Forensic Chemistry
Forensic Chemistry
Forensic Chemistry
Is the branch of chemistry, which deals with the application of chemical principles in the
solution of problems that arise in connection with the administration of justice. It is
chemistry applied in the elucidation of legal problems. It is used in
courts of law.
BLOOD- has been called the circulating tissue of the body. It is referred to as highly
complex mixture of cells, enzymes, proteins, and inorganic substances.
Normally, pH is 7.35- 7.45.
COMPOSITIONS OF BLOOD
1.(45% ) formed elements or the solid materials consisting chiefly of cells namely:
a.Red Blood Cells (erythrocytes) around 4 -5 millions of red cell per cc of blood.
b.White Blood Cells (leukocytes)
c.Blood Platelets (thrombocytes)
2. (55%) plasma- the fluid or liquid portion of blood where the cells are suspended. It
is principally composed of:
a.Water- 90%
b.Solid- 10% (largely protein in nature and consists of albumen, several globulins and
fibrinogen)
PLASMA- The yellowish fluid of blood in which numerous blood corpuscles are
suspended. Astro-yellow liquid formed when blood to which oxalate has been added to
prevent clotting is allowed to strand.
SERUM- a straw-yellow liquid formed when clotted blood is allowed to stand for
sometime and the clot contracts.
Determine whether the stain contains blood or another substance. Determines whether
visible stains do or do not contain blood.
a. Benzidine test or Benzidine Color Test
b. Phenolphthalein Test (Kastle- Meyer Test)
c. Guaiacum Test (Van Deen Dya’s/ Schonbein’s Test)
d. Leucomalachite Green Test
e. Luminol Test
PHYSICAL EXAMINATIONS:
1.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; hemoglobin is
transformed to-hemoglobin or hematin. If the stain has been kept in damp places for a
long time; hemoglobin is transformed to hematin.
2. Heat test: Solution of the blood stain when heated will impart a muddy precipitate.
3. Luminescence test: Stains on dark fabric mixed with mud, paint, etc. emit bluish
white luminescence in a dark room when sprayed with one of the two solutions:
CHEMICAL EXAMINATIONS:
1.Benzidine test: A piece of white filter paper is pressed firmly on the suspected stain.
Benzidine reagent is dropped on the paper, then 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 the 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. It is not conclusive like the
benzidine test because potato skin, iron rust, cheese, blue and indigo may give a
positive reaction to the test. The guaiacum test is positive up to 1:5,000 dilution.
5. Leucomalachite Green test This test which was recommended by Adler in 1904 is
quite useful, but it is not so sensitive as the benzidine test. It depends upon the fact that
leucomalachite green is oxidized to malachite green with a bluish-green or
peacock-blue color by hydrogen peroxide solution.
MICRO-CHEMICAL TESTS
1. Hemochromogen crystal or Takayama test A fragment of the suspected material is
placed on a slide glass and a drop of hemochromogen 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 hemoglobin
BIOLOGIC EXAMINATIONS
1.Precipitin test -This test is to determine whether the blood is of human origin or not.
The following common substances, if present in the extract, will precipitate the anti-
serum and thus produce the semblance of a positive reaction:
a. Soap - This is of special importance in washed stains.
b. Rancid oil-is found sometimes in pillow cases and head dresses. The soluble
fraction of the oil prevents frothing yet the extract gives a sharp and quick reaction.
Two errors are introduced, namely:
(1) False positive reaction from the rancid oil itself.
(2) Failure of foam test and danger of group reaction— monkey blood in high
concentration will act likehuman blood.
c. Alum- This is used as a household remedy in gargles and mouth washes and may
stain any article.
d. Tannin and allied substances derived from vegetable tissue. The stain of saliva of
the betel chewer not only looks like bloodstain but by virtue of the tannin present in most
of the constituents of the prepared betel it also acts on the antisera. Stains on leather
and plant tissue should be removed by applying moist filter paper to the surface
(leather contains tannin).
Value of the precipitin test - If positive result is obtained, we can tell in a more or less
conclusive way that the blood stain is of human origin; although anthropoid ape may
give the same result. The same test and technique may be made to determine whether
muscles, secretions, bones and other body fluids are of human origin or not.
Certain materials like alcohol, formaldehyde, corrosive , lysol, creoline, carbolic acid,
acids and alkalies destroy the property of blood to react with precipitin.
2.Blood grouping 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.
Two methods may be utilized and both should be employed in the examination:
a. Detection of agglutinins (antibodies)
b. Detection of agglutinogens (antigen)
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 notthe child of the alleged parents.
Age of the Blood Stains: When blood is exposed to the atmosphere or some
other influences, its hemoglobin is converted to meth-hemoglobin or hematin. The color
is changed from red to reddish-brown. The presence of acid accelerates the formation
of hematin. These changes take place in warm weather within 24 hours. Blood of
one week old and that of six weeks may not present a difference in physical and
chemical properties
1.Arterial Blood:
a. Bright scarlet in color.
b. Leaves the blood vessel with pressure.
c. High oxygen contents.
2. Venous Blood:
a. Dark red in color.
b. Does not spill far from the wound.
c. Low oxygen content.
3. Menstrual Blood:
a. Does not clot.
b. Acidic in reaction owing to mixture with vaginal mucous.
c. On microscopic examination, there are vaginal epithelial cells.
d. Contains large number of Deoderlein's bacillus.
5. Child's Blood:
a. At birth, it is thin and soft compared with that of adult.
b. Red blood cells are nucleated and exhibit greater fragility.
c. Red blood cells count more than in adult.
IDENTIFICATION OF HAIR AND FIBERS
5. Jute Fibers- Fibers are quite smooth without either longitudinal or transverse
markings. The fibers have typical large cavities which are not uniform but vary
with the degree of contraction of the walls of the fibers which are lignified.
The ends are blunt.
2. Length of the hair: Hair from the scalp grows 2.5 cms. a month. Beard hair grows
at the rate of 0.4 millimeter a day.
3. Color of the hair- The color of the hair may be black, blonde or brunette. Hair
from older persons may be white or gray. The hair may be artificially colored by
bismuth, lead or silver salts. It may be bleached by addition of hydrogen peroxide,
chlorine or diluted hydrochloric or nitric acid.
4. Does the Hair Belong to a Male or a Female? -In many instances it is quite
impossible to state the sex from the hair, but certain points may be worthy mention: Hair
on the scalp of male are shorter, thicker and more wiry than that of female's. Eyebrow
hair of a male is generally long and more wiry than that of a female's.
Estimations of Age Based on the Hair- This is quite difficult and the examiner
hesitates in giving his opinion. However, there are some points of distinction:Hair of
children are fine, short, deficient of pigments and, as a rule, devoid of medulla. At
the adolescent age, hair may appear at the pubic. Hair on the scalp becomes long,
wiry, and thick. In the case of older persons, the color is usually white or gray, with
marked absorption of pigments and degenerative changes.
FLUID AND SPERMATOZOA The semen is the viscid, albuminous fluid with faint
grayish-yellow color, having the characteristic fishy odor, and containing spermatozoa,
epithelial cells, lecithin bodies and other substances.
The following specimens may be examined for seminal fluid and spermatozoa
1. Wearing apparel of the victim and of the alleged accused.
2. Vaginal smear from the victim.
3. Stains on the body of the victim and of the accused.
4. Stains found at the site of the commission of the offense.
PROCEDURE:
1. Gross Examinations:
a. Inspection by means of the naked eye or with the use of the hand lens: The stain is
grayish-white to faint yellow in color. Infabrics, the area occupied is slightly
depressed. It usually has a maplike contour with silvery appearance of the surface. It is
hardened with shiny borders.
Value of the Test- This test is produced by the action of iodine on choline, a natural
base found in many cells. It is not a specific test for spermatic fluid. The test is not a
proof of seminal fluid but only of the presence of some vegetable or animal substance.
A positive result is merely a presumptive evidence of seminal fluid; a negative result
means, in all probability, it is not that of the seminal fluid.
B. Barberio's Test-To the spermatic stain on the slide glass, a saturated solution
(alcoholic or aqueous) of picric acid is added. The preparation is placed under the
microscope and needleshape crystals with yellow color is produced. Some allege
that this test is specific for spermatic fluid. The reaction probably depends-on the
presence of spermatic secretion.
C. Acid Phosphatase test: The semen produces a very high acid phosphatase
activity as compared with other body fluids (saliva, perspiration, urine, etc.) and
common vegetable and fruit juice stains.The method of estimating the activity of a stain
onclothings or other materials is to extract with distilled water and perform the acid
phosphatase determination on the filtered extract
Dr. Hankin's Method: The fabric with seminal stain is boiled with tannin solution
before dissolving in a solution of potassium cyanide so as to render the spermatozoa
capable of removal. The fabric is then placed on a slide, teased with carbol-fuchsin.
This is examined with a medium power lens
4. Biological Examinations
a. Precipitin Test (Biological test of Farnum): This is a test to determine whether the
semen is of human origin or not. A rabbit is immunized with human semen for four
to six weeks. After a time the blood is drawn and the blood serum is taken and its
potency made at different dilutions This is used for the test of unknown semen in the
same way as blood precipitin is done.The presence of a white ring at the point of
contact between unknown semen extract and the anti-human semen serum shows that
the unknown is of human origin.
How long after sexual intercourse can spermatozoa be found in the vaginal canal?
Authorities differ in their opinions in this respect:
1.but, there is every reason to believe that the life of the effective sperm in the
maternal passage is very short, probably less than thirty hours" (Taylor's Principles
and Practice of Medical Jurisprudence by K. Simpson, 12th ed.. Vol 2, p. 32).
2. "There has been a great number of observations this latter point, and the evidence
points to a comparatively short life of the sperm in the female tract and the period
appears shorter with the number of observations. It is a present belief that the life of the
sperm in the vagina is a matter of hours and its total life in the female tract is a matter of
two or three days at most" (Forensic Medicine by S. Smith, 10th ed., p. 311).
3. that spermatozoa may survive less than two hours in the vagina, but they live as
long as forty- three hours both in the cervix and uterus where the secretion are more
favorable" (Medical Jurisprudence & Toxicology by J. Glaister, 12th ed., p. 325).
5. "Fertilization of the ovum does not necessarily occur immediately after coitus, as it
is known that spermatozoa can remain alive in the upper recesses of the vagina for
more than two weeks" (Coy\ p.246).
6. "Some observers have reported finding spermatozoa in the vagina after seven
hours while others have reported finding them as long as 48 hours after
intercourse" (Homicide Investigation byL. Snyder, 1st ed., p. 327).
3.Color Test:
b. Barbiturates: (1) Dillie Koppanyi test — (1% cobalt acetate in methanol is first
added to the suspected material followed by 5% isoprophylamine in methanol). A
violet-blue color is produced. This is a valuable screening test for barbiturates.
c.Marijuana: (1)Duquenois-Levine test — Solution A is a mixture of 2% vanillin and
1% acetaldehyde in ethyl alcohol; solution B is concentrated hydrochloric acid; and
solutions A,B and C are added respectively to the suspected material. A positive result
is shown by purple color in the chloroform layer.
d.LSD: (1) Van Urk test (1% p-dimethylaminobenzaldehyde and 10% concentrated
hydrochloric acid in ethyl alcohol). This reagent turns blue-purple in the presence of
LSD. However, owing to the extremely small quantities of LSD in illicit preparations, this
test is difficult to conduct under field conditions.
4. Chromatography:
a. Thin layer chromatography.
b. Gas chromatography. In both methods the drug is separated from the diluent
while providing for its identification.
A habitual drunkard-is one who excessively uses intoxicating drink. Habit should be
actual and confirmed, but it is not necessary that it be continuous or of daily
occurrence.It lessens individual resistance to evil thought and undermines will power,
making its victim a potential evil doer.
2. Distilled Liquor — Distilled liquors are alcoholic beverages produced from distillate
of wines, distilled from grains or starch solution or distillate from aromatic substances.
In distilled beverages the alcohol contents are expressed in proofs. "Proof" is
approximately twice the percentage of alcohol by volume. The sole purpose of the
distillation process is to increase the concentration of alcohol in the finished product.
This is necessary because fermentation ceases when the alcohol concentration is
approximately 12% to 14% by volume.
3. Malt Liquors — Alcoholic beverages brewed from malt or from a mixture of malt
and malt substitute, like rye, and may contain other cereal grains and starchy
saccharine matters. A characteristic bitter flavor is imparted by the addition of hops.
The amount of alcohol need not be stated in the label.
1.Stage of Excitement — This develops a few minutes after the initial dose of
alcoholic drink has been absorbed and has reached the central nervous system. It is
characterized by a feeling of wellbeing and slight excitation. The actions, speech
and emotion are less strained. Self-confidence develops, as well as blunting of self-
criticism, self-consciousness and self-control.
3. Stage of Narcosis or Coma — The person passes into a deep sleep and may
only respond to strong stimuli. Pupils are dilated, breathing is slow and stertorous,
pupils are dilated and reflexes, abolished. Death may ensue from paralysis of the
cardiac or respiratory center.
Degree of Intoxication:
Slight Inebriation — There is flushing of the face, with exaggerated mood, but a
person is able to control his behavior. He shows no signs of mental impairment,
incoordination of movement and difficulty of speech.
Very Drunk, "Dead drunk" — The mind is confused and disoriented. There is difficulty
in speech and marked motor incoordination and often walking is impossible.
2.Let the subject stand straight with one foot ahead of the other so that the toes of
one foot touch the heel of the other. This will remove the brace to prevent side sway. If
drunk, there is more likelihood that the subject will sway sidewise and fall. The test is
repeated after the subject is free from the effect of alcohol and make a comparison
of stability.
3.Let the subject sit comfortably in a desk and get samples of his handwriting.
Compare these writings, with those taken when he is free from the effects of alcohol.
4.Let the subject bend down and pick up a small object from the floor. If he stumbles,
then his nervous system is not stable and that he may be drunk.
5.Let the subject walk straight forward to a comer of a room and rapidly turn around
without stopping. Tell him to walk back. You will notice that the subject may have
uncertainty of steps, side steps, or he staggers while making the turn and in walking.
1.Persons with blood alcohol below 0.05% are not considered intoxicated.
2.The majority of persons (80-90%) with blood alcohol levels between 0.1% and
0.15% will have their faculties so impaired as to render them unfit to drive motor cars
with reasonable safety.
3.The majority of young people who are not habitual drinkers will be intoxicated to
the extent of staggering when the blood level is about 0.15%.
4.The majority of all persons (80-90%) including habitual drinkers will be intoxicated to
the extent of staggering when the blood alcohol level is approximately 0.2%.
5.The majority of persons will be in a coma when the blood alcohol level is
approximately 0.5%
The American Medical Association and the National Safety Council of the United
States recommended the following presumptive limits of intoxication:
1.Persons who have 0.05% alcohol or less in their blood are presumed to be
uninfluenced by an alcohol.
2.Persons who have 0.05% to 0.10% alcohol in the blood are considered to possibly be
under the influence of alcohol.
3.Blood alcohol level of 0.10% to 0.15% or more gives rise to the presumption that the
person is drunk.
1.Analysis of Blood: Analysis of the blood is probably the most widely accepted way to
determine the concentration of alcohol in the body. It is a direct method of estimation
although the subject may refuse blood extraction for such analysis.
2. Analysis of the Breath: The concentration of alcohol in the blood can be determined
indirectly by making a quantitative determination of alcohol in the respired air. The basis
of the analysis is that there is a constant ratio between the concentration of alcohol in
the blood stream and in the alveolar air.
3.Analysis of the Urine: Urine as a specimen for alcohol determination has not gained
widespread use because of variability in the different periods of alcohol intake.
POISONOUS SUBSTANCES
A.Gaseous Poisons -(Poisons Present in the Gaseous State): Carbon dioxide; Carbon
monoxide; Hydrocarbons; Hydrogen sulfide; Sulfur dioxide; the Oxides of nitrogen
(Nitrous oxide, Nitric acid and Nitrogen dioxide); war gases.
B. Inorganic Poisons:
Corrosives- (Poisons characterized principally by an intense and destructive action —
a few organic corrosives are included in this group for the sake of completeness):
a.Acid; Mineral and Organic: Sulfuric acid; Hydrochloric acid; Nitric acid; Oxalic acid;
Acetic acid.
b. Alkaline Corrosives: Potassium hydroxide; Sodium hydroxide; Calcium oxide;
Ammonium hydroxide.
c. Halogens: Chlorine; Bromine; Iodine; Fluorine
d. Corrosive Metallic Salts: Silver; Zinc.
e. Organic Corrosives: Phenol; Pyrogallol; Formaldehyde.
(These chemicals are protoplasmic irritants, but their chief action is the deleterious
effect produced after absorption into the system.)
A.Heavy Metals: Phosphorus; Antimony; Arsenic; Bismuth; Mercury; Lead; Radioactive
substances; Thallium; Gold; Osmium; Platinum; Nickel; Chromium; Tin; Vanadium.
b. Inorganic Salts: Alum; Alkaline earths; Magnesium sulfate; Lithium salts; Potassium
salts; Boric acid and borax; Tellurium; Sodium silicate.
C. Organic Poisons:
1.Volatile Poisons (Volatile liquids or easily sublimated solids many of which are
irritants; their chief action occurs after absorption)
2.Alkaloidal Poisons: (These substances are toxic principles of plants which have a
characteristic action on some parts of the central nervous system; they are a well-
defined group)
D. Miscellaneous Poisons: (Associated with botulism; food poisoning; mushroom
poisoning; snake venom poisoning
1.Food Poisoning: Toxic substances in the food; Abnormal hypersensitivity to normal
constituents of food.
2.Poisonous Plants.
3.Poisonous Animals and Their Poisons: Arachnids;Centipedes; Insects; Caterpillars;
Vertebrates.
4.Biological Products.
IRRESPIRABLE GASES
4.Gas Chromatograph.
5.Infra-red Analysis.
Detection: The offensive odor may be recognized even if the dilute gas is one part in
10,000 part of air. A piece of filter paper moistened with lead acetate will turn black if it
was brought in
contact with the stomach or other organs containing the gas.
HYDROGEN CYANIDE: Hydrogen cyanide is one of the most toxic and rapid acting
gases. It is formed by addition of acid to potassium or sodium salt of cyanide. It is
naturally found in leaves of cherry-laurel, in bitter almond, in kernels of common cherry,
plum, peaches, in ordinary bamboo shoots, and in certain oil seed and beans. These
plants contain a crystalline glucoside known as amygdalin which, in the presence of
water and natural enzyme, called emulsin, is readily decomposed into hydrocyanic acid,
glucose and benzaldehyde.
SULFUR DIOXIDE:
Sulfur dioxide is a colorless gas, which is heavier than air and with pungent odor. It is
employed as a disinfectant, as a bleaching agent, a powerful reducing agent, and found
usually in eruption of volcanoes.
The gas produces irritation of the respiratory passage,
thus causes sneezing, coughing, spasm of the glottis and suffocation.