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CSI - Application of Forensic Chemistry

Introduction Definition of Forensic Chemistry
Forensic science is the scientific method of gathering and examining information about
the past. The most prestigious and well-known use for forensic science is within the
field of crime scene investigation, in which the field is called upon to identify trace
materials and substances, in addition to analysing samples from crime scenes and areas
of criminal suspicion to aid law enforcement officers/organisations.
Forensic scientists provide objective scientific evidence for use in courts of law to
support the prosecution or defence in criminal and civil investigations. Primarily, their
concerns focus on trace material associated with crimes. This follows the established
key principle known as Locards Exchange Principle, that states whenever someone
enters or exits an environment, something physical is added to and removed from the
scene

This principle can be summarised by simply stating that every contact leaves a
trace that could potentially be linked to a criminal suspect.
Chemistry is a wide-ranging science concerned with the synthesis, structures,
dynamics, properties and transformations of all types of materials organic, inorganic
and biological.

At its core, Chemistry is about applying the scientific method to examine
interactions between all types of substances.
A chemists role is to search for and discover information about matter, and research
how this new information can be applied in the world. Chemists are also known for
designing and developing instruments and equipment used to aid chemistry studies.
Forensic chemistry is a fusion of these two fields. It is the application of chemistry to
crime scene investigation, law enforcement and, less ordinarily, the failure of products
or processes. Forensic chemistry is unique among chemical sciences in that its
research, practice, and presentation must meet the needs of both the scientific and the
legal communities.
There are many different analytical methods that can be used to identify chemical
changes that have occurred during an incident (criminal or not), and so help to
reconstruct the sequence of events leading to the incident. Many disciplines and
techniques used in forensic chemistry for crime scene investigation will be detailed
within this essay.




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Collecting Evidence Samples
Before any analysis can take place, evidence from the crime scene must be collected.
Evidence must be evaluated to see if it is worth the valuable time and money it will take
to analyse. Examples of samples that may be collected at a crime scene include but are
not limited to biological evidence (e.g. blood, body fluids, hair, and other tissues), latent
print evidence (e.g. fingerprints), trace evidence (e.g. fibres, soil, glass fragments) and
drug evidence. Chemical analysis can help forensic scientists identify substances and, by
extension, suspects in a criminal case.
It is important for a forensic scientist to not inadvertently damage or destroy evidence.
As a result, crime scenes are photographed and documented before any sampled are
directly collected. When collecting trace samples, forensic scientists utilise swabs and
other tools to collect potential evidence. Once samples are collected, depending on their
nature, they are sent to an appropriate department such as a forensic laboratory. It is
here that chemical analysis takes place.
Methods of Chemical Analysis
For effective use within the field of crime scene investigation, it is necessary to separate
and identify potentially several individual components of an unknown substance or
mixture. One method that is at a chemists disposal is the use of a gas chromatograph-
mass spectrometer (GC-MS). This technique is known as a tandem or hyphenated
technique as it consists of two instruments attached together physically.
The first section is the gas chromatograph. This portion separates the chemical mixture
into pulses of pure chemicals, with chemical separation based on each substances
volatility (or ease at which they evaporate into a gas). Simplified, the gas
chromatograph is a temperature-controlled oven containing a (typically) coiled,
polymer-coated glass column, that can be anywhere between 1 40m in length.
The technique involves a small volume (a few microlitres) of an unknown sample that
has been dissolved in an organic solvent being quickly injected into the hot column
(with temperatures ranging from 40C to 320C). Components with high volatility are
vaporized first by the heat of the oven and are forced toward the end of the column by
the constant flow of an inert (non-reactive) carrier gas. This carrier gas tends to be
helium or a similar gas. Different substances eventually emerge from the end of the
column (are eluted) in differing amounts of time. The time taken for a substance to
travel through the column is known as the retention time. This retention time can be
compared with the retention time of standard molecules that have been eluted using the
same method and settings (e.g. temperature program, carrier gas, rate of flow of carrier
gas, etc.). Though this method can provide a reasonable indication as to which
chemicals are being dealt with, this stage of the process is primarily for chemical
separation.

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To provide an accurate and reliable identification of the chemical components of the
sample, the eluent (matter that has been eluted) is fed into the mass spectrometer
section. The mass spectrometer bombards the sample with fast-moving electrons,
causing the atoms within the sample to be ionised. These ions are accelerated by an
electric field, and are separated (or filtered) as a result of their mass causing the ions to
travel at different speeds. Ions with a small mass travel much faster than ions with a
large mass. The fast-moving ions then come into contact with an ion detector which
records the time taken for the ions to travel through the mass spectrometer. This
provides a fragmentation pattern in the form of a mass spectrum, which is used to
identify the elements within the sample, similar to how a humans fingerprint is used to
identify a person. The mass spectrum provides us with data relating to the abundance of
ions with certain masses. Using the periodic table, we can determine what elements are
present.
Another instrument used for precise identification of compounds is an Infrared
Spectrometer. Within an IR spectrometer, the sample is bombarded with infrared
radiation. If you project a range of IR frequencies individually through a sample of an
organic compound, some of the frequencies are absorbed by the compound. A detector
on the other side of the IR spectrometer shows that some of the frequencies passed
through the compound with close to no loss, but other frequencies were strongly
absorbed. How much of a particular frequency gets through the compound is measured
as the percentage transmittance.
Each frequency of light (including infra-red) has a specific energy. If an individual
frequency of light is being absorbed as it passes through the compound under
investigation, it must mean that its energy is being transferred to the compound.
Energies in infra-red radiation correspond to the energies involved in bond vibrations.
Bonds in compounds are vibrating all the time. If you project exactly the right amount of
energy onto a bond, you can force it into a higher state of vibration. Different bonds
need different amounts of energy, and hence difference frequencies of infra-red
radiation. As a result, we can use the frequency that a bond is absorbing large amount of
infra-red radiation at to identify the bond.
An IR spectrum charts the amount of light absorbed vs the wavelength. This is
represented typically by Wavenumber (1/Wavelength in cm, cm-1) on the x axis and
Percentage Transmittance (%) on the y axis. Similar to the GC-MS, this spectrum can be
compared to that of a known sample, providing evidence for the identification of a
compound.




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A third type of spectroscopy is known as Ultraviolet-visible-near infrared spectroscopy
(UV-visible-NIR spectroscopy). This technique is used to compare known and
questioned sampled of trace evidence such as fibres/strands, and is also used in the
analysis of inks and papers of questioned documents. Considering that any sample
analysed using this technique is not altered in any way, this technique is regarded as
non-destructive.
Drug Chemistry
In forensic drug chemistry, scientists use their findings to help investigators pursue
legal action against individual(s) suspected of a drug-related crime. The goal of forensic
drug chemistry is to determine whether the sample submitted to the forensic laboratory
contains an illegal substance. Based on the results of the analysis, law enforcement can
pursue criminal charges and the court can determine appropriate sentencing.
To analyse drug samples, the submitted samples are weighed and the sample weights
are recorded. The first stage of drug analysis is known as presumptive
testing/screening, which determine the general characteristics of the sample material to
narrow down the range of confirmatory tests that will be used. Examples of
presumptive tests include:
Microscopic analysis - looks at the structure of the material to make a
preliminary estimate.
Microcrystalline test - involves dissolving a small amount of the sample in a
solution and letting the material form crystals. Different substances crystallise in
different but specific manners, allowing analysts to identify components of a
sample by viewing the crystals under polarised light.
UV Spectroscopy - Explained previously
Once presumptive testing/screening is complete, forensic scientists move on to
confirmatory testing. This generally requires two steps, with the first step being to
separate the compounds using gas/liquid chromatography, capillary electrophoresis, or
wet chemistry. Capillary electrophoresis involves using an electrical field to separate
the components of a substance inside a capillary tube. Molecules move toward the
positive or negative charges places at either end of the tube and analysis can measure
the speed and direction at which they move to compare against references. Wet
chemistry involves the use of liquid solvents to separate components of compounds.
Once the first step of confirmatory testing is complete, the compounds are generally
identified using mass spectrometry or infrared spectroscopy, as explained previously.




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Blood Chemistry
Though part of forensic science involves bloodstain pattern analysis (explaining and
determining the projectile motion of blood after wounds/impacts), chemistry can be
used to identify traces of blood where it is not obvious. These blood traces can also be
compared to blood references from suspects in a criminal case using chemical analysis.
Similar to testing for drugs, testing for drugs involves presumptive tests and
confirmatory tests. An example of a presumptive test is the Luminol test. This method,
frequently used in forensic chemistry, involves using a chemical known as luminol
(C8H7N3O2, a derivative of phthalic acids) which reacts with metal cations, and hence
detects traces of blood. Luminol is mixed with a dilute solution of hydrogen peroxide,
which is spread carefully in places where it is thought blood traces exist.
Within the chemistry behind the process, the iron in the haemoglobin within the blood
catalyses an oxidation reaction in which the luminol gains oxygen atoms while losing
nitrogen and hydrogen. The compound product of the reaction is the anion 3-
aminophthalate. The electrons with this compound are in an excited state. Blue light
(blue luminescence) is emitted as energy is released when the electrons return to their
ground state.
An example of a confirmatory test for blood is the Takayama test. After applying a
specific solution developed by Takayama, haemochromogen crystals form by treating a
small amount of blood. These crystals are observable under a microscope and appear
as pink rhomboid crystals. This test requires a relatively large amount of sample,
however (0.1mg Haemoglobin).
Toxicology
Toxicology is the study of the adverse effects of chemicals on living organisms. Forensic
toxicology improves upon this, and applies methods and disciplines involved in
toxicology to detect and interpret signs of drugs and poisons. This is useful in crime
scene investigation as traces of illicit substances can be found within bodily fluid
samples and used as evidence in court.
Forensic toxicologists employ a large number of analytical techniques to determine the
drugs or poisons relevant to a case investigation. Though toxicology techniques can be
as simple as paper chromatography, most commonly used drug screening tests involve
immunoassay techniques. Immunoassays are laboratory tests that use antibodies to
detect a reaction with specific substances. They utilise the specific 3D nature of
antibodies and how they only bind with specifically shaped substances to form
antibody-antigen complexes. This makes antibodies useful for testing for specific
macromolecules (sometimes referred to as analytes), which are in many cases a protein.


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Immunoassays come in many different formats (e.g. ELISA (enzyme-linked
immunosorbent assay)), but are based on the premise that the antibodies will still be
detectable (generally by some kind of chemical marker, e.g. an enzyme) if they have
binded to an antigen that is complementary to the shape of their binding site.
Immunoassay screening tests are designed to detect whether a sample is positive or
negative for the targeted drug.
For those samples that give positive screening results, confirmation tests should be
performed, preferably using mass spectrometry. Specific immunoassay tests are
available for many drug classes, including all illicit drugs related to drug abuse.
Future of Forensic Chemistry
The techniques explained above are modern practices used for crime scene
investigation; however this does not mean that scientists havent innovated further. A
new bioassay carried out by Evgeny Katz, in collaboration with Jan Halmek, claims that
analysis of two biomarkers, creatine kinase and lactate dehydrogenase, in the blood of
people of Caucasian and Africa-American ethnicity allows clear distinction between the
two ethnicities. This could potentially mean identifying the ethnicity of a suspect from
blood samples at a crime scene. Ashley Hall (University of Nebraska-Lincoln) states that
this type of biological eyewitness would be a welcome improvement to current
forensic investigations. The group that conducted this research hope to be able to
distinguish between gender, age, and other key characteristics in future. It is noted how
fingerprint and DNA analysis took time to be accepted as evidence, so the same is likely
to happen for biomarkers.
Complementing this previous work directly, Evgeny Katz and other fellow scientists
conducted a new investigation for distinguishing between genders from blood samples.
A biocatalytic assay was carried out on the presence of creatine kinase and alanine
transaminase within blood samples, combined with the knowledge of the
concentrations of these two enzymes in the blood of healthy male and female adults.
The analysis, once again, allowed clear discrimination of samples corresponding to
male/female groups.
As with all biological samples, the levels of biomarkers will vary between individuals,
and illness or disease may have a huge effect, leading to a third possible outcome: that
no gender can be determined. However, Katz says, the information about the health
problems might be also important for the criminal investigation.
It is clear there is a rising interest in biomarkers for their uses in identification, and
scientists will most likely expand even further on this ground-breaking area of forensic
chemistry.



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