MODULE 2 BLOOD AND BLOODSTAINS

INTRODUCTION

The significance of blood and bloodstains as evidence in crimes of violence is obvious such
that we need not place emphasis on this. The test for the identification of blood is employed as
an important part of the routine investigation in many case of violent death. The specimen usually
submitted is fresh blood or fluid blood, dried blood and clotted blood. Very often it is brought to
the laboratory in the form of dried red or brown stains on weapons, clothing or other objects.

At the end of this chapter, the students must be able to:

1. Give the importance of blood in Criminal Investigation
2. Identify what particular cases are related to blood.
3. determine the laboratory examination being performed to identify its presence

PRE-COMPETENCY CHECKLIST

Self-checkup

LEARNING RESOURCES

1. CBSUA Website
2. Supplementary Online Learning Materials
https://www.healthline.com/health/blood-typing#followup
https://www.bswhealth.com/patient-tools/blood-center/Pages/blood-type-genetics-
and-compatibility.aspx

EXPLORE (TASKS/ACTIVITIES)

Lecture/Discussion

Reading of Supplementary Online Learning Materials

DISCUSSION BOARD

Blood is a viscous fluid formed of cellular element suspended in plasma.

The cellular element is composed of:
1. Erythrocytes (red blood cells)
2. Leucocytes (white blood cells)
3. Thrombocytes (Platelets)

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Plasma is a viscous, translucent, yellowish fluid composed of
1. water (90%)
2. proteins (7%)
3. organic salts (1%)
4. and organic compound (2%) such as amino acids, lipids, and vitamins.
The total blood volume in human is about 5 L (depending on body size).
Outside the blood vessels, blood undergoes a complex reaction called coagulation or
clot formation, which plays an important role in repairing damaged blood vessels and
preventing blood loss.
Erythrocytes and blood platelets perform their functions inside the blood vessels, whereas
leukocytes reside temporarily in the blood vessels and then leave the blood stream through the
capillary walls and venules to enter either the connective tissues or lymphoid tissues.
The ratio of erythrocytes to the total blood volume is about 43% and is known as
hematocrit.

Blood film showing red blood cells and different types of white blood cells

COMPOSITION OF PLASMA
A. Water: constitutes 90% of plasma volume.
B. Solutes: constitutes 10% of plasma. and include plasma proteins and other organic
compounds as well as inorganic salts.
1. Plasma proteins.
Plasma contains a rich variety of soluble proteins, 7% by volume. Important examples
include:
a. Albumin. This is the most abundant plasma protein (3.5-5 g/dL of blood) and is
mainly responsible for maintaining the osmotic pressure of blood.

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 b. Globulins (Alpha, beta, and gamma globulins) are globular proteins dissolved in the
plasma. The gamma globulins include the antibodies, or immunoglobulins, synthesized
by plasma cells.
c. Blood coagulation proteins: such as prothrombin, fibrinogen which is converted
into fibrin during clot formation. Fibrinogen is synthesized and secreted by the liver.
2. Organic compounds. They include nutrients such as amino acids and glucose,
vitamins, and a variety of regulatory peptides, steroid hormones, and lipids.
3. Inorganic salts. They constitute 0.9% of plasma volume, include blood electrolytes
such as sodium, potassium, and calcium salts.
4. Serum: portion of plasma that separates from coagulum after clotting. When
anticoagulants (heparin, citrate, etc) are added, blood samples can be separated by
centrifugation into 3 major fractions: -

The erythrocytes constitute the densest fraction and end up at the bottom of the tube.
The percentage of packed erythrocytes per unit volume of blood is termed hematocrit.
In adults, normal hematocrit values vary from 35 to 50% and are sex dependent.
Leukocytes are less dense and less numerous (about 1 % of blood volume) and form a
thin white or grayish layer over the erythrocytes (buffy coat).
On top of the buffy coat is a thin layer of platelets.

The least dense is the clear layer of plasma, which constitutes 42 - 47% of the blood
and overlies the buffy coat.

THE CELLULAR OR FORMED ELEMENTS OF BLOOD

A- Erythrocytes (RBC)

RBCs are structurally and functionally specialized to transport oxygen from the lungs to
other tissues. Their cytoplasm contains the oxygen binding protein hemoglobin. Mature RBCs lack
nuclei and cytoplasmic organelles, which they lose during differentiation. Mature erythrocytes
therefore have a limited lifespan (120 days) in the circulation before they are removed by
macrophages in the spleen and bone marrow.
Erythrocytes (RBC) are anucleated corpuscles (nucleated in embryonic and fetal mammals
and in other vertebrates). They are biconcave disks about ~7μm in diameter, 2μm thick at its
rim and less than 1μm at its center. They contain hemoglobin, which fills almost the entire
cytoplasm.

Erythrocytes are elastic and can withstand deformation. Their number is about 4.5-5 million/mm³,
the number is more in males than in females.

The lifespan of an erythrocyte in the bloodstream is 100-120 days i.e. about 5×10¹¹ erythrocytes
are formed/ destroyed each day.

Erythron (census): whole mass of RBCs & their precursors in bone marrow.

RBCs are considered as 'yardstick' for estimating dimensions of other cells in sections, (the
diameters of relative cells are about 7μm).

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Anisocytosis: refers to the presence of a high percentage of RBCs with great variations in size.
Those larger than 9μm in diameter are termed macrocytes, and those smaller than 6μm are
termed microcytes.
Nuclear fragments: in some diseases nuclear fragments or Howell-Joliy bodies remain in
some mature RBCs. When these form circular filaments they are termed Cabot rings.
Reticulocytes are immature RBCs released from bone marrow; normally estimated ~ 1% of
circulating RBCs. Reticulocytes contain a small amount of residual RER and ribosomes, and are
stained with supravital staining (brilliant cresyl blue) in fresh blood film.
Reticulocytosis means increase reticulocyte count which indicate an increased demand for
oxygen carrying capacity (eg, from loss of RBCs due to hemorrhage or anemia).

B- White blood cells (WBC) or leucocytes

Leukocytes can be subdivided into:

a. granular leukocytes (neutrophils, basophils and eosiniphils)

b. non-granular leukocytes (monocytes and lymphocytes).

In healthy individuals the total number of circulating leukocyte is about 4000-

10.000/mm³. Increase the leucocytes count above the upper range is called leuocytosis; which
occurs in infection, inflammatory conditions, and in leukemia. Whereas, decrease the count below
the lower range is called leucopenia; which occurs in excessive exposure to X-ray and after
prolonged treatment with steroids.

Differential Cell Count: Blood is also studied by spreading a drop on a slide to produce
a single layer of cells (blood smear). The cells are stained, differentiated by type, and counted to
reveal disease-related changes in their relative numbers. The smears are usually stained with dye
mixtures (eg, Wright's or Giemsa) containing eosin and methylene blue. Blood cells and their
components exhibit 4 major staining properties that allow the cell types to be distinguished:
Basophilia: is affinity for methylene blue. Basophilic structures stain purple to black.

Azurophilia: is affinity for the oxidation products of methylene blue called azures.
Azurophilic structures stain reddish-purple.

Eosinophilia, or acidophilia: is affinity for eosin. Eosinophilic structures stain pink to

orange.

Neutrophilia: is affinity for a complex of dyes (originally thought to be neutral) in the

mixture. Neutrophilic structures stain pink.

The differential leukocytic count is the percentage of each type of WBCs in blood,
and would typically produce the following cell frequencies:
~ 60% neutrophils (50% - 70%)
~ 4% eosinophils (>0% - 5%)
~ 0.5-1% basophils (>0% - 2%)
~ 5% monocytes (1% - 9%)

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~ 40% lymphocytes (20% - 40%)
Changes in their relative numbers indicate that something abnormal is happening in
individual.
A larger than usual number of neutrophils (neutrophilia) would indicate e.g. an acute or
chronic infection.
The number of basophils and eosinophils may increase ( eosinophilia or basophilia) as a
consequence of e.g. allergic disorders.
Granular Leukocytes - Granular leukocytes are all approximately the same size (12-15
μm in diameter). - Their nuclei are lobulated and nucleoli cannot be seen. The number of nuclear
lobes varies according to cell type.
All granulocytes are motile. The term granulocyte refers to the presence of granules in
the cytoplasm of these cells. The granules correspond to secretory vesicles and lysosomes.
Specific granules are the granules which are only found in one particular type of granulocytes. 1-
Neutrophils
Neutrophils (polymorphonuclear granulocytes): one of granular leucocytes that have a
very characteristic nucleus. It is divided into 3-5 lobes, which are connected together by thin
strands of chromatin. The number of lobes increases with cell age. Up to 7 lobes can be found in
very old neutrophils (hypersegmented cells). Barr body is a drumstick chromosome or condensed
chromatin visible in about 3% peripheral blood of females. Neutrophils contain all the organelles
that make up a typical cell. In addition to the usual complement of organelles, they also contain
three types of granules that stain "neutral", hence the term neutrophil

Types of neutrophilic granules:

a. Primary (azurophilic granules): large Lysosomes contain hydrolytic enzymes such as
myeloperoxidase and acid phosphatase that have microbicidal effect.
b. Secondary (specific) granules: smaller, twice as numerous; alkaline phosphatase =
phosphasome (light fraction of membranes)
c. Tertiary: contain gelatinase; enhance phagocytosis

Neutrophilia: Means increased number of neutrophils in circulation as in acute bacterial

infection, tissue injury and malignancy.
Neutropenia: Means decreased number of neutrophils in circulation as in viral infection,
chronic bacterial infection such as typhoid fever and tuberculosis.
Functions:
Their lifespan is only about one week inside blood vessels, and then pass to the connective
tissue where they last for another 1-4 days. Neutrophils play a central role in inflammatory
processes. Neutrophils are the first wave of cells invading infection sires. Receptors in their plasma
membrane allow them to recognize foreign bodies, e.g. bacteria, and tissue debris, which begin
to phagocytose and destroy. The phagocytotic activity of neurophils is further stimulated if
invading microorganisms are "tagged" with antibodies (or opsonised). Dead neutrophils and
tissue debris are the major components of pus.

2- Eosinophils
Eosinophils are small 12-18μ in diameter, slightly larger than neutrophils. Eosinophils
represent 1-6% of the total white cell count, and show diurnal variation (greatest in morning;
least in afternoon). Their nuclei usually have two lobes that hidden by the numerous granules,

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which cover almost all of the cytoplasm. As the term "eosinophil" indicates, these granules are
stained red or pink with eosin or other similar dyes.

Eosinophils contain some large rounded vesicles (~1 μm) in their cytoplasm. The specific
granules contain lysosomal enzymes, and electron-dense, proteinaceous crystal composed of
major basic protein (MBP). Smaller granules: contain aryl sulfatase and acid phosphatase. Life
span: Eosinophils circulate in blood for 3 to 8 hours before migrating to connective tissue where
they last for 10-12 days. Eosinophilia: Increase the number of eosinophils above the normal as
in parasitic disease; and in allergic disorders.

Functions
1. Contain receptors for IgE which stimulates the immune system.
2. Their granules contain histaminase and arylsufatase enzymes that break down histamine
and leukotrienes.
3. Major Basic Protein, which can function as a cytotoxin, and involved in the response of
the body against parasitic infections.
4. Produce eosinophil-derived-inhibitor, which inhibits mast cell degranulation.

3- Basophils
Small granulocytes with a diameter of 8 to 10μm. Their nucleus is bilobed which hidden
by the large cytoplasmic granules. Their number is ~1% of differential white cell count. The
granules are not as numerous as those in eosinophils and have metachromatic stainability with
toluidine blue. Their specific granules (about 0.5 μm) appear quite dark in EM pictures. They
contain heparin, histamine lysosomal enzymes and leukotrienes. Their cell membrane contains
receptors for IgE (produced in response to allergens); that triggers rapid exocytosis of granular
contents (degranulation).

Functions
1. Heparin and histamine are vasoactive substances, dilate the blood vessels, make vessel
2. walls more permeable and prevent blood coagulation.
3. They facilitate the access of lymphocytes and other antibodies to the site of infection.

Non-granular leukocytes
1- Monocytes
Monocytes are large cells, 12-18μm in diameter; represent 2-10 % of the differential
cell count. Monocytes are highly motile and phagocytic cells; i.e. they are the precursor of tissue
phagocytes that migrate into tissues. Their nucleus less dense than lymphocytes; deeply
indented, kidney or C-shaped. Their cytoplasm is pale grayish blue with small pink to purple
stained lysosomal granules, and contain cytoplasmic vacuoles (frosted glass). Monocytes contain
granules (visible in the EM) which are similar to the primary granules of neutrophils, i.e.
Lysosomes containing acid phosphatase, aryl granules. They contain also secondary granules of
unknown function.
Functions

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 1. Once monocytes enter the connective tissue they differentiate into macrophages that
phagocytose microorganisms, tissue debris and the dead neutrophils.
2. Monocytes also give rise to mononuclear phagocytic system: which include histiocytes;
multinucleate giant cells; hepatic macrophages (Kupffer ) cells; microglia of CNS;
macrophages (Langerhans cells) of skin; antigen-presenting cells (APCs) of lymphoid
organs; and osteoclasts of bone.
3. Monocytosis: increase numbers of monocytes more than 8% as in lymphoma and
monocytic leukemia; subacute bacterial endocarditis; some chronic infection and malaria.

2- Lymphocytes
Lymphocytes represent 20 to 40% of the differential white cell count. There are two
structural types:

1. Small lymphocytes: ~5μm in diameter, and represent 3% of lymphocytes inperipheral

blood. Most small lymphocytes in the blood stream belong to either the group of B-
lymphocytes (~5%) or the group of T-lymphocytes (~90%).
2. Large lymphocytes: 9 to 15μm in diameter, possibly natural killer cells; possibly dividing
lymphocytes. The cells are rounded with densely stained nuclei, small amount of pale
basophilic cytoplasm with free ribosomes; short microivilli (seen in EM) more numerous
on B-lymphocytes than T lymphocytes. Unless they become activated, the two groups
cannot easily be distinguished using routine light or electron microscopy.

Only blood lymphocyte capable of division outside the bone marrow.

Functional types of lymphocytes:

o B-lymphocytes: responsible for humoral immune response and produce antibodies.

o T- lymphocytes: responsible for cell mediated immune response.

o T- helper lymphocytes

o T- suppressor lymphocytes

o T- memory lymphocytes

o Cytotoxic T- lymphocytes (Killer cells)

o Natural killer lymphocytes

Functions
- Upon exposure to antigens B-lymphocytes differentiate into antibody producing plasma cells
that produce antibodies which directed against foreign antigen.

- T-lymphocytes represent the "cellular arm" of the immune response (cytotoxic T cells) and may
attack foreign cells, cancer cells and cells infected by e.g. a virus.

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- Lymphocytosis: increased numbers above the normal as in viral infection, chronic bacterial
infection such as typhoid fever and tuberculosis; lymphoma and lymphocytic leukemia.

- Lymphopenia: decrease the number of lymphocytes less than 20% as in AIDS, and in aplastic
anemia.

C- Blood Platelets (Thrombocytes)

Blood platelets or thromhacytes, are the smallest formed elements in the blood. They are
cytoplasmic fragments of very large thrombocyte (megakaryocytes) that are found in the bone
marrow. Their number is 150,000 - 400,000/mm³, with a lifespan of about 8 days, and appear in
clumps in blood smears. They are rounded or oval, biconvex discs, 1.5 to 3.5μm in diameter. The
cytoplasm is divided into two zones: an outer hyalomere, and an inner granulomere, which
contains a few mitochondria, glycogen granules and a variety of purple granules. The hyalomere
contains cytoskeletal fibers, which include actin and myosin filaments. Different types of vesicles
contain either serotonin (electron-dense delta granules) or compounds important for blood
coagulation (alpha granules), they also contain platelet-derived growth factor (PDGF) which may
play a role in the repair of damaged tissue. Their cytoplasm is purple-staining, granular;
organelles concentrated toward the center; granules constitute about 20% of the volume. The
glycocalyx is rich in glycosaminoglycans and is associated with adhesion, the major functional
characteristic of platelets. Platelets have an important physical role in plugging wounds, and they
contribute to the cascade of molecular interactions among the various clotting factors dissolved
in the plasma.

The Clot and Serum: Clotted blood consists of 2 parts:

a. the clot, or thrombus, which includes the formed elements and some of the proteins
dissolved in the plasma,
b. the serum, a clear yellow liquid that is similar to plasma except that it lacks fibrinogen and
contains more serotonin.

Clotting Factors:
Clotting involves a cascade of molecular interactions among several plasma proteins and
ions (clotting factors I - XIII). The cascade can be initiated by 2 converging pathways, each of
which results in the conversion of fibrinogen into fibrin by the enzyme thrombin. Other factors
act as promoters and accelerators of the clotting process or help stabilize the fibrin once it has
formed. An inherited abnormality in factor VIII results in the clotting disorder known as
hemophilia.

The Role of Platelets:

1. Primary aggregation. Platelets in the damaged region attach to collagen revealed by the
discontinuity in the vessel wall, forming a platelet plug.
2. Secondary aggregation. Platelets in the plug release the contents of their alpha and delta
granules. This release of serotonin explains the higher concentration of serotonin in serum
than in plasma. Serotonin, a vasoconstrictor, restricts blood flow to the damaged area by
causing contraction of vascular smooth muscle.
3. Blood coagulation. Platelets release fibrinogen in addition to that normally found in the
plasma. The fibrinogen is converted by the clotting factor cascade into fibrin, which forms a

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 dense fibrous mat to which more platelets and other blood cells attach, forming a clot and
plugging the opening in the blood vessel wall.
4. Clot Retraction: The clot (thrombus) initially bulges into the vessel lumen, but later it
contracts and condenses through the interactions of thrombosthenin (a contractile protein)
and platelet actin, myosin, and ATP. Clot Removal: As the vessel wall heals and the
protection afforded by the clot is no longer needed, the clot is removed by the enzyme
plasmin, Plasmin is formed by the action of plasminogen activators (from endothelial cells) on
the plasma proenzyme plasminogen (from the liver). Enzymes released by the lambda
granules (lysosomes) of the platelets also aid in clot digestion

Cell Type Main Products Main Functions

Erythrocyte Hemoglobin CO2 and O2 transport
Leukocytes Specific granules and Phagocytosis of bacteria
modified lysosomes
(azurophilic granules)
Neutrophil
Eosinophil Specific granules,
Defense against parasitic
pharmacologically active
helminths; modulation of
substances inflammatory processes
Basophil Specific granules containing
Release of histamine and
histamine and heparin other inflammation
mediators
Monocyte Granules with lysosomal Generation of mononuclear-
enzymes phagocyte system cells in
tissues; phagocytosis and
digestion of protozoa and
virus and senescent cells
B lymphocyte Immunoglobulins Generation of antibody-
producing terminal cells
(plasma cells)
T lymphocyte Substances that kill cells. Killing of virus-infected cells
Substances that control the
activity of other leukocytes
(interleukins)
Natural killer cell (lacks T Attacks virus-infected cells Killing of some tumor and
and B cell markers) and cancer cells without virus-infected cells
previous stimulation
Platelet Blood-clotting factors Clotting of blood

The lifespan of an erythrocyte in the bloodstream is 100-120 days i.e. about 5×10¹¹
erythrocytes are formed/ destroyed each day.

Erythron (census): whole mass of RBCs & their precursors in bone marrow.

RBCs are considered as 'yardstick' for estimating dimensions of other cells in sections, (the
diameters of relative cells are about 7μm).

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Plasmalemma and stroma The stroma is composed of proteins such as spectrin that are
associated with the inner surface of the plasmalemma; it maintains the biconcave shape of the
RBC. The external surface of the plasmalemma is covered by a carbohydrate-rich glycocalyx,
which contains genetically determined antigens that allow blood types (including A, B, 0, AB
groups) to be distinguished.

Hemoglobin Each hemoglobin molecule consists of polypeptide subunits, each includes an iron-
containing heme group. Hemoglobin (Hb) exists in a different forms, distinguishable on the basis
of the amino acid sequence of their subunits. In humans, only 3 forms are considered normal in
postnatal life: HbAI constitutes 97%, HbA2 2%, and HbF 1% of the hemoglobin of healthy adults.
HbF makes up around 80% of the hemoglobin of newborns, however; this proportion gradually
decreases until normal adult levels are reached at about 8 months of age. HbS is an abnormal
form of HbA that is found in patients with sickle cell anemia. Unlike HbA, HbS becomes insoluble
at low oxygen tensions and crystallizes into inflexible rods that deform the RBCs, giving them the
characteristic sickle shape. When the rigid sickled cells pass through narrow capillaries, they
cannot bend as normal RBCs do. They may become trapped, obstructing blood flow through the
capillary, or rupture, decreasing the number of RBCs available for oxygen transport (anemia).

Abnormalities of RBCs: usually named as anemia that are due to changes in shape, number,
or hemoglobin content.

Importance of the Study of the Blood

1. As circumstantial or corroborative evidence against or in favor of the perpetrator.

2. As evidence in case of disputed parentage.
3. As evidence in the determination of the cause of death and the length of victim survived
the attack.
4. As evidence in the determination of the direction of the case of victim or the assailant.
5. As evidence in the determination of the origin of the flow of blood.
6. As evidence in the determination of the approximate time the crime was committed.

Problems in the study of the Blood

1. Where blood has to be searched?
In the collection of bloodstains, usually attention is directed to clothing and
weapons. The investigator should also look for the bloodstain under the fingernails, linings
of the pockets, seams and the folds of the garments of the suspect, under the edges of
the table, etc.
2. Collection, preservation, packing and transportation of specimen suspected to
contain blood.
Blood offers little resistance to decomposition. It undergoes a rapid change in its
character with the passage of time as process of clotting and drying commences almost
immediately on exposure to air. Sodium fluoride may be added to preserve if for a week
at room temperature or infinitely in a refrigerator. Between 40 - 50°C is the ideal
preserving temperature for blood and other perishable specimens. Collection of
bloodstains should be done as soon as possible. Mere washing of garment/clothing
removes the blood.

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GUIDING PRINCIPLES IN COLLECTION, PACKING, PRESERVATION AND
IDENTIFICATION OF BLOODSTAINED EVIDENCES

A. LIQUID BLOOD
Before collection, make careful notes describing the exact location and condition of
any blood or bloodstains you may find. Note down the general color and condition of
the bloodstains. If the stains has a bright red color and conditions of the bloodstains
becomes older; the red color change to dark brown color.

1. A) Collect the blood sample by picking up with a clean medicine dropper and place in
a test tube containing Sodium Fluoride.
B) Cover the test tube with stopper and seal.
C) Label the test tube bearing the initials or the name of the investigator and date of
the collection and seal. Label maybe in the form of paper pasted on the container or
a tag tied to the object.

2. A) in the absence of sodium fluoride, put the blood on a microscope slide or soak them
in pieces of blotting paper and let them dry carefully without any heat.
B) Place the dried stain in a pillow or white mailing envelope and seal by placing
masking tape across the flap of the envelope and label them by inscribing thereon the
case information.

B. BLOOD STAINS ON WALLS AND FLOOR (solid, immovable objects)

1. Place a clean sheet of paper below the surface bearing the stain.
2. With the use of a razor blade or clean knife, loosen and scrape the dried material into
the paper. Crust should be removed without breaking if possible.
3. Shake the powdered stain to the center of the paper and then fold. Scrapings removed
from the different areas of the walls and floors are separately placed on the paper and
labeled as to the location where found.
4. Seal the folded paper thoroughly with cellophane tape and individually place inside a
white mailing envelope, seal with masking tape and affix the initials of the investigator,
date and other information.

C. BLOODSTAINS ON CLOTHING AND FABRICS

1. Encircle with chalk or any materials stained areas found on the clothing. If clothing is
wet, let it dry completely at room temperature in secure, well-ventilated room. Do
not place in direct sunlight or heat.
2. Place identifying marks directly on the cloth, as far away from the stained areas as
possible.
3. If buttons are present, attach a string tag bearing information regarding the case.
4. Wrap and pack the clothing separately in a close paper container such as a paper
bag.
5. Place the bag containing the items in the coldest, driest facility possible until it sent
to the laboratory.

D. BLOODSTAINS ON SMOOTH/GLAZED OBJECTS

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 1. With the use of separate cottons, moistened with saline solution, swab alleged
bloodstains found on different areas of the surface.
2. Dry the cotton thoroughly at room temperature and place the swabbing separately in
a white mailing envelope.
3. Seal the envelope with masking tape by placing across the flap and label by inscribing
thereon the location where the stain was found; initial of the investigator and the
date and time the sample was collected.

E. BLOODSTAINS ON HARD OBJECTS:

1. Place identifying marks directly on the recovered object, as far away from the stained
area as possible. Provide a label or string, tag bearing the following information:
a. Case number
b. Date and time the specimen was found
c. Name and description of the specimen
d. Location and time of recovery
e. Signature of investigator
f. Name of witness to the discovery
2. Dry wet objects at room temperature. Avoid exposure to direct sunlight or heat.
After drying, wrap the object in closed container and proceed as in number one

3. Does the stain contain blood or another substances?

The examination of the specimen should determine if the stain is blood, if it is animal or
human blood, what blood groups are present.

The criminalist must be prepared to answer the following questions when examining dried blood:

1. Is it blood?
2. From what species did the blood originate?
3. If the blood is of human origin, how closely can it be associated to a particular
individual?

Wet blood has more value than dried blood because more tests can be run. For example,
alcohol and drug content can be determined from wet blood only. Blood begins to dry after 3-5
minutes of exposure to air. As it dries, it changes color towards brown and black. Blood at the
crime scene can be in the form of pools, drops, smears, or crusts. Pools of blood obviously have
more evidentiary value in obtaining a wet sample.

Blood Analysis
Drops of blood tell the height and angle from which the blood fell. The forensic science of
blood spatter analysis says that blood which fell perpendicular to the floor from a distance of 0-2
feet would make a circular drop with slightly frayed edges. Drops from a higher distance would
have more pronounced tendrils fraying off the edges (a sunburst pattern). A blood smear on the
wall or floor tells the direction of force of the blow. The direction of force is always in the
direction towards the tail, or smaller end, of the smear, or splatter. In other words, the largest
area of the smear is the point of origin (a wave cast-off pattern). Blood crusts need to be tested
with crystalline methods to make sure it's blood.

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Refrigerated red blood cells have a shelf life of about 42 days, and the serum containing white
blood cells can be refrigerated much longer, almost up to a year. DNA can be extracted from
blood (if white blood cells which always contain a nucleus are present), and also from sperm,
bone marrow, tooth pulp, and hair roots. Blood, however, is commonly used in DNA testing, as
per the following steps:
1. Blood samples are collected from the victim, defendant, and crime scene.
2. White blood cells are separated from red blood cells
3. DNA is extracted from the nuclei of white blood cells
4. A restrictive enzyme is used to cut fragments of the DNA strand
5. DNA fragments are put into a bed of gel with electrodes at either end
6. Electric current sorts DNA fragments by length.
7. An absorbent blotter soaks up the imprint; it is radioactively treated, and an X-ray
photograph (called an autoradiograph) is produced.

Stain Patterns of Blood

Surface texture is of paramount importance. In general, the harder and less porous the
surface, the less spatter results. The direction of travel of blood striking an object may be
discerned because the pointed end of a bloodstain always faces its direction of travel. The impact
angle of blood on a flat surface can be determined by measuring the degree of circular distortion.
At right angles the blood drop is circular, as the angle decreases, the stain becomes elongated.
The origin of a blood spatter in a two-dimensional configuration can be established by drawing
straight lines through the long axis of several individual bloodstains. The intersection or point of
convergence of the lines represents the origin point.

Illustration of stain convergence on a two-dimensional plane. Convergence represents the

point from which the stains emanated. Courtesy Judith Bunker, J. L. Bunker & Assoc., Ocoee, FL

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 Categories of Blood Stains

a. Passive

Passive bloodstains are drops created or formed by the force of gravity acting alone.

b. Transfer

A transfer bloodstain is created when a wet, bloody surface comes in contact with
a secondary surface. A recognizable image of all or a portion of the original surface may
be observed in the pattern, as in the case of a bloody hand or footwear.

c. Projected

Projected bloodstains are created when an exposed blood source is subjected to an action
or force, greater than the force of gravity. (Internally or Externally produced). The size, shape,
and number of resulting stains will depend, primarily, on the amount of force utilized to strike the
blood source.

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• Arterial Spurt/Gush

Bloodstain pattern(s) resulting from blood exiting the body under pressure from a breached
artery
• Cast-off Stains
Blood released or thrown from a blood-bearing object in motion
• Impact Spatter
Blood stain patterns created when a blood source receives a blow or force resulting in the
random dispersion of smaller drops of blood.
Velocity affects stain pattern

Low Velocity Medium Velocity

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High Velocity

Directionality of Bloodstains

When a droplet of blood strikes a surface perpendicular (90 degrees) the resulting
bloodstain will be circular. That being the length and width of the stain will be equal. Blood
that strikes a surface at an angle less than 90 degrees will be elongated or have a tear
drop shape. Directionality is usually obvious as the pointed end of the bloodstain ( tail ) will
always point in the direction of travel.

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 SIN < = Width (a) 1.5cm
Length (c) 3.0cm

SIN < = 0.5

< = 30 degrees

The determination of blood is best made by means of a preliminary color test.

CHRONOLOGICAL TEST FOR BLOOD

1. PRELIMINARY TEST
It determines whether the stains contain blood or another substance. It is used to
demonstrate the presence of blood. It determines whether visible stains do or do not contain
blood.

a. BENZIDINE TEST
An extremely sensitive test that can be applied to minute stain. For many years,
the most commonly used preliminary test for blood. Its use has generally been discontinued,
as it is known carcinogen. A very delicate test and will detect blood when present in dilution
of 1:300, 000 parts. The Benzidine test never fails to detect blood even when very old,
decomposed stain with all sorts of contamination is examined. This test is more sensitive than
the guaiacum test and is valuable as a negative result. If the stain reacts negatively it is not
blood. The positive result is an indication that blood may be present.
Reagent: a. Benzidine Solution (a small amount of powdered benzidine dissolved in
glacial acetic acid)
b. 3% solution of hydrogen peroxide
Procedure: Place a small fragment/portion of the stained material on a filter paper.
Add a drop of benzidine solution and then a drop of hydrogen peroxide solution.
Positive Result: intense blue color appears immediately
Limitation of the Test: Benzidine test is not specific test for blood. Positive result may
be obtained from the substances as sputum, pus, nasal secretion, plant juices, formalin, clay
and gum. The reaction is weaker and produce faint coloration.

b. PHENOLPHTHALEIN TEST
An alternative test to Benzidine test. It can detect blood in a dilution of
1:80,000,000 parts. A positive result with this test is highly indicative of blood. The negative
result is therefore valuable and is conclusive as to the absence of blood.
Reagent: a. Phenolphthalein solution (1 to 2 grams of phenolphthalein to 100 mL of
a 25% potassium hydroxide in water added with one gram of zinc powder heated until
colorless)
b. 3% solution of hydrogen peroxide

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 Procedure: Place a small fragment/portion of the stained material on a filter paper.
Add a drop of phenolphthalein solution and then a drop of hydrogen peroxide.
Positive Result: Rose color develops deep pink /permanganate color.
Limitation of the Test: The test is also given by the copper salts, potatoes and
horseradish.

c. GUAIACUM TEST
A fairly delicate test showing the presence of fresh blood in a solution of 1:50,000
dilution. It may not react to very old stains.
Reagent: a. Fresh tincture of guaiac resin (few lumps this to 95% alcohol, then filter.)
b. 3% hydrogen peroxide solution or few drops of turpentine
Procedure: Place a small piece of the stained fabric on a porcelain dish. Soak with fresh
tincture of guaiac. Add a few drops of hydrogen peroxide.
Positive Result: Beautiful blue color that appears immediately.
Limitation of the Test: The test also reacts with saliva, pus, bile, milk, rust, iron salts,
cheese, gluten, potatoes, perspirations, and other oxidizing substances.

d. LEUCOMALACHITE GREEN TEST

This test is not as sensitive as the benzidine test.
Reagent: a. Leucomalachite Green Solution (1 gram leucomalachite green dissolved
in 48 mL glacial acetic acid and diluted to 250 mL water)
b. 3% hydrogen peroxide
Procedure: Place a small piece of a stained fabric on a filter paper. Add a drop of
leucomalachite green solution and after a few seconds add a drop of hydrogen peroxide
Positive Results: Malachite green or bluish green

e. LUMINOL TEST
An important presumptive identification for blood. The reaction of luminal with
blood results in the production of light rather than color. By spraying luminal reagent onto a
suspected item, large areas can be quickly screened for the presence of bloodstains. The
sprayed object must be located in a darkened area while being viewed for the emission of
light. Luminol test is extremely sensitive test. It is capable of detecting bloodstains diluted
up to 10, 000 times. Luminol is known to destroy many important blood factors necessary for
the forensic characterization of blood, so its use should be limited only to seeking out blood
invisible to the naked eye.
Positive Result: Luminescence or emission of light

PRINCIPLE INVOLVED IN THE FOUR PRELIMINARY TEST FOR BLOOD

The Peroxidase present in the hemoglobin acts as carrier of Oxygen from the Hydrogen
Peroxide to the active ingredients of the reagents (benzidine, guaiac, phenolphthalein and
leucomalachite) and produces the characteristic colored compounds by oxidation.
Peroxidase – is an enzyme that accelerates the oxidation of several classes of organic
compounds by peroxide.

2. CONFIRMATORY TEST
The actual proof that a stain is blood consists of establishing the presence of the
characteristics blood pigment hemoglobin or one of its derivatives. Hemoglobin is the red
coloring matter of the RBC.

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 The three confirmatory tests for blood that determine whether stain is really blood are:
a. THE MICROSCOPIC TEST FOR BLOOD
It is useful for the demonstration and mensuration of blood corpuscles for making the
distinction between mammalian, avian, piscine and reptilian blood for the investigation
of menstrual, lochial and nasal charges.
Method of Microscopic Examination
1) Take two small fragments of the dried blood.
2) Place each fragment on separate slides with a drop of 0.9% salt solution.
3) The slides are placed in a covered dish to prevent evaporation and the
preparation allowed to stand for 1-2 hours.
4) One of the slides is examined as wet preparation.
5) The other preparation is spread evenly over the slide, allowed to dry and
stained by:
a) Fix preparation in absolute methyl alcohol for 3 minutes. Stain in a
0.5% aqueous solution of eosin for 1-3 minutes. Loffer’s methylene
blue is added for 1-3 minutes. Eosin stains the red blood cells, while
methylene blue stains the nuclei.
b) Fix smear with methyl or ethyl alcohol for 3 minutes. Pour off alcohol
and flood smear with Geimsa’s stain. Stain for 15 minutes, cover to
prevent evaporation, wash in water and dry.
c) Wright’s stain – the smear is flooded with the stain and allowed to stand
for a minute. Distilled water is added until a metallic scum forms on the
surface. Let stand for 3 minutes, wash with water and dry.

Visible Results:
1. Mammalian Red Blood Cells – circular, biconcave discs with nucleus. Appear as
characteristics non-nucleated discs.

Exception is camel and closely related animal such as llama whose red blood cells are oval
without nucleus

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2. Birds, Fish and Reptile Red Blood Cells – large, oval and nucleated

Bird’s RBC Fish RBC Reptile RBC

3. Amphibian Red Blood Cells – larger than mammals, oval and nucleated

4. Lamprey and Eel Red Blood Cells – circular and nucleated

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 b. MICROCHEMICAL TEST OR MICROCRYSTALLINE TEST
The identification of blood can be made more specific if microchemical or
microcrystalline test is applied or performed. Takayama test and Teichmann test are
the most popular ones.

THREE MICROCHEMICAL AND MICROCRYSTALLINE TEST FOR BLOOD

1) TEICHMANN HAEMIN REACTION OR TEICMANN TEST OR HAEMIN CRYSTAL

TEST

The Test depend on the addition of the specific chemicals to the blood so that
characteristic crystals with hemoglobin will be formed.
Reagent: Sodium chloride, glacial acetic acid
Procedure: Place a minute fragment of the stain on a glass slide. Add a small
crystal of sodium chloride and 2 to 3 drops of acetic acid. Place cover slip and heat
gently over a small flame to evaporate the acid. Cool. Examine under the high-power
objective.
Positive Result: Dark brown rhombic crystal of haemin or haematin chloride
arranged singly or in cluster.
Limitation of the Test: The test is also given by indigo-dyed fabrics. If the stain is
old or washed or is changed by chemical reagents, the crystals are not formed. The
addition of too much salt or presence or moisture in the acid or over- heating of the
slide may result in failure.

2) ACETONE-HAEMIN TEST
The test depends on the addition of specific chemicals to the blood so that
characteristics crystal with hemoglobin will be formed.
Reagent: Acetone, dilute acetic acid or oxalic acid
Procedure: Place dried stain on a glass slide and cover with cover slip
with a needle interposed to prevent direct contact of the cover slip with a cover slip
with the slide. Add a drop of acetone then a drop of acetic acid.
Positive result: small, dark diachronic acicular crystal of acetone haemin

3) HAEMOCHROMOGEN TEST OR TAKAYAMA TEST

A delicate test for the presence of hemoglobin. The test depends on the addition of
specific chemicals to the blood so that characteristic crystals of hemoglobin derivatives
will be formed.

Reagent: Takayama reagent (3 mL of 10% sodium hydroxide, 33 mL of

pyridine, 3 cc of saturated glucose solution and diluted with 7 mL water)
Procedure: Place a small piece of suspected material on a glass slide. Add a
drop of Takayama reagent. Cover with a glass slip.
Positive result: Large rhombic crystal of a salmon pink color arranged in cluster,
sheaves and other forms that appear within 1 to 6 minutes when viewed under the
low power objective. To hasten result heat may be applied.

c. SPECTROSCOPIC TEST

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 The most delicate and reliable test for the determination of the presence of blood in
both old and recent stains. This test is performed by means of an applied instrument
known as Spectroscope, an optical instrument for forming and examining spectra.
Procedure: Dissolved bloodstain in water or saline solution. Place in small
chamber (glass) with parallel sides so arranged that the rays of light will pass directly
through it. The chamber is placed in the spectroscope and the instrument is so
adjusted that the spectrum is clearly visible.
Positive Result: Upon absorbing some of the rays from the spectrum, it is produced
characteristic dark colored bands, which vary with the type of blood pigment. Example:
Oxyhemoglobin is marked by two bands, hemoglobin-broad band, carboxyhemoglobin
its spectrum like oxyhemoglobin.
Principle involved in the Spectroscopic Test: The absorption properties of
translucent colored fluids can be observed on the solar spectrum.
3. PRECIPITIN TEST
The precipitin Test is the standard test used to determine whether the stain/blood
is of human or animal origin. The precipitin test is very sensitive and requires only a small
amount of blood for testing. Human bloodstain dried for as long as 10-15 years and longer
may still give a positive precipitin reaction. Even extracts of tissues from mummies four
to five years old have given positive reaction with the test. Experience has shown that
human bloodstain diluted by washing in water and left with only a faint color may still
yield a positive precipitin reaction.
Reagent: Precipitin/antiserum
Procedure: Scrape off bloodstain if is on the hard object. Powder the scrapings and
extract with saline solution. Allow mixture to stand overnight. Centrifuge to clean the
solution. Dilute with saline solution. Layer an extract of the bloodstain on top of the human
antiserum/precipitin in a capillary tube.
Positive Result:
o Development of a white cloudy line at the contact point of the fluids that appears
immediately or within one or two minutes.
o Human Blood, or for that matter, any protein of human origin in the extract will react
specifically with antibodies present in the serum as shown by the formation of cloudy
ring or band at the interface of the two liquids.
Principle involved in the Precipitin Test
When a rabbit is injected with a human blood serum or whole human blood the
precipitin that develops in its serum will react with the protein of human blood serum,
other human body fluid and other human tissue extracts. The reaction is a specific one
and if positive, will identify blood proteins or any other protein as human origin.
Limitation of the Test
The precipitin reacts not only with blood proteins but also with other body proteins as
those in saliva, semen, mucus and other body fluids. For this reason, the test does not
identify specifically human body but only a protein material from the specific animal type.
In order that a conclusion of human blood is arrived the precipitin must be corroborated
by supplementary chemical, microscopic or spectroscopic tests.
The specificity and delicacy of the precipitin reaction is great, but the reaction may be
inhibited or even destroy by a number of factors. Chemicals like acid, alkalis, alcohols,
cresols, formaldehyde, corrosive sublimate or other germicide may alter blood to such
extent that the reaction cannot be formed. Heat had the same effect. Fluid blood loses its
power of reacting with antiserum if it is heated from 60 to 90°C, while dried blood may

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 stand 150°C. Rust and postmortem decomposition may react with it poorly. Old stains
may be identified after a long period of time.

4. BLOOD GROUPING TEST OF FRESH BLOOD

If the specimen is human blood, the next question is, did it come from the victim,
the accused or from other persons? So the origin of blood or bloodstain will be determined
by the identification of the blood groups to which it belongs, in short to what blood group
does it belong? This identification is carried out on both fresh blood and bloodstains.
Human blood of all races can be divided into definite groups.

The reciprocal relationship between antigens on the red blood cells and antibodies
in the serum is known as Landsteiner’s law. Karl Landsteiner suggested that the
phenomenon was not pathology, as was the prevalent thought at the time, but was a
physiological phenomenon due to the unique nature of the individual’s blood.

It states that:

1. If an agglutinogen is present in the red cells of a blood, the corresponding

agglutinin must be absent from the plasma.
2. If an agglutinogen is absent in the red cells of a blood, the corresponding
agglutinin must be present in the plasma.

Applicability of the law:

• The first law holds true for all types of blood grouping.
• The second law is a fact for ABO blood groups.
• The Rh,M,N and other blood groups do not follow the second part of landsteiner’s
law.

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 Short biography About Karl Landsteiner:

Karl Landsteiner

• Karl Landsteiner (1868-1943), the Austrian-born American immunologist was the

only child of Leopold Landsteiner, a prominent Austrian journalist and editor, and
Fanny Hess Landsteiner.
• Karl Landsteiner, has been titled as the “father of immunology”.
• He discovered the blood groups (ABO) and framed the Landsteiner’s Law by 1904.
• In 1930 he received the Nobel Prize for his discovery.
• Continued to research on Blood groups and types till his death.

Blood typing is a test that determines a person’s blood type. The test is essential
if you need a blood transfusion or are planning to donate blood. Not all blood types are
compatible, so it’s important to know your blood group. Receiving blood that’s
incompatible with your blood type could trigger a dangerous immune response.

The blood types

Your blood type is determined by what kind of antigens your red blood cells have
on the surface. Antigens are substances that help your body differentiate between its own
cells and foreign, potentially dangerous ones. If your body thinks a cell is foreign, it will
set out to destroy it.

The ABO blood typing system groups your blood into one of four categories:

• Type A has the A antigen.

• Type B has the B antigen.
• Type AB has both A and B antigens.
• Type O has neither A nor B antigens.

If blood with antigens that you don’t have enters your system, your body will
create antibodies against it. However, some people can still safely receive blood that isn’t

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 their blood type. As long as the blood they receive doesn’t have any antigens that mark it
as foreign, their bodies won’t attack it.

In other words, donations work as follows:

• O: Type O individuals can donate blood to anyone, because their blood has no
antigens. However, they can only receive blood from other type O individuals (because
blood with any antigens is seen as foreign).
• A: Type A individuals can donate to other type A individuals and type AB individuals.
Type A individuals can receive blood only from other type A individuals and type O
individuals.
• B: Type B individuals can donate blood to other B individuals and AB individuals. Type
B individuals can receive blood only from type B individuals and type O individuals.
• AB: Type AB individuals can give blood only to other AB individuals, but can receive
blood of any type.

Blood types are further organized by Rh factor:

• Rh-positive: People with Rh-positive blood have Rh antigens on the surface of their
red blood cells. People with Rh-positive blood can receive Rh-positive or Rh-negative
blood.
• Rh-negative: People with Rh-negative blood do not have Rh antigens. People with
Rh-negative blood can receive only blood that is also Rh-negative.

Together, the ABO and Rh grouping systems yield your complete blood type. There
are eight possible types: O-positive, O-negative, A-positive, A-negative, B-positive, B-
negative, AB-positive, and AB-negative. While type O-negative has long been considered
a universal donor, more recent research suggests that additional antibodies are sometimes
present and may cause serious reactions during a transfusion.

Austrian Karl Landsteiner discovered blood types in 1901. Before that, blood
transfusions were risky and potentially lethal. Landsteiner made the process much safer,
and he was awarded the Nobel Prize for his work.

Why blood typing is done

Blood typing is done prior to a blood transfusion or when classifying a person’s

blood for donation. Blood typing is a fast and easy way to ensure that you receive the
right kind of blood during surgery or after an injury. If you’re given incompatible blood, it
can lead to blood clumping, or agglutination, which can be fatal.

Blood typing is especially important for pregnant women. If the mother is Rh-
negative and the father is Rh-positive, the child will likely be Rh-positive. In these cases,
the mother needs to receive a drug called RhoGAM. This drug will keep her body from
forming antibodies that may attack the baby’s blood cells if their blood becomes mixed,
which often happens during pregnancy.

Blood Type Compatibility

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 Your ABO blood type is based on the presence or absence of the A and B antigens
on your red blood cells. The A blood type has only the A antigen and the B blood type has
only the B antigen. The AB blood type has both A and B antigens, and the O blood type
has neither A nor B antigen.

By the time you are six months old, you naturally develop antibodies against the
antigens your red blood cells lack. For instance, a person with A blood type will have anti-
B antibodies, and a person with B blood type will have anti-A antibodies. If you have type
A blood, you cannot receive B blood because your body's anti-B antibodies will fight the
B blood's B antigens. It is crucial we have all blood types available to our patients.

Can receive

O- O+ B- B+ A- A+ AB- AB+
Blood Type
O- Yes
O+ Yes Yes
B- Yes Yes
B+ Yes Yes Yes Yes
A- Yes Yes
A+ Yes Yes Yes Yes
AB- Yes Yes Yes Yes
AB+ Yes Yes Yes Yes Yes Yes Yes Yes

Beinsten’s Theory of Blood Group Inheritance

Beinsten’s theory postulates the presence of three allelic genes A, B and O.

According to him, the blood group of any individual is determined by combinations of A,
B, and O in a particular pair of chromosomes. One gene is derived from the father and
the other gene from the mother. Genes A and B are dominant over gene O. A and B
determine the presence of the corresponding agglutinogens, while O determines their
absence. The possible combination of this three genes arranged in pairs give rise to six
different genotypes corresponding to the four phenotypes or the blood groups. There are
ten different matings possible between the four blood groups.

ABO Blood Type

Everyone has ABO blood type (A, B, AB, or O) and an Rh factor (positive or
negative). Just like eye or hair color, our blood type is inherited from our parents.

Each biological parent donates one of two ABO genes to their child. The A and B
genes are dominant and the O gene is recessive. For example, if an O gene is paired with
an A gene, the blood type will be A.

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 For instance, a parent with O blood with 2 O genes and a parent with A blood with
2 A genes will have an A blood type child with one A gene and one O gene.

• AA = A blood type
• AO = A blood type
• BB = B blood type
• BO = B blood type
• OO = O blood type
• AB = AB blood type

Rh Factor

The Rh factor is simply a protein that is found on the covering of the red blood
cells. If your red blood cells have this protein, you are Rh positive. If your blood cells don't
have this protein, you are Rh negative.

Just as everyone inherits ABO genes, every person inherits one Rh factor gene
from each parent. The Rh-positive gene is the dominant gene when paired with an Rh-
negative gene.

Negative /
Parent Genes Positive / Postive Negative / Positive
Negative

Negative / Negative Neg/Neg Neg/Pos Neg/Neg, Neg/Pos

Positive / Positive Neg/Pos Pos/Pos Neg/Pos, Pos/Pos

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 Negative /
Parent Genes Positive / Postive Negative / Positive
Negative

Neg/Neg, Neg/Pos,
Negative / Positive Neg/Neg, Neg/Pos Neg/Pos, Pos/Pos
Pos/Pos

• Neg/Neg = Negative Rh factor

• Neg/Pos = Postive Rh factor
• Pos/Pos = Positive Rh factor

The MN System of Blood Group

In 1927, Lansteiner and Levine discovered two new agglutinogens in human red blood
cells that define three types of blood, namely Type M, Type N, Type MN. These are independent
of the agglutinogens A and B. The human sera, however, do not contain natural for M. The
agglutinins can be demonstrated only by hetero-agglutination reaction with appropriate immune
rabbit sera. So anti-M and anti-N were produce by heter-agglutination reaction and just like anti-
A and anti-B, it determines the presence of agglutinogen M and N in the red blood cells.

Identification of Blood Type with Known Anti-M and Anti-N

Anti-M+ Whole Anti-N+ Whole Antigen present Type

Blood Blood
+ - M M
- + N N
+ + M and N MN

Inheritance of the Three M-N Types

Six different mating are possible between the three blood types. Types MN is always
heterozygous. The heredity of agglutinogens M and N according to Landsteiner and Levine
depends upon a single pair of allelic genes M and N which give rise to the three genotypes MN,
M and N respectively

Type of Parent Type of children possible Impossible

MxN M N, MN
M x MN M, MN N
MxN MN M, N
NxN N M, MN
N x MN N, MN M
MN x MN M, N, MN None

Grouping of Dried Bloodstains

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 Absorption technique or absorption-elution technique is an indirect grouping technique of
bloodstains and it depends on the detection of agglutinogen in the dried blood.

In dried blood grouping one cannot use the direct method as used in grouping of fresh
blood because in direct grouping, the identification of A and B antigens is accomplished by directly
reacting the blood with Anti-A and Anti-B serum. In dried blood, the red blood cells are already
ruptured due to dying, leaving no cells in the stain to be agglutinated. However, although the
cells may have disintegrated, the antigens present on their surface remain and are still identifiable
by indirect means.

Identification of Bloodstain with Known Cells (Absorption Technique)

A Cells + Anti-A+ B cells + Anti B+ Antibody present Blood Group

Bloodstains Bloodstains
+ - Anti-A B
- + Anti-B A
+ + Anti-A and B O
- - Neither Anti-A or AB
Anti-B

Importance of Blood Group Data

Questions of illegitimacy and relationship in many cases may be solved by means of the
Blood groups as determined by the agglutinogens A, B, M and N.

1. Determination of whether a man accused of fathering a child out of wedlock could or

not be its parents.
2. Determination of whether a child born of a married woman could or could not have
been fathered by her legal spouse.
3. Determination of whether a child could or could not belong to a given set of parents
in the case of accidental interchange of infants in hospital.
4. Determination of whether a child who has been lost and later recovered after a long
interval could or could not belong to a given set of parents

POST-COMPETENCY CHECKLIST (FORMATIVE ASSESSMENT)

online quiz

FORENSIC 3 -FORENSIC CHEMISTRY AND TOXICOLOGY RCPP 1st Semester 2020-201 29