Fundamentals of Toxicology

1) What is toxicology and it's various branches

Definition of Toxicology
Toxicology is the branch of science that deals with the study of harmful effects of toxicants on
living organisms, particularly humans. It includes the study of source, nature, dose, mechanism
of action, symptoms, fatal effects, detection and management of poisons.
The word toxicology is derived from the Greek words “Toxicon” (arrow poison) and “Logos”
(study).
• Father of Toxicology: Paracelsus (stated that “The dose makes the poison”)
• Father of Modern / Forensic Toxicology: Mathieu Joseph Bonaventure Orfila (Author of
Traité des Poisons, 1812)
Toxic substances entering the body from outside are called xenobiotics, whereas toxic substances
produced within the body are known as endobiotics.

Branches of Toxicology
Toxicology is divided into the following major branches:
1. Environmental Toxicology
Deals with the study of toxic substances present in the environment such as air, water, soil
and food, and their effects on humans, animals and plants.
2. Occupational Toxicology
Concerned with health hazards due to exposure to toxic chemicals in workplaces and
industries, such as pesticides, heavy metals and industrial solvents.
3. Forensic Toxicology
Application of toxicological principles for legal and criminal investigations, especially in
cases of poisoning, drug abuse, suspicious deaths and medico-legal cases.
4. Regulatory Toxicology
Involves evaluation and regulation of chemicals, drugs, food additives and pesticides to
ensure public safety and compliance with legal standards.
5. Food Toxicology
Deals with toxic substances present in food, food adulterants, contaminants, preservatives
and food poisoning.
6. Analytical Toxicology
Concerned with qualitative and quantitative analysis of toxic substances in biological
and non-biological samples using laboratory techniques.
 7. Clinical Toxicology
Focuses on diagnosis, treatment and management of poisoning cases in living patients.

2) History

The history of toxicology dates back to ancient civilizations where poisons were used for hunting,
warfare, execution and assassination. Over time, systematic study and classification of poisons
developed, leading to modern toxicology.

1. Ancient Period
• Rig Veda (12th century BC)
One of the earliest Indian texts that describes several plant poisons and their effects.
• Ebers Papyrus (c. 1500 BC)
An ancient Egyptian medical document containing information on poisons such as hemlock,
aconite, opium, and metals like arsenic, lead, copper and antimony.
• Greek Period
• Theophrastus, a student of Aristotle, described poisonous plants in De Historia
Plantarum.
• Dioscorides, a Greek physician, gave the first systematic classification of poisons
into:
• Plant poisons
• Animal poisons
• Mineral poisons
• Socrates (470–399 BC) was executed using hemlock, demonstrating state use of
poison.

2. Mithridatic Period
• King Mithridates VI of Pontus experimented with poisons and antidotes on prisoners.
• He developed a universal antidote known as Mithridatium.
• The practice of gradually administering small doses of poison to develop resistance is called
Mithridatism.

3. Medieval Period
• Poisons were widely used for political assassinations, especially arsenic, known as
“poudre de succession” (powder of inheritance).
• Knowledge remained largely empirical with little scientific analysis.
4. Modern Period
• Paracelsus (1493–1541)
Regarded as the Father of Toxicology. He introduced the fundamental principle:
“The dose makes the poison.”

• Mathieu Joseph Bonaventure Orfila (1787–1853)

Known as the Father of Modern / Forensic Toxicology.
• Authored Traité des Poisons (1812), a landmark work on detection of poisons from
human organs.
• Established toxicology as a scientific and medico-legal discipline.
• James Marsh (1836)
Developed the Marsh test for detection of arsenic, which became crucial in forensic
investigations.

5. Development of Forensic Toxicology

With advancements in chemistry and analytical techniques during the 19th and 20th centuries,
toxicology evolved into a scientific, analytical and legal discipline, forming the basis of modern
forensic toxicology.

3) Scope and significance of toxicology

Scope of Toxicology
The scope of toxicology is very broad and covers the study, detection, prevention and management
of toxic substances affecting living organisms. The major areas included in its scope are:
1. Study of Toxic Substances
Toxicology deals with the study of toxicants, toxins, xenobiotics and endobiotics, their
source, chemical nature, dose, metabolism and mechanism of action.
2. Detection and Analysis of Poisons
It includes qualitative and quantitative analysis of poisons in biological samples such as
blood, urine, viscera and tissues, which is essential in forensic investigations.
3. Clinical Management of Poisoning
Toxicology plays an important role in diagnosis, treatment and prognosis of poisoning
cases in living patients.
4. Forensic and Medico-legal Investigations
It assists in determining the cause of death, manner of death, time since exposure and
interpretation of toxicological findings in courts of law.
5. Environmental and Occupational Safety
Toxicology helps in assessing the harmful effects of pollutants, industrial chemicals and
pesticides on humans, animals and ecosystems.
 6. Regulatory and Public Health Applications
It aids in framing safety standards, permissible exposure limits and regulations for drugs,
food additives, cosmetics and chemicals.

Significance of Toxicology
1. Medicolegal Importance
Toxicology is crucial in cases of suicidal, homicidal, accidental and stupefying poisoning,
helping in legal decision-making.
2. Public Health Protection
It helps in preventing food poisoning, drug toxicity, adulteration and environmental
contamination, thereby safeguarding public health.
3. Crime Detection and Justice Delivery
Toxicological evidence supports criminal investigations and provides expert testimony in
courts.
4. Drug Safety and Development
Toxicology ensures the safe use of medicines, helps determine toxic doses and prevents
adverse drug reactions.
5. Occupational Health
It protects workers from harmful exposures in industries and workplaces.
6. Scientific Advancement
Advances in analytical and forensic toxicology contribute to better detection methods and
accurate interpretation of poisoning cases.

4) What is a poison & ideal poison characterisitics

Definition of Poison
A poison is any substance which, when introduced into the living body by ingestion, inhalation,
injection or absorption through skin, in sufficient quantity, is capable of producing ill-health or
death by its local or systemic action.
According to toxicological principles, any substance can act as a poison depending on the dose,
as stated by Paracelsus:
“The dose makes the poison.”

Characteristics of an Ideal Poison

An ideal poison is a hypothetical concept in forensic toxicology. The characteristics of an ideal
poison include:
 1. Colourless
It should not produce any visible colour change when mixed with food or drink.
2. Odourless
It should not emit any smell that may arouse suspicion.
3. Tasteless
It should not alter the taste of food or beverages.
4. Highly Potent
A very small quantity should be sufficient to produce death.
5. Easily Administered
It should be easily soluble in food, water or drinks.
6. Rapid and Certain Action
It should act quickly and cause death without prolonged suffering.
7. Undetectable by Routine Tests
It should be difficult to detect by conventional chemical or toxicological analysis.
8. Easily Decomposable in the Body
It should not leave traces in viscera or tissues after death.
9. Non-irritant
It should not cause immediate irritation or vomiting, which may alert the victim.
[Link] Specific Post-mortem Changes
It should not produce characteristic post-mortem findings.

5) Types of poisoning

Types of Poisoning
Poisoning may be classified on the basis of manner of death and dosage or duration of exposure.
This classification is of great importance in forensic and medico-legal investigations.

A. Types of Poisoning Based on Manner of Death

1. Suicidal Poisoning
• Poison is taken deliberately by an individual to end their own life.
• It is the most common type of poisoning, especially in India.
• Common poisons used include pesticides, drugs, corrosives and household chemicals.
2. Homicidal Poisoning
• Poison is administered with the intention of causing death of another person.
• Comparatively rare due to difficulty in administration and detection.
• Common homicidal poisons include arsenic, cyanide and thallium.

3. Accidental Poisoning
• Occurs without intention, due to negligence, ignorance or mishandling.
• Common in children, industrial workers, food poisoning cases and animal bites.

4. Stupefying Poisoning
• Poison is administered to render the victim unconscious or helpless for committing crimes
like robbery or sexual assault.
• Commonly used substances include alcohol, datura and sedative-hypnotics.

5. Abortifacient Poisoning
• Poison is administered to induce abortion or miscarriage.
• May result in serious injury or death of the mother.
• Commonly used abortifacient substances include ergot, quinine, lead and herbal
preparations.
• Has significant medico-legal importance.

B. Types of Poisoning Based on Dosage / Duration of Exposure

6. Hyperacute (Fulminant) Poisoning
• Caused by very large doses of poison.
• Produces rapid and severe symptoms, often leading to death within minutes or hours.
• Examples include cyanide and organophosphorus compounds.

7. Acute Poisoning
• Results from a single exposure or large dose of poison.
• Produces symptoms within a short period of time.
8. Sub-acute Poisoning
• Caused by repeated exposure to small doses over days or weeks.
• Symptoms are less severe than acute poisoning but more pronounced than chronic
poisoning.

9. Chronic Poisoning
• Results from prolonged intake of small doses over months or years.
• Produces cumulative toxic effects.
• Common examples include poisoning by lead, mercury and arsenic.

6) Roots of administration

Routes of Administration of Poison

The route of administration refers to the path by which a poison enters the body. It plays an
important role in determining the absorption, onset of action and severity of poisoning, and has
significant medico-legal importance.

1. Topical Route
• In this route, the poison is absorbed through the skin (cutaneously).
• Absorption may occur through intact or damaged skin.
• Commonly seen with pesticides, organic solvents and corrosive substances.
• Produces local as well as systemic effects.

2. Enteral Route
In this route, the poison enters the body through body cavities or passages connected externally.
It includes:
• Oral route – ingestion of poison through mouth
• Nasal route – absorption through nasal mucosa
• Ocular route – absorption through conjunctiva
• Aural (otic) route – absorption through external auditory canal
• Urethral route – administration through urethra
 • Rectal route – absorption through rectal mucosa
This route is commonly involved in accidental, suicidal and stupefying poisoning.

3. Parenteral Route
• In this route, the poison is absorbed directly through body systems, bypassing the
gastrointestinal tract.
• It mainly includes injections such as intravenous, intramuscular and subcutaneous routes.
• Produces rapid and severe toxic effects.
• Seen in drug abuse, homicidal poisoning and venom injection.

7) Toxins VS Toxicants

Basis Toxins Toxicants

Toxins are naturally occurring Toxicants are man-made or synthetic
Definition poisonous substances produced by poisonous substances or chemicals
living organisms. harmful to living organisms.
Produced by plants, animals, fungi or Usually derived from industrial,
Source
microorganisms. agricultural or laboratory sources.
Nature Biological in origin. Chemical or synthetic in nature.
Usually enter the body through bite, Enter through ingestion, inhalation,
Mode of Entry
sting, ingestion or contact. injection or skin absorption.
Snake venom, scorpion venom,
Pesticides, heavy metals, industrial
Examples botulinum toxin, mycotoxins, plant
chemicals, drugs of abuse.
toxins.
Study Related
Study of toxins is called Toxinology. Studied under general toxicology.
Term
Important in animal bites, food Important in suicidal, homicidal,
Forensic
poisoning and biological poisoning accidental and occupational poisoning
Importance
cases. cases.

8) Poisons VS Venom

Basis Poison Venom

A poison is any substance which, when
Venom is a toxic secretion produced
introduced into the body in sufficient quantity,
Definition by animals which is actively injected
can cause ill-health or death by its local or
into the body of another organism.
systemic action.
Source May be natural or synthetic in origin. Always biological in origin, produced by animals.
Mode of Enters the body by ingestion, inhalation, Enters the body only by injection
Entry injection, absorption through skin or mucosa. through bite or sting.
Route of Usually passive (swallowed, inhaled or Always active (bite or sting
Administration absorbed). mechanism).
Arsenic, cyanide, pesticides, drugs, Snake venom, scorpion venom, spider venom,
Examples
corrosives. bee venom.
Speed of Action depends on dose, route and Usually rapid acting due to direct entry into
Action chemical nature. circulation or tissues.
Common in suicidal, homicidal,
Forensic Important in cases of snakebite, scorpion
accidental and stupefying
Importance sting and animal envenomation.
poisoning.
Antidote/ Treated using antidotes, supportive Treated mainly with specific
Management therapy and decontamination. antivenom and supportive care.

9) Toxin VS Toxicant VS Poison VS Venom

Basis Toxin Toxicant Poison Venom

Naturally
Man-made or Any substance
occurring Toxic secretion
synthetic which, when
poisonous produced by animals
chemical introduced into the
Definition substances and actively
substances that body in sufficient
produced by injected into
cause harm to quantity, can cause
living another organism.
living organisms. ill-health or death.
organisms.
Always biological,
Synthetic / May be natural or
Origin Biological origin. produced by
industrial origin. synthetic.
animals.
Industries,
Plants, animals, Includes toxins,
laboratories,
Source bacteria, fungi, toxicants and Venomous animals.
agriculture,
algae. venoms.
pharmaceuticals.
Ingestion,
Ingestion, Ingestion, inhalation,
inhalation, Only by bite or
Mode of Entry contact, bite or injection, skin or
injection or skin sting (injection).
sting. mucosal absorption.
absorption.
Route of Passive or active Always active
Usually passive. Usually passive.
Administration depending on type. injection.
Botulinum toxin,
Pesticides, heavy Arsenic, cyanide, Snake venom,
mycotoxins, plant
Examples metals, industrial pesticides, drugs, scorpion venom, bee
toxins, snake
chemicals, drugs. corrosives. venom.
toxin.
Core subject of Studied under
Study is called Studied under
Related Term Forensic Venomology /
Toxinology. Toxicology.
Toxicology. Toxinology.
Forensic Important in Important in Broad medico-legal Important in
Importance biological and suicidal, importance in all snakebite and
 Basis Toxin Toxicant Poison Venom
homicidal, animal
food poisoning accidental and envenomation cases.
poisoning cases.
cases. occupational
poisoning.

10) Drug paraphernalia

Drug paraphernalia refers to all equipment, materials and accessories used for the preparation,
administration, concealment or consumption of drugs of abuse. The presence of drug
paraphernalia is an important indicator of drug abuse or dependence in forensic investigations.

Types of Drug Paraphernalia

1. Paraphernalia for Drug Preparation
Used to prepare drugs before consumption.
• Spoons
• Bottle caps
• Burnt foils
• Mixing containers
• Water for dissolving drugs
Example: Heating heroin on spoon before injection.

2. Paraphernalia for Drug Administration

Used to introduce drugs into the body.
• Injection devices: syringes, needles, tourniquets
• Smoking devices: pipes, chillums, bongs
• Snorting devices: rolled currency notes, straws
• Oral devices: capsules, tablets

3. Paraphernalia for Drug Storage and Concealment

Used to store or hide drugs.
• Small plastic packets (pudiya)
• Aluminum foil
 • Matchboxes
• Cigarette packets
• Plastic vials

4. Paraphernalia for Inhalation

Used mainly for volatile substance abuse.
• Glue tubes
• Plastic bags
• Cloth soaked with solvent
• Aerosol cans

5. Paraphernalia Indicating Chronic Drug Abuse

Suggest long-term or habitual use.
• Multiple used syringes
• Scar marks or track marks
• Burnt spoons and foils
• Drug residue on instruments

Medico-Legal Importance of Drug Paraphernalia

1. Helps in establishing drug abuse or dependence
2. Assists in identifying the type of drug and route of administration
3. Useful evidence in cases under NDPS Act
4. Supports cause and manner of death in suspected overdose cases
5. Aids forensic toxicologists during scene investigation

11) Role of forensic toxicologist

A forensic toxicologist is an individual who collects, analyses and prepares reports of suspected
or unknown toxicants for use in medico-legal and criminal investigations.
The role of a forensic toxicologist is crucial in cases of poisoning, drug abuse and suspicious
deaths.
Roles of a Forensic Toxicologist
1. Collection and Examination of Samples
Collects and examines biological and non-biological samples such as viscera, blood, urine
and tissues suspected to contain toxic substances.
2. Study of Autopsy Report
Carefully reads and correlates autopsy/post-mortem findings with toxicological results.
3. Qualitative Analysis of Toxicants
Performs qualitative analysis to identify the nature of the poison or toxic substance present.
4. Quantitative Analysis of Toxicants
Carries out quantitative analysis to determine the amount or concentration of the toxicant.
5. Determination of Lethal Dose and Lethal Time
Helps in assessing the lethal dosage and time required to cause death, which is important
for medico-legal interpretation.
6. Preparation of Toxicology Report
Prepares a scientific, accurate and legally valid toxicology report based on analytical
findings.
7. Expert Opinion to Investigating Agencies
Provides expert opinion to investigating officers and medical practitioners during inquiry.
8. Expert Testimony in Court
Acts as an expert witness and presents toxicological evidence in the court of law.

12) Classification of poisons

Poisons can be classified based on their mode of action and nature of poison. This classification
helps in understanding the toxic effects, diagnosis and medico-legal importance.

I. Classification Based on Mode of Action

1. Corrosive Poisons
These poisons destroy or corrode tissues at the site of contact.
Examples:
• Acids
• Organic: Acetic acid (CH₃COOH), formic acid, oxalic acid
• Inorganic: Sulphuric acid (H₂SO₄), nitric acid (HNO₃), hydrochloric acid (HCl)
• Alkalis: Sodium, magnesium, calcium
2. Irritant Poisons
Cause irritation, inflammation, pain, redness and tissue damage.
Inorganic irritants:
• Metallic: Arsenic, lead, bismuth, mercury
• Non-metallic: Chlorine, halogens
Organic irritants:
• Plant based: Ricin
• Animal based: Scorpion venom, snake venom, bee venom
• Mechanical: Diamond dust, powdered glass

3. Systemic Poisons
Act after absorption and affect specific organs.

A. Neurotic Poisons (Act on CNS)

• Spinal: Nux vomica, gelsemium
• Peripheral: Curare, conium
• Cerebral:
• Somniferous: Opioids, sleeping pills
• Inebriants: Alcohols, anaesthetics, sedatives
• Deliriants: Datura, cannabis, belladonna

B. Cardiac Poisons
• Act on cardiovascular system
• Examples: Aconite, oleander, digitalis

C. Asphyxiant Poisons
• Act on respiratory system
• Examples: Carbon monoxide, tear gas, nitrous oxide, sodium isocyanate

4. Miscellaneous Poisons
• Antihistamines
• Antibiotics
• Amphetamines
 • Antipyretics
• Tranquilizers

II. Classification Based on Nature of Poison

1. Gaseous Poisons
• Hydrogen sulphide (H₂S)
• Carbon monoxide (CO)
• Sodium isocyanate
• Nitrous oxide

2. Volatile Inorganic Poisons

• Arsine gas
• Phosgene gas
• Phosphine gas

3. Volatile Organic Poisons

• Alcohol
• Ethanol
• Methanol

4. Non-Volatile Inorganic Poisons

• Anions: Halides, chlorides, sulphates
• Cations: Heavy metals

5. Non-Volatile Organic Poisons

• Neutral: Pesticides
• Organophosphates
• Organochlorines (DDT)
• Carbamates
• Pyrethroids
 • Acidic: Barbiturates, sulpha drugs, acidic drugs, aspirin
• Basic: Alkaloids, benzodiazepines

6. Plant Poisons
• Aconite
• Castor seeds

7. Miscellaneous
• Mechanical: Diamond dust, powdered glass
• Food: Mycotoxins
• Animal: Venoms

13) Laws related to poisons

Several laws have been enacted in India to regulate the manufacture, sale, possession and misuse
of poisons and drugs and to deal with their medico-legal consequences.

1. Opium Act, 1857

• One of the earliest laws related to poisons.
• Regulated the cultivation and use of opium.
• Now abolished.

2. Poisons Act, 1919

• Regulates the import, sale, possession and transport of poisons.
• Empowers State Governments to frame rules regarding licensing and safe handling of
poisons.

3. Drugs and Cosmetics Act, 1940

• Regulates the import, manufacture, sale and distribution of drugs and cosmetics.
• Aims to prevent the sale of spurious and adulterated drugs.
• Ensures quality, safety and efficacy of medicinal products.
4. Drugs and Cosmetics Rules, 1945
• Framed under the Drugs and Cosmetics Act.
• Regulates over-the-counter drugs, testing of drugs and functioning of laboratories.
• Establishment of Central Drugs Laboratory and Drug Advisory Board.

5. Pharmacy Act, 1948

• Regulates the profession of pharmacy.
• Establishes State Pharmacy Councils.
• Ensures that drugs are dispensed only by qualified pharmacists.

6. Drugs and Magic Remedies (Objectionable Advertisements) Act, 1954

• Prohibits misleading advertisements related to drugs and magical cures.
• Prevents false claims regarding treatment of diseases.

7. NDPS Act, 1985 (Narcotic Drugs and Psychotropic Substances Act)

• Controls narcotic drugs and psychotropic substances such as opium, cannabis and coca
leaves.
• Offences under this Act are non-bailable.
• Regulates cultivation, manufacture, possession, sale and transport of narcotic drugs.

8. Relevant Sections of Bharatiya Nyaya Sanhita (BNS) / BNSS

• Section 123 BNS: Causing hurt by means of poison
• Section 286 BNS: Negligent conduct with respect to poisonous substances
• Section 325 BNS: Killing or poisoning animals
• Section 194 BNSS: Police inquiry and reporting in cases of suicide

14) Medicolegal aspects

Poisoning has significant medico-legal importance as it may be involved in suicidal, homicidal,

accidental or other unlawful acts. Proper medico-legal evaluation is essential for justice delivery.
1. Manner of Poisoning
Poisoning may occur in different medico-legal manners:
• Suicidal poisoning – most common
• Homicidal poisoning – rare
• Accidental poisoning – common in children and occupational exposure
• Stupefying poisoning – used to facilitate crimes
• Abortifacient poisoning – used to induce abortion
Determination of manner of death is crucial in legal investigation.

2. Cause of Death
• Poisoning may be the direct cause of death
• Or may act as a contributory cause
• Toxicological findings must be correlated with clinical history and post-mortem findings

3. Legal Duty of Medical Practitioner

• All suspected poisoning cases must be treated as medico-legal cases
• Doctor must:
• Provide life-saving treatment first
• Inform the police
• Maintain proper medical records

4. Post-Mortem Examination
• Autopsy is required in fatal poisoning cases
• Proper collection and preservation of viscera is essential
• Helps in determining cause and manner of death

5. Toxicological Analysis
• Qualitative and quantitative analysis of poisons
• Helps establish:
• Type of poison
• Dose
 • Time since administration

6. Expert Opinion
• Forensic toxicologist provides expert opinion based on analysis
• Plays a key role in interpretation of findings
• Assists the court in understanding scientific evidence

7. Relevant Legal Provisions

• Section 123 BNS – Causing hurt by means of poison
• Section 286 BNS – Negligent conduct with respect to poisonous substances
• Section 325 BNS – Killing or poisoning animals
• Section 194 BNSS – Police inquiry and reporting in suicide cases
• Poisons Act, 1919 – Regulation of poisons
• NDPS Act, 1985 – Narcotic and psychotropic substances

8. Court of Law
• Toxicological report is produced as documentary evidence
• Forensic toxicologist may be examined as an expert witness
• Findings help in deciding criminal responsibility

15) Nodal agencies and stakeholders

Nodal agencies are organizations that play a key role in the prevention, detection, analysis and
management of poisoning cases, and provide support in medico-legal investigations.

1. Poison Information Centre (Poison Control Centre)

• Provides medical advice and treatment guidance in poisoning cases.
• Offers free and immediate consultation, often through telephone services.
• First Poison Information Centre was established in Netherlands (1940).
• In India, the National Poisons Information Centre (NPIC) was established at AIIMS,
New Delhi in 1995.
2. Forensic Science Laboratory (FSL) / Central Forensic Science Laboratory
(CFSL)
• Responsible for analysis of biological and non-biological samples in poisoning cases.
• Conducts qualitative and quantitative toxicological analysis.
• Provides expert opinion and reports to investigating agencies and courts.

3. TIAFT (The International Association of Forensic Toxicologists)

• An international organization promoting research and development in forensic toxicology.
• Encourages scientific exchange, training and standardization of toxicological practices.

4. SOFT–AAFS
• SOFT (Society of Forensic Toxicologists)
• AAFS (American Academy of Forensic Sciences)
• These bodies provide guidelines for forensic toxicology laboratories, quality assurance
and professional development.

5. FSSAI (Food Safety and Standards Authority of India)

• Regulates food safety and food adulteration.
• Plays an important role in food poisoning cases and toxic contamination of food items.

6. Stakeholders in Forensic Toxicology

• Forensic toxicologists
• Investigating officers
• Government medical practitioners
• Professionals of forensic medicine
These stakeholders work together to ensure proper investigation, analysis and legal action in
poisoning cases.

16) Liquid liquid extraction and nernst’s distribution law

Liquid–Liquid Extraction (LLE)
Liquid–liquid extraction is a separation technique used in forensic toxicology to isolate poisons
or drugs from biological samples by distributing them between two immiscible liquid phases,
usually an aqueous phase and an organic solvent.
It is commonly used for extracting toxicants from blood, urine, gastric contents and viscera prior
to analysis.

Principle of LLE
The principle of liquid–liquid extraction is based on Nernst’s distribution law, according to which
a solute distributes itself between two immiscible solvents in a constant ratio at a given temperature.

Nernst’s Distribution Law

Statement:
When a solute is added to two immiscible solvents, it distributes itself between the two solvents in
such a way that the ratio of its concentrations in the two solvents remains constant at
equilibrium, provided the solute exists in the same molecular form in both solvents.
Mathematically,
C1C2=K\frac{C_1}{C_2} = KC2C1=K
Where:
• C1C_1C1 = concentration of solute in organic solvent
• C2C_2C2 = concentration of solute in aqueous solvent
• KKK = partition coefficient

Role of pH in LLE
• Weak acidic poisons are better extracted in acidic medium
• Weak basic poisons are better extracted in alkaline medium
This ensures the toxicant remains in unionized form, increasing extraction efficiency.

Procedure of LLE
1. Biological sample is adjusted to suitable pH.
2. An organic solvent is added.
3. The mixture is shaken thoroughly.
4. Two layers are allowed to separate.
 5. Organic layer containing the toxicant is collected.
6. Solvent is evaporated and residue is analysed.

Applications in Forensic Toxicology

• Extraction of alkaloids, barbiturates, pesticides and drugs
• Used as a preliminary step before instrumental analysis
• Helpful in qualitative and quantitative toxicological analysis

Advantages
• Simple and economical
• Applicable to a wide range of poisons
• No sophisticated instruments required

Limitations
• Not suitable for highly polar compounds
• Emulsion formation may occur
• Time-consuming compared to modern techniques

17) Toxicant is any substance embedded in the matrix,

Matrix is any substance in which analyte of interest is embedded.

18) Instruments used in detection of poisons

• Atomic Absorbtion Spectroscopy (AAS)

• Atomic Emission Spectroscpoy (AES)
• Inductively Coupled Plasma – Mass Spectroscopy (ICP-MS)
• X-Ray Flourescence (XRF)
• Energy Dispersive X-Ray Flourescence (EDSRF)
• Neutron Activation Analysis (NAA)
• Scanning Electron Microscope (SEM)
19) Poison detection/extraction

• Wet digestion ( Heavy metals)

• Dry digestion (Heavy metals)
• Distillation (volatile poison extraction)