniature sensors forhigh-resolution measurement of surfacestress associated with the formation ofaffinity complexes on the sensitizedsurface of micro-cantilevers

• characterize the sensitivity and specificityof cocaine detection with MC-basedsensors functionalized with suitablereceptor layers of aptamer molecules.

• functionalize micro-cantilevers with anaptamer capable of sensitive and specificdetection of cocaine molecules

METHODOLOGY

In this project, a proof-of-concept isdeveloped for the sensing approach bybuilding an MC-based sensor functionalizedwith aptamer molecules for sensitive andspecific detection of cocaine molecules.


A detailed design and fabrication plan wasdeveloped to assemble the MC-basedsensor for high resolution measurement ofsurface stress. Detailed design consisted oftwo micro-mechanical cantilevers (a sensing/reference pair), two microlens arrays (MLA1+ MLA2), fiber optic cables, micro-actuatorsand alignment accessories.

Figure 2. Alkanethiol SAM formation.

Model systems were developed and experi-ments on Alkanethiol SAM Formation andDNA Hybridization were conducted. They Figure 1. Aptamer based MC sensor.

22

demonstrated sensor performance in ambi-ent and aqueous environments, and vali-dated the functional capability of a DNAaptamer in the micro-cantilever.

Figure 3. DNA hybridization.

The ability of the aptamer to bind with co-caine by colorimetric displacement assayand fluorescence assay using FluorescenceResonance Energy Transfer (FRET) wasalso established. For these assays, thesynthetic oligonucleotide anti-cocaineaptamer (5� - GGGAGACAAGGAAAATCCTTCAATGAAGTGGGTCGACA-3�) was used.

Figure 4. FRET Quenching results.

TECHNOLOGY BENEFITS

Development of MC-based sensors symbol-ize important breakthroughs for forensicscience because of the tremendous sensitiv-ity that can be achieved by aptamer-coatedmicro-cantilevers and the specificity impartedby aptamers for identification of chemicalspecies at concentrations of parts-per-billion.

23

Such sensitivity revolutionizes forensic analy-sis of controlled substances, explosives, toxicspecies, biological molecules, and DNAmatching required for crime-scene identifica-tion.

COLLABORATION

This project is a collaborative effort betweenIowa State University and the Michigan StatePolice. The PIs designed and developed theMC-based sensor, while the Michigan StatePolice contributed its expertise in controlledsubstance identification for sensor develop-ment.

DISSEMINATION

Sensor design and initial research findingswere presented at an SPIE symposium andat the MFRC annual meeting. Upon comple-tion of the project, research results will bepublished in a peer-reviewed forensic sciencepublication. A final technical report will alsobe posted on the MFRC web site.


• Shrotriya, P., Kang, K., Marquardt, J.�Surface stress sensors for detection ofchemical and biological species.� SPIE14th International Symposium on SmartStructures and Materials NondestructiveEvaluation and Health Monitoring, March2007, San Diego, CA.

• Shrotriya, P., Nilsen-Hamilton, M.,Sachau, S., Kang, K. �MicromechanicalCantilever (MC)-Based Sensors forChemical Species Detection�. MidwestForensics Resource Center AnnualMeeting, April 2008, Bemidji, MN.

Percent change in fluorescence

-80 -60 -40 -20 0 20 40 60 80 100 120

10

100

1000

2000

2500

Coca

ine

[mM

]

% quenching

15 Minute10 Minute5 Minute1 Minute

IMPLEMENTATION

This project established proof-of-conceptfor chemical species detection by MC-based sensors. The PIs utilized the resultsof this research to seek additional fundingfrom NIJ to develop a portable MicroElectron Mechanical Systems (MEMS)sensors. An award for funding was recentlymade.

CONTACTS


Pranav ShrotriyaDepartment of Mechanical Engineering2025 Black Engineering BuildingIowa State UniversityAmes, IA 50011-2161Phone: [email protected]

Marit Nilsen-HamiltonDepartment of Biochemistry, Biophysicsand Molecular Biology3206 Molecular Biology BuildingIowa State UniversityAmes, IA 50011Phone: [email protected]

24

Optimization of Headspace-Solid Phase Microextraction forOrganic Impurity Profiling of Ilicit MDMA Tablets


Current profiling of illicit methylenedioxy methamphetamine (MDMA) tabletsis based on physical and chemicalcharacterization of tablets. As tablets withthe same physical characteristics do notnecessarily have the same chemicalcomposition, chemical profiling hasbecome the preferred method used bylaw enforcement agencies.

Conventionally, organic impurities areextracted using liquid-liquid extraction(LLE) procedures and then analyzed bygas chromatography-massspectrometry (GC-MS) to generate animpurity profile. Although the success ofthis technique has been documented inthe literature, the procedure has a num-ber of disadvantages, including:relatively large sample masses arerequired; trace impurities may bemasked by the controlled substancewhen present at high concentrations; anduse of organic solvents may requireexpensive disposal.


In this research, an alternative extractionprocedure is developed and optimizedbased on headspace-solid phasemicroextraction (HS-SPME). In HS-SPME, a polymeric fiber is exposed tothe headspace of a liquid or solidsample. Impurities absorb and/or adsorbto the fiber and are pre-concentrated onthe fiber during the extraction procedure.The fiber is then removed from thesample vial and inserted into the heatedinlet of the GC, where impurities aredesorbed directly from the fiber and

Figure 1. Analyst inserting SPME fiber into GC inlet to desorb impurities.

HS-SPME reduces or eliminates the use oforganic solvents and associated disposalcosts. With the correct choice of fiber coating,impurities can be preferentially extracted overthe illicit substance.

METHODOLOGY

In this study, HS-SPME procedures for theextraction of organic impurities from illicitMDMA tablets are developed. Two differentfiber types are investigated, a statisticalexperimental design procedure is used tooptimize the extraction time and temperaturefor each. The optimal fiber type is assessedbased on the number of impurities extracted,as well as repeatability and reproducibility ofthe extraction. The optimized HS-SPME isthen compared to a conventional LLEprocedure, in terms of the number and level ofimpurities extracted, repeatability, and

25

reproducibility of the two extractionprocedures.


A HS-SPME procedure for the extraction oforganic impurities from tablets containingMDMA was developed and optimized. TheSPME fiber was exposed to the headspaceof a buffer solution containing the solidMDMA sample.

Figure 2. HS-SPME extraction from carbonate buffer solution of impurities from a homogenized batch of MDMA tablets.

Relative standard deviations were typicallyless than 15% for the early eluting impurities(retention time less than 14 minutes), usingan extraction time of 30 minutes at anextraction temperature of 60°C.

The optimized HS-SPME procedure wascompared to a conventional LLE procedurein terms of the number of impuritiesextracted as well as the repeatability andreproducibility of the extraction. In general,only 12 impurities were extracted using theLLE procedure, compared to 16-22 impuritiesusing the HS-SPME procedures, depending

on extraction time and extractiontemperature.

Figure 3. Liquid-liquid extraction of impurities from a homogenized batch of MDMA tablets.

Impurity profiles of MDMA tablets from fivedifferent exhibits were obtained using theHS-SPME procedure, with analysis by GC-MS. All five exhibits were differentiatedbased on the impurity profiles obtainedusing the optimized HS-SPME procedurefor impurity extraction. This development issignificant as it demonstrated the potentialof HS-SPME in organic profiling; a conceptnot previously realized.

While the MFRC funded part of theresearch has been completed, the researchis ongoing in the laboratory usingdepartmental and other fundingmechanisms.

TECHNOLOGY BENEFITS

HS-SPME offers several advantages overconventional LLE procedures. They include:improved organic impurity extraction, andinformative impurity profiles coupled withobjective comparisons. As such, they allowimpurity profiling to become a more usefultool for forensic practitioners.

RT: 0.65 - 25.03

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NL:1.73E8TIC F: MS LLE_ext_6_02

RT: 0.37 - 24.96

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8.63 22.1211.176.095.473.11 6.312.13

NL:1.62E8TIC F: MS rr_11

26

COLLABORATION

This project is a collaboration betweenMichigan State University and the MichiganState Police Crime Laboratory. The collabora-tor provided the MDMA tablets.

DISSEMINATION

Results of the work were disseminated atthe Midwestern Association of ForensicScientists Fall Meeting 2007. A posterpresentation was also made at the AmericanAcademy of Forensic Sciences, February2008, in Washington, D.C. Currently, amanuscript is being prepared for submissionto the Journal of Forensic Sciences. Resultsof the work will also be disseminated in afinal technical report on the MFRC web site.


• Meisinger, S. and Waddell, R.�Optimization of Headspace � SolidPhase Microextraction (HS-SPME) forOrganic Impurity Profiling of SeizedMDMA Tablets.� Midwestern Associationof Forensic Scientists Fall Meeting,September 2007, Traverse City, MI.

• Meisinger, S. and Waddell, R.�Comparison of Extraction Procedures forOrganic Impurity Profiling of SeizedMDMA Tablets.� American Academy ofForensic Sciences 60th Annual Meeting,February 2008, Washington, D. C.

IMPLEMENTATION

The Drug Enforcement Administration hasshown an interest in the research and askedto be kept informed of progress. Currently,the methodology is still under developmentand opportunities for implementation are notyet available.

CONTACTS


Ruth J.H. Waddell-SmithForensic Science Program560 Baker HallMichigan State UniversityEast Lansing, MI 48824Phone: [email protected]

27

The Temporal Fate of Drugs in Decomposing Tissues*


Interpreting drug concentrations in decom-posed remains is difficult. Postmortemtissues that are routinely collected andanalyzed to provide the largest comparativedatabase for interpretation (blood, urine,vitreous humor) are frequently lost in theearly stages of putrefaction.

When drugs are found in weatheredtissues, there is currently little or noinformation available to help guide thetoxicologist (and subsequently thepathologist) in evaluating whether a drug ordrugs played any significant role in causingthe death.


This study monitors the concentration ofdrugs, in multiple tissues and organs,during whole body decomposition.

Specific questions addressed are:

• How does the concentration of drugschange during tissue decomposition?

• How long do drugs and metabolitespersist in tissues at detectable levels?

• Which collection sites are most usefulfor detecting drug exposure andinterpreting results?

The study uses Sus domestica, thedomestic pig. Pigs are selected becausetheir size is comparable to humans allowinghigh-level dosing and sequential samplingof multiple tissues. Also, their physiology issimilar to humans. Specifically, their diges-tive and cardiovascular physiology allow

for drug absorption and distribution thatclosely mimics those of humans.

Figure 1. Animal research enclosure site.

METHODOLOGY

The concentration of 16 drugs in tissue fromdecomposing pigs weighing 120-150 lbs., ismonitored in blood, brain, liver, kidney,muscle, maggots, and soil during one week.

Pigs are divided into groups (n=5) with eachgroup receiving four drugs. Drug cocktailsprepared from pills (capsules or tablets) areobtained as pharmaceutical formulations.

Drugs tested include: morphine, amitriptyline,citalopram, diazepam, methadone, fluoxetine,doxepin, acetaminophen, oxycodone, diphen-hydramine, venlafaxine, carisoprodal,verapamil, zolpidem, and propoxyphene.

Sacrifice of pigs (using intracardiac pentobar-bital) is four hours after dosing (gavage), withtissue collection at 4, 24, 48, 96, and 168hours post-dosing. Samples are frozen untilassay.

* Approval to perform animal research was obtained from the Ohio State University Institutional Laboratory Animal Care and Use Committee.

28

Detection and quantitation of drugs isachieved through Solid Phase Extraction(SPE) followed by Gas Chromatography-Mass Spectrometry (GC-MS) analysis. Mor-phine is derivatized with heptafluorbutyricanhydride and assayed by Selective IonMonitoring (SIM).


Twenty-four pigs were obtained for testing.Nine of which were sacrificed in the initialattempt: five for testing and four as controls.Control pigs were included to determinewhether total organ weight changed as aresult of necrospy.

The initial attempt to dose the pigs throughtheir food was unsatisfactory. The amountof drugs consumed was of very low levelsand varied from animal to animal. It wasdecided that drugs needed to be adminis-tered by gavage as slurry.

Figure 2. Dosing of pigs allowed to live four hours after gavage.

It was also found that the rate ofdecomposition was much faster thananticipated. Brain and kidneys were notavailable after forty-eight hours, whilemuscle and liver persisted for one week.

Figure 3. Results of the four drugs in liver tissue collected for two weeks.

Concentrations of drugs analyzed typicallyincreased with time during decomposition.This was most pronounced (and statisticallysignificant) for amitriptyline. It was concludedthat attempting to interpret drug levels indecomposed bodies may lead to incorrectconclusions about cause and manner ofdeath.

TECHNOLOGY BENEFITS

This investigation provides importantinsights into the fate of drugs in varioustissues over an extended period of decay.The information gained allows pathologists,toxicologists, and criminologists to interpretcase results more accurately therebyproviding a more complete conclusionregarding cause of death and fate of victims.

COLLABORATION

This is a collaborative project between theFranklin County Coroner�s Office (FCCO)and the Ohio State University (OSU).Pathologists from both institutions assistedin the collection of tissues. Toxicology wasperformed at the FCCO, while a biostatisti-cian performed statistical analysis of resultsat OSU. Approval to perform animal re-search (the site of decomposition was leasedfrom OSU) was obtained at OSU.

Average Drug Concentration Over Time

0.0050.00

100.00150.00200.00250.00300.00350.00

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entr

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AmitriptylineCitalopramDiazepam

29

DISSEMINATION

Presentations of research findings will bemade at the Society of Forensic Toxicology,October 2008, Phoenix, AZ . A manuscript iscurrently being prepared for submittal to theJournal of Forensic Sciences. The project andits research findings will also be disseminatedin a final technical report posted on the MFRCweb site.


• Wyman, J., Dean, D., Yinger, R.,Simmons, A., Brobst, D., Bissel, M.,Silveira, F., Shott, R., Ohr, J., Tatarek, N.,and Lewis, B. �The Temporal Fate ofDrugs in Decomposing Porcine Tissue.�Paper submitted to the Society of ForensicToxicologists (SOFT), October, 2008,Phoenix, AZ.

IMPLEMENTATION

As a result of this work, the Franklin CountyCoroner�s Office in Columbus, OH, no longertries to interpret therapeutic, toxic, and lethaldrug concentrations in decomposing bodies.

Some of the findings from this study wereincorporated in presentations on �TheToxicology of Suicide� given at theSociety for the Scientific Detection of Crime(Columbus, OH; March 17, 2008) and at theInternational Association of Coroners andMedical Examiners Annual Conference(Cincinnati, OH: June 12, 2008).

A medical examiner from the city and countyof San Francisco, CA, also expressed aninterest in the research results for possibleapplication.

CONTACTS


John F. WymanFranklin County Coroner�s Office520 King AvenueColumbus, OH 43201Phone: [email protected]

Dorothy DeanSummit County Medical Examiner�s Office85 North Summit StreetAkron, OH 44308Phone: 330-643-2102

Michael BissellDepartment of PathologyN 337 Doan Hall410 West 10th AvenueThe Ohio State UniversityColumbus, OH 43210

30

Ultra-Fast Gradient Elution HPLC as a High Throughput, HighInformation Content Screening Tool for Drugs of Abuse


Gradient elution high performance liquidchromatography (HPLC) with diode arraydetection is a common method used forscreening and even identifying drugs ofabuse in biological samples.

Although the technique has tremendouschemical selectivity and can produce pre-cise retention time data under well-con-trolled conditions, the technology is quiteslow, requiring 20-30 minutes per run.

As the sample load in forensic laboratoriescontinues to increase, using HPLC tech-niques to deliver analytical sample judge-ment in a timely fashion is becoming moredifficult.


In a previously funded project, the principalinvestigators developed an ultra-fast gradi-ent elution reversed-phase HPLC with diodearray detection as a fast tool for screeningsamples for regulated intoxicants. Througha combination of precise retentionmeasurements and chemometric analysis ofthe spectro-chromatograms, both the speedand selectively of older HPLC methodscould be significantly improved. This projectfocuses on enhancing these improvementsfor drugs of abuse.

METHODOLOGY

To improve the selectivity of the previouslydeveloped rapid liquid chromatographyintoxicant screening method, a secondary�orthogonal� column with different selectivitythan the standard (primary) column is used.Analytes that cannot be identified in

the first separation (due to incomplete chro-matographic or spectroscopic resolution) arenow subjected to a second, orthogonal ultra-fast liquid chromatography separation result-ing in a dramatic reduction in the number ofindistinguishable analytes at very low cost intime.

Figure 1. Two Column Approach

To improve the speed of the liquidchromatography method, recently commerci-alized very small stationary phases that usethe �core-shell� concept are applied. Bytesting a commercial reversed (C18-type)phase and a novel hydrophobically-assistedcation exchange phase, it may be possible todo reproducible gradient elution chromato-graphy in as little as 1 minute, whileretaining resolution on longer time scales.

To optimize chromatographic resolution, anewly developed general purposeoptimization scheme is applied. Thismay result in an increase in peak capacity ofgradient elution liquid chromatography. Byusing real samples whose compositions havebeen pre-assessed by accepted methods, thesensitivity and specificity of the methodologycan be evaluated.

Sample Collection and

Screening Method

Generate List of Candidate Drugs

Stop � No further analysis

NO

YES

Confirmation Method

Run sample on second,

orthogonal column

31


Based on Fs values, twelve columns wereselected to test the Snyder-Dolan reversedphase selectivity classification scheme forpredicting similarities and differences in thestationary phase behavior of representativedrugs.

Table 1. The twelve columns for this study.

The first three groups were each comprisedof reversed phases that were nearlyidentical in terms of phase parameterswhile the phases in the fourth groupswere extremely different.

All columns were checked with 16 standardsolutes and then tested with twenty-fivehighly varigated drug substances, inlcudingbenzodiazepines, antihistamines, andantidepressants under both isocratic andgradient conditions. The retention times ofall drugs on each column were measuredand used to compare column selectivity.

Results were consistent with those obtainedwith the 16 standard solutes, therebyverifying the value of the Snyder-Dolan

model of reversed-phase selectivity trianglefor predicting the similarity and differencesof phase behavior for a wide variety ofdrugs.

Figure 3. Stationary phase selectivity (S-B-C) triangle plot.

It was found that the Snyder-Dolandatabase of 350 columns can be used toreliably choose similar and very differentcolumns for drugs while the selectivitytriangle model based on Snyder�sparameters can be used for selectingmaximally orthogonal columns for drugs. Afew promising candidate columns for use ina two-column screening approach wereidentified, and the PI is currently working on�wedding� the orthogonality of the conven-tional C18-type phase and the SO3-HC-C8 phase with the high speed of therecently introduced micro-particulate core-shell particles. The distributor of thesematerials is supplying underivatizedmaterial to facilitate transfer of the SO3-HC-C8 phase to the new type of particle.

The PI is also studying the sensitivity andspecificity of the methodology by evaluatingreal samples whose composition have been

Set Type Column A

Column B

Column C

Column D

Fs (vs. SB C18)

I (Similar) Zorbax SB C18 -Agilent

ACE 5 C18 -Mac Mod

Discovery C18

-Supelco

Alltima HP C18 -Alltech

8.1~13.6

II (Similar)

Beta Basic Phenyl

-Thermo

Protosil 60-5

Phenyl -

Bischoff

Zorbax Phenyl -Agilent

18.1~29.4

III (Similar)

Hypersil Prism C18

-Thermo

Bonus RP

-Agilent

255.1, 266.7

IV (Different)

ZirChrom PS -

ZirChrom

Nova Pak CN HP-60 -Waters

Inertsil CN -GL

Science

48.8-230.5

32

pre-assessed by LC-MS.

TECHNOLOGY BENEFITS

This work will vastly improve the screening fordrugs in blood, urine, and other tissuesamples. It will also make a dramatic impactin forensic toxicology.

COLLABORATION

This project is a collaborative effort betweenthe University of Minnesota, and the Minne-sota Bureau of Criminal Apprehension Foren-sic Science Laboratory. The collaboration hasconsisted of forensic scientists from the BCAForensic Science Laboratory visiting the PI�slaboratory and attending group meetings. Inreturn, the PI and his group visited the BCA towork on specific project aspects.

DISSEMINATION

Information on and data from the project werepresented at professional meetings andconferences, including the American Acad-emy of Forensic Science, the HPLC Sympo-sium, and the Midwest Forensics ResourceCenter Annual Meeting.

Additionally, research information and findingswill be published in such journals as Analyti-cal Chemsitry, the Journal of Chromatogra-phy, the Journal of Analytical Toxicology, andthe Journal of Forensic Sciences. A finaltechnical report on the project and its findingswill also be posted on the MFRC web site.


• Stoll, D., Wenzhe Fan, Paek, C., YuZhang, Carr, P. �Development of a RapidHPLC-Based Screening Approach forIntoxicants-Selection of OrthogonalColumns to Maximize IdentificationPower.� Midwest Forensics Resource

Center Annual Meeting, April 2008,Bemidji, MN.

• Wenzhe Fan, Carr, P., Schellinger, A.,Pritts, W. �Application of Snyder-DolanClassification Scheme to the Selectionof Orthogonal Columns forPharmaceutical Analysis.� 32nd

International Symposium on HighPerformance Liquid PhaseSeparations and Related Techniques,May 2008, Baltimore, MD.

• Wenzhe Fan, Schellinger, A, Carr, P.�Development of a Snyder-DolanBased Approach to PickingOrthogonal Phases for Fast Screeningof Illicit Drugs and Impurity Profiling ofPharmaceuticals. PittsburghConference on Analytical Chemistryand Applied Spectroscopy (Pittcon),March 2009, Chicago, IL.

IMPLEMENTATION

After demonstrating adequate predictivecapabilities, the methodology will betransferred to the BCA where thetechnique will be used to establishperformance statistics.

CONTACTS


Peter CarrDepartment of Chemistry207 Pleasant Street SEUniversity of MinnesotaMinneapolis, MN 55455Phone: [email protected]

33

A Steganalyzer Package for Forensic Applications


The use of steganographic software forhiding information in image files is becomingmore widespread for illicit purposes. Whilesteganalysis algorithms are abound in theacademic literature, there are few softwareprograms that address the needs of localpolice departments who perform computerforensic functions for steganalysis.

Currently available steganalyzer packagesare very expensive and require extensivetraining of forensic personnel. This researchproject attempts to make a very limited butpractical steganalysis software programavailable to a local crime laboratory fortesting and evaluation.


In a previously funded project, proof-of-concept on the use of an artificial neuralnetwork for wavelet steganalysis (ANNTS)was demonstrated. The software developedsimply scanned images for hidden content. Itexhibited good steganalysis detection ratesand ease of use by non-specialists.

Specific aims of this project are to make thesoftware available for practical use in com-puter forensics. This is achieved by:

• enabling more images to be scanned andinspected

• adding two more targeted detectiontechniques for images embedded with S-Tools and JP Hide & Seek

• extending the capabilities of ANNTS bydeveloping a detection technique forimage data

METHODOLOGY

The current capacity of ANNTS is to scan asingle image at a time. In this project, thecode will be modified so that a file folder ofimage data can be inspected, and theresults be written to a file. In addition, thesoftware user interface will be slightlymodified to make the meaning of labelswritten on screen clearer.

The current version of ANNTS can identifyimages embedded with the freely availablejsteg-jpeg and F5. In this project, two moredetection techniques will be added: Theseare two widely used embedding techniquesavailable from the nearly 100 freeware atsegoarchive.com.

The current capabilities of ANNTS will beextended by developing a blind (or general)detection technique that is based on thedual-tree complex wavelet transform (DT-CWT) and a partially ordered Markovmodel (POMM) for image data. The DT-CWT transform is a recently developedtransform, developed as a more generalvision of the traditional wavelet transform.To date, neither the transform nor thePOMM has been used for steganalysis.Yet, they show great promise as blinddetection techniques.


This is a new project that has not startedyet.

TECHNOLOGY BENEFITS

The research will provide crime laborato-ries with a software package that

34

addresses basic stenography needs forforensic applications. Currently, few crimelaboratories have these capabilities.

COLLABORATION

This project is a collaborative effort betweenIowa State University and the Iowa Division ofCriminal Investigation (DCI). The softwareprogram will be developed at Iowa StateUniversity and tested and evaluated by DCI.

DISSEMINATION

Project findings will be disseminated throughpresentations at regional and national confer-ences, including the Association of DigitalForensics Security and Law, the Digital Infor-mation Hiding Workshop, and (if possible) theInternet Crimes Against Children (ICAC)conference. A manuscript on the researchand its findings will also be prepared forsubmission to the Journal of Digital ForensicPractice, while a technical report on theproject will be posted on the MFRC web site.


This is a new project with no publications orpresentations to date.

IMPLEMENTATION

Once the software package is developed, itwill be beta-tested and evaluated. DCI per-sonnel will receive instruction for use as wellas a user�s manual. Feedback will be used tomake adjustments to the program.

CONTACTS


Jennifer DavidsonDepartment of Mathematics/Department of Electrical and ComputerEngineeringIowa State UniversityAmes, IA 50011Phone: [email protected]

Technology Evaluator

Gerard MeyersIowa ICAC Task Force CommanderIowa Division of Criminal Investigation2006 South Ankeny BoulevardAnkeny, IA 50021Phone: 515-965-7404

35

Application of a Crystal Orientation Method to ForensicPhysical Matching of Metal Surfaces along a Fracture Line


Examples of metal fragments found at acrime scene include knife tips, pieces fromweapons, pry tools, or automobile partssuch as antennas or trim pieces. A physicalmatch between two surfaces is routinelyviewed as definitive proof that the fracturesurfaces were generated from the samepart. However, in the Summer 2005 issueof the Association of Firearm and ToolmarkExaminers (AFTE) Journal, Katterwe statedthat there were current �trial challenges� tothe practice of fracture matchingexaminations by toolmark examiners. Todate, there has been little systematic effortto establish a basis for the uniqueness of aphysical match of surfaces.


Conventional matching of the fracturesurfaces utilizes physical characteristics ofthe fracture such as shape, color and otherfeatures. While useful, these featuresunfortunately do not lend themselves to anobjective or quantitative evaluation of theuniqueness of a fracture. The objective ofthis study is to provide an additionalmethod for establishing characteristics of afracture based on atomic arrangements.

Metals are composed of crystals whoseatoms are arranged in regular longperiodicity. The atomic arrangement ofthese atoms is precisely known and can bedetermined with a high degree of accuracy.By probing the arrangement of atoms andcombinations of metal crystals within andalong a metal, a determination can bemade that two separate pieces of metal areindeed from a single piece.

METHODOLOGY

The study tests if the surface crystalorientations of the fractured crystals acrossthe fracture plane for two surfaces are infact unique and constitute a reliable,quantitative measure to determine if the twopieces sharing the fracture plane at onetime belonged to the same piece. This isachieved by examining the uniqueness ofcrystal orientations within the requirementsnecessary for accurate determination ofcrystallography using Electron Back ScatterDefraction (EBSD).

Figure 1. Grid placement for point-to-origin misorientation vector with 5° angular separation between vectors.

EBSD is a point-to-point (grain-to-grain)measurement on the surface of the metalthat can be used to determine theorientation of individual crystals. EBSD�sbasic operation collects electron back-scattered patterns using a specializedcollector on the scanning electronmicroscope. Orientation patterns aregenerated by sample interaction with theelectron beam.

36


Metal fractures of two Fe-based alloys wereexamined: a 304 standard stainless steeland an iron-gallium alloy. The study showedthat, in the case of intragranular fracture,pieces of grain can be associated acrossthe fracture surface, and that it is feasible toassociate two fracture pieces withsequence of grain orientations.

It also showed that it is possible to use thesequence of the difference in orientationbetween grains (misorientation) along alength on one side of the fracture surface toassociate the other side of fracture surface.

Figure 2. Orientation imaging microscopy maps of the top and bottom of the steel sample including the fracture edge.

Orientation angles for grains andmisorientation values between grains werecalculated and the resulting values werecompared for grains across fracturesurfaces. For iron gallium samples, it wasshown that it is feasible to use surfacecrystal orientation to associate one fractureedge to another.

Regarding the uniqueness of crystal orien-tations, it was estimated that for a se-quence of six randomly oriented grains

along a fracture, the resulting probabilitythat the same sequence would occur againis 2-31.

Table 1. Three orientation Euler angles for grain parts of two sides of fracture.

Finally, it was found that besidescrystallographic orientation, anotherparameter which has potential for beingused in combination with orientation isgrain size and shape along the fractureline. This is a facet which could beexamined in future work.

TECHNOLOGY BENEFITS

The outcome of this study is a workablemeasure of the orientation of crystals alongthe fracture line of many materials,including metal fractures of knives, prytools, and car antennas. The data collectedstrengthens the scientific foundation of thefracture match practice with data to beginthe task of estimating error rates.

COLLABORATION

This work was conducted at the AmesLaboratory in collaboration with Jim Kreiser,a retired toolmark examiner from the IllinoisState Police. Kreiser performed thepreliminary match of the test materials priorto the EBSD measurements. The principalinvestigators generated the EBSDexperimental data.

Edge Color Angle

Φ1 Angle Φ

Angle Φ2

Bottom Pink 341° 29° 56° Purple 162° 16° 47° Brown 202° 19° 3° Top Pink 343° 28° 53° Purple 169° 11° 38° Brown 198° 19° 6°

37

DISSEMINATION

Results of the feasibility study werepresented at the Midwest ForensicsResource Center Annual Meeting, April2007, in Madison, WI. A poster of the resultswas presented, and discussions withforensic professionals were conducted, atthe IAI meeting, July 2007, in San Diego,CA. Research results will also be presentedas a paper submitted to a forensic journal.Finally, the project and its findings will bedisseminated in a technical report posted onthe MFRC web site.


• Lograsso, B., Lograsso, T., Glamm, R.�Crystallographic Descriptors of a MetalSurface along a Fracture Line.� Posterpresentation at the InternationalAssociation of Identification, InternationalEducational Conference, July 2007, SanDiego, CA.

• Lograsso, B., Lograsso, T., Glamm, R.�Application of Crystal OrientationMethod to Fracture Evaluation.� Oralpresentation at the Midwest ForensicsResource Center Annual Meeting, April2007, Madison, WI.

• Lograsso, B., Lograsso, T. �Application ofa Crystal Orientation Method forMatching Surfaces Along a FractureLine.� Accepted for the Proceedings ofIMECE08, October/November 2008,Boston, MA.

• Bingham, M., Nordman, D., Vardman, S.�A Tractable Class of Distributions forRotations in 3 Dimensions and SomeApplications to Measured CrystalOrientations.� Submitted to the Journal ofAmerican Statistical Association.

IMPLEMENTATION

The purpose of this study was to evaluatethe feasibility of using surface crystalorientation to associate or differentiate metalfracture fragments. Additional research isneeded to determine error rates associatedwith performing physical matches.

CONTACTS


Barbara LograssoSchool of Technology426 EERC BuildingMichigan Technological UniversityHoughton, MI 49931Phone: [email protected]

Thomas LograssoAmes Laboratory126 Metals DevelopmentAmes, IA 50011Phone: [email protected]

38

High-Speed Digital Video Analysis of Bloodstain PatternFormation from Common Bloodletting Mechanisms


Bloodstains form on two dimensionalsurfaces. As a result, bloodstain patternanalysis is a study of the aftermath of ablood transfer event. From the size, shape,and distribution of bloodstains, inferencescan be drawn regarding the mechanismthat gave rise to their formation.

To date, most investigators rely on patternobservation and laboratory simulation torecognize and classify bloodstain patterns.Relatively little has been documentedabout the dynamics of the blood transferevent. Yet, understanding the dynamics ofblood transfer is critical to the sound inter-pretation of the resultant bloodstain pattern.


The objective of this study is to analyze anddocument the dynamics of common bloodtransfer and to compile a set of video clipsof common blood transfer events that canbe used by instructors in a bloodstainpattern course.

Specific aims of the study are to analyze:

• the dynamics of the collision between ablood drop and a hard surface

• the dynamics of the collision between ablood drop and the surface of a fabricas well as the cast-off from a movingbloodied object

• the impact splatter from beating,kicking, and stomping

• the impact splatter from shooting

METHODOLOGY

The study aims to document the three-dimensional dynamics of blood transferevents. To achieve this, a series ofexperiments are designed to maximizeavailable light, provide high contrast images,and where possible calibrate the scale of theexperiment.

For each experiment, digital video sequencesare captured, stored and backed up onportable hard drives. Bloodstain patternscreated in each experiment are collected andpreserved.

Figure 1. A blood drop falling onto a hard surface.

A wide range of target surfaces, such ascardboard, building materials, carpet andvarious types of clothing, is used for thedisposition of blood and bloodsplatter.

Experiments are set up using standardlaboratory equipment and some speciallybuilt devices for producing and capturingblood splattered by different mechanisms.

Human blood is drawn from laboratorydonors and is used within 72 hours for allexperiments. Blood is stored and refrigerated

39

when not in use and is heated to roomtemperature during use.

High Speed Digital Video (HSDV) is usedto analyze and document the dynamics ofcommon blood transfer events. HSDVcameras are ideally suited to see,measure, and understand high speedprocess offering outstanding image qualityat frame rates up to several thousand persecond.


Over 500 video sequences were collected.Most of the blood events studies were high-ly complex and variable. The collection ofvideo sequences only represents a set ofexamples of the events in question.Yet, the sequences provided valuable dataon the mechanism of blood events.

The following events were documented:

• passive drops formation from fivedifferent objects

• passive drops of three different volumesfree falling from 1-9 meters.

Figure 2. Blood drop landing on a cement tile

• passive drops from different heightsimpacting different surfaces

• passive drops impacting three differentsurfaces at known angles from 10° to 90°

• impact splatter resulting from blooddripping into blood

• impact splatter resulting from the impact offive instruments on bloodied surfaces

Figure 3. Blood droplets blown back by gases from firearm

• impact splatter resulting from thestomping, slapping, and flicking of blood

• formation of impact splatter stains oncotton fabric at various angles

• gunshot induced bloodsplatter producedby three different caliber handguns at upto four distances from the blood source

• swing cast-off from five different objects

• cessation cast-off

• large volumes of blood dripping from fourheights

• large volumes of blood projected from fourheights and at two angles

• projected blood from coughing andexhaling through the nose and mouth

40

• contact blood staining produced byfingers, various fabrics, and hair

AVI files, using the Cinepak codec compres-sion were prepared from a representativeset of the video clips. The files can be playedby media players such as Windows MediaPlayer. Alternatively, the Photron FastcamViewer (PFV) can be used if frame-by-framecontrol is required. The PFV can be down-loaded free from the internet.

TECHNOLOGY BENEFITS

In the past, bloodstain pattern analysts weretrained in bloodstain pattern analysis (BPA)primarily by studying the aftermath of blood-letting events. The recording of major blood-letting events by high speed digital video,and making these videos available to theforensic community, enhances the ability ofthe bloodstain pattern analyst to understandthe dynamics of blood transfer that gave riseto bloodstain pattern.

The study of how blood is deposited inspecific events will assist in thedifferentiation of closely related events. Assuch, it will contribute to the underlyingscience behind BPA methods and thevalidation of current bloodstain patternrecognition methods. Finally, the foundationsof BPA opinion evidence in court will bestrengthened by this study. The technologywill enhance the expert witness� opinion andhelp to satisfy the continued demand bycourts to use the most advancedtechnologies available.

COLLABORATION

This project is a collaborative effort betweenstaff from the Minnesota Bureau of CriminalApprehension, the Institute of EnvironmentalScience and Research in New Zealand, anda forensic consultant. The advantage of this

collaboration is a broader international per-spective of the research need and a connec-tion with a research program that has anestablished network of partnerships.

DISSEMINATION

Research results will be presented at re-gional, national, and international forensicscience meetings and seminars. One ormore papers will also be prepared for submit-tal to suitable, peer-reviewed forensic jour-nals. In addition, a full in-house report of theoutcomes of the project will be made avail-able to the Minnesota BCA Forensic Labora-tory and the ESR in New Zealand. Finally, atechnical report on the project and its resultswill be posted on the MFRC web site.


• Laber, T., Epstein, B., Taylor, M.�Bloodstain Formation by CommonBloodletting Mechanisms.� MidwestForensics Resource Center AnnualMeeting, April 2008, Bemidji, MN.

IMPLEMENTATION

A series of video clips were preparedillustrating the formation of the mainbloodstain pattern studied. The video clipsare available to forensic science educatorsand BPA instructors through the MFRC.

CONTACTS


Terry LaberMinnesota Department of Public SafetyBureau of Criminal Apprehension1430 Maryland Avenue, EastSt. Paul, MN 55106-2801Phone: [email protected]

41

Michael TaylorInstitute of Environmental Scienceand Research27 Creyke RoadChristchurch, NZPhone:[email protected]

Bart EpsteinForensic Consultant4520 Sedum LaneEdina, MN [email protected]

42

CD-ROM Based Digital Information Database on Pipe andTubing Utilized in Improvised Explosive Devices


The rise in terrorist bombings has made theidentification of bomb materials a priority forforensic scientists. The volume of evidenceto be analyzed and the importance of test-ing for preventative and investigative pur-poses have increased the need for toolswhich can aid in the identification of specifictubing material used for bombs.

Currently, available information on tubingmaterials and their manufacturers is eitheroutdated or incomplete. Additionally, thedata that has been gathered is either inprinted format or in handwritten notes. Nodigital captures of this information arecurrently available.


The objectives of this project are to docu-ment:

• all known types of pipes and tubing

• worldwide pipe and tubingmanufacturers

• manufacturing identification marks onpipes and tubing

• companies that import pipes and tubinginto the United States

• companies that regularly stock or marketeach type of pipe and tubing

All of this information is placed into asearchable database containing a self-running platform (Filemaker) which does notrequire the user to own the software.

METHODOLOGY

The first part of the project targetsdocumenting as many manufacturers ofpipes and tubing as can be found. This isdone primarily by internet searches. Thenext step is to contact themanufacturers to request informationregarding any identification markings.

Samples of as many types as possible ofpipe and tubing are purchased. This in-cludes purchasing samples in variousregions of the U.S. The samples are mea-sured, digitally photographed and all dataentered into the database.

Additionally, companies that import pipesand tubing into the U.S. are identified anddocumented, as well as companies thatcommercially sell pipes and tubing.

Figure 1. Pipe end cap made be a U.S. company.


Currently, the database contains more than250 entries from 25 countries and consists

43

of the following information fields:

� Company Name� Contact Information� City/State� Zip Code� Country� Telephone� Fax� Logo Description� Type of Pipe� Miscellaneous Information� Website Information

Figure 2. Pipe Bomb Database.

Figure 2. Pipe Bomb Database.

In addition, there are four fields for anyphotos that can be found or produced forthe individual pipes as well as drawings ofany logos or characteristic markings.

At this time, the database deals only withboth steel and plastic pipe, but there areupdates planned to include other types aswell, such as brass and copper.

TECHNOLOGY BENEFITS

The availability of a searchable databasegreatly reduces the amount of time andeffort that is required by forensic scientiststo process bomb samples.

COLLABORATION

This project is a collaboration between theWestern Law Enforcement Training Center(WFLETC), the Missouri State HighwayPatrol, the Lake County Crime Laboratory,and the Midwest Forensics Resource Cen-ter (MFRC).

Personnel from the WFLETC, Lake CountyRegional Crime Laboratory, and theMissouri State Highway Patrol CrimeLaboratory conducted internet searches andmade direct contact with manufacturers,distributors, and retailers of pipe and tubingthat can be used to construct pipe bombs.

The data obtained from these contacts wasforwarded to the WFLETC, whichpurchased all samples of pipe and tubing,digitally documented the samples, anddeveloped the database. The equipmentneeded to digitally document the pipe/tubingwas housed at the WFLETC.

The MFRC compiled all information into asearachable CD format, and helpeddistribute the CDs to the forensiccommunity.

44

DISSEMINATION

The product, as a beta version, was distrib-uted to several state and federal agencies(including the FBI and TSA), at various StateDepartment ATA courses, at forensic confer-ences nationwide, and at the InternationalAssociation of Bomb Technicians and Investi-gators. A copy of the database has also beenposted on the MFRC web site.


• Crippin, J., �Pipe and Tubing Database.�Presented at WFLETC explosivesclasses, Pueblo, CO.

• Crippin, J., �Pipe and Tubing Database.�Presented at State Department post-blastinvestigation classes

.• Crippin, J., �Pipe and Tubing Database.�Presented at American Academy ofForensic Science 59th Annual Meeting,February 2007, San Antonio, TX.

IMPLEMENTATION

The final CD-ROM product is intended fordistribution nationwide to all levels of lawenforcement. This includes both theinvestigative and forensic laboratory levels.To date, TWGFEX/NCFS and Tripwire haveexpressed an interest in placing the databaseas an online source for investigators.

CONTACTS


Jamie CrippinWestern Forensic Law EnforcementTraining Center1007 Candytuft BoulevardPueblo, CO 81001-4901Phone: [email protected]

David GreenLake County Crime Laboratory235 Fairgrounds RoadPainesville, OH 44077Phone: [email protected]

Will RandleMissouri State Highway Patrol CrimeLaboratory1510 East Elm StreetJefferson City, MO 65101Phone: [email protected]

45

Quantification of the Individual Characteristics of theHuman Dentition


Daubert vs. Merrell Dow Pharmaceuticalsestablished several factors for theadmissibility of scientific evidence. Onefactor not available to ForensicOdontologists is the ability to scientificallyquantify the occurrence of specific dentalcharacteristics in the population. Another isthe error rate in the analysis of bitemarks.Without these abilities, an Odontologist�sconclusions lack a scientific basis forprobability expressions and are limited tosubjective opinions.


This project targets the development of adataset which defines the frequency that agiven set of dental characteristics occurs inthe general population.

Specifically, the project aims to:

• develop a computer assistedmethodology for analyzing, measuringand recording the measurement ofdental characteristics

• establish a dataset of six dentalcharacteristics which define theirdistribution in the general population

• investigate and calculate the probabilitythat any two of six dentalcharacteristics are the same

• establish the feasibility of developing adatabase that can be used to define thefrequency a given set of dentalcharacteristics occurs in a population

METHODOLOGY

Dental samples (n=400) are obtainedrandomly from clinic patients. Theserepresent a diverse population composedof Caucasian, Black, Asian, and Hispanicmales, age 18 to 44. The samplepopulation mirrors the national population.

Exemplars are selected from the samplepopulation, scanned as digital images,archived as read-only images andduplicated to create working files for eachinvestigator. Using Adobe Photoshop CS2,and an automated software program deve-loped at Marquette University, data aregathered and measurements recorded.

Recorded information includes incisorwidth, width of the dental arch, degree ofincisor rotation, presence of diastemas(spaces), missing teeth, and accidentaldamage. Statistical calculations are thenperformed on all characteristics.

Figure 1. Graphical association model for the mandibular dental arch.

Graphical Association ModelFor Mandibular Dental Arch

LI = Lateral IncisorCI = Central Incisor

Numbers denote Pair-wisePearson Correlation betweenMeasurements

ArchWidth

Left LI # 23

Angle

Left LI # 23 Width

Left CI # 24 Width

Left CI # 24

Angle

Right CI# 25

Angle

Right CI # 25 Width

Right LI # 26 Width

Right LI# 26

Angle

0.63

0.29 0.21

0.33

0.430.48

0.16

0.43 0.44

0.44 0.43

46

Since tooth position does not occurindependently, a study is also conducted ofthe correlation of jaw width and tooth widthon position, to determine tooth position.Pairwise Pearson Product Momentcorrelations are computed to describeassociations between each pair ofmeasurements.


A protocol was established to assureaccuracy and repeated reliability of thescanner, computer, and software. Theprotocol consisted of making imprints,selecting imprints, scanning imprints asdigital images, archiving the images asread-only images in .psd format, andduplicating the images as working files.

Figure 2. An automated program to read pixels which were inserted into the area to be measured. Each pixel had a different greencolor value from 1 to 250. The software recognized the distance and the degree of rotation.

Adobe Photoshop CS2, along with theautomated software developed byMarquette University, were used to performexemplar measurements. Quality control ofthe measurements was achieved by

comparing measurements read by theautomated computer software program withmeasurements taken manually using themeasure tool in Photoshop. Finalcalculations were performed using SASStatistical Analysis Software.

Research findings indicate that less than5% of the samples have a mandibular archwidth of less than 2.375 centimeters. Usingthe Wald-method, the 95% confidence levelfor the proportion of adult males having amandibular arch width of less than 2.2centimeters ranges from <0.1% to 2.3%. Itwas also found that approximately 50% ofthe patterns exhibited at least one outlieramong the six characteristics measured. Analpha level of 0.05 was used throughout todenote statistical significance.

Table 1. Proportion of exemplars with outlying measurements in mandibular jaw.

The findings established statisticalevidence that quantification of dentalcharacteristics can be accomplished.Ongoing work focuses on refining andvalidating the data derived using the thirddimension to obtain in a morediscriminatory fashion the width of eachindividual tooth.

Collector Radmer Johnson

Number of Exemplars 416 410

Number of Outlying Traits n (%) n (%)

0 205 (49.3) 204 (49.8)

1 109 (26.2) 102 (24.9)

2 61 (14.7) 61 (14.9)

3 22 (5.3) 23 (5.6)

4 9 (2.2) 13 (3.2)

47

The protocol used in this study of dentalcharacteristics showed potentialapplications for analysis of bitemarks, aswell as pattern analysis in actual casework.

TECHNOLOGY BENEFITS

The ability to scientifically express theprobable linkage between a humanbitemark and a suspect establishescredibility and admissibility of bitemarkanalysis. The expansion of this data into adatabase reduces, if not eliminates, thereliance on opinion assessments. It alsodiscourages theanalysis of indistinct bitemark patterns,since the characteristics necessary toapply the database must be clearlyobservable.

COLLABORATION

The project is a collaboration between theWisconsin Department of Justice CrimeLaboratory � Milwaukee and the Universityof Marquette. Partnering with theWisconsin Department of Justice CrimeLaboratory provided two imagingspecialists who helped set up the protocoland calibrate and test the hardware foraccuracy and reliability. It also assures thatthe methodology conforms to theguidelines of the Scientific WorkingGroup on Imaging Technology (SWGIT)and will be applicable to forensic analysis.

DISSEMINATION

Research findings were disseminatedthrough various presentations andpublications and have also beenhighlighted online (www.sciencentral.com),on TV, in several newspapers, and in twoWisconsin Department of Justicepublications. A paper on the research andits findings was recently accepted forpublication by the Journal of Forensic

Identification. A final technicalreport on the project will be posted on theMFRC web site.


• Johnson, L.T., Radmer, T. �Quantificationof the Individual Characteristics of theHuman Dentition: A Preliminary Report.�Oral presentation at the AmericanAcademy of Forensic Science, 58th

Annual Meeting, February 2006, Seattle,WA.

• Johnson, L.T., Radmer, T. �DentalScience Assists Criminal Justice.� Oralpresentation at the American Academy ofForensic Science, 59th Annual Meeting,February 2007, San Antonio, TX.

• Johnson, L.T., Radmer, T. �DentalScience Assists Criminal Justice.� OralPresentation at the InternationalAssociation for Identification,International Educational Conference,July 2007, San Diego, CA.

• Johnson, L.T, Radmer, T. �Quantificationof the Individual Characteristics of theHuman Dentition.� Poster presentation atthe International Association forIdentification, International EducationalConference, July 2007, San Diego, CA.

• Johnson, L.T., Wirtz, T., Radmer, T. �TheVerdict is in: Can Dental Characteristicsbe Quantified?� Oral presentation at theAmerican Academy of ForensicsSciences 60th Annual Meeting, February2008, Washington, D.C.

• Johnson, L.T., �A Methodology for theQuantification of IndividualCharacteristics of the Human Dentition:Methodology.� Article accepted forpublication in the Journal of ForensicIdentification, July/August 2008.

48

IMPLEMENTATION

The technology was made available to threeWisconsin State Crime Laboratories for betatesting. Imaging specialists in Milwaukee,Madison, and Wausau will use thetechnology in the analysis of patternedinjuries.

CONTACTS


L. Thomas JohnsonSchool of DentistryP. O. Box 1881Marquette UniversityMilwaukee, Wisconsin 53201-1881Phone: [email protected]

Technology Users

Ronald GroffyForensic Scientist, ImagingWisconsin State Crime Laboratory - MadisonPhone: 608-266-2031

Aaron MatsonForensic Scientist, ImagingWisconsin State Crime Laboratory - MadisonPhone: 414-328-7500

49

Spectral Analysis of the 3D Fracture Surfaces for EnhancedMatching


Examples of metal fragments found at acrime scene include knife tips, pieces fromweapons, pry tools, or automobile partssuch as antennas or trim pieces.

Physical pattern evidence using both classand individual fragment characteristics isused to link suspects to crime scenes andcrime victims. Individual characteristicsmake objects or substances unique evenamong members of the same class.

There is a need for a method to measurethe nature and characteristics of materialswith a known error rate that can be appliedto performing physical matches.


The overall objective of this study is toevaluate whether surface topographymeasurements can be used to associatemetal fracture fragments. As such, it testsif the fracture feature distribution signa-tures for two surfaces are in fact unique.Also, that they can be relied on to deter-mine if the two pieces sharing the fracturesurface were at one time a single piece.

Specifically, the project aims to:

• develop a testing protocol to improvethe power of the match by utilizing 3Dtopographic data of the fracture surface

• identify proper magnifications andlength scales for a benchmarkspecimen to provide unique fracturesignatures that ascertain the twopieces share the same fractureevent

Figure 1. Forensic samples supplied by the collaborator for investigations

METHODOLOGY

A typical knife material (SS-440C) is used ina flexural three point bend test to determineproof-of-concept. Once proof-of-concept isestablished, hard plastic and glass couponsare tested in addition to metal.

The pairs of fractal surfaces are analyzed bya non-contrast 3D optical interferometer.Interferometer measurements are based onthe principle that when light is reflected fromtwo opposing surfaces, the reflected beamsinterferes with one another to generateinterference fringes which can be used todetermine the distance between two sur-faces.

The collection of displacement measure-ments are then used to acquire surfaceheight topographic maps from the pairs offractal surfaces. These height topographic

50

Figure 2. Measurements and analyses for two fractal surfaces showing the difference in frequency as a function of the norm frequency.

An implementation methodology wasproposed to compare two fracture sur-faces. The methodology consists of sixsteps: 1) select regions of interest andperform 3D surface topography usingdifferent magnifications; 2) calculatespectral frequencies for each surfaceprofile; 3) divide spectra from two surfacesinto zones; 4) examine the difference infrequency (DIF) as a function of the normfrequency; 5) determine the baseline formatch; 6) for each image, compare theDIF relative to the baseline DIF for match-ing surfaces and no-match surfaces.

TECHNOLOGY BENEFITS

The study tests the core assumption that ifthe pattern matches, the surfaces wereoriginally together because a fracturesurface is stochastically complex andtherefore unique. As such, it strengthensthe scientific foundation of the fracturematch practice with data to begin the taskof estimating error rates.

COLLABORATION

This project is a collaborative effort be-tween Iowa State University, Michigan

maps are quantized using spectral analysisfor a broken chisel fracture surface.

The first step of the analysis is to identify thenumber of main features on each image andtheir populations. For a pair of fracturedsurfaces the population of these features arecompared via autocorrelation to provide thequality of the match.

Two main issues or challenges are ad-dressed. 1) the proper magnification and thesize of the data set to identify the propersurface topography population to identifyeach unique feature, 2) the directionality ofthe long wave length features of crackingsuch as river marks.


For the featured knife, a 1D analysis wasconducted along lines on the sample sur-faces. The spectral analysis of the dataresulted in distributions for a range of spatialfrequencies, giving the likelihood of featuretypes (river marks, grain size, dimples) onthe fracture surface. The information wasused to train the 2D full spectral analysis ofthe surface.

The collection of displacement measure-ments was used to acquire surface heighttopographic maps from the pairs of fractalsurfaces for comparison. A Matlab-programwas generated to read the surface heighttopographic maps and to perform the dataquantization match analysis.

The height topographic were quantizedusing spectral analysis involving 2D FastFourier Transform algorithms to decomposethe topographic height data into a finitenumber of frequencies. The comparison wascarried out on a preselected range of thesefrequencies, guided by the 1D measure-ments.

51

IMPLEMENTATION

The Johnson County Sheriff�s CrimeLaboratory is critically assessingimplementation of the test methodology.

CONTACTS


Ashraf BastawrosDepartment of Aerospace Engineering2271 Howe HallIowa State UniversityAmes, IA 50011Phone: [email protected]

Barbara LograssoSchool of Technology426 EERC BuildingMichigan Technological UniversityHoughton, MI 49931-1235Phone: [email protected]

Jeremiah MorrisJohnson County Sheriff�s CrimeLaboratory6000 LamarMission, KS 66202Phone: [email protected]

Technological University, and the JohnsonCounty Sheriff�s Crime Laboratory.

The principal investigators generated thepreliminary samples and experimental dataand analysis. Jeremiah Morris, from theJohnson County Sheriff�s Crime Laboratory,generated simulated forensic metal fracturesamples of the test materials prior to the 3DSpectral Analysis. He also collaborated on thetesting protocol.

DISSEMINATION

The PIs disseminated their findings at apresentation of the International Associationfor Identification (IAI). A paper was alsosubmitted for presentation at the MidwesternAssociation of Forensic Scientists. Currently,a manuscript is being prepared for submittalto a peer-reviewed forensic science journal.The project and its results will also be postedon the MFRC web site in the form of a finaltechnical report.


• Bastawros, A., Lograsso, B., Morris, J.�Spectral Analysis of the 3D FractureSurfaces for Enhanced Matching.� Posterpresentation at the InternationalAssociation for Identification, InternationalEducational Conference, July 2007, SanDiego, CA.

• Bastawros, A., Hui Wang, Lograsso, B.,Morris, J. �Spectral Analysis of the 3DFracture Surfaces for EnhancedMatching.� Oral presentation at the 37th

Annual Midwestern Association ofForensic Scientist Meeting, October 2008,Des Moines, IA.

52

A Method for Lifting Bloody ImpressionsUsing a Lifting Strip Containing Titanium Dioxide


Bloody impressions are of great importanceto the forensic science community as theyare frequently encountered at crime scenesand on evidence submitted to the laboratoryfor analysis. A variety of techniques arecurrently available for enhancing bloodyfingerprints on some non-porous, semi-porous and porous surfaces.

Yet, these require different enhancementprocesses depending on the surface porosityand background color. In addition, many ofthe processes require using toxic chemicalreagents and need to be conducted in alaboratory setting. Since impression evidencein blood cannot always be removed from thecrime scene for analysis in a laboratorysetting, many potentially identifiableimpressions can only be photographed andnot enhanced.


In an earlier study, a lifting strip containingtitanium dioxide was successful in lifting andenhancing bloody fingerprints from severalnonporous and semi-porous surfaces ofcontrasting colors. The lifting strips workedbest when activated with a surfactant. Afterdrying, the strip could easily be removedfrom the surface, lifting the bloody fingerprintonto a contrasting white background forexamination.

METHODOLOGY

The primary objective of this study is toimprove upon methods used in the initialresearch by utilizing the lifting strip containingtitanium dioxide to lift bloody fingerprints,

palm prints, foot prints, and footwearimpressions.

To achieve this, the study will incorporateexperiments on

• pure proteinaceous blood samples andblood samples of diluted proteinaceousmaterial with water

• a larger variety of non-porous, semi-porous, and porous surfaces, includingglass, drywall, wood, ceramic tile, andlinoleum

• aged impressions

• larger lifting strips conducive to liftingbloody palm prints, foot prints, andfootwear impressions

• application of the lifting strip

• new reagents, including methanol andwater, Kodak Photo-Flo 200 with water,Citron detergent with water, and acombination of Kodak-Flo 200 withCitron detergent and water

• air drying lifting strips after activation

• fingerprint enhancement techniques

• presumptive DNA testing on liftedimpressions to determine if DNA isdestroyed during the lifting process


This is a new project that has not startedwork yet.

53

TECHNOLOGY BENEFITS

Immovable objects from crime scenes cannotbe brought back to the laboratory foranalysis. Because of this, many potentiallyidentifiable impressions may only bephotographed and not enhanced therebycomplicating the latent print identificationprocess. With the use of the new uniformlifting strip, these problems can be alleviatedwhile still providing optimal qualityimpressions for analysis.

COLLABORATION

This is a collaborative effort between theNorthville Policy Department and the LatentPrint, DNA, and Trace units of the MichiganState Police Northville Forensic ScienceLaboratory.

DISSEMINATION

The results of this study will be disseminatedthrough presentations at forensic scienceconferences and at trade shows for the lawenforcement community. Research findingswill also be published in forensic journalsand as a technical report posted on theMFRC web site.


This is a new research project with nopublications or presentations to date.

IMPLEMENTATION

Upon completion of the research, theMichigan State Police Northville CrimeLaboratory plans to field test the lifting stripwith intent to incorporate this method intodepartment policies and procedures state-wide. The analyst is also working with theLynn Peavy Company to discussmanufacturing the product. If manufactured,the product will be marketed to law

enforcement agencies nationwide throughforensic catalogs and web sites.

CONTACTS


Jessica ZarateNorthville City Police DepartmentNorthville, MI 48167Phone: [email protected]

Charles MordenNorthville Forensic Science Laboratory42145 West Seven Mile RoadMichigan State PoliceNorthville, MI 48167Phone: [email protected]

54

Testing for Potential Contextual Bias during the VerificationStage of the ACE-V Methodology when Conducting

Latent Print ExaminationsFORENSIC TECHNOLOGY NEED

Recent Daubert challenges to fingerprintevidence and the Analysis, Comparison,Evaluation, and Verification (ACE-V) meth-odology have questioned the reliability ofACE-V methodology as it is currently appliedby experts. Until recently, no research hadbeen conducted to test the effects of poten-tially biasing context information upon finger-print experts. A recent study performed by agroup in England suggests that experts arebiased by context information. However,concerns have been raised about the studydesign, limited number of participants, andthe lack of control group. This study exploreshow expert opinion varies under differentexperimental conditions.


Objectives of the project are to investigatepotential contextual bias during the verifica-tion stage of ACE-V. When verifying theconclusions of a latent print examiner, theverifier typically knows the conclusions ofthe initial examiner. A primary question thatneeds to be answered is: does this priorknowledge alone influence the verifier�sconclusion? A secondary question is: doesthe identity and reputation of the initialexaminer, coupled with prior knowledge oftheir conclusion, influence the verifier�sconclusion.

METHODOLOGY

These questions were answered through acontrolled experiment with experts and non-experts. Experiments involving experts wereconducted at the International Associationfor Identification (IAI) Annual Educational

Conference in Boston, MA. Experimentswith non-experts were conducted at a localcommunity college. The principal investiga-tor (PI) instructs an introductory forensicscience course, and during three semes-ters, a class of about 30 students partici-pated in the non-expert experiments.

Each set of expert and non-expert partici-pants was divided randomly into threegroups. Each group was given a set of sixlatent fingerprints for which they wereinstructed to report two variables for mea-surement. The first variable was the conclu-sion. The second variable was the amountof information used to reach the conclu-sion. Statistical analyses were performedon the data collected.

The materials for each group consisted of aseries of latent print comparisons of varyingdifficulty. The latent prints and knownexemplars for comparison were derivedfrom a sample of prints whose sourceswere known to the researchers.

The expert portion of the experiment wasconducted at the IAI conference in Boston,MA, July 2-7, 2006. Forty-three volunteersparticipated, randomly divided into threeseparate groups.

Group A received a series of six latent printcomparisons for which they were instructedto provide an expert opinion. This was thecontrol group: no contextual informationwas provided. Group B was given the samesix comparisons as the first group, but thistime provided with a conclusion that wassaid to have been reported by an unnamedexpert. Group C was given the same sixcomparisons as the previous two groups,but was presented with conclusions by Pat

55

Wertheim, an internationally recognizedexpert.

The same protocol was followed in experi-ments involving non-experts. The experi-ment with Group B participants (low bias)was conducted in May 2006, while GroupC (high bias) and Group A (control) experi-ments were conducted in September 2007,and January 2007, respectively. Duringthese experiments data were collected andentered into a database. Statistical analy-ses on the data were performed at theUniversity of Lausanne, Switzerland.


Results showed that fingerprint expertswere influenced by contextual informationduring fingerprints comparisons, but nottowards making errors. Instead, fingerprintexperts under the biasing conditions pro-vided significantly fewer definitive anderroneous conclusions than the controlgroup. They tended to provide opinionsthat were inconclusive.

Figure 1. Pooled expert trials assessing images from the same source.

Novice participants were more influencedby the bias conditions and did tend to makeincorrect judgments, especially whenprompted towards an incorrect response bythe bias prompt. This was not the case with

Pooled "Same Source" Trials (Q1-Q4-Q5)

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the fingerprint experts.

Experience (in terms of years of experiencein fingerprint examination) was not shown tobe a significant factor for specialists whenassessing images from the same source, butwas a factor when assessing images fromdifferent sources.

Figure 2. Trail Q4 Opinion vs. the number of corresponding minutiae reported by the expert.

Significant variation was observed for finger-print experts when asked to count the num-ber of minutiae in agreement for trials wherethe images originated from the same source.The number of minutiae in agreement asreported by the specialist was a significantfactor in claiming the images to be a defini-tive match.

TECHNOLOGY BENEFITS

The results of this experiment providesvaluable insight on the effects of potentiallybiasing context information when presentduring the verification of latent print examina-tions. Strong critical voices have called for acomplete overhauling of the identification

56

methodology but presently there is limitedresearch evidence to support this.

The major recommendation that hasemerged is to use a blind testing scheme fordifficult examinations, particularly thoseresulting in an exclusion. Also, when examin-ers are placed in an �alert� state, they tend tobecome more conservative. Therefore,quality control schemes which achieve this�alert� state, such as selection of cases foraudits and random checks, are also encour-aged.

COLLABORATION

This project is a collaboration between theMinnesota Bureau of Criminal Apprehension(BCA), the University of Lausanne Switzer-land (UNIL), and the Arizona Department ofPublic Safety Crime Laboratory.

PI Glenn Langenburg is currently a CertifiedLatent Print Examiner at the BCA. He is alsoa candidate in forensic science at UNIL. Thisproject represents a portion of his thesis worktowards completion of the Ph.D.

DISSEMINATION

The results of this study were presented atseveral national and international meetings,conferences, and symposia. Results of theresearch were also presented at UNIL as partof a thesis defense. A paper on the researchand its findings was accepted for publicationin the Journal of Forensic Sciences. A copy ofthe final technical report on the project and itsfindings will be posted on the MFRC web site.


• Langenburg, G. �Testing for thePotential Effects of Contextual Biasduring the Verification Stage of the

Latent Fingerprint Comparisons.�International Fingerprint ResearchGroup Symposium, March 2007,Canberra, Australia.

• Langenburg, G., �Experiments with theACE-V Process of FingermarkComparisons-including a ContextualBias Study during the VerificationStage,� 30th Anniversary of theCanadian Identification Society, July2007, Montreal, Canada.

• Langenburg, G., Champod, C.,Wertheim, P. �Testing for PotentialContextual Bias during the VerificationStage of the ACE-V Methodology whenConducting Latent Print Examinations.�Presentation at the InternationalAssociation for Identification,International Education Conference,July 2007, San Diego, CA.

• Langenburg, G., �Fingerprint Issues forLatent Examiners.� 42nd WisconsinAssociation for Identification,Educational Conference, March 2008,Wauwatosa, WI.

• Langenburg, G., �Testing for PotentialContextual Bias during the VerificationStage of the ACE-V Methodologywhen Conducting Latent PrintExaminations.� Midwest ForensicsResource Center Annual Meeting,April 2008, Bemidji, MN.

• Langenburg, G., Dror, I., and Srihari, S.�Whose Print Is It Really: DoesConfirmation Affect the Answer?�National Institute of JusticeConference, July 2008, Arlington, VA.

• Langenburg, G., Champod, C.,Wertheim, P. �Testing for PotentialContextual Bias Effects During theVerification Stage of the ACE-V

57

Methodology when ConductingFingerprint Comparisons.� Paperaccepted for publication in theJournal of Forensic Sciences, May2009.

IMPLEMENTATION

The Minnesota BCA is currently writingSOPs that reflect some of therecommendations made. Research findingsand recommendations were alsoincorporated in the course �Advanced ACE-V Applications for Latent Print Examiners�.

The Hennepin County Sheriff�s Office, NewYork State Police, and Los Angeles Policeand Sheriff�s Departments have allrequested copies of the SOPs to considerduring their policy drafting.

CONTACTS


Glenn LangenburgMinnesota Department of Public SafetyBureau of Criminal Apprehension1430 Maryland Avenue, EastSt. Paul, MN 55106Phone: [email protected]

Christophe ChampodUniversity of Lausanne, E.S.C.BCH, CH-1015Lausanne, SwitzerlandPhone: [email protected]

Pat WertheimArizona Department of PublicSafety Crime Laboratory6401 South Tucson BoulevardTucson, AZ 85706Phone: [email protected]

58

Testing DNA Samples for Population of Origin


A large portion of annual DNA criminal casesare �unsub cases� with completely unknowninformation about the perpetrator. For thesethe crime laboratory can provide CombinedDNA Index System (CODIS) DNA profiles.

If the profile is not in the database, the DNAinformation does not provide immediateassistance to field investigators. For suchcases, a forensic technique called �popula-tion test� can be used.


The �population test� uses available equip-ment to provide information about the popula-tion of origin of the DNA sample contributor.The biological basis for the test resides infrequency patterns of a few single nucleotidepolymorphisms (SNPs).

For most SNPs, allele frequencies amongdiverse populations are similar. However, fora small fraction of SNPs, they are quitedivergent. A collection of the most extreme ofthese allele frequencies, called ancestralinformative markers (AIMs), has been identi-fied and characterized in nine populations.

METHODOLOGY

Specific aims of this project are to:

• identify a subset of AIMs that will besufficiently powerful for the populationtest

• assemble components of the populationtest, including chemistry and analysistools, and validate them by characterizingadditional sets of DNAs

• assist collaborators in implementing thepopulation test by providing components,protocols, and technical advice

• publicize the population test to the widerforensic science community


A set of sixteen AIMs was identified for usein the population test. They were selectedbased on: TagMan assay availability,NCBI/dbSNP database identification,international HapMap project genotyping,autosomal distribution in the genome; andhigh frequency divergence between two ormore populations of different continentalorigin.

Figure 1. Number of AIMs required.

Extensive in silico simulations were perfor-med on the sixteen AIMs to establish thetheoretical framework for the populationtest. On the appropriate scale, each of 1000simulated individuals from a European-derived population (EUA) could be clearlyexcluded from African (AFR), East Asian(EAS), American Indian (AMI), and fromAfrican American populations.

Accuracy of Panel Prediction By Number of AIMs

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However, they could not always beexcluded from the South Asian (SAS)population or from the Puerto Ricanpopulation. Also, the test did not tell whichpopulation the sample came from. Yet itcould exclude many populations(particularly those of different continentalorigin) as the source of the sample.

Figure 2. Population test results for 1000 simulated European-derived samples.

The HapMap project database was minedto confirm how the population test wouldperform on humans of known populationorigin. Genotypes were obtained for theselected AIMs in the 90 CEPH trios ofEuropean descent, 90 Yoruba trios fromNigeria, 45 unrelated Han Chinese fromBeijing, and 45 unrelated Japanese fromTokyo.

For each of the individuals except one,populations of origin on different continentswere excluded. For the exception, many ofthe genotypes were missing, demonstratingone complexity of actual data sets. Withmissing data, the population test is conser-vative, possibly failing to exclude somepopulations.

TagMan reagents were assembled andtested, confirming the same genotypes asthe HagMan project. Previously, TagMangenotyping has been used to analyze a few

EUA-1,000 simulated samples

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SNPs for many DNA samples. To make thetest more efficient, technology was developedto analyze 16 SNPs/AIMs for 1- 6 DNAsamples in one regiment. The software forthis process was developed as well as thesoftware to calculate how likely a genotypedsample is from any of the nine referencepopulations.

TECHNOLOGY BENEFITS

The test used equipment available in mostforensic laboratories to provide informationabout the DNA sample contributor�s ancestry.Although the test could not tell which popula-tion the sample came from, it could excludepopulations of different continental origin asthe source of the sample.

COLLABORATION

This project is a collaborative effort betweenthe School of Medicine at Washington Uni-versity, the Missouri State Highway PatrolCrime Laboratory, the St. Louis County CrimeLaboratory, and the St. Charles County CrimeLaboratory.

DISSEMINATION

A publication on the research and its findingsis in preparation. A final technical report onthe project and the project�s findings will beposted on the MFRC web site.


This work was briefly described in a newsand opinion feature article in BioScience,�Crime Scene Genetics: Transforming Foren-sic Science through Molecular Technolo-gies.�, Volume 58, Issue 6 (484-489).

Protocols, training materials, the list of AIMs,and data generated as part of the project areavailable online (http://snp.wustl.edu).

60

IMPLEMENTATION

Discussions are being held with the threecollaborators to implement the test at theircrime laboratories.

CONTACTS


Raymond MillerDepartment of GeneticsSchool of MedicineCampus Box 82324566 Scott AvenueWashington UniversitySt. Louis, MO 63110Phone: [email protected]

61

Midwest Forensics Resource Center

Ames Laboratory130 Spedding Hall

Ames, IA 50011-3020515-296-6372 fax: -4748

[email protected]://www.mfrc.ameslab.gov/

Documents

Midwest Forensics Resource Center - ameslab.gov · biochemistry offers opportunities for developing microscopic analytical devices with unique characteristics. This project develops