Submitted To: Vasundhara Devi

Submitted by:
Chaitanya Srivastava (12104010)
Tanuj Kumar (12104025)
Abhishek Khurana (12104027)
Abhishek Marwaha (12104015)

Forensic Toxicology
Forensic toxicology is the use of toxicology and other disciplines such
as analytical chemistry, pharmacology and clinical chemistry to aid
medical or legal investigation of death, poisoning, and drug use. The
primary concern for forensic toxicology is not the legal outcome of the
toxicological investigation or the technology utilized, but rather the
obtaining and interpreting of the results. A toxicological analysis can be
done to various kinds of samples. A forensic toxicologist must consider
the context of an investigation, in particular any physical symptoms
recorded, and any evidence collected at a crime scene that may narrow
the search, such as pill bottles, powders, trace residue, and any
available chemicals. Provided with this information and samples with
which to work, the forensic toxicologist must determine which toxic
substances are present, in what concentrations, and the probable
effect of those chemicals on the person.
Determining the substance ingested is often complicated by the body's
natural processes as it is rare for a chemical to remain in its original
form once in the body. For example: heroin is almost
immediately metabolized into another substance and further
to morphine, making detailed investigation into factors such as
injection marks and chemical purity necessary to confirm diagnosis. The
substance may also have been diluted by its dispersal through the
body; while a pill or other regulated dose of a drug may
have grams or milligrams of the active constituent, an individual sample
under investigation may only contain micrograms or nanograms.

Forensic toxicology is utilized in most areas of a forensic investigation,

though is particularly vital in suspicious deaths involving the suspected
intake of some toxic substance. Drug testing methods employed are
often used in cases of rape in which the victim has been slipped some
kind of date-rape drug, rendering them unable to fight off their attacker
while they are being sexually assaulted. Unfortunately many such drugs
leave the system quickly.

An examination of the crime scene can often give investigators a clue as

to what may have been ingested by the victim. Such evidence may
include pill bottles, powders, trace residues and any other chemicals
present.

The application of poisons has been a common method of murder for

hundreds of years, though methods of detection have advanced in
recent years. Initially one or more screening tests will be conducted in
order to gain a general idea of what the toxin is. Common screening
tests include physical tests (boiling point, refractive index), crystal test
(treatment with a chemical reagent to produce crystals), chemical spot
tests (treatment with color-changing reagents), and chromatography
(method of separating substance components). Following this,
confirmation tests, such as gas chromatography and mass spectrometry
may be conducted. Various chromatography methods are commonly
used to separate the various compounds of a sample, allowing for the
substance to be identified. Immunoassays can also be used to identify a
poisonous substance by identifying the antibodies produced for the
foreign substance.

However this technique is still being developed and so is not widely

used. In cases involving a corpse with little soft tissue present, insect
analysis may be beneficial. Carrion feeding insects will most likely have
ingested any toxic substances whilst feeding on the corpse. By
dissecting and extracted the ingested tissue, further analysis may be
able to identify the substance.

Hair is potentially an idea recording medium for long-term drug abuse.

As chemicals are absorbed into the bloodstream they are transferred
into the hair, where they may be stored. This can even provide a rough
timeline of drug intake. Head hair grows at an approximate rate of 1 to
1.5cm a month, therefore cross-sections may be able to provide an
estimation of when a substance was ingested.

Despite these techniques, the identification of toxic substances it is

rarely straightforward. The bodys natural processes often complicate a
substance, altering the chemicals original form. For example, after
ingestion, heroin is almost immediately metabolized into another
substance and then metabolized further into morphine. Urine and
blood samples are most commonly extracted for drug testing, though
other bodily fluids and organs will suffice. The brain, liver and spleen
are commonly used organs for this. While gastric contents are useful
for the detection of undigested pills or liquids, such an analysis may not
be useful hours after ingestion.

Though there is a vast array of toxin substances available, a selection

are particularly common. Below are some regular poisons and their
symptoms.
Acids: Characteristic burns around the mouth, lips and nose.
Aniline: Darkened skin around the face and neck.
Arsenic: Severe diarrhea.
Atropine: Dilated pupils.
Bases: Burns around the mouth, lips and nose.
Carbolic acid: Strong odor of disinfectant.
Carbon monoxide: Cherry red skin.
Cyanide: Quick death, red skin, and a distinctive odour of almonds.
Food poisoning: Severe vomiting, diarrhea and abdominal pain.
Metallic compounds: Diarrhea, vomiting and abdominal pain.
Nicotine: Convulsions.
Opiates: Pupil contraction.
Oxalic acid: Strong garlic odour.
Sodium fluoride: Convulsions.
Strychnine: Convulsions, lockjaw, a high pulse, and dark face and neck

IMP DEFINITIONS
Toxin: A poisonous substance produced during the metabolism
and growth of certain microorganisms and some higher plant and
animal species
Dose: Dose makes the poison is a fundamental principle of
toxicology, but the actual study of toxicology is much more
nuanced and complex than this simple statement suggests.

Toxicology: The study of the nature, effects, and detection of

poisons and the treatment of poisoning.

An intoxicant, such as alcohol or carbon monoxide,

requires that you ingest a large amount to be lethal.
A true poison, such as cyanide, requires only a very small
amount.

Various Samples
Stomach contents
Digestion stops at the moment of death. Liver the toxin sponge of
your body can reflect levels of toxins that even the blood may not
reveal. Vitreous Humor eyeball fluidvery slow to decay so will retain
toxins even longer than most other organs. Insects toxins can
accumulate (bioaccumulation) in the bodies of insects that feed off of
decomposed bodies

Urine Sample
A urine sample is urine that has come from the bladder and can be
provided or taken post-mortem. Urine is less likely to be infected with
viruses such as HIV or Hepatitis B than blood samples.[1] Many drugs
have a higher concentration and can remain for much longer in urine
than blood. Collection of urine samples can be taken in a noninvasive
way which does not require professionals for collection. Urine is used
for qualitative analysis as it cannot give any indication of impairment
due to the fact that drug presence in urine only indicates prior
exposure.

Blood Sample
A blood sample of approximately 10 ml (0.35 imp fl oz; 0.34 US fl oz) is
usually sufficient to screen and confirm most common toxic substances.
A blood sample provides the toxicologist with a profile of the substance
that the subject was influenced by at the time of collection; for this
reason, it is the sample of choice for measuring blood alcohol
content in drunk driving cases.

Hair sample
Hair is capable of recording medium to long-term or high dosage
substance abuse. Chemicals in the bloodstream may be transferred to
the growing hair and stored in the follicle, providing a rough timeline of
drug intake events. Head hair grows at rate of approximately 1 to
1.5 cm a month, and so cross sections from different sections of the
follicle can give estimates as to when a substance was ingested. Testing
for drugs in hair is not standard throughout the population. The darker
and coarser the hair the more drug that will be found in the hair.If two
people consumed the same amount of drugs, the person with the
darker and coarser hair will have more drug in their hair than the
lighter haired person when tested. This raises issues of possible racial
bias in substance tests with hair samples.[3]

Other
Other bodily fluids and organs may provide samples, particularly
samples collected during an autopsy. A common autopsy sample is
the gastric contents of the deceased, which can be useful for detecting
undigested pills or liquids that were ingested prior to death. In highly
decomposed bodies, traditional samples may no longer be available.
The vitreous humour from the eye may be used, as the fibrous layer of
the eyeball and the eye socket of the skull protects the sample from
trauma and adulteration. Other common organs used for toxicology are
the brain, liver, and spleen.
The inspection of the contents of the stomach must be part of every
postmortem examination if possible because it may provide qualitative
information concerning the nature of the last meal and the presence of
abnormal constituents. Using it as a guide to the time of death,
however, is theoretically unsound and presents many practical
difficulties, although it may have limited applicability in some
exceptional instances. Generally, using stomach contents as a guide to
time of death involves an unacceptable degree of imprecision and is
thus liable to mislead the investigator and the court. Characteristic cell
types from food plants can be used to identify a victim's last meal;
knowledge about which can be useful in determining the victim's
whereabouts or actions prior to death (Bock and Norris, 1997). Some of
these cell types include (Dickison, 2000):

sclereids (pears)
starch grains (potatoes and other tubers)
raphide crystals (pineapple)
druse crystals (citrus, beets, spinach)
silica bodies (cereal grasses and bamboos)

In a case where a young woman had been stabbed to death, witnesses

reported that she had eaten her last meal at a particular fast food
restaurant. However, her stomach contents did not match the limited
menu of the restaurant, leading investigators to conclude that she had
eaten at some point after being seen in the restaurant. The
investigation led to the apprehension of a man whom the victim knew,
and with whom she had shared her actual final meal (Dickison, 2000).
Time since death can be approximated by the state of digestion of the
stomach contents. It normally takes at least a couple of hours for food
to pass from the stomach to the small intestine; a meal still largely in
the stomach implies death shortly after eating, while an empty or
nearly-empty stomach suggests a longer time period between eating
and death (Batten, 1995). However, there are numerous mitigating
factors to take into account: the extent to which the food had been
chewed, the amount of fat and protein present, physical activity
undertaken by the victim prior to death, mood of the victim,
physiological variation from person to person. All these factors affect
the rate at which food passes through the digestive tract. Pathologists
are generally hesitant to base a precise time of death on the evidence
of stomach contents alone.

Detection and classification

Detection of drugs and pharmaceuticals in biological samples is usually
done by an initial screening and then a confirmation of the
compound(s), which may include a quantitation of the compound(s).
The screening and confirmation are usually, but not necessarily, done
with different analytical methods. Every analytical method used in
forensic toxicology should be carefully tested by performing a
validation of the method to ensure correct and indisputable results at
all times. A testing laboratory involved in forensic toxicology should
adhere to a quality programme to ensure the best possible results and
safety of any individual.
The choice of method for testing is highly dependent on what kind of
substance one expects to find and the material on which the testing is
performed. Biological samples are more complex to analyze because of
factors such as the matrix effect and the metabolism and conjugation of
the target compounds.

Gas chromatography
Gas-liquid chromatography is
volatile organic compounds.

of

particular

use

in

examining

Detection of metals
The compounds suspected of containing a metal are traditionally
analyzed by the destruction of the organic matrix by chemical or
thermal oxidation. This leaves the metal to be identified and quantified
in the inorganic residue, and it can be detected using such methods as
the Reinsch
test,
emission spectroscopy or X-ray
diffraction.
Unfortunately, while this identifies the metals present it removes the
original compound, and so hinders efforts to determine what may have
been ingested. The toxic effects of various metallic compounds can vary
considerably.

Detection of Alcohol:
The analysis of alcohol exemplifies the primary objective of forensic
toxicologythe detection and isolation of drugs in the body for the
purpose of determining their influence on human behavior. Alcohol, or
ethyl alcohol, is a colorless liquid normally diluted with water and
consumed as a beverage. Like any depressant, alcohol principally
effects the central nervous system, particularly the brain.Alcohol
appears in the blood within minutes after it has been taken by mouth
and slowly increases in concentration while it is being absorbed from
the stomach and the small intestine into the bloodstream. When all the
alcohol has been absorbed, a maximum alcohol level is reached in the
blood; and the post-absorption period begins. Then the alcohol
concentration slowly decreases until a zero level is again
reached.Factors such as time taken to consume the drink, the alcohol
content, the amount consumed, and food present in the stomach
determine the rate at which alcohol is absorbed. Elimination of alcohol
throughout the body is accomplished through oxidation and excretion.

Oxidation takes place almost entirely in the liver, while alcohol is

excreted unchanged in the breath, urine, and perspiration. The extent
to which an individual may be under the influence of alcohol is usually
determined by either measuring the quantity of alcohol present in the
blood system or by measuring the alcohol content in the
breath.Experimental evidence has verified that the amount of alcohol
exhaled in the breath is in direct proportion to the blood concentration.

Alcohol & Circulatory System:

Humans have a closed circulatory system consisting of a heart, arteries,
veins, and capillaries.Alcohol is absorbed from the stomach and small
intestines into the blood stream.Alcohol is carried to the liver where
the process of its destruction starts.Blood, carrying alcohol, moves to
the heart and is pumped to the lungs.In the lungs, carbon dioxide and
alcohol leave the blood and oxygen enters the blood in the air sacs
known as alveoli.Then the carbon dioxide and alcohol are exhaled
during breathing.

Breath Testers
Breath testers operate on the fact that at 34C, the ratio of alcohol in
the blood to alcohol in alveolar breath is approximately 2,100 to 1.
Breath testers that operate on the principle of infrared light absorption
are becoming increasingly popular within the law enforcement
community.Many types of breath testers are designed to analyze a set
volume of breath. The captured breath is exposed to infrared light. Its
the degree of the interaction of the light with alcohol in the captured
breath sample that allows the instrument to measure a blood alcohol
concentration in breath.Some breath testing devices also use fuel cells.

Nonvolatile organic substances

Drugs,
both
prescribed
and
illicit, pesticides,
natural
products, pollutants and industrial compounds are some of the most
common nonvolatile compounds encountered. Screening methods
include thin-layer chromatography, gas-liquid chromatography and
immunoassay. For complete legal identification, a second confirmatory
test is usually also required. The trend today is to use liquid
chromatography tandem mass spectrometry, preceded with sample
workup as liquid-liquid extraction or solid phase extraction. Older
methods include: spot test (see Pill testing), typically the Marquis
Reagent, Mecke Reagent, and Froehde's reagent for opiates, Marquis
Reagent and Simon's
reagent for amphetamine,
methamphetamine and other analogs, like MDMA, the Scott's test for
cocaine, and the modified Duquenois reagent for marijuana and
other cannabinoids. For compounds that don't have a common spot
test, like benzodiazepines, another test may be used, typically mass
spectrometry, or spectrophotometry.

Branches of Toxicology
Clinical Toxicology
Medical Toxicology
Forensic Toxicology
Industrial Toxicology
Environmental Toxicology
Biochemical and Molecular Toxicology
Product Development Toxicology
Regulatory Toxicology

Common poisons:
Cyanide one of the most lethal chemicals knownused for
executionscauses a bright cherry red blood Strychnine rat
poisonscauses so much pain that it is rarely used in suicide Ethylene
glycol antifreezea favorite (deadly) beverage among alcoholics when
they cant get ethanol Heavy metals No, not Metallica, Megadeath or
Anthrax but they are interesting names.arsenic, mercury and lead
Insulin lifesaving for diabetics but deadly overdoses Corrosive
chemicals strong alkalis (lyeNaOH) or acids (HCl, H2SO4)burn the
mouth, esophagus, and stomach

Important Factors
Dose makes the poison
The concentration of the chemical
The threshold required to exert a particular effect
How the interaction takes place
Which factors are of importance in each exposure scenario.

Role of the Toxicologist:

Beyond the analysis of alcohol, the toxicologist is confronted with a
maze of drugs and poisons.
The toxicologist is originally presented with body fluids and/or organs
and is normally requested to examine them for the presence of drugs
and poisons.
Without supportive evidence, such as the victims symptoms, a
postmortem pathological examination, or an examination of the
victims personal effects, the toxicologist is forced to use general
screening procedures with the hope of narrowing thousands of
possibilities to one.
In addition, the toxicologist is not dealing with drugs at the
concentration levels found in powders and pills, having been dissipated
and distributed throughout the body.
Furthermore, the body is an active chemistry laboratory as few
substances enter and completely leave the body in the same chemical
state.
Last, when and if the toxicologist has surmounted all of these obstacles,
he or she must be prepared to assess the toxicity of the drug or poison.

The Analytical Scheme

The forensic toxicologist must devise an analytical scheme that will
successfully detect, isolate, and specifically identify toxic drug
substances.
Once the drug has been extracted from appropriate biological fluids,
tissues, and organs, the forensic toxicologist can proceed to identify the
drug substance present.
Drug extraction is generally based on a large number of drugs being
either acidic or basic.
The strategy used for identifying abused drugs entails a two-step
approach: screening and confirmation.

The Screening Step

A screening test is normally employed to provide the analyst with quick
insight into the likelihood that a specimen contains a drug substance.
Positive results arising from a screening test are considered to be
tentative at best and must be verified with a confirmation test.
The most widely used screening tests are thin-layer chromatography,
gas chromatography, and immunoassay.

The Confirmation Step

Gas chromatography/mass spectrometry is generally accepted as the
confirmation test of choice.
The GC separates the sample into its components, while the MS
represents a unique fingerprint pattern that can be used for
identification.
Once the drug is extracted and identified, the toxicologist may be
required to provide an opinion on the drugs effect on an individuals
natural performance or physical state.

Significance of Findings
Once a drug is found and identified, the toxicologist assesses its
influence on the behavior of the individual.
For many drugs, blood concentration levels are readily
determined and can be used to estimate the pharmacological
effects of the drug on the individual.
Often, when dealing with a living person, the toxicologist has the
added benefit of knowing what a police officer may have
observed about an individuals behavior and motor skills.

Detecting Drugs in Hair

Drugs present in blood diffuse through the capillary walls into the
base of the hair and become permanently entrapped in the hairs
hardening protein structure.
As the hair continues to grow, the drugs location on the hair shaft
becomes a historical marker for delineating drug intake.
Given that the average human head hair grows at the rate of 1
centimeter per month, analyzing segments of hair for drug
content may define the timeline for drug use.
The chronology of drug intake may be distorted by drugs
penetrating the hairs surface as a result of environmental
exposure, or drugs may enter the hairs surface through sweat.

Biotransformation
When a chemical enters the body, the body reacts by breaking it down
in order to eliminate it. So if you have injected something like heroin,
the body will break it down into the morphine originally used to
produce it. Hunting for heroin is futile but if you find morphine you
have found signs of heroin. These products are called metabolites.
Ingested toxins show up in the stomach, intestines, or liver Inhaled
gases are concentrated in the lungs.Toxins that are injected
intramuscularly concentrate themselves around the injection site.
Drugs that are given intravenously are directly absorbed into the blood
bypassing the stomach and liver. Concentrations are found throughout
the body, are low in the stomach and liver and high in the blood
stream.

Nondrug Poisons
Heavy metals such as arsenic, bismuth, antimony, mercury, and
thallium are only occasionally encountered because severe
environmental protection regulations restrict their availability to the
general public.
Carbon monoxide is one of the most common poisons encountered in a
forensic laboratory.
To measure the concentration of carbon monoxide in the blood
spectrophotometric
methods
determine
the
amount
of
carboxyhemoglobin relative to oxyhemoglobin or total hemoglobin; or
a volume of blood can be treated with a reagent to liberate the carbon
monoxide, which is then measured by gas chromatography.