Lesson 3 FORENSIC SEROLOGY Blood Stain Examination

FORENSIC
SEROLOGY:BLOODSTAIN
EXAMINATION
INTRODUCTION
•
• In 1901, Karl Landsteiner announced one of the
most significant discoveries of this century-the
typing of blood. He recognized that all human
blood was not the same. Instead, he found blood
to distinguishable by its group or type. He came
out with the classification system that we
presently call the A-B-O system.
NATURE OF BLOOD
• Components of Blood
• Blood refers to a highly complex
mixture of cells, enzymes proteins, and
inorganic substances
• The fluid portion of the blood is called
PLASMA. It is composed principally of
water and accounts for 55% of blood
content
• Suspended in the plasma are solid
materials consisting chiefly of cells-that
is red-blood cells, white blood cells and
platelets
NATURE OF BLOOD
• The solid portion of blood accounts for 45% of its

content. When a blood has the opportunity to clot
and allowed to stand for sometimes a pale
yellowish liquid known as SERUM would be left
• Serum contains proteins known as antibodies
• White blood cells assist with the defense against
foreign substances and infection. The nuclei of the
white blood cells are the sources of DNA in the
blood
• Red blood cells transport oxygen from the lungs
to all the cells in the body and return carbon
dioxide to the lungs for expiration.
FUNCTION OF BLOOD
▪ Like all body tissues blood serves multiple

functions necessary for life as defense against
infection, gas exchange and distribution of
nutrients. To fulfill all these functions has
different types of cells suspended in plasma.
▪ All cells that comprise the blood are produced
in the bone marrow. This is in the spongy
tissue of the flat bones (skull, vertebrae,
sternum, iliac crests) and in the medullar
canals of long bones (femur, humerus).
FUNCTION OF BLOOD
▪ The blood is a renewable tissue of the human

body; this means that the bone marrow is making,
throughout life, blood cells because they have a
limited life span.
▪ This health “factory” in certain situations can
increase production depending on the needs. For
example, before bleeding increases up to seven
times the production of red blood cells to an
infection increases the production of white blood
cells.
•
•
BLOOD… IMPORTANT
▪ Volume of the human blood is about 8% of the

total body weight.
▪ Male has 5 to 6 liters of blood
▪ Female has 4-5 L of blood
▪ blood makes up around 7% of the weight of a
human body.
• While blood clotting is meant for good, it can
become dangerous. If blood clots in blood
vessels of heart, one can experience heart
attack. Similarly, blood clot in brain can lead to
STROKE.
BLOOD….
▪ Average blood pressure will always

vary from one person to another
despite the fact that standard
considered BP is 112/64 mmHg.
▪ Though we are familiar with
common blood types A, B, AB and O,
which is a part of simplified ABO
system, there are actually around 30
different recognized blood groups
or blood types.
BLOOD…
• Blood Loss:
▪ 40% loss of blood, internally or/and
externally is required to produce irreversible
shock (death)
•
▪ Blood loss of 1.5 liter, internally or externally,
is required to cause incapacitation
FORENSIC CHARACTERIZATION OF BLOODSTAINS
• The criminalist must be prepared to answer

the following questions when examining a
dried blood:
▪ Is it blood?
▪ From what species did the blood originate?
▪ If the blood is of human origin, how closely
can it be associated to a particular
individual?
IDENTIFICATION OF BLOOD
▪ Presumptive or Preliminary Test for

Blood
o When positive, allows a qualified
conclusion that blood is present in a
tested sample
o When negative, the test often helps to
eliminate stains that need not receive
further consideration
IMPORTANCE OF THE STUDY OF BLOOD
▪ As circumstantial or corroborative evidence

against or in favor of the perpetrator of the crime
▪ Blood typing and DNA examination can be used to
match and compare the parent and the child
▪ Determination of the origin of blood flow: position
of the stain relative to the explanation of the
suspect
o If blood comes from the nose, mucus of hairs from nostrils may
be present
o Blood from the lungs will show numerous bubbles
o Stomach blood is brownish color due to the action of strong
acid in the stomach
o Blood due to rape contains semen, or hairs from the suspect
o Menstrual blood contains plenty of epithelial cells from the
vagina
IMPORTANCE OF THE STUDY OF BLOOD
▪ Determination of the cause of death and the

length of time the victim survived the attack
▪ Determination of the approximate time of the
crime committed using the color and age of
bloodstain
o Vein blood is darker in color
o Arterial blood is bright red
o Blood clot/drying usually takes from 12 to 36
hours depending on the size of the clot and
other external conditions. As the stain
continues to dry, the stain becomes darker in
color, until completely dried with dark brown ARTERIAL BLOOD AND VENOUS BLOOD
appearance (10 10 12 days old)
COLOR CHANGE IN BLOOD
• Color change (in blood) is due to the

conversion of red blood pigment
hemoglobin into methaemoglobin and
haematin
BLOOD EXAMINATION
• ACTUAL EXAMINATION OF BLOOD

• Determining if the specimen is Blood
• A preliminary examination and Confirmatory
examination is done to determine that it is actually
blood
•
• BLOOD CHARACTERIZATION/TEST FOR BLOOD
• Typical forensic examiner experienced questions such
as the identity of blood, origin and how closely it can be
associated to a particular individual.
PRELIMINARY EXAMINATION OR SCREENING TEST
FOR BLOOD
Chemical test Procedure + result: Color reaction
Benzidine test Add 3 drops of hydrogen peroxide to bloodstain then Intense blue color
Benzidine reagent
Phenolphthalein test Add a few drops of ethanol, then a few drops of Deep pink color or sample turn violet
phenolphthalein and finally a few drops of hydrogen peroxide
are dipped onto the sample
Schonbein’s test guaiacum test A drop of tincture of guiac, turpertine and ether, hydrogen Blue coloration
peroxide
Luminol test A spray reagent is used to test for blood even blood is not Production of light (luminescence) bright
visible under ordinary light. This is viewed under ultra violet blue
light
Hemastix Test strip used for field test of blood by moistening with Color band are present
distilled water and placed in contact with blood
CONFIRMATORY TEST WHETHER A STAIN IS BLOOD
• MICROSCOPIC EXAMINATION
• examination of menstrual blood

corpuscles to differentiate mammal
blood to other animals
CONFIRMATORY TEST
• SPECTROSCOPIC METHOD
o Spectrophotometric procedures are seldom
used at the present time in forensic
analyses.
o This type of examination is based on the
identification of hemoglobin and its
derivatives through their specific absorption
spectra.
CONFIRMATORY TEST
• CHEMICAL TEST
o When blood dries to form a bloodstain, the cells are
destroyed and their contents released into the
surrounding environment. More than 250 proteins,
enzymes, and other compounds have been found in the
red blood cell, mostly in the soluble portion of the
erythrocytes.
o The predominant erythrocyte protein is hemoglobin
(Hb). More than 100 variants of hemoglobin have been
described. Identification of blood in stains by means of
chemical methods is based on the detection of heme
or its derivatives in the stain sample. Such tests can be
classified under one of two categories: catalytic tests
and crystal tests.
CHEMICAL METHODS
o The general presumptive
• Catalytic tests (screening or presumptive
test reaction is:
tests) All catalytic blood tests depend on an
H202 + reduced reagent
oxidation reaction in which an oxidant, for
example, hydrogen peroxide, oxidizes a
(color 1) <— H20 +
colorless material, such as phenol-phthalin oxidized reagent (color 2).
or tetramethylbenzidine, to a colored one.
Alternately, 3-amino-phthalhydrazide
(luminol), a colorless material, can be
oxidized to a product which luminesces.
CHEMICAL METHODS
• The heme group of hemoglobin exhibits a peroxidase-like activity which may catalyze the breakdown of
hydrogen peroxide. The majority of tests which have been devised for the forensic identification of blood
are based on the peroxide-mediated oxidation of leukomalachite green, phenolphthalin, o-tolidine, luminol,
tetramethylbenzidine, fluorescein, and other less commonly used compounds. At one time, benzi-dine and
its derivatives were widely used as the color reagent in screening tests for blood. However, due to the
carcinogenic nature of these compounds and the health risks involved in their use, laboratories no longer
use these types of chemical reagents.
• The tests most commonly employed in modern crime scene procedures are phenolphthalin,
leukomalachite green, luminol and tetramethylbenzidine. Reaction schemes for some of these common
chemical reagents are shown in Fig. 2. All of these chemicals are highly sensitive to minute traces of
hemoglobin and its derivatives, but all suffer from the occurrence of false positive reactions with some of
the following materials: catalases, peroxidases, cytochromes, strong oxidizing agents and metallic salts.
TESTS DONE FOR CONFIRMATION
Chemical test Procedure Positive result
Takayama test: Add Takayama reagent to stain Large rhombic crystals with salmon color
Test for Hemoglobin
Teichmann test Salt and acetic acid is added to stain and acid is allowed to Reddish brown rhombic crystal
Hemin crystal test evaporate
Acetone-Haemin/or Wagenhaar Acetone and oxalic acid small dark dichroic crystals
test
•THANK YOU FOR
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FORENSIC

• The solid portion of blood accounts for 45% of its

▪ Like all body tissues blood serves multiple

▪ The blood is a renewable tissue of the human

▪ Volume of the human blood is about 8% of the

▪ Average blood pressure will always

• The criminalist must be prepared to answer

▪ Presumptive or Preliminary Test for

▪ As circumstantial or corroborative evidence

▪ Determination of the cause of death and the

• Color change (in blood) is due to the

• ACTUAL EXAMINATION OF BLOOD

• examination of menstrual blood

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