Forensic Chemistry Handbook ebook

Table of Contents

Title Page

Copyright

Preface

Contributors

Chapter 1: Forensic Environmental Chemistry

1.1 Introduction

1.2 Chemical Fingerprinting

1.3 Spatial Association of Environmental Incidents

References

Chapter 2: Principles and Issues in Forensic Analysis of Explosives

2.1 Introduction

2.2 Sample Collection

2.3 Packaging

2.4 Sorting

2.5 Documentation

2.6 Environmental Control and Monitoring

2.7 Storage

2.8 Analysis

2.9 Records

2.10 Quality Assurance

2.11 Safety and Other Issues

Conclusion

References

Chapter 3: Analysis of Fire Debris

3.1 Introduction

3.2 Evolution of Separation Techniques

3.3 Evolution of Analytical Techniques

3.4 Evolution of Standard Methods

3.5 Isolating the Residue

3.6 Analyzing the Isolated ILR

3.7 Reporting Procedures

3.8 Record Keeping

3.9 Quality Assurance

Conclusion

References

Chapter 4: Forensic Examination of Soils

4.1 Introduction

4.2 Murder and the Pond

4.3 Oil Slicks and Sands

4.4 Medical Link

4.5 Examination Methods

4.6 Chemical Methods

4.7 Looking Ahead

References

Chapter 5: Analysis of Paint Evidence

5.1 Introduction

5.2 Paint Chemistry and Color Science

5.3 Types of Paint

5.4 Paint Evidence Interpretation Considerations

5.5 Analytical Methods

5.6 Examples

References

Chapter 6: Analysis Techniques Used for the Forensic Examination of Writing and Printing Inks

6.1 Introduction

6.2 Ink

6.3 Ink Analysis

6.4 Office Machine Systems

Conclusion

References

Chapter 7: The Role of Vibrational Spectroscopy in Forensic Chemistry

7.1 Introduction to Vibrational Spectroscopy

7.2 Infrared Spectroscopy

7.3 Infrared Sampling Techniques

7.4 Raman Spectroscopy

7.5 Raman Spectroscopic Techniques

7.6 Applications of Vibrational Spectroscopy in Forensic Analysis

References

Chapter 8: Forensic Serology

8.1 Introduction

8.2 Identification of Blood

8.3 Species Identification

8.4 Identification of Semen

8.5 Identification of Saliva

References

Chapter 9: Forensic DNA Analysis

9.1 Introduction

9.2 Methodology

9.3 Problems Encountered in STR Analysis

9.4 Methodology for mtDNA Analysis

References

Chapter 10: Current and Future Uses of DNA Microarrays in Forensic Science

10.1 Introduction

10.2 What is a DNA Microarray?

10.3 DNA Microarrays in Toxicogenomics

10.4 Detection of Microorganisms Using Microarrays

10.5 Probing Human Genomes by DNA Microarrays

Conclusion

References

Chapter 11: Date-Rape Drugs with Emphasis on GHB

11.1 Introduction

11.2 Molecular Mechanisms of Action

11.3 Societal Context of Date-Rape Agents

11.4 Metabolism Fundamentals

11.5 Biosynthesis of Endogenous GHB

11.6 Absorption and Distribution of Ingested GHB

11.7 Initial Catabolism of GHB

11.8 Chemistry of GHB and Related Metabolites not Requiring Enzymes

11.9 Experimental Equilibrium Constants for Redox Reactions of GHB

11.10 Estimated Equilibrium Constants for Redox Reactions of GHB in Vivo

11.11 Different Perspectives on Turnover of Endogenous GHB Are Consistent

11.12 Disposition of Succinic Semialdehyde

11.13 Conversion of Prodrugs to GHB and Related Metabolites

11.14 Subcellular Compartmentalization of GHB-Related Compounds

11.15 Comparative Catabolism of Ethanol, 1,4-Butanediol, Fatty Acids, and GHB

11.16 Catabolism of MDMA, Flunitrazepam, and Ketamine

11.17 Detection of Date-Rape Drugs

11.18 Special Circumstances of GHB

11.19 Considerations During Development of Field Tests

11.20 Development of an Enzymatic Test for GHB

Conclusion

References

Chapter 12: Forensic and Clinical Issues in Alcohol Analysis

12.1 Introduction

12.2 Blood Alcohol Concentration

12.3 Alcohol Impairment and Driving Skills

12.4 Field Sobriety Tests

12.5 Blood Alcohol Measurements

References

Chapter 13: Fundamental Issues of Postmortem Toxicology

13.1 Introduction

13.2 Tissue and Fluid Specimens

13.3 Specimen Collection and Storage

13.4 Extraction Procedures

13.5 Analytical Techniques

13.6 Interpretation

Conclusion

References

Chapter 14: Entomotoxicology: Drugs, Toxins, and Insects

14.1 Introduction

14.2 The Fly and Forensic Science

References

Color Plates

Index

Copyright © 2012 by John Wiley & Sons. All rights reserved.

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Library of Congress Cataloging-in-Publication Data:

Forensic chemistry handbook / edited by Lawrence Kobilinsky.

p. cm.

Includes index.

ISBN 978-0-471-73954-8 (cloth)

1. Chemistry, Forensic–Handbooks, manuals, etc. 2. Forensic sciences–

Handbooks, manuals, etc. 3. Criminal investigation–Handbooks, manuals,

etc. I. Kobilinsky, Lawrence.

HV8073.F5595 2011

363.25′62–dc22

2010053071

ePDF ISBN: 978-1-118-06222-7

oBook ISBN: 978-1-118-06224-1

ePub ISBN: 978-1-118-06223-4

Preface

In February 2009 a report entitled Strengthening Forensic Science in the United States: A Path Forward was issued by the National Research Council (NRC) of the National Academy of Sciences. The committee members who wrote the report included scientists, judges, lawyers, statisticians, and forensic scientists. The authors of the report recognized that there is an ongoing need to assure that evidence analysis is held to the highest standards and that what is reported in writing and in testimony must be reliable and credible. Forensic science is a large umbrella science consisting of many subdisciplines, including serology, forensic DNA analysis, toxicology, document examination, hair and fiber analysis, arson investigation, firearms and toolmarks, explosives analysis, blood spatter pattern analysis, digital evidence, impression evidence, forensic pathology, forensic anthropology, forensic odontology, and others. Crime scene personnel are trained to identify and collect biological and physical evidence for subsequent laboratory analysis. This evidence can shed light on events leading up to and during the commission of a crime. Often, evidence collected at a crime scene can be associated with either a victim or a suspect. Criminals can often be linked to the crime scene and/or to the victim. At the same time, those falsely accused can be excluded or exonerated based on reliable analysis of physical evidence. Following scientific analysis, the criminalist writes a report and will testify in a court of law. The testimony of an expert witness must be unbiased, accurate, and based on a sound scientific foundation. The admissibility of novel scientific evidence must be determined prior to expert testimony. The use of a novel scientific method to analyze evidence and the subsequent testimony that describes the results of such testing can be challenged for a variety of reasons.

The Strengthening Forensic Science (SFS) report recognized that not all forensic disciplines were at the same level with respect to standards and reliability. This problem is based in part on insufficient funding for forensic research and lack of oversight by any national organization. Although ASCLD-LAB accredits forensic laboratories, American Board of Criminalistics (ABC) certifies forensic analysts, and the National Institute of Justice provides funding for some forensic research projects, techniques and principles used in many forensic disciplines are not at the same level of reliability as that achieved by DNA scientists. The science of using DNA for human identification dates back to the work of Alec Jeffries, who applied restriction fragment length polymorphism analysis for this purpose in the mid-1980s. This major breakthrough was complemented by a second breakthrough technique, polymerase chain reaction, developed by Kary Mullis. He used this in vitro method to amplify a relatively small number of template nucleic acid molecules (derived from biological evidence) into billions of copies for subsequent identification of the DNA donor. These procedures are considered highly reliable and are admissible in all courts in the United States.

The SFS report spells out 13 recommendations that if accomplished would certainly expand funded research and establish the reliability of non-DNA forensic evidence analysis.

The full SFS report, which describes all of the above, is available through the National Academies Press, 500 Fifth Street N.W., Washington, DC 20001.

Some of the issues faced by forensic experts who write official reports and present courtroom testimony about their analysis are (1) whether or not the techniques of the discipline are founded on a sound scientific foundation, so that the experimental findings are accurate and reliable; (2) whether or not there is any significant possibility of human error or analyst bias that could potentially taint the results; and (3) whether or not rigorous standards have been established for interpreting the results.

The authors of the SFS report believe that with the exception of DNA, no subdiscipline of forensic science is sufficiently reliable that a unique identification can be made of physical evidence: for example, “This bullet was fired from this gun and from no other gun in the world”; or “This fingerprint was left by this person and by no other person”; or “This tire track impression was made by this tire and by no other tire”. What is the statistical basis for such a statement? What is the measurement error rate? What is the human error rate? How should conclusions be expressed in a report or in testimony?

With all of this in mind, I invite you to read through the various chapters in this book and keep in mind that the subject matter is about science and technology and not about art. Keep in mind how the expert comes to a conclusion and how that conclusion is reported. What is the statistical basis for the expert's report and testimony? Can the expert testify to a high degree of scientific certainty that the questioned and known specimens have a common origin? I leave that to the reader to decide.

This handbook begins with a review of forensic environmental chemistry which involves the use of trace chemical techniques for investigating environmental spills in an effort to determine if there is any civil or criminal liability. The field can be broken down into two broad areas based on the techniques used to determine liability: chemical fingerprinting and spatial association. In chemical fingerprinting, complex mixtures of chemicals or chemical isotopes are used to associate a spill or environmental release with a specific source. In spatial association, a geographical information system and geochemical techniques are used to attribute the location of a contaminant with a possible source in physical space.

Chapter 2 addresses the principles and issues that exist in the forensic analysis of explosives. It lays out the foundation for proper handling of evidence, which is critical to identifying and convicting the criminal. Evidence at the scene of an explosion, especially a large explosion, offers some unique challenges. Basic principles of evidence collection, handling, storing, and identifying various explosives are discussed herein.

Chapter 3 is a review of arson and fire debris analysis. The isolation and identification of ignitable liquid residues (ILRs) from fire debris is a critically important aspect of arson investigation. This chapter covers common techniques for the isolation and identification of ILRs. Analytical procedures have become more sensitive, and results of testing play a very important role during litigation in a criminal or civil court. Quality control is an important component in fire debris analysis. Reports of findings should be written in a scientific manner describing the fire under investigation, evidence handling, a description of the evidence and where it was collected, the isolation procedure and what testing was done and with what kind of equipment, observations made, and conclusions, with a discussion of the meaning of the results.

Chapter 4 reviews the forensic examination of soils. Soils and sediment are excellent sources of trace evidence in both criminal and civil cases because there are an almost unlimited number of identifiable soil types based on the content of rocks, minerals, glasses, and human-made particles and chemicals. Forensic examination commonly identifies the original geographic location of soils associated with a crime, thus assisting an investigation. Studies of soil and related material samples associated with a suspect and crime scene can produce evidence that the samples had or did not have a common source, thus indicating whether or not a suspect was ever at a particular location. Gathering intelligence for criminal and civil investigations, as well as gem and art fraud studies, often use the methods of forensic geology

Chapter 5 deals with the analysis of paint evidence. Paint and coatings often appear in criminal, civil, and art-authenticity investigations. This chapter reviews the current methodologies and approaches used by forensic paint examiners to analyze this type of physical evidence as well as the problems that they may encounter. Fragments of multilayered in-service paint are one of the most complex types of materials encountered in the forensic science laboratory. They consist of both organic and inorganic components heterogeneously distributed in very small samples, often on the order of only 1 square millimeter. These characteristics dictate the requirements of the analytical chemistry approaches to be used, and they can present a formidable challenge to the forensic analyst responsible for classification of the materials and an evaluation of their evidential significance. Several case examples are presented to illustrate these concepts.

Chapter 6 describes analytical techniques used for the forensic examination of writing and printing inks. The analysis and identification of writing and printing inks and toners are generally very important in document examination, especially when used in conjunction with a reference library. Inks can be differentiated based on the chemistry of colorants, solvents, resins, and additives. Instrumental analysis, including GC–MS, HPLC, and FT–IR and Raman spectroscopy, can often be used following visual examination, microscopic observation, and thin-layer chromatography. Analysis of toners can be performed with XRF, SEM–EDS, or pyrolysis GC. Although chemical analysis of materials used to create documents can provide vast amounts of relevant information and strongly support associations between questioned and known materials, in nearly all cases, the data obtained will not support a conclusion that identifies a particular writing instrument of printing device.

Chapter 7 describes the role of vibrational spectroscopy in forensic chemistry. Spectroscopy is the study of the interaction of electromagnetic radiation with matter to determine the molecular structure of a solid sample or one dissolved in a specific solvent. This interaction depends on the intrinsic properties of the sample material and can be classified by the energy of the probing electromagnetic radiation. Energy can be in the form of ultraviolet, visible, or infrared light, as well as other forms of energy. Infrared spectroscopy is a good technique to use to identify such fibers as acrylics, nylons, or polyesters or paints or alkyds, acrylics, or nitrocellulose. The size of the sample may require the use of microscopic infrared spectroscopy, and the nature of the sample may indicate the use of external reflection spectroscopy or attenuated total reflectance spectroscopy. These techniques are reviewed as well as related methods of sample identification.

Chapter 8 discusses the important science of forensic serology, an important area of modern forensic science. The primary activity of the forensic serologist is the identification of bodily fluids, as these fluid stains are commonly associated with violent criminal cases. Proving the presence of blood, semen, saliva, and so on, can often confirm alleged violent acts.

Chapter 9 reviews the field of forensic DNA analysis. It describes how DNA became a valuable forensic tool in identifying the source of physical evidence left at a crime scene. The use of restriction fragment length polymorphism analysis in the mid-1980s was replaced by the use of the polymerase chain reaction (PCR) method, which is more sensitive, requiring far less high-molecular-weight DNA, uses less hazardous materials, and is faster and more economical. PCR-STR-based genetic profile typing methods have improved in sensitivity over the past 20 years and have become a basic tool in the crime lab. Where nuclear DNA is insufficient to generate a full genetic profile, mitochondrial DNA can sometimes be used to provide identifying information. Also described are low-copy-number procedures and the typing of single-nucleotide polymorphisms within the human genome.

Chapter 10 reviews current and future uses of DNA microarrays in forensic science. DNA microarrays have revolutionized basic research in molecular and cellular biology, biochemistry, and genetics. Through hybridization of labeled probes, this high-throughput technology allows the screening of tens or even hundreds of thousands of data points in a single run. The technology is most advanced with nucleic acids, but protein and antibody microarrays are coming of age as well. Because of the unique ability to screen for large numbers of molecules, such as DNA sequences, simultaneously, the potential utility to forensic investigations is tremendous. Indeed, progress has been made demonstrating that microarrays are powerful tools for use in the forensic laboratory. As the technology matures and associated costs come down, the day that microarray analysis becomes a routine part of the forensic toolkit draws nearer.

Chapter 11 reviews the problem of date-rape drugs such as MDMA, flunitrazepam, and ketamine, with an emphasis on GHB. Recreational, predatory, and lethal doses, metabolism, and diagnostic metabolites are described. Similarities to and differences from the effects and metabolism of ethanol are also discussed. The advantages of field tests to detect date-rape drugs, and limitations of antibodies and advantages of enzymes for field testing, are discussed. The development of a rapid enzymatic test for the detection of GHB is described.

Chapter 12 covers forensic and clinical issues in alcohol analysis. Ethanol, a clear volatile liquid that is soluble in water and has a characteristic taste and odor, is a central nervous system (CNS) depressant and causes most of its effects on the body by depressing brain function. CNS depression is correlated directly with the concentration of alcohol in the blood (BAC). The Estimation of a person's blood alcohol concentration is based on important parameters such as body weight, ethanol concentration of the beverage consumed and number of beverages consumed, and length of time and pattern of the drinking. Because men and women have different body water amounts (men average 68% and women 55%), there are differences between the ethanol concentration achieved in men and women of similar weight for the same amount of alcohol. Various methods are described that can help to determine BAC in the field as well as in the laboratory.

Chapter 13 discusses fundamental issues of postmortem toxicology. The basic principles of forensic postmortem toxicology are presented. This chapter covers the acquisition and usefulness of different specimens, current analytical techniques, and the interpretation of findings. Special problems associated with the interpretation of drug levels include the conditions of the specimens and the effects of postmortem redistribution, postmortem drug changes, pharmacogenomics, drug interactions, and embalming fluid.

Chapter 14 reviews a field of growing importance in forensic science, entomotoxicology: drugs, toxins, and insects. Forensic entomology is gaining widespread acceptance within the forensic sciences as one method of estimating a portion of the postmortem interval by utilizing the time of insect colonization of a body, also known as the period of insect activity. Additionally, insect evidence can be utilized as alternative toxicology samples in cases where no other viable specimens exist. This subfield, known as entomotoxicology, can provide useful qualitative information to investigators as to the presence of drugs in the tissues at the time of larval feeding. The presence of drugs can alter the developmental period of the insects and should always be taken into consideration by the forensic entomologist. The relationship between toxicology and forensic entomology is also examined.

Lawrence Kobilinsky

Contributors

Chapter 1

Forensic Environmental Chemistry

Anthony Carpi and Andrew J. Schweighardt

John Jay College of Criminal Justice, The City University of New York, New York

Summary

Forensic environmental chemistry involves the use of trace chemical techniques for investigating environmental spills in an effort to determine civil or criminal liability. The field can be broken down into two broad areas based on the techniques used to determine liability: chemical fingerprinting and spatial association. In chemical fingerprinting, complex mixtures of chemicals or chemical isotopes are used to associate a spill or environmental release with a source. In spatial association, geographical information systems and geochemical techniques are used to attribute the location of a contaminant with a possible source in physical space.

1.1 Introduction

As technology for trace chemical analysis has expanded in recent decades, so has its application to criminal and civil casework. This has transformed traditional forensic investigations and has expanded their applicability to less traditional areas, such as those involving environmental crimes. Prior to 1950, environmental law in the United States was based on tort and property law and was applied to a very limited number of incidents. Driven by growing environmental awareness in the 1950s and 1960s, the U.S. Congress passed the first Clean Air Act in 1963. This was followed by a slow but steady string of further developments, including the founding of the Environmental Protection Agency (EPA) in 1970 and the passage of the Clean Water Act in 1972, the Endangered Species Act in 1973, and the Comprehensive Environmental Response, Compensation, and Liability Act (Superfund) in 1980. International law began to address environmental issues with the signing of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) in 1975 and other international treaties. These early milestones have been bolstered by recent amendments, new agencies, and renewed funding, all of which make up a series of laws and regulations that define criminal practices and govern civil liability cases involving the environment. Increased legislation and improved enforcement have led to a significant decrease in easily identifiable environmental disasters, such as when the Cuyahoga River in Cleveland, Ohio burst into flames in 1969 as a result of industrial discharge. As these visible issues have diminished, environmental scientists have found themselves faced with questions that are more difficult to identify and are more intractable in nature. This has led, in turn, to advances in the investigative techniques used to investigate environmental crimes.

It is impossible to pinpoint the exact birth date of forensic environmental science. However, one source attributes the origin of the term environmental forensics to the scientific contractor Battelle in the late 1990s (Haddad, 2004). One of the company's specialties is forensic environmental chemistry, and the company provides services in hydrocarbon fingerprinting, contamination identification, and product identification. Regardless of when the field was named, most sources would agree that the field began gathering momentum about 30 years ago. Since that time, various subdivisions have emerged. Some of these divisions have their roots in diverse areas such as geology, toxicology, biology, physics, and chemistry. As such, the term environmental forensics might be considered a misnomer for two reasons. The first is the tendency of the word forensics to be semantically confusing, because it has no real meaning when used in this context. The second is the loss of the word science, for this serves as a necessary reminder of the field's vast and diverse capabilities, spanning across not just one but many sciences.

The term environmental forensics is often misapplied to what should rightfully be called forensic environmental chemistry. For example, environmental forensics has been defined as “the systematic investigation of a contaminated site or an event that has impacted the environment,” a definition that is clearly biased toward the chemistry perspective (Stout et al., 1998). The broad capabilities of the field are unnecessarily simplified to the question: Who caused the contamination, and when did it occur? (Ram et al., 1999). Surely this is not the only question that environmental forensics is capable of answering. Nevertheless, this mindset has persisted because it is acknowledged and reaffirmed repeatedly. Many of the shortfalls of the earlier definitions of environmental forensics have been identified and amended in subsequent definitions. Many of these revisions offer a more generic, all-inclusive definition. One source defines forensic environmental science simply as “litigation science” (Murphy, 2000); another as “environmental ‘detective work’ … operating at the interface junction points of several main sciences including chemistry and biochemistry, biology, geology and hydrogeology, physics, statistics, and modeling” (Petrisor, 2005). Vives-Rego (2004) defines it not just as the environmental application of chemistry, biology, and geology, but as “science and the art of deduction.” Finally, Carpi and Mital (2000) define it as “the scientific investigation of a criminal or civil offense against the environment.” These updated definitions more accurately reflect the capabilities of forensic environmental science beyond the chemistry realm. In particular, the definition provided by Carpi and Mital (2000) specifically includes the use of DNA to solve crimes perpetrated against wildlife and plant life. In this chapter we focus on the specific subarea of forensic environmental chemistry and leave to another source the broader description of the methods and techniques that apply to environmental forensics.

However one chooses to define this growing field, one thing is certain: Forensic environmental science is filling the significant niche left void by forensic science and environmental science. Due in large part to its close association with the core sciences, forensic environmental science has experienced significant growth since its inception, especially in recent years. Aside from technological achievements in the past 30 years, several important advances have helped propel forensic environmental science from a burgeoning offshoot of forensic science to a scientific discipline in its own right. One such advancement was the founding of the journal Environmental Forensics in 2000 (Taylor & Francis, London). Although research pertaining to forensic environmental science occurred before the journal existed, the journal can be credited with offering a place for environmental research that falls under the forensic science umbrella. Thus, Environmental Forensics provides a forum to facilitate the exchange of information, ideas, and investigations unique to forensic environmental science (Wenning and Simmons, 2000).

Forensic environmental science has become such a diverse field that it is difficult to find a single work that adequately covers all its subdisciplines. The literature on the subject that enjoys the most success does so because it focuses on a specific area of forensic environmental science. As such, in this chapter we focus on forensic environmental chemistry. Our aim is to elaborate on several key areas of forensic environmental chemistry, perhaps where other resources have been unable to or have failed to do so. In particular, we focus on chemical fingerprinting and its subsidiaries, such as hydrocarbon fingerprinting, isotope fingerprinting, and complex mixture fingerprinting. Chemical fingerprinting attempts to individualize a chemical and trace it back to its origin. This technique has become increasingly important not only to identify that a chemical spill has indeed occurred, but also to identify the party responsible. We also focus on spatial analysis for the purpose of source attribution. Several cases are discussed that are illustrative of the capabilities of spatial analysis and chemical fingerprinting as they pertain to forensic environmental chemistry.

1.2 Chemical Fingerprinting

Chemical fingerprinting is a subsidiary of forensic environmental chemistry that examines the constituents of a mixture for the purpose of creating a unique chemical signature that can be used to attribute the chemicals to their source. At one time it was sufficient to arrive at a generic classification and quantitation of the chemical mixture so that appropriate remediation measures could be designed and implemented. However, modern analytical techniques that are focused on individualizing and associating a mixture with a source have become increasingly popular, both for liability reasons and because of the recognition and attempt to apportion liability when multiple and/or temporally distant parties may be responsible for chemical contamination. The main objectives of chemical fingerprinting are to characterize, quantitate, and individualize a chemical mixture (Alimi et al., 2003). In this section we provide the reader with a review of some of the constituents of a mixture that are useful for assembling a chemical fingerprint as well as the techniques used to screen for these constituents. The efficacy of these analytes and of detection techniques are evaluated by illustrating their application in several cases.

1.2.1 Hydrocarbon Mixtures

The majority of chemical spills involve hydrocarbon mixtures; as a result, many techniques are tailored for these mixtures (Sauer and Uhler, 1994). Early techniques were used simply to quantify the total petroleum hydrocarbon concentration, but modern techniques must be capable of quantification as well as identification and individualization (Zemo et al., 1995). The latter two are especially important for litigation purposes. However, identification and individualization may also provide for the design of a more effective remediation plan that accounts for dispersal, weathering, and degradation of the chemical mixture (Zemo et al., 1995).

Petroleum hydrocarbon mixtures may be broadly classified into three general groups. Petrogenic hydrocarbons are present in crude oil or its refined products. Pyrogenic hydrocarbons are the combusted remnants of petrogenic hydrocarbons and other by-products. Biogenic hydrocarbons are those that arise from more recent natural processes: for example, swamp gas or the volatile hydrocarbon mixtures released by decaying plant or animal tissue exposed to anaerobic conditions. Within each of these three broad groups, hydrocarbons are generally separated into three types: saturated aliphatics (alkanes), unsaturated aliphatics (alkenes, etc.), and aromatic hydrocarbons. Aromatic hydrocarbons include both light petroleum products [e.g., benzene, toluene, ethylbenzene, and xylenes (BTEX)] and heavier products such as polycyclic aromatic hydrocarbons.

Analytical techniques such as gas chromatography are usually adequate for differentiating among petrogenic, pyrogenic, and biogenic hydrocarbon mixtures because of the unique ratios of alkanes, alkenes, and aromatic structures that can be expected in these mixtures. Furthermore, gas chromatography can be used to differentiate different grades of petrogenic hydrocarbons because crude mixtures have a variety of hydrocarbon components (i.e., unresolved complex mixtures), which often present themselves as a “hump” on a chromatogram, whereas more refined mixtures have less variety in their components. Retention time for various petrogenic compounds is affected by the structure of these compounds: for example, gasoline elutes first (C4 to C12), along with Stoddard solvents (C7 to C12), which are followed by middle distillate fuels (C10 to C24), and crude mixtures (up to C40) (Zemo et al., 1995). Crude oil mixtures contain a diverse array of hydrocarbons and, on average, are comprised of 15 to 60% paraffins and isoparaffins, 30 to 60% naphthenes, and 3 to 30% aromatics, with the remainder of the mixture being composed of asphaltenes and various trace compounds (Bruce and Schmidt, 1994). Pyrogenic hydrocarbon mixtures can be recognized on the chromatogram because large molecules undergo combustion first, leaving behind a disproportion of smaller molecules. However, pyrogenic compounds are more difficult to attribute to a source because the chemical signature is further removed from the original petrogenic source (Bruce and Schmidt, 1994). Steranes and hopanes are often used as target analytes when the focus of a study is biogenic hydrocarbons, because these analytes are more resistant to many more forms of weathering than are other biogenic components (Alimi et al., 2003).

Although it is worthwhile to classify a mixture as petrogenic, pyrogenic, or biogenic in origin, this is commonly not enough. To arrive at a unique chemical signature, the analysis must extend beyond identifying the class characteristics of a mixture. Modern methods often involve the examination of ancillary components of a mixture, such as dyes, additives, stable isotopes, radioactive isotopes, biomarkers, polycyclic aromatic hydrocarbons (PAHs). PAH homologs, and metabolized PAHs. It is customary to screen for many of these analytes with the intent of providing the most comprehensive chemical signature possible. Before selecting a suite of analytes, it is wise first to consider if these analytes may already have been present at a location (due to a prior contamination or natural processes) and if these analytes are highly susceptible to degradation. Indeed, the characteristics of a good chemical marker are that it is resistant to degradation and that it can uniquely identify the hydrocarbons released from other sources (Sauer and Uhler, 1994).

The first step in confirming hydrocarbon contamination is accomplished by screening for saturated hydrocarbon molecules such as pristane and phytane. These are isoparaffins that are resistant to degradation and are highly indicative of hydrocarbon contamination (Sauer and Uhler, 1994). Pristane and phytane usually represent themselves to the right of C17 and C18 peaks on a chromatogram. Fresh hydrocarbon mixtures have prominent C17 and C18 peaks in relation to pristane and phytane peaks, whereas the converse is true for degraded mixtures (Bruce and Schmidt, 1994; Morrison, 2000b). Due to the proportionality between the ratios of these compounds and the extent of degradation, the ratios of pristane and phytane to the C17 and C18 peaks are often used to estimate the degree of weathering.

1.2.2 Polycyclic Aromatic Hydrocarbons

Polycyclic aromatic hydrocarbons (PAHs) are hydrocarbon compounds with two to six rings. Homologs of the PAH compounds may be similar to the parent compound except that they are substituted for by one or more alkyl groups. The ratio of two PAHs to two other PAHs is sometimes expressed in double-ratio plots, in which certain regions of the plot are diagnostic for one source or another. It is also becoming increasingly common to screen for metabolized PAHs (as well as BTEX compounds), whose structures differ predictably from the original PAH. PAHs are often very useful in studies involving weathered mixtures, because the complex structure of PAHs makes them more resistant to degradation. The rate of degradation is proportional to the complexity of the ring structure, with the compounds having the fewest number of rings degrading first (Alimi et al., 2003). Some target parent PAH compounds and their alkyl homologs are shown in Table 1.1.

Table 1.1 Target Parent PAH Compounds and Their Alkyl Homologs

Source: Alimi et al. (2003).

One of the most prominent applications of PAH analysis has been the study of the Exxon Valdez oil spill. The spill occurred when the tanker hull was punctured as it ran aground on March 24, 1989, releasing some 10.8 million gallons of oil into Prince William Sound, Alaska. The oil released was dispersed by water currents and a windstorm that followed the spill a few days later, and concern was raised over the dispersal of the oil into adjacent bodies of water (Galt et al., 1991). Many of these concerns were seemingly corroborated by the detection of oil in neighboring bays. However, it was speculated that some of the oil detected in these neighboring waters may have been from biogenic sources, petrogenic sources from previous spills, or pyrogenic sources from hydrocarbons that had previously undergone combustion. A massive effort was mounted to identify the extent to which Exxon was responsible for oil detected in these adjacent waters.

The study immediately focused on components of oil that were the most resistant to degradation, such as PAHs and biomarkers. A substantial part of the investigation focused on evaluating the effects of weathering on the Exxon Valdez cargo if it was to be accurately differentiated from other sources (Figure 1.1). As expected, lighter components of the oil matrix were preferentially lost to weathering. With the effects of weathering understood, the investigation then turned to PAH analysis. Two PAHs that were focused on for distinguishing different crude mixtures were phenanthrenes and dibenzothiophenes; chrysenes were used to differentiate crude from refined mixtures because chrysenes are removed during the refining process (Boehm et al., 1997). The ratios of the PAH compounds to one another were particularly useful because the concentrations of the PAHs will change with weathering; however, the ratio of one PAH to another generally remains constant (Boehm et al., 1997). In this case, researchers created a double-ratio plot comparing dibenzothiophenes to phenanthrenes in order to distinguish PAHs of the Exxon Valdez spill from PAHs of other sources (Figure 1.2). As seen in the figure, the double-ratio plot showed distinct clustering of oil samples from different sources, allowing a differentiation to be made. When the PAHs in neighboring bays were analyzed, some were attributed to the Exxon Valdez spill, but many were found to have originated from other sources, both natural and anthropogenic (Boehm et al., 1998).

PAHs have also been used to study contamination at former manufactured gas plant (MGP) facilities. Prior to the use of natural gas, MGPs made coal gas to use as fuel. Former MGP sites are evaluated for contamination by screening for PAHs that would have been introduced to the environment as coal tar, which is a by-product of the coal gas manufacturing process. This can sometimes be a difficult task because the sites often contain PAHs that may be unrelated to the MGP, having been introduced via other natural and anthropogenic avenues. The investigations are further complicated because similar PAH signatures are obtained for MGP coal tar residues and background residues. Although the composition of the PAHs contained in MGPs and background sources may be similar, the PAH ratios and patterns (i.e., petrogenic or pyrogenic) can be used to differentiate PAHs from different sources. One study examined the ratios and patterns of PAHs for the purpose of distinguishing MGP PAHs from background PAHs in soil samples collected in and around a stream near an MGP (Costa et al., 2004).

PAHs may be present either as unsubstituted parent compounds or as a substituted alkyl homolog (see Table 1.1 for examples). Petrogenic patterns of PAHs are recognized because they contain a bell-shaped distribution of the parent PAH and its homologs where concentration of the single- or double-substituted homologs are highest, and concentrations decrease as one moves in either direction toward the unsubstituted parent or toward the complex, multisubstituted homolog (see Figure 1.4 for an example). Pyrogenic patterns of PAHs are recognized because they contain a distribution in which the parent PAH is more abundant, due to preferential combustion of the substituted homologs. Researchers observed a pyrogenic pattern in PAH residues derived from the MGP site in question (Figure 1.3), but samples collected from an adjacent stream indicated a mix of petrogenic and pyrogenic PAHs (Figure 1.4).

The researchers then turned to high-molecular-weight PAH ratios to determine if the pyrogenic pattern observed in the stream was from weathered MGP residues or from recent background contamination. Several PAHs were chosen to create double-ratio plots in which certain sections of the plot were diagnostic for either the MGP, background sources, or a mix of the two. A comparison of samples from the streambed surface (Figure 1.5) and samples from the streambed subsurface (Figure 1.6) indicated that most of the surface (i.e., newer) PAHs were derived from background sources, whereas most of the subsurface (i.e., older) PAHs were derived from the MGP site. Results of studies such as this can help to draw attention to other potential sources of contamination in order achieve the most efficacious remediation effort.

1.2.3 Biomarkers

Biomarkers such as steranes and hopanes are hydrocarbon remnants of deceased organisms that are useful in chemical fingerprinting because they are extremely resistant to weathering (Alimi et al., 2003). Thus, biomarkers can often be useful to individualize a hydrocarbon mixture when saturated hydrocarbons and PAHs have already been degraded (Sauer and Uhler, 1994). A comprehensive list of biomarkers that are useful in hydrocarbon mixture studies is provided by Alimi et al. (2003).

1.2.4 Additives

Inorganic compounds are often added to hydrocarbon mixtures to serve as antiknock agents, octane boosters, corrosion inhibitors, and anti-icers (Kaplan, 2003). Additives are not present in crude mixtures, of course, so their presence is indicative of a refined mixture (Bruce and Schmidt, 1994). Because refining practices and additives change over time, and since these changes have been well documented, the presence of particular additives in a hydrocarbon mixture can be highly indicative of a certain time frame during which a sample was produced. For example, lead was first added to gasoline in 1923, and its concentration in gasoline decreased steadily until it was phased out in U.S. automobile fuels in 1995 (Kaplan, 2003). Other gasoline additives that have predictably appeared and disappeared throughout history are methylcyclopentadienyl manganese tricarbonyl (MMT) and methyl tert-butyl ether (MTBE). The chronology of some popular additives has been thoroughly documented in several sources (Morrison, 2000a,b; Kaplan, 2003). Some additives, such as lead, have been used over large time frames, but the concentration of lead in gasoline has varied predictably over the years. Although this can be used to arrive at a reasonable estimate of time of manufacture of the hydrocarbon mixture in question, it is not an infallible method because additive concentrations are often reported based on a pooled standard, which ignores batch-to-batch variation (Morrison, 2000a).

The use of additives for dating a release can be complicated by the fact that additives may be discontinued in certain countries or for certain applications, but may still be used in others. Additives that are supposedly absent in a mixture may also be present in very dilute amounts. The utility of additives in dating manufacture or release dates is greater than their capacity to individualize a mixture. This is because many companies often purchase additives from the same manufacturer. These additives are then added unaltered into various hydrocarbon mixtures, so many different mixtures may have the same additives present (Morrison, 2000a). Further complications when screening for old additives may be encountered because these compounds often contain oxygen, which contributes to their rapid weathering over time (Morrison, 2000b).

1.2.5 Isotopes

When complex hydrocarbon mixtures cannot be identified by analysis of stable components, the mixtures can be identified by analysis of stable isotopes within the mixture. Stable isotopes are often analyzed with respect to one another. In other words, the ratio of one stable isotope to another stable isotope within the same mixture can often be unique, thereby allowing for the creation of an isotope signature. In contrast to stable isotopes, unstable isotopes decay predictably such that the degree of decay can be correlated with the age of the mixture. Analysis of unstable isotope decay is often referred to as a long-term method because it is capable of estimating release dates thousands of years prior. Unstable isotopes are also useful because their decay is independent of environmental factors such as weathering (Kaplan, 2003).

Isotopes can be useful in chemical fingerprinting in two ways. The ratio of two isotopes can be compared as a means of individualization because no two mixtures will have exactly the same ratio of two isotopes. Carbon and lead isotope ratios are commonly used for source identification. Radioactive isotopes are also useful for dating a release because these isotopes have known rates of decay that are independent of environmental conditions.

Carbon isotopes were used in one study to determine the origin of soil gas methane near the site of a prior gasoline spill (Lundegard et al., 2000). The investigation was triggered by the detection of high methane levels near a service station where approximately 80,000 gallons of gasoline had been spilled 20 years earlier. Initially, it was speculated that the methane was due to the bacterial degradation of the gasoline, but the investigators were considering other possibilities. Suspicion was raised because high levels of methane were detected outside the original gasoline plume, and in some cases the levels detected outside the plume were higher than those within the plume (Figure 1.7).

The initial hypothesis of methane generation by bacterial degradation of the gasoline was also challenged because this is not a common degradation pathway. For gasoline to be fermented to methane, it would first have to be converted to the necessary precursor compounds for methanogenesis by fermentation (Lundegard et al., 2000). Although the generation of methane via this pathway is possible, the investigators were considering more plausible origins of the methane that, coincidentally, were unrelated to the gasoline spill. One of the potential origins considered was the biodegradation of organic matter.

The methanogenesis of petrogenic compounds can be distinguished from that of organic compounds from biogenic origins through the use of 13C, which is a stable carbon isotope. Differentiating the methanogenesis of petrogenic and organic compounds is accomplished based on the idea that older, petrogenic compounds have lower quantities of 13C isotopes than does newer organic matter. The process of methanogenesis significantly reduces the amount of 13C present in the original organic matter, but the 13C in the nascent methane remains stable regardless of environmental conditions (Lundegard et al., 2000). The study indicated that wood fill from beneath the service station site and gasoline from within the original plume had indistinguishable 13C quantities.

Another way of differentiating methane from petrogenic and biogenic sources is through the use of 14C, which is a naturally occurring radioactive isotope of carbon taken up by all living organisms. The age of the source from which the methane was formed can be predicted because 14C has a half-life of about 5700 years, and therefore it will still be detectable in methane formed from biogenic organic matter less than 50,000 years old. The hypothesis that methane originated from the degradation of biogenic organic matter was corroborated by the 14C analysis, which indicated that the highest 14C levels were detected outside the original plume (Figure 1.8). The level of 14C in petrogenic hydrocarbons is zero, so the researchers concluded that the methanogenesis must be of biogenic origins. This hypothesis was further supported because a review of the site history indicated that the area consisted of organic fill, including wood and sawdust.

1.2.6 Tracers

When none of the analytes discussed previously are amenable to the case at hand, techniques that rely on tracers can sometimes be used for forensic tracking of environmental chemicals. A tracer can be any molecule that is diagnostic of one source but not others. Sometimes, multiple tracers are used to augment the significance of the results. One study used organic tracers to determine the origin of gas- and particle-phase air pollutants in two California cities (Schauer and Cass, 2000). The objective of the study was to determine the primary source(s) of air pollutants in Fresno and Bakersfield, California. The results and chemical composition of samples collected at the two locations were compared to those collected at a remote site located at the Kern Wildlife Refuge and distant from anthropogenic sources of air pollutants. Previous tests for air pollutants have used generic compounds to draw connections between air pollutants and their sources (Harley et al., 1992). However, some of the analytes used in these other studies are not exclusive to a particular source. The researchers in the California study aimed to develop a more accurate method for tracing the origin of the air pollutants. Atmospheric samples in the two cities were collected, as well as single-source control samples that consisted of combustion emissions from gasoline-powered motor vehicles, diesel engines, hardwood combustion, softwood combustion, and meat-cooking operations. Tracers that were unique to sources and those that were common between multiple sources were chosen both to fingerprint and then apportion emissions in particulate samples with mixed origins. Further criteria used to choose tracers were (1) that they were not selectively removed from the environment, and (2) that they were not formed by atmospheric reactions to any significant extent. The researchers used direct measurements of these tracer compounds to draw conclusions about the source of particulate pollutants in the areas indicated.

Specific tracers were used to apportion the results obtained from specific sources. For example, the compound levoglucosan was found to be specific to wood combustion, so the concentration of levoglucosan in proportion to other constituents in a mixed sample could be used to apportion the contribution of wood combustion to particulate loading in an area (see Figure 1.9). Based on the concentrations of the other tracers in the samples, the relative contributions of each source (e.g., automobiles, wood combustion) were apportioned (Figure 1.10). Low levels of pollutants derived from anthropogenic sources at the remote site were noteworthy. The researchers concluded that local anthropogenic emissions (particularly automobile exhaust) were responsible for the majority of air pollutants in the two urban environments, whereas naturally occurring dusts were primary contributors at the remote site.

Inorganic compounds can also be used for forensic chemistry purposes. One study used metal tracers to identify dust that resulted from the collapse of the World Trade Center (WTC) buildings (Scott et al., 2007). The analysis of particulate matter arising from this catastrophic event has been an area of great interest because there are significant health implications associated with inhalation of the dust. Substantial amounts of the dust were transferred from the collapse to nearby buildings, so the objective of the research was to develop a method based on metal tracer detection to determine which buildings were most affected, and for those that were severely affected, to determine if appropriate remediation efforts had been undertaken.

Techniques employed included screening for human-made vitreous fibers, as well as trace metals, including As, Cd, Cr, Cu, Pb, Mn, Ni, V, and Zn. Trace metal detection was found to be more applicable to the identification of WTC dust because the atmosphere and buildings around the WTC were probed routinely for these metals after the collapse (Scott et al., 2007). Although these metals can originate from other sources as well, the researchers expected trace metals to be detected in quantities and ratios that were unique to WTC dust.

Concentrations of the nine metals in WTC dust as reported by four studies were compared to concentrations in background dust collected from Arizona. A discriminant analysis model was used to classify each sample as having originated from WTC or background dust based on the relative concentrations of the nine metals (Figure 1.11). The analysis indicated that WTC dust had elevated levels of Cr and Mn and low levels of As, Cd, and Cu compared to background dust. The researchers were able to demonstrate that trace metals could be used to distinguish pure WTC dust from background dust with 94% accuracy; however, mixed dust samples had lower levels of accuracy (Scott et al., 2007).

1.2.7 Methods of Detection

One of the most widely used techniques in chemical fingerprinting for hydrocarbons is gas chromatography. This is based on the concept that each compound has a unique structure and will therefore be retained differentially in the gas chromatograph before being eluted. As long as other parameters (e.g., temperature, column length, column packing) are held constant, any differences in retention time can be attributed to the structure of the compound (Bruce and Schmidt, 1994). Mixtures contain many different compounds, so a gas chromatogram represents a chemical fingerprint of all the chemical constituents in a mixture. Gas chromatography is often combined with other techniques to achieve a more detailed analysis. For example, a gas chromatograph is commonly used as a preliminary separation technique that is followed by detection using mass spectroscopy. Some researchers even use two-dimensional gas chromatography (GC × GC) to achieve superior resolution (Gaines et al., 1999). A good review of the literature focusing on these techniques is provided by Suggs et al. (2002). The potential weaknesses and vulnerabilities of these techniques are discussed by Morrison (2000b).

Although many of the aforementioned techniques are highly effective, they often have a deleterious impact on the sample. That is, substantial portions of the sample are often destroyed in the course of the analysis. Sample destruction may not be a major concern in other disciplines, but evidence is sometimes limited in forensic investigations, and what little sample may be available often attains the status of a precious and rare commodity. Another example of when the destruction of a sample is avoided is when the sample itself is, quite literally, a rare commodity, such as an archaeological treasure or artifact. For samples of limited quantity or prized value, less invasive methods of analysis are often sought.

One study that warranted the use of a minimally invasive technique involved the analysis of ancient tools made of obsidian (Tykot, 2002). The purpose of the analysis was to evaluate the Mediterranean sources and trade routes of obsidian tools without damaging them. To bolster the results and to compensate for the potential weaknesses of certain techniques, this study relied on a series of methods, including scanning electron microscopy (SEM), x-ray fluorescence (XRF), neutron activation analysis (NAA), and inductively coupled plasma mass spectroscopy (ICP-MS). The elemental compositions indicated by the four techniques were used to construct possible sources and distributions of obsidian. The results helped lend credence to the theory of a vast distribution network for the tools rather than the lone source theory that was once promulgated.

1.2.8 Weathering

When screening for various analytes, one variable that must be kept in mind is weathering. This is the process by which the chemical signature of a mixture is altered due to evaporation, dispersal, biodegradation, or oxidation of certain components of the mixture. Short-chain hydrocarbons are most vulnerable to weathering mainly because their simple structure makes them susceptible to degradation, particularly to biodegradation (Alimi et al., 2003). Compounds with more complex structures are generally more resistant to weathering. Weathering generally occurs at predictable rates such that the age of the mixture can be estimated accurately based on the relative amount of weathering of short-chain hydrocarbons to larger, more resistant molecules. However, weathering can vary because of site-specific differences in environmental conditions (Morrison, 2000b). The compounds that exhibit the greatest longevity are generally the most useful for estimating the degree of weathering and therefore the age of a particular compound. Isotopes, BTEX compounds, PAHs, and biomarkers have all been used with varying degrees of success for determining the extent of weathering of hydrocarbon samples.