FORENSIC CHEMISTRY & TOXICOLOGY

FORENSIC CHEMISTRY

That 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 well-settled that this science has an advantage over extracted confession, eye witnesses and other
circumstantial evidence. Thus, it is regarded as the highest form of uncontestable and conclusive piece of evidence
with the utmost legal significance.

Forensic Toxicology
Deals with the examination of human organs, food samples and water and gastric contents for the purpose
of detecting the presence of poisonous substance, its dosage, effects, and treatment.

SCOPE OF FORENSIC CHEMISTRY

 It includes the chemical side of criminal investigation

 It includes the analysis of any material the quality of which may give rise to legal proceeding.
 It is not limited to purely chemical questions involved in legal proceedings.
 It has invaded other branches of forensic sciences notably legal medicine, ballistics, questioned documents,
dactyloscopy, and photography

Practice of forensic Chemistry

The work of forensic chemist is divided into 4 stages namely:
1. Collection or reception of the Specimen
A. Sufficiency of sample
B. Standard comparation
C. Maintenance of individuality
D. Labeling and sealing

2. The Actual examnination

3. The communication of the result of the examination

4. Court appearance

What are the GOLDEN RULES in the practice of Forensic Chemistry?

In and out of the Crime Scene:

 Go Slowly

 Be thorough

 Take note consult others

 Use imagination

 Avoid complicated theories

Failure to consider the golden rules will contribute to destruction of evidence/specimen. The other reasons of
forensic disaster are:
 Improper packing of specimen
 Failure of identification of specimen
 Improper, precaution use in transmitting the specimen.
 Lack of precautions to prevent tampering of the specimen.

DEVELOPMENT OF SCIENTIFIC CRIME LABORATORY IN THE PHILIPPINES

FEBRUARY 14, 1829- Massacre occurred in Chicago. Then a group of public-minded individuals was responsible
for the establishments of scientific crime laboratory which today has marked it place in the historical annals of
police science.

MARCH 31, 1876- Creation of ‘’Medicos Titulares” by virtue of Royal Decree No. 188 of Spain. For every
province , Forensic Physician was assigned to perfoem public sanitary duties and at the same time provide medico-
legal aids to the administration of justice.

DECEMBER 15, 1884, - Governor General Joaquin Javellar creatd a committee to study the mineral waters of
Luzon and appointed Anacleto del Rosario as Chemist.

SEPTEMBER 13, 1887 – Establishment of “ Laboratorio Municipal de Manila “ under the inspection of the
“Direction General de Administacion Civil ” and the control of the “Gobiernode Provincias”. The function of the
laboratory was to make analysis not only of food , water , and other from the standpoint of public health and legal
medicine but also of specimen for clinical purposes.

1894 – “Laboratorio medico legal” was created under the dependency if the judicial branch of the government and
the laboratory functioning under the direction of a physician and assisted by pharmacist- chemist.

1895- Antonio Luna established a clinical laboratory where some original works in chemistry were done.
Unfortunately , all prospects of the advancement of the scientific laboratiories were paralyzed as a result of the
revolution in 1896.

1899 – The first scientific laboratory was established in a small building on the banks of Pasig River with Lt. R.P.
Strong of the US army in charge.

1901 – actual scientific work began under the initiative if Dean C. Worcester by virtue of act no. 156 approved by
the Civil Commission.

July 01, 1901 – The bureau of Government Laboratories was created for the purpose of performing biological and
chemical examinations as well as for the production of vaccines and sera.

MARCH 11, 1915 - The Department of Legal Medicine was created pursuant to the resolution of the Board of
Regents. The Department took charge of the courses of legal medicines and its branches in the College of Medicine
and Law.

OCTOBER 14, 1924 – Through the passage of Act No. 3043 by the Philippine Legislature, the same Department
of Legal Medicine became the branch of the Department of Justice and the same time the integral part of the
University of the Philippines.
December 01, 1946 – The “Division of Investigation” was created under the Commonwealth Act no. 181 with twi
medico-legal officers and a chemist. The Medico Legal Section of Division of Investigation started the definite
movement towards the creation of scientific crime detection laboratory.

SEPTEMBER 1945 – The national Bureau of Investigation (NBI) was organized with the Division of Investigation
as the nucleus. The chemical laboratory of Medico-Legal Section was expanded into a forensic Chemistry Division.

PHYSICAL EVIDENCE
 Are articles and materials which are found in connection with an investigation and which aid in
establishing the identity of the perpetrator of the circumstances under which the crime was committed or which in
general assist in the prosecution of the criminal. It encompasses any and all objects that can establish that crime has
been committed or can provide a link between a crime and its victims or a crime and its perpetrator.

EVIDENCE – evidence is the proof of allegation, it is means sanctioned by the law of ascertaining in a judicial
proceeding the truth respecting the matter of fact. ( rule 128, revised rule on evidence)

Evidence may be (a) direct; (b) indirect ,which includes circumstantial evidence ; (c) hearsay.

FORMS OF SCIENTIFIC EVIDENCE

1. Real or Autoptic Evidence – is that evidence which addressed to the senses of the court.

2. Testimonial Evidence – an expert may be called on the witness stand to answer all questions propounded by
both parties in the case.

3. Documentary Evidence – ay written evidence presented by an expert in the court which is relevant to the
subject matter in dispute and not excluded by the rules Court. Formal written report , expert opinion ,
certificates and dispositions are included this group.

WITNESS
A witness in court may be an ordinary witness or expert witness

Under the law, an ordinary witness must have the following qualifications:

1. He must have the organ and power to perceive.

2. Perceiving can make known to his perception to others.
3. He does not fail in any of the exception provided for by law, 123 rules of court

The opinion of a witness regarding a question of science, art , trade, where he is skilled therein,
may be receive in evidence. This kind of evidence is called the testimony of an expert witness.

ORDINARY WITNESS EXPERT WITNESS

 Can state what his senses have perceive State what he has perceive and also give his
opinion , deduction or conclusions to his
perception

May not skilled on the line he is testifying Must skilled in the art, science or trade hes is
testifying

Cannot testify on things or facts not perceive exept Testify on things hes has not seen by hid opinion,
thise provided by the law deduction ot conclutions on the statements of facts.

BLOOD AND BLOOD STAINS

What is the importance of studying blood?

 As circumstance or corroborative evidence against or in favor of the perpetrator of the crime.

 As evidence in case of disputed percentage
 As evidence in the determination of the cause of death and the length of time the victim survived the attack.
 Determination of the direction of escape of the victim or the assailant
 Determination of the origin of the flow of blood
 As evidence in the determination of the approximate time the crime was committed.

What is BLOOD?

Blood has been called the circulating tissue of the body. It is refereed to as a highly complex mixture of
cells, enzymes, proteins, and inorganic substances. It is the red fluid of the blood vessels. Blood is opaque. On the
treatment with either, water or other reagents becomes transparent lake color. It is finally alkaline. Normally pH is
7.35 – 7.45.

Composition of Blood
Water ---- 90%
Solid ----- 10% ( largely protein in nature and consist of albumen, several globulin’s and fibrinogen.

(45%) formed elements or the solid materials consisting chiefly of cells namely:

Red Blood Cells or RBC (ERYTHROCYTES) around 4 – 5 millions of red cell per cc. of blood.
White Blood Cells or WBC (LEUKOCYTES)
Blood Platelets (THROMBOCYTES)

(55%) PLASMA – The fluid or liquid portion of blood where the cells are suspended. It is principally composed of:

PLASMA is the yellowish fluid of blood in which numerous blood corpuscles are suspended. A straw-yellow liquid
formed when blood to which oxalate has been added to prevent clotting is allowed to strand.

SERUM is a straw – yellow liquid formed when clotted blood is allowed to stand for sometime and the clot
contracts.

Problems in the Study of Blood

Blood is difficult to be searched, the collection, preservation, packing and transportation of specimen suspected to
contain blood is another. Blood offers little resistance to decomposition. It undergoes a rapid charge in its character
with the passage of time as process of clotting and drying commences almost immediately on exposure to air.
Sodium fluoride maybe added to blood to preserve it for a week at room temperature or indefinitely in a refrigerator.
Between 40 – 50 degrees centigrade is the ideal preserving temperature for blood and other perishable specimens.
Collection of blood stains should be done as soon as possible, mere washing of garments/clotting removes the blood.

Blood Collection

FLUID BLOOD are usually collected from victims of crimes of violence, parent and child in case of disputed
parentage.

DRIED BLOOD OR BLOOD STAINS are collected from smooth surface like walls, finished floors, table tops,
hard surface like axe, hammer, knives, stones, crowbars, glazed surface like glass, tiles, automobiles, bulky objects
like blackboard, linoleum sheets, doors, window frames, clothing, and blood absorbed by the soil

Blood Examination

PRELIMINARY TEST - determine whether the stain contains blood or another substance. Determines whether
visible stains do or do not contain blood. It is used to demonstrate the presence of blood.
CONFIRMATORY TEST - determines whether bloodstain really contains blood. Test that positively identifies
blood.
PRECIPITIN TEST- determines whether blood is a human or non-human origin, and if non human, the specific
animal family from which it originated.
BLOOD GROUPING TEST - determines the blood group of human

THE PRELIMINARY TEST FOR BLOOD (COLOR TEST)

Benzidine Test or Benzidine Color Test

Phenolphthalein Test ( also known as Kastle – Meyer Test)
Guaiacum Test (Van Deen Test, Day’s or Schonbein’s Test)
Leucomalachite Green Test
Luminol Test

Benzidine Test

This is an extremely sensitive test that can be applied to minute stain. For many years the most commonly
used preliminary test for blood. The Benzidine test never fails to detect blood even when very old, decomposed stain
with all shorts of contamination is examined. The positive result is only indicative that the blood maybe present.

REAGENT: Benzidine solution ( small amount of powdered benzidine dissolved in glacial acetic acid)
and 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 drop of hydrogen peroxide solution.
POSITIVE RESULTS: Intense blue color produced immediately
LIMITATION: Benzidine test is not a specific test for blood. Positive results maybe obtained from
substances as sputum, pus, nasal secretion, plant juices, formalin, clay, gun. The reaction is weaker and produces
faint coloration.

Phenolphtalein Test

This is an alternative test to benzidine test. It can detect blood in a dilution of 1:80,000,000 parts. A
positive results with this test is highly indicative of blood. The negative result is, therefore, valuable and is
conclusive as to the absence of blood.

REAGENTS: Phenolphthalein solution (1 – 2 grams phenolphthalein to 100 ml of a 25% KOH in water

added with one gram zinc powder heated until colorless) and 3% solution of hydrogen peroxide.
PROCEDURE: Place a small fragment/portion of the stained materia on a filter paper. Add a drop of
phenolphthalein solution and then a drop of hydrogen peroxide solution.
POSITIVE RESULT: Rose color develops or deep pink color or permanganate coor.
LIMITATION: Test is also given by copper salts, potatoes and horseradish.

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.

REAGENTS: Fresh tincture of guaiac resin (Few lumps of this to 95% alcohol, then filter) and 3% of
hydrogen peroxide or few drops of turpentine.
PROCEDURE: Place a small piece of the stained fabric on porcelain dish. Soak with fresh tincture of
guaiac. Add a few drops of hydrogen peroxide.
POSITIVE RESULTS: Beautiful blue color that appears immediately.
LIMITATION: The test also reacts with salvia, pus, bile, milk, rust, iron salts, cheese, gluten, potatoes, perspiration
and other oxidizing substances.
Leucomalachite Green Test

This is a test not as sensitive as the benzidine test

REAGENT: Leucomalachite Green solution ( 1 gram leucomalachite green dissolved in 48 ml. glacial
acetic acid and diluted to 250ml. water) and 3% hydrogen peroxide.
PROCEDURE: A small piece of the stained fabric on a filter paper. Add a drop of leucomalachite green
solution and after a few seconds add drop of 3% hydrogen peroxide.
POSITIVE RESULTS: Malachite green or bluish green

Take Note – The principle involved in blood testing is that the peroxidase present in 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.

Hemoglobin is the red coloring matter of the red blood cells of the blood.

Luminous Test

It is an important presumptive identification test for blood. The reaction of luminol with blood results in the
production of light rather than color. By spraying luminol reagent onto a suspect 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. (LUMINESCENCE). Luminol test is extremely sensitive test. It is capable of detecting
bloodstains diluted up to 10,000X. 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.

THE CONFIRMATORY TEST FOR BLOOD

The actual proof that stain is blood consists of establishing the presence of the characteristic of the red blood cells of
the blood.

The three (3) confirmatory tests for blood are:

Microscopic Test - Useful for the demonstration of blood corpuscles for making the distinction between
mammalian, avian, piscine, and reptilian blood and for the investigation of menstrual, lochial and nasal charges. In
short it differentiates mammalian, avian, piscine and reptilian blood.

Take Note: The Mammalian red blood cells are circular, biconcave disc without nucleus birds, fish and reptiles
red blood cells larger, oval and nucleated amphibians-animal living on land breeding in water. Red blood cells
are larger than mammals, oval and nucleated.

Microchemical Test – also known as Microcrystalline test which include Teichmann Hemin Reaction/Teichman
Test/Haemin Crystal Test, Haemochromogen crystal Test or Takayama Test, Acetone-Haemin Test. One of the two
popular microchemical test is the Takayama Test, a delicate test for the presence of hemoglobin.
PROCEDURE: Place a small piece of suspected material on a glass slide. Add 2 – 3 drops of Takayama
reagent. Cover with glass slip.
POSITIVE RESULTS: Large rhombic crystals of a salmon pink color arranged in clusters, sheaves and
other forms that appears within to 6 minutes when viewed under the low power objectives. To hasten result heat
maybe applied.
REAGENT: Takeyama reagent (3 cc. of 10% NaOH, 33 cc. pyridine, 3 cc. of saturated glucose solution
and diluted with 7 ml. of water.

Spectroscope Test – is the almost delicate and reliable test for the determination of the presence of blood in both
old recent stains. This is performed by means of an optical instrument known as SPECTROSCOPE.

THE PRECIPITIN TEST

It is the standard test used to determine whether the stain/blood is of human or animal origin

Reagent: Precipitin/antiserum

PROCEDURE: Scrape off blood stain if on hard material. Powder the scrapings and exact with saline
solution. If the stain is cloth, paper or similar material, cut a small portion and then place in a test tube and add
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: A white cloudy line or ring or band at the contact points of the fluid that appears
immediately or within one or two minutes.
LIMITATION: The precipitin reacts not only with blood proteins but also with other body proteins as
those as saliva, semen, mucus and other body fluids.

THE BLOOD GROUPING AND BLOOD TESTING

The Four Blood Groups

1) Group “O”
2) Group “A”
3) Group “B”
4) Group “AB”
Agglutinogen or Antigen

These are characteristic chemical structures or “principles” that the found on the surface of each red blood
cells which stimulates the production of agglutinins or antibodies. There are two different agglutinogens or antigens
classified as AGGLUTINOGEN A OR ANTIGEN A AND AGGLUTINOGEN B OR ANTIGEN B.

Antibody or Agglutinin

These are properties or “principles” contained in the serum which cause agglutination or clumping together of the
red blood cells. They are antitoxic substances within the body which reacts when confronted with a specific antigen
to protect the system. There are two different agglutinins classified as Anti-A and Anti-B. Agglutinins are
demonstrable in about 50% of newly born infants.

We have the four groups because of the presence of absence of two antigens A and B in the RBC and two
agglutinins Anti-A and Anti-B in the serum.

BLOOD GROUP ANTIGEN/AGGLUTINOGEN ANTIBODIES/AGGLUTINIES

PRESENT IN THE RBC PRESENT IN THE SERUM

A A ANTI-B

B B ANTI-A

AB A&B NO A & NO B or NONE

NO A & NO B or NONE ANTI-A & ANTI-B

(+) Means agglutination or clumping of RBC

(−) Means absence of agglutination or no clumping of RBC

The Blood Typing (M-N System) of Blood

There are two agglutinogens in human red cells which defines three types of blood. Namely: Type M, Type
N, and Type MN.

(+) Means agglutination

(−) Means absence of agglutination

Inheritance of Blood Groups

Knowledge of genetics will make it easier to understand the principle involved in the inheritance of blood
groups. The inheritance of blood group is predetermined by the presence and absence of two facts or GENES called
Gene A and Gene B.

GENES - any of the complex chemical units in the chromosomes by which hereditary characters are transmitted,
responsible for the transmission of hereditary characteristics. They occur in pair. There are two genes or factors
called gene A and gene B. these are found in the chromosomes. Since chromosomes go in pair, each of which carries
or fails to carry one of these genes. An individual’s called genotypes, where O represents the absence in the
chromosomes of either the A or B gene.

PHENOTYPES – the term used to denote the expression of the inherited characteristic as found in the individual.
Actually the blood groups
GENOTYPES - Are paired genes.

Application of Blood Group Data

Questions of illegitimacy and relationships in may cause maybe solved by means of the blood groups as determined
by the agglutinogens A, B, M, and N.
Determination of whether a man accused of fathering a child out of wedlock could or could not be its parent.
Determination of whether a child born of a married woman could or could not have been fathered by her legal
spouse.
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 a hospital.
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.