The Important Test for blood found in the crime scene. That blood accrues the Preperstive Test and Comfanmatory Test to Confirm the found blood sample is blood or not.

1. Blood Analysis
“FORENSIC BIOLOGY AND SEROLOGY”
Ved Prakash Meena

2. Jens
Martensson
Content
1. Introduction
2. Nature of Blood
3. Blood Grouping
4. Crime Scene
5. Collection of Blood Stain
6. Blood Analysis
7. Role Of Blood In Forensic
8. Conclusion
9. References
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Introduction
• Blood is a fluid connective tissues
• It circulates continually around
the body , allowing constant
communication between tissues
distant from each other .
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Function Of Blood
• It transport oxygen from the lungs to
the tissues and carbon dioxide from the
tissues to the lungs for extraction
• Nutrient from the elementary tract to
the tissues and cell waste to the
excretory organs principally the
kidneys
• Hormones secreted by endocrine
glands to their target glands and
tissues
• Transport protective substance e.g.
antibiotics to area of infection and
clotting factor that stop bleeding
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Composition of blood
• Blood is composed of a clear , straw
colored watery fluid called plasma in
between several type of blood cell are
suspended
• Plasma normally consist of 55% of the
volume of blood the remain 45% is
accounted for by the cellular fraction
of the blood
• The two fraction of blood is that is
blood cell and plasma can be
separated by the centrifugation or by
gravity when blood is allowed to stand
because cell is heavier than plasma
Nature of Blood

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Plasma
•Plasma consist of 90 to 92 % of water and have
some suspended substance that are
•Plasma protein
•Inorganic salts
•Nutrients , principally from digested foods
• waste materials
•Hormones
•Gases

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Plasma protein:- Make 7% of plasma they remain into the
blood due to there size and created osmotic pressure of blood
which keep plasma with in circulation.
Albumin:- Most abundant plasma proteins about 60 % of total
and have function to maintain osmotic pressure and also act as
carrier molecule for free fatty acid and some drugs and steroids.
Globulin:- antibodies transportation of some hormones and
mineral salts eg. thyroglobulin carry the hormone thyroxin
inhibition of some protolytic enzymes.
Clotting factors :-These are responsible for clotting of blood
most abundant factor is fibrinogen.
Electrolytes :-The ph of blood is maintain between 7.35 to 7.45
by involving buffer system.
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Nutrient :- The product of digestion e.g. Glucose, amino acid, fatty acid
and glycerol .
Waste products:-Urea ,Creatinine and uric acid are the waste products of
protein metabolism formed in the liver and carried in the blood to the
kidney.
Hormones:-Chemical messenger synthesized by endocrine gland.
Gasses:-Oxygen , carbon dioxide and nitrogen are transported round the
body dissolved in plasma.

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Blood cell
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3 types of blood cells are present
•Erythrocytes also called red blood cells
•Platelets are also called Thrombocytes
•Leukocytes also called white blood cells

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RED BLOOD CELL
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•It is biconcave no nucleus, and diameter is about 7 micrometer.
•The main function Is gas transport like oxygen and carbon dioxide they are
flexible so they can squeeze through narrow capillaries and contain no
intracellular organelles leaving more room for hemoglobin.
•Produce in red bone marrow process of development of red blood cells from
stem cells takes about 7 days called erythropoiesis life span o in the
circulation is about 120 days.

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White blood cell
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•An important function in defense and
immunity these are the largest blood cell
but contain only about 1% of the blood
volume they have nuclei and granule in
their cytoplasm .
•Granulocytes are Neutrophils,
Eosinophils, Basophils
•Agranulocytes are Monocytes and
Lymphocytes

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Platelets
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•They contain a variety of substance that
promote blood clotting
•Normal blood platelet count is about
200x 10*9 /1
•And 350 x 10 *9/1 200000 to
35000/mm*3

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Blood Grouping
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•Individuals have different type of antigens on the faces of their red blood
cells these antigens which are inherited determined the individual blood
group.
•Individuals make antibodies to these antigen but not to there own type of
antigen.
• this reaction would be fatal while transfusion .

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• About 55% of the population has either A
type of antigen and b type of antigen or
AB type on their red cell surface remaining
45 % neither A nor B that is O
• Blood group, A individual cannot make
anti-A so do not have their antibodies
blood group AB people neither make anti-
A nor make anti-B antibodies. They are
universal recipient.
• Group O neither has antigen A nor B so
act as a universal donor.
ABO System
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• About 85 % of people have this antigen they are rhesus positive and do not make
anti rhesus antibodies the remaining 15% have no rhesus antigen they are negative
Rh-negative are capable of making anti rhesus antibodies but are stimulated to do
so only in certain circumstances e.g. in pregnancy
RHESUS System
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Wet VS. Dry Blood:-
• Wet blood is more significant than dried blood
because the forensic scientist can perform more
tests in order to gain insight to the happenings of
the crime. (i.e.. alcohol and drug content can be
determined from wet blood only).
• Blood begins to dry after 3 to 5 minutes of
exposure to air.
• As it dries, it changes color from deep red to
brown and black.
• Blood can be categorized into pools, drops,
smears, or crusts.
Crime Scene
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1. Murder
• Blood on the weapon can be matched against the blood of the victim.
• Bloodstains on the clothing or person of the accused can be matched with the blood of
the victim.
• Hair roots found on weapons can be matched with the blood of the victim and accused.
2. Accidental deaths( RTA etc)
• Bloodstains on the tire, radiator grill, and other parts of the offending vehicle may
connect the victim and it.
3. Sexual crimes
• Bloodstains on the thigh and private parts of the victims
4. Blood of pregnancy/abortion
• Presence of chorionic gonadotropin ( early stages)
• Heat –stable alkaline phosphates( late stages)
Nature of Crime
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Liquid / Fresh Stain:-
• A clean white filter paper or a piece of clean white cloth; a control filter paper
should be also sent if the object is porous, obtain a portion of the unstained area
as well.
Old and Dried Stain:-
• Should be carefully scraped if on an immovable object.
• Scrapings placed in clean containers or envelopes sealed and labeled.
• On clothes or fibers... Scrapped off or a fragment of the material cut, collected in
paper bags, sealed & labeled.
• Dried blood stains on weapons, garments, etc. can be left intact and the entire
object submitted.
• Tape lifting bloodstains; If fingerprints present
Collection of Blood Stain
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1. Presumptive Tests :-
• Luminol Test
• Phenolphthalein Test
2. Confirmatory Tests:-
• Takayama Test
• Teichmann’s test
• ABAcard HemaTrace test strips
• Microscopic test
Blood Analysis
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Luminol Test
•A chemical compound, known as Luminol, is used in solution
or sprayed onto suspected surfaces.
•This compound gives a strong blue fluorescence when viewed
with a UV light.
•The Luminol reacts with hematin, a substance formed as
bloodstains age, and produces a luminescence which is best
observed in the dark.
•The luminescence lasts for several minutes and can be
photographed.
•Aged bloodstains tend to give more intense and longer-lasting
luminescence than fresh blood, and can be re-sprayed with
Luminol to be viewed again

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• Also known as the Kastle Meyer Test
• A Phenolphthalein solution is used to show
the possible presence of blood based upon a
peroxides reaction of hemoglobin which
produces a pink color.
Phenolphthalein Test
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• It is also known as Haemochromogen test.
• This test determines if the substance in question is
blood.
• The application of a specific solution developed by
Takayama, Haemochromogen crystals form by
treating a small amount of blood or a stain fragment.
• The crystals are observable under a microscope and
look like salmon-pink rhomboid crystals.
• It works by gently heating heme (dark red, non-
protein, iron-coating part of hemoglobin,
C34H32FeN4O4) and adding it to pyridine
(C5H5N) when there is some kind of sugar present,
such as glucose, in order to create pyridine
ferroprotoporphyrin crystals.
• This test produces brown crystals
Takayama Test
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Takayama reagent’s –
Saturated solution of glucose
Pyridine solution
NaoH (sodium hydroxide solution)
Glacial Acetic Acid

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• This test determines if there is blood present on
suspected dried blood stains.
• The hematin crystals begin to form when heated
blood is mixed with glacial acetic acid where there is
salt.
• The positive result is caused by the combination of a
halogen with ferroprotoporphyrin (red-brown to blue-
black crystalline salt containing iron).
• This test produces pink crystals that can reach up to
10 micrometers in size.
Teichmann’s test
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Teichmann’s reagent-
KCl, KBr
Glacial Acetic

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• HemaTrace test strips are used to detect blood by indentifying the presence of
human hemoglobin.
• The test strip contains an antihuman hemoglobin antibody.
• A blood sample is applied to the bottom of the test strip. If human hemoglobin is
present, then a mobile antibody-antigen complex will be formed.
• This complex will then migrate through the test strip to a test window.
• This window will indicate if there is a positive result for human hemoglobin with
a pink dye band.
• An immuno chromographic one-step test for the detection of human blood.
• HemaTrace uses anti-human hemoglobin (Hb) antibodies to provide a means of
detection for the presence of human blood.
• Is coated on test strip and has a detection limit of 0.07 ug Hb.
ABAcard HemaTrace test strips
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Microscopic Method of examination is
having paramount importance relating
to identification of blood stain.
• Determination of the species of origin
by nucleated RBC’s along with the cell
structure.
• Sex determination by examining
chromatin bodies in Leucocytes.
• Detection of blood related pathological
condition.
Microscopic test
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• The blood is important evidence found at crime scene ,that leads to
further investigation of various crimes.
• Blood examination has important role in solving suicidal ,accidental
and homicidal cases.
• Various test are performed to identify and confirms blood stain of
human or animal origin.
• Identification of paternity and maternity cases by blood grouping.
• Species and origin of blood detection given indication of various
animals.
• Determination of age of blood stains.
Role of Blood in Forensic
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• Blood is an important evidence in crime scene and must be carefully detected
and collected.
• Blood screening is used generally to screen out the suspects not involve and to
provide a more solid evidence to determine the criminals.
Conclusion
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• U.S. Department of Justice, Sourcebook in Forensic Serology, Immunology, and
Biochemistry
• Establishment of a Fast and Accurate Proteomic Method for Body Fluid/Cell
Type Identification
• NC State Crime Lab Technical Procedures Manual: Body Fluid Identification
• Science Service Justice. http://www.nfstc.org/pdi/Subject02/pdi_s02_m02_0
2_b.htm
Reference
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34. Ved Prakash Meena
M.Sc.- Forensic Science (2020 -2022)
Chandigarh University
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