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The word “Toxicology” is derived from the Greek word “Toxicon”
which was used as a poisonous substance in arrowheads.
Science embodying the knowledge, source, character, fatal effect,
lethal dose analysis of poisons and the remedial measures.
Poison is defined as the substance, which is capable of producing
injury or death when absorbed. Appropriate dosages can
differentiate poison and also the remedial measures
• All chemicals can produce injury or death under certain
conditions. Hence, a poison can be defined as a substance
that is capable of producing detrimental effects on a
living organism.
• The toxicologist is specially a trained expert to examine
the role of such substances and their adverse effects.
• Forensic toxicology emerged as the hybrid of analytical
Chemistry and toxic principle effects.
• Forensic toxicologists are also primarily concerned with
the medico - legal aspects of the harmful effects of
chemicals on human and animals
BIOLOGICAL: Viscera, blood, urine, saliva, stomach contents, intestinal
contents, gastric, lavage, vomit, brain matter, stool, fecal matter, bone, nails,
hair, skin.
NON BIOLOGICAL MATRICES: Water, remnants or traces of poison in small
container, food and food products, milk and milk products, fruits, vegetables,
tea, coffee, cooked materials, drinks, cereals, pulses, wines, etc.
VISCERA: Internal organs viz. liver, kidney, stomach, intestine, gall bladder,
uterus, heart, lungs, brain etc.
IN CASE OF DEATH IN CASE OF LIVING OTHER MATERIALS
Stomach with contents Blood Tablets
Small intestine with contents Urine Suspected bottles with liquids
Liver Stomach wash Powders
Spleen Other suspected items
Kidney
Brain
Blood
Bones
Hair
Nail
• Exhibits collected on the crime scene are packed, sealed and sent to the
laboratory for examination along with a mention of what information is requisite.
• Theses evidence are received by the lab and given a special number of
examinations according to chronology of receiving.
• Opening of parcels for examination, their examination and report writing after
examination was described and demonstrated.
• brief description of how records are maintained was also mentioned.
PARCEL CONTENTS
• Brief history of the case under investigation: a copy of the FIR
• Certificate by forwarding authority mentioning police station, under section, name of forwarding
officer
 Description of the exhibits: Describing all the exhibits in the parcel in detail, how & when they are
collected & by whom, & the source of the Exhibits & with which seal the parcels are sealed.
• Letter describing the Queries & nature of examination required
• Certificate by the forwarding authority
• The sample seals with which the parcels & sub-parcels containing the exhibits are sealed,
• An additional MLC (medico legal report) by the doctor’s examination of the victim,
• Other documents like Road certificate & HQ forwarding letter.
Extraction (separation) of active constituent i.e. poisons in biological and non-biological
matrices.
Stripping or purification of active constituent thus separated.
Identification or Rapid Screening test through TLC and Chromatographic Techniques.
Quantitation, if required.
Interpretation/conclusion/Examination report.
It includes:
1. Solvent Extraction:
2. Distillation
3. Dialysis
4. Absorption
5. Chemical Digestion
6. Chromatography
Tests performed:
Extraction Procedure:
50-100 gms. of Tissue or Viscera in distillation flask
About 5 ml of H2SO4 or H3PO4 is added
steam distillation
distillate collected in AgNO3
Ammonium -Molybdate Test Procedure:
2 ml of extract taken
2 ml of ammonium- molybdate added to extract
A few drops of conc, HNO3 is added and warmed
“CANARY YELLOW “ color observed.
Ammonium – Molybdate Benezidine Test:
1 drop of acidic solution of extract is placed on filter paper
1 drop of ammonium -molybdate is added followed by 0.05% Benezidine
acetate in acetic acid
Paper is exposed to ammonia vapour
“BLUE STAIN” developes.
• Any substance or mixture of substances intended for preventing, destroying or controlling any pest,
including vectors of human or animal disease.
• Powerful inhibitors of ACETYLCHOLINESTERASES.
• Acetylcholine is produced at the myoneural junction, and acts as a chemical signal transmitter at
synapses.
• Common Mode of suicide in India.
 ORGANOPHOSPHOROUS :
• DDVP ( Dichlorovos) ,Phosphamidon, Mevinphos, Methyl Parathion, Fenitrothion,
• Fenthion, Chlorpyriphos, Quinalphos, Diazinon, Dimethoate, Malathion, etc.
 ORGANO CHLORO PESTICIDES:
Includes- Dichlorodiphenyltrichloroethane ( DDT ), Lindane, Benzene
hexachloride group ( BHC ), Chlordane ,etc.
 CARBAMATE INSECTICIDES:
Includes- N-methyl carbamates e.g. Carbaryl .
EXTRACTION FROM BLOOD:
10 ml blood + 10 ml of 10% Sodium Tungstate solution + 5ml of 1N
Sulphuric Acid
shake for 2 min.
filter the contents and wash residue with 0.1 N H2SO4
Extract the Filtrate with n- hexane
collect the hexane layer, pass through anhydrous sodium sulphate and evaporate to 1 ml.
Air dry the sample and leave overnight.
IDENTIFICATION OF PESTICIDES BY TLC:
 ORGANOPHOSPHOROUS PESTICIDES:
MOBILE PHASE : Hexane : Acetone ( 4: 1 ), Benzene : Methanol (6:4)
STATIONARY PHASE: Silica Gel G
CONTROL USED: DDVP, Malathion, Prathion etc.
SPRAYING REAGENT: Palladium chloride.
COLOUR OF SPOT: “ yellow-red “ for DDVP , “Brown” for
Malathion and Parathion
 ORGANOCHLORO PESTICIDES:
MOBILE PHASE: Hexane : Acetone (4:1)
STATIONARY PHASE: Silica Gel G
CONTROL USED: Endosulphan , Endrin , Quinone phosphate , Phosphomidon
SPRAYING REAGENT : ZnCl2 –Diphenylamine solution
COLOUR OF SPOT: “Bluish- Green”
 CARBAMATE PESTICIDES:
MOBILE PHASE: Hexane : Acetone (4:1)
STATIONARY PHASE : Silica Gel G
CONTROL USED: Propoxur ( Baygon ) , Carbaryl , Carbofuran.
SPRAYING REAGENT: Fast Blue B
COLOUR OF SPOT: “Red/ Violet”
A drug can be defined as a “natural or synthetic substance that is used to produce physiological or psychological
effects in humans or other animals ”.
EXTRACTION PROCEDURE:
FROM VISCERA:
 “ AMMONIUM SULFATE METHOD” :
visceral material + 10 ml of 5% caetic acid + Sodium Ammonium Sulphate in conical flask
kept on water bath
( deproteinisation )
cool and filtered
STEP 2:
Extract taken in separating funnel + 50 ml Diethyl Ether
gently shaken for 2 min.
allowed to stand
Now 2 layers get separated
aqueous layer taken in another flask
Organic layer is filtered with sodium sulphate and taken for test
Step3.
Aqueous layer is made alkaline By adding ammonia solution + Same amount of Chloroform &
Ether (1:1) is taken in separating Funnel
↓(gently shaken for 20 min.&
↓ allowed to stand
Now both layers get separated
↓
Aqeous layer is decant off and Organic layer is filtered through anhydrous Sodium sulphate
organic layer used for basic drug
Step4.
Now both acidic and basic drugs are used are allowed to perform TLC along with
standards.
Acidic Drugs:
Standard for acidic
drugs
Solvent system Spray Reagent
I
dentification colour
Phenobarbitone Chloroform :Acetone (9:1) FPN Purple violet colour
Phenargon Chloroform :Acetone (9:1) Zwiker Violet colour
Basic Drugs:
Standard for basic drugs Solvent system Spray Reagent Identification colour
Alprazolam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed
by iodoplatinate
Pink color
Lorazepam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed
by iodoplatinate
Pink color
Diazepam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed
by iodoplatinate
Pink color
Nitrazepam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed
by iodoplatinate
Pink color
Phenargan Chloroform:Methanol(9:1) Dragendorff‟s reagent followed
by iodoplatinate
Pink color
 EXTRACTION FROM BLOOD :
Blood sample + sodium tungstate + sulphuric acid
↓(for deproteinisation)
Filter & use filtrate for solvent extraction
Now apply step 2, 3 & 4 for drug analysis.
From Gastric lavage & urine, apply step 2, 3& 4 directly for drug analysis. Before applying step 2 made
it acidic
Isolation:
By acid-distillation:
50-100 gms. of viscera (properly minced) or Blood brought to the consistency of a thin gruel by
adding 3-5 times of distilled water.
aciDified with tartaric or sulphuric acid and submitted to steam distillation.
(The condenser and the receiving flask should be well cooled with ice especially during the hot
season, the outlet of the condenser being dipped in a little water.)
A few pieces of porcelain dish is added in flask to prevent bumping.
Distillate is collected in conical flask for chemical testing.
“DICHROMATE TEST” :
1ml of distillate + 1ml of potassium dichromate + 1ml of sulphuric acid
(drop wise through wall)
↓
BLUE/GREEN ring will formed
“IODOFORM TEST”:
1ml of distillate + few drops of 10% NaOH+ iodine solution
↓warm on water bath
Solution color changes to brown
↓add few drops of 10% NaOH + iodine sol.
↓& warm
Solution color changes from brown to yellow
↓cooled & observed under microscope
Characteristic Hexagonal crystals are formed
“IMAGE OF IODOFORM CRYSTAL OBSERVED” :
Toxicological analysis of visceral sample

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Toxicological analysis of visceral sample

  • 1.
  • 2. The word “Toxicology” is derived from the Greek word “Toxicon” which was used as a poisonous substance in arrowheads. Science embodying the knowledge, source, character, fatal effect, lethal dose analysis of poisons and the remedial measures. Poison is defined as the substance, which is capable of producing injury or death when absorbed. Appropriate dosages can differentiate poison and also the remedial measures
  • 3. • All chemicals can produce injury or death under certain conditions. Hence, a poison can be defined as a substance that is capable of producing detrimental effects on a living organism. • The toxicologist is specially a trained expert to examine the role of such substances and their adverse effects. • Forensic toxicology emerged as the hybrid of analytical Chemistry and toxic principle effects. • Forensic toxicologists are also primarily concerned with the medico - legal aspects of the harmful effects of chemicals on human and animals
  • 4. BIOLOGICAL: Viscera, blood, urine, saliva, stomach contents, intestinal contents, gastric, lavage, vomit, brain matter, stool, fecal matter, bone, nails, hair, skin. NON BIOLOGICAL MATRICES: Water, remnants or traces of poison in small container, food and food products, milk and milk products, fruits, vegetables, tea, coffee, cooked materials, drinks, cereals, pulses, wines, etc. VISCERA: Internal organs viz. liver, kidney, stomach, intestine, gall bladder, uterus, heart, lungs, brain etc.
  • 5. IN CASE OF DEATH IN CASE OF LIVING OTHER MATERIALS Stomach with contents Blood Tablets Small intestine with contents Urine Suspected bottles with liquids Liver Stomach wash Powders Spleen Other suspected items Kidney Brain Blood Bones Hair Nail
  • 6. • Exhibits collected on the crime scene are packed, sealed and sent to the laboratory for examination along with a mention of what information is requisite. • Theses evidence are received by the lab and given a special number of examinations according to chronology of receiving. • Opening of parcels for examination, their examination and report writing after examination was described and demonstrated. • brief description of how records are maintained was also mentioned.
  • 7. PARCEL CONTENTS • Brief history of the case under investigation: a copy of the FIR • Certificate by forwarding authority mentioning police station, under section, name of forwarding officer  Description of the exhibits: Describing all the exhibits in the parcel in detail, how & when they are collected & by whom, & the source of the Exhibits & with which seal the parcels are sealed. • Letter describing the Queries & nature of examination required • Certificate by the forwarding authority • The sample seals with which the parcels & sub-parcels containing the exhibits are sealed, • An additional MLC (medico legal report) by the doctor’s examination of the victim, • Other documents like Road certificate & HQ forwarding letter.
  • 8. Extraction (separation) of active constituent i.e. poisons in biological and non-biological matrices. Stripping or purification of active constituent thus separated. Identification or Rapid Screening test through TLC and Chromatographic Techniques. Quantitation, if required. Interpretation/conclusion/Examination report.
  • 9. It includes: 1. Solvent Extraction: 2. Distillation 3. Dialysis 4. Absorption 5. Chemical Digestion 6. Chromatography
  • 10. Tests performed: Extraction Procedure: 50-100 gms. of Tissue or Viscera in distillation flask About 5 ml of H2SO4 or H3PO4 is added steam distillation distillate collected in AgNO3
  • 11. Ammonium -Molybdate Test Procedure: 2 ml of extract taken 2 ml of ammonium- molybdate added to extract A few drops of conc, HNO3 is added and warmed “CANARY YELLOW “ color observed.
  • 12. Ammonium – Molybdate Benezidine Test: 1 drop of acidic solution of extract is placed on filter paper 1 drop of ammonium -molybdate is added followed by 0.05% Benezidine acetate in acetic acid Paper is exposed to ammonia vapour “BLUE STAIN” developes.
  • 13. • Any substance or mixture of substances intended for preventing, destroying or controlling any pest, including vectors of human or animal disease. • Powerful inhibitors of ACETYLCHOLINESTERASES. • Acetylcholine is produced at the myoneural junction, and acts as a chemical signal transmitter at synapses. • Common Mode of suicide in India.  ORGANOPHOSPHOROUS : • DDVP ( Dichlorovos) ,Phosphamidon, Mevinphos, Methyl Parathion, Fenitrothion, • Fenthion, Chlorpyriphos, Quinalphos, Diazinon, Dimethoate, Malathion, etc.
  • 14.  ORGANO CHLORO PESTICIDES: Includes- Dichlorodiphenyltrichloroethane ( DDT ), Lindane, Benzene hexachloride group ( BHC ), Chlordane ,etc.  CARBAMATE INSECTICIDES: Includes- N-methyl carbamates e.g. Carbaryl .
  • 15. EXTRACTION FROM BLOOD: 10 ml blood + 10 ml of 10% Sodium Tungstate solution + 5ml of 1N Sulphuric Acid shake for 2 min. filter the contents and wash residue with 0.1 N H2SO4 Extract the Filtrate with n- hexane collect the hexane layer, pass through anhydrous sodium sulphate and evaporate to 1 ml. Air dry the sample and leave overnight.
  • 16. IDENTIFICATION OF PESTICIDES BY TLC:  ORGANOPHOSPHOROUS PESTICIDES: MOBILE PHASE : Hexane : Acetone ( 4: 1 ), Benzene : Methanol (6:4) STATIONARY PHASE: Silica Gel G CONTROL USED: DDVP, Malathion, Prathion etc. SPRAYING REAGENT: Palladium chloride. COLOUR OF SPOT: “ yellow-red “ for DDVP , “Brown” for Malathion and Parathion
  • 17.  ORGANOCHLORO PESTICIDES: MOBILE PHASE: Hexane : Acetone (4:1) STATIONARY PHASE: Silica Gel G CONTROL USED: Endosulphan , Endrin , Quinone phosphate , Phosphomidon SPRAYING REAGENT : ZnCl2 –Diphenylamine solution COLOUR OF SPOT: “Bluish- Green”  CARBAMATE PESTICIDES: MOBILE PHASE: Hexane : Acetone (4:1) STATIONARY PHASE : Silica Gel G CONTROL USED: Propoxur ( Baygon ) , Carbaryl , Carbofuran. SPRAYING REAGENT: Fast Blue B COLOUR OF SPOT: “Red/ Violet”
  • 18. A drug can be defined as a “natural or synthetic substance that is used to produce physiological or psychological effects in humans or other animals ”. EXTRACTION PROCEDURE: FROM VISCERA:  “ AMMONIUM SULFATE METHOD” : visceral material + 10 ml of 5% caetic acid + Sodium Ammonium Sulphate in conical flask kept on water bath ( deproteinisation ) cool and filtered
  • 19. STEP 2: Extract taken in separating funnel + 50 ml Diethyl Ether gently shaken for 2 min. allowed to stand Now 2 layers get separated aqueous layer taken in another flask Organic layer is filtered with sodium sulphate and taken for test
  • 20. Step3. Aqueous layer is made alkaline By adding ammonia solution + Same amount of Chloroform & Ether (1:1) is taken in separating Funnel ↓(gently shaken for 20 min.& ↓ allowed to stand Now both layers get separated ↓ Aqeous layer is decant off and Organic layer is filtered through anhydrous Sodium sulphate organic layer used for basic drug
  • 21. Step4. Now both acidic and basic drugs are used are allowed to perform TLC along with standards. Acidic Drugs: Standard for acidic drugs Solvent system Spray Reagent I dentification colour Phenobarbitone Chloroform :Acetone (9:1) FPN Purple violet colour Phenargon Chloroform :Acetone (9:1) Zwiker Violet colour
  • 22. Basic Drugs: Standard for basic drugs Solvent system Spray Reagent Identification colour Alprazolam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed by iodoplatinate Pink color Lorazepam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed by iodoplatinate Pink color Diazepam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed by iodoplatinate Pink color Nitrazepam Chloroform:Methanol(9:1) Dragendorff‟s reagent followed by iodoplatinate Pink color Phenargan Chloroform:Methanol(9:1) Dragendorff‟s reagent followed by iodoplatinate Pink color
  • 23.  EXTRACTION FROM BLOOD : Blood sample + sodium tungstate + sulphuric acid ↓(for deproteinisation) Filter & use filtrate for solvent extraction Now apply step 2, 3 & 4 for drug analysis. From Gastric lavage & urine, apply step 2, 3& 4 directly for drug analysis. Before applying step 2 made it acidic
  • 24. Isolation: By acid-distillation: 50-100 gms. of viscera (properly minced) or Blood brought to the consistency of a thin gruel by adding 3-5 times of distilled water. aciDified with tartaric or sulphuric acid and submitted to steam distillation. (The condenser and the receiving flask should be well cooled with ice especially during the hot season, the outlet of the condenser being dipped in a little water.) A few pieces of porcelain dish is added in flask to prevent bumping. Distillate is collected in conical flask for chemical testing.
  • 25. “DICHROMATE TEST” : 1ml of distillate + 1ml of potassium dichromate + 1ml of sulphuric acid (drop wise through wall) ↓ BLUE/GREEN ring will formed
  • 26. “IODOFORM TEST”: 1ml of distillate + few drops of 10% NaOH+ iodine solution ↓warm on water bath Solution color changes to brown ↓add few drops of 10% NaOH + iodine sol. ↓& warm Solution color changes from brown to yellow ↓cooled & observed under microscope Characteristic Hexagonal crystals are formed
  • 27. “IMAGE OF IODOFORM CRYSTAL OBSERVED” :