Forensic Toxicology:

Forensic science is the application of science to those criminal and civil laws that are enforced by police
agencies in the criminal justice system.
Forensic chemistry is the application of chemistry to law enforcement.
Forensic serology analysis refers to screening of evidence for body fluids.
Steps of Forensic work:
⮚ Collection of pieces of evidence from the crime scenes to be analyzed.
⮚ Identification of body fluids in the collected pieces of evidence.
⮚ Analysis of the detected body fluid.
The types of evidence are the items on which body fluids might be present such as:
•clothing •blood evidence (swabbing) • cigarettes • cups & bottles • gum • toothbrushes
These items usually provide enough DNA for a profile to be established or contain a detectable amount of
toxins or drugs that might have caused toxicity.
Why do we analyze body fluids?
⮚ They are a source of DNA (saliva, blood, semen, vaginal secretions….)
⮚ Toxins and drugs are detectable in some body fluids (blood, urine…etc.)
Other types of evidence that are analyzed by forensic chemist:
⮚ Hair samples, Skin, Autopsy samples (brain, liver, kidney, uterus….)

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 Identification of body fluids in the evidence:
❖ Identification of Semen:
➢ Important in cases of sexual assault.
➢ The composition of semen can be simplified into: seminal fluid and spermatozoa.
▪ Seminal fluid is a protein-rich fluid originating from prostate & seminal vesicles.
▪ Spermatozoa (sperm) are the male gametes, or sex cells, produced in the testis.
➢ semen samples contain Acid phosphatase (AP) enzyme.
• Presumptive tests:
• Confirmatory test:
1- Ultraviolet Light 2- Acid Phosphatase (AP) Screening (Brentamine test)
• It is used to pre-screen The most commonly used presumptive test for the detection of seminal
evidence for AP presence before fluid (Why?)
doing the Brentamine test. • The detection of AP is only presumptive because other body fluids
• Semen stains have the (blood, saliva, urine, vaginal secretions) might also give a positive
tendency to fluoresce more reaction.
intensely than most other body • However, the amount of AP in seminal fluid is greater than that found
fluids when excited with in other tissues.
ultraviolet (UV) light. Principle of Brentamine test:
• Fluorescing areas of an item • AP is identified by using the Brentamine reagent (α-napthyl
can be identified and AP tested. phosphate & diazo blue dye placed onto an item where seminal fluid is
present, the tested area quickly changes to a purple color).
• Because the test for AP is very sensitive, a lack of color change may
indicate that no seminal fluid is present
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1- Microscopic Identification of Spermatozoa: 2- Protein Confirmation of Semen:
• To confirm presumptively +ve AP, microscopic • If the semen belongs to a male with a defect
detection of spermatozoa or chemical detection of a in the male reproductive system, spermatozoa
semen specific protein is done. may not be present in the semen.
• In these cases, to confirm seminal fluid, the
presence of a protein specific to semen
(prostate-specific antigen PSA, p30) is done
by antibody–antigen complex detected by
electrophoresis.

❖ Identification of Blood:
➢ Analysis of blood evidence helps establishing which individual have been bleeding & the
manner in which blood was deposited.
➢ The study of the size, shape, volume and distribution of bloodstains found at crime scenes
and the mechanism's which may have caused them.
➢ Arterial bleeding: Blood comes in spurts
➢ Venous bleeding: Blood comes in continuous flow
➢ Capillary bleeding: Blood comes in the form of oozing

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Size and Shape of Drops:
Standard drop Rapid bleeding gives Shaking/movement
Size is 50ul (0.05ml)
slightly larger drop casts off smaller drops

Texture of
Surface

Smooth Textured

1) Naked eye examination of blood:

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C- Antemortem postmortem
More hemorrhage Less hemorrhage
Sprouting present Sprouting absent
Elastic clots present No clots
Color not easy to wash Color easy to wash
2) Chemical examination:
- These are preliminary test (to exclude presence of blood)
A) Catalytic Color Tests: 1- Benzidine (Adler Test):
• Applying a solution of chromogen to a sample The addition of one drop of benzidine reagent to the
followed by oxidizing agent (H2O2), produces color. sample produces blue to dark blue color which may
• The catalyst is actually the peroxidase-like activity turn to brown with blood, pus, faeces, nasal and
of the heme group of hemoglobin present in the red bronchial secretions
blood cells. 2- Phenolphthalein test (Kastle-Meyer test):
Hemoglobin accelerates oxidation of Phenolphthalein
reagent producing pink color.
3- Leucomalachite Green:
under acidic conditions, is catalyzed by heme to
produce a green color
B) Tests Using Fluorescence:
• Useful even if a blood stain was cleaned up or washed.
• They involve spraying a chemical mixture on a suspected blood-stained area and observing the result.
Luminol
• Luminol reacts in a similar fashion as the color tests producing a blue-white light where blood is present.
• Outlines and details are often visible for up to 30 seconds before additional spraying is required (result in
stain pattern diffusion).

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C) Microscopic Examination
(More conclusive, performed on a slide):
1- Techiman test:
(Glacial acetic acid and a crystal of NaCl, and gently heated = Yellowish to red to
brownish black rhombic crystals in clusters are formed with blood stains

2- Takayama test:
Addition of Takayama’s reagent to a sample produces feathery pink
crystals within 3 minutes with blood

❖ Identification of Saliva:
➢ Detection of saliva is useful in many types of criminal cases.
➢ The detection of amylase enzyme in saliva is the most widely utilized presumptive method.
➢ Amylase is found in a variety of body fluids (saliva, blood, urine, sweat, tears, semen, breast milk, feces,
and vaginal secretions) but is more concentrated in saliva than in other bodily fluids.

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 ❖ Hair Evidence:
A hair consists of two portions:
➢ a shaft projecting from the skin and
➢ a root that is embedded within the skin.
➢ The hair shaft contains large amounts of the protein keratin which is resistant to decay and there are few
animals capable of digesting it.
➢ Consequently, hair remains long after the soft body tissues have decomposed.
➢ Loss of hair may be passive (falling out naturally) or hairs may be transferred between people during
vigorous body contact.
➢ The hair root extends down into the dermis where it is surrounded by the hair follicle and the hair bulb.
➢ A hair with its follicle attached therefore provides a good source of DNA.
Importance of hair examination as an evidence:
➢ The forensic examination of human hairs involves the identification of questioned hairs and the
comparison to a known hair.
➢ Hair examination involves determination whether that hair originated from an animal or human.
Assimilation of drugs, poisons & explosives:
➢ Drugs such as methadone and poisons such as lead and arsenic can be sequestered in hair and detected
long after the last dose was administered and even after death.
➢ Because hair grows at a constant rate, the amount of the chemical along the length of a hair can indicate
when it was administered.
➢ It is possible to detect traces of explosives in hair, even after it has been washed.

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 ❖ DNA Testing
➢ After evidentiary items have been screened and +ve samples identified, DNA analysis can begin.
➢ DNA molecules are found in almost every cell in a person’s body.
➢ Each person’s DNA is unique, except in identical twins.
➢ DNA is the same in every cell in a person’s body throughout life.
Steps of DNA analysis:
➢ DNA Extraction
➢ DNA Quantification (gel electrophoresis, PCR….)
➢ DNA profiling
DNA analysis of Short Tandem Repeat (STR): Mitochondrial DNA Sequencing (mtDNA)
➢ STRs are repetitive sequences of DNA (2 -5 base pairs ➢ Each mitochondria have its own DNA
in length). (mtDNA) which is only inherited maternally;
➢ Forensic STR analysis determines the number of therefore, mtDNA is not unique to one person.
tetranucleotide (four base) or pentanucleotide (five base) ➢ (Each individual will share the same mtDNA
repeats at specific locations (loci) on the DNA strand. sequence with their mother, siblings, and other
➢ Profiles from evidence are compared to profiles from maternal relatives), so mtDNA is not as
known individuals to conclude whether specific discriminating as nuclear DNA analysis.
individuals have contributed to the DNA on evidentiary ➢ mtDNA is circular in shape so it is more stable
items. over time than the linear nuclear DNA.
So mtDNA can be utilized in cases involving:
➢ Skeletonized remains or old biological
Y-Chromosome STR Analysis
samples.
➢ This focuses on the Y-chromosome (in males) in sexual ➢ Cases of mass disaster where remains may be
assault cases. subjected to harsh conditions, such as salt
➢ Similar to mtDNA, the Y-chromosome is inherited water, charring, or other elemental conditions
uniparentally, meaning it is passed from father to son. that degrade DNA
➢ Therefore, male relatives will have the same Y-profile as
other male members of their family.
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