BLOOD

Has been called the circulating tissue of the body. It is referred to as highly complex mixture of
cells, enzymes, proteins and organic substances. It is the red fluid of the blood vessels. Blood is
opaque on treatment with either, water or other reagents becomes transparent and assumes
lake color. It is faintly alkaline. Normal PH is 7.35 to 7.45.
COMPOSITION OF BLOOD
1.) 45% formed elements or the solid materials consisting chiefly of cells.
A) RED BLOOD CELLS OR ERYTHROCYTES – contains haemoglobin and carry oxygen
to various cells in the body. RBC are circular, biconcave disc and have rounded edges.
FUNCTION > they help oxygenate the body.
B) WHITE BLOOD CELLS OR LEUKOCYTES
Are masses of nucleated protoplasm it depends the body from invading microorganisms.
FUNCTION >protect the body from invading germs and helps in allergic response
C) BLOOD PLATELES OR THROMBOCYTES
Cells that are produced by the bone marrow necessary for proper clotting of blood.
Normally responsible for the retraction of blood clot.
FUNCTION > help in clotting of blood
2) 55% Plasma
The fluid or portion of blood where the
cells are suspended. It is principally
composed of:
1. WATER (90%)
2. SOLID(10%
Largely protein in nature, and consists of albumen, several globulins and fibrinogen.

ALBUMEN – the most abundant protein in the blood. It binds with many drugs.

GLOBULINS - have an important role in the immune

mechanism of the body. The globulins carry drugs as well as sex and thyroid hormones, lipids
and iron.
FIBRINOGEN - the soluble precursors of fibrin which forms blood clot.
PLASMA - the yellowish fluids of the blood in which numerous blood corpuscles
suspended. A straw – yellow liquid formed when blood to which an oxalate has been
added to prevent clotting is allowed to stand.
SERUM - a straw-yellow liquid formed when clotted blood is allowed to stand for sometimes
and the blood contracts.
IDENTIFICATION OF BLOOD AND BLOOD STAINS

LEGAL IMPORTANCE OF THE STUDY OF BLOOD

1. For disputed parentage (maternity and paternity)

 a. Disputed paternity may arise:
(1) When the wife committed adultery and the husband denied to the father of the
child.
(2) When a child was born out of lawful wedlock and the mother claimed someone to
be the father but he vehemently denied it
(3) In a claim for support or right of succession of the alleged illegitimate child
b. Disputed maternity may arise:
(1) In a case of allegation of interchange of children in a hospital or nursery home,
either accidentally or deliberately.
(2) In a case of wayward or stray children being claimed by two or more women
(3) For ownership of dead fetus or newly born child found in street trash.
2. Circumstantial or corroborative evidence against or in favor of the perpetrator of a crime
3. Determination of the origin of the flow of blood
4. Determination of the cause of death
5. Determination of the place of the commission of crime
6. Determination of the approximate time the crime was committed
7. Determination of the escape of the victim or the assailant
8. Determination of presence of diseases.

QUESTIONS TO BE ANSWERED IN THE EXAMINATION OF BLOOD

• Determine whether the stain is due to blood
• If due to blood, determine whether it is human origin or not.
• Does it belong to the person in question?
• If it is of human origin, to what group does it belong?
• The manner, degree and condition of the article which have been stained.
• Age of the stain.

STAGES IN THE EXAMINATION OF BLOOD

1. PRELIMINARY OR SCREENING TEST
A. PHYSICAL EXAMINATION
1.a SOLUBILITY TEST:
Recent blood shed is soluble in saline solution and imparts a bright red color.
Stains which have been exposed to air become dry; haemoglobin is transformed
to meth-haemoglobin or haematin. If the stain has been kept in damp places for a
long time, haemoglobin is transformed to haematin.
1.b Heat test – solution of the blood stain when heated will impart muddy
precipitate.
1.c Luminescence Test - Stains on dark fabric mixed with mud, paint, etc. emit
bluish-white luminescence in a dark room when sprayed with luminol.

B. CHEMICAL EXAMINATION
1. Saline extract of the blood stain plus ammonia will give a brownish tinge due to
the information of alkaline haematin.
2. BENZIDINE TEST:
 A piece of white filter paper is pressed firmly on the suspected stain. Benzidine
reagent is dropped on the paper, the followed by drops of active hydrogen
peroxide. A positive result will show blue color.
3. GUAIACUM TEST (Van Deen’s Dyas’ or Schombein’s test).
To a white filter paper pressed and rubbed on the surface of the stain, the solution
of alcoholic tincture of guaiacum is added and then hydrogen peroxide or ozonic
ether is applied by drops. If blood is present, a blue color is imparted by the
mixture. 4. PHENOLPHTHALEIN TEST
A drop of the Kastle-Meyer’s reagent is dropped on a white filter paper with the
stain and left for at least 10 seconds. A positive result will show a pink color after
the addition of hydrogen peroxide
5. LEUCOMALACHITE GREEN TEST -This test is not so sensitive as the
benzidine test. It depends upon the fact that leucomalachite green is oxidized to
malachite green will result in a bluish-green or peacock-blue color by hydrogen
peroxide solution.
2. CONFIRMATORY TEST
A. MICROSCOPIC EXAMINATION
Saline extract of the stain is examined under the microscope. Note the presence of
red blood cells, leucocytes, epithelial cells and microorganisms. The presence of
red blood cells will conclusively show that the stain is blood. By microscopic
examination, we can differentiate the origin or the part of the body it came from.
Menstrual blood will show abundance of vaginal epithelial cells and Doe-derlein’s
bacilli.

B.MICRO-CHEMICAL TEST
1. HEMOCHROMOGEN CRYSTAL OR TAKAYAMA TEST:
A fragment of the suspected material is placed on a slide glass and a drop of
haemochromagen reagent is added. A cover glass is placed on top and heated
gradually for a time, then examined under the microscope. Crystals varying from
salmon color to dark brown and pink and which are irregular rhomboids or in
clusters, may be seen. This test is positive to any substance containing
haemoglobin.
2.TEICHMANN’S BLOOD CRYSTALS OR HEMIN CRYSTALS TEST:
On the microscopic slide is placed fragments of the stain and a drop of water with
trace of sodium chloride added. Add glacial acetic acid and evaporate to dryness
under a cover slip. Dark brown rhombic prisms of chloride of haematin are formed.
This is considered as the best of the micro-chemical test.
3. ACETONE-HAEMIN OF WAGENHAAR TEST:
A particle of dried stain or a fiber of the stained fabric is placed on a glass slide and
covered with a cover slip with a needle interposed to prevent direct contact of the
cover slip with the slide. A drop of acetone is run under the cover slip so that the
material is surrounded and a drop of diluted oxalic or acetic acid is then added.
When examined under high power microscope, small dark, dichroic acicular
crystals of acetone-haemin are seen.
4. SPECTROSCOPIC EXAMINATIONS:
This examination depends on the principle that blood pigments have the power to
absorb light of certain wave length and produce certain characteristic absorption
bands on the spectrum.
 3. ANALYSIS OR BIOLOGICAL EXAMINATION- This test is to determine whether the
blood is of human origin or not.
PINCIPLE OF THE TEST:
By injecting an animal, usually a rabbit, with defibrinated blood of unrelated animal, an
anti-serum is produced in the blood of the animal injected. The serum of this animal
injected is capable specifically of precipitating the serum of the unrelated animal whose
blood serum has been injected. However, closely related animal may also give the same
response.

4. BLOOD GROUPINGS

PRINCIPLE OF THE TEST:

All human beings have their blood belonging to any of the four principal blood groups. A
normal suspension of human red blood cells when mixed with its own serum or serum of
a similar group will make the red blood cells suspension remain even. But if suspended
in the serum of another group, the red blood cells clump with one another and this is
called agglutination. The red blood cells contain agglutinogens and the serum contains
agglutinins.

VALUE OF THE TEST

It may solve disputed parentage (paternity or maternity). A positive result is not
conclusive that the one in question is the offspring, but a negative result is conclusive
that he is not a child of the alleged parents.

IDENTIFICATION OF BLOOD GROUP WITH KNOWN SERUM (ANTI-A and ANTI-B)

ANTI-A + ANTI -B + Anti gen Present Blood Type

Fresh Blood Fresh Blood
+ - A A
- + B B
+ + Both A and B AB
- - Neither A and B O

ANTI-A + ANTI -B + Blood Type

Fresh Blood Fresh Blood
+ - A A
- + B B
+ + Both A and B AB
 - - Neither A and B O
IDENTIFICATION OF BLOOD GROUP WITH KNOWN Antigen (A-Cell and B-Cell)

ANTI-A + ANTI -B + Blood Type

Fresh Blood Fresh Blood Antigen present
+ - A A
- + B B
+ + Both A and B AB
- - Neither A and B O

INHERITANCE PATTERN FROM PARENTS TO OFFSPRING

GROUP OF GROUP OF EXCLUSION

PARENTS CHILDREN CASES
OxO O A, B, AB
OxA O,A B, AB
OxB O, A, AB
B
AxA O,A B, AB
AxB O, A, B, AB
BxB O, A, AB
B
O x AB A,B O, AB
A x AB A, B, AB O
B x AB A, B, AB O
AB x AB A, B, AB O