Post on 16-Jan-2016
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FORENSIC MEDICINE COURSE
FORENSIC TOXICOLOGY
&
FORENSIC ALCOHOLOGY
KRZYSZTOF S. BOROWIAK
HISTORICAL ROOTS OF TOXICOLOGY
FORENSIC TOXICOLOGYAs medicine science was develope in the end of
XVIII C. (London, Vien, Berlin). The first Forensic Department in Poland was founded in
Krakow in the 1805
CLINICAL TOXICOLOGY
First Poisining Centre in
Washington D.C.
(1957)
OCCUPATIONAL AND
INDUSTRIAL
TOXICOLOGY
60. of XX C.
EXPERIMENTAL
TOXICOLOGY
- Geno
- Immuno
- new products
FORENSIC CLASSIFICATION OF TOXINS
POISONS
VOLAITABLE WITH WATER STEAM
Detectioned with:
GC, GC-MS or Spectrometry
ORGANIC NOT VOLAITABLE SUBSTANCES ISOLATED WITH SOLID 0PHASE OR LIQUID EXTRACTION, detectioned with GC, HPLC, Immuno-methods, spectrometry
NON-ORGANIC SUBSTANCES (METALS) WHEN MINERALISATION IS NECCESSARY, DETECTION WITH AAS
FORENSIC CLASSIFICATION OF TOXINS (2)
POISONS
EFFECTS CAN BE OBSERVED IN AUTHOPSY
ANATHOMOPATOLOGICAL CHANGES ARE NOT OBSERVED
Blood toxins:
- blocking oxygen transport or metabolism
Heavy metals
With characteristic odourCorrosive
and irritants
•Drug of abuse
• chemicals
•Drugs and pharmaceutics
• others
MOST FREQUENT POISONS DETECTED IN THE POST MORTEM SAMPLES
DRUGS
ALCOHOLS
DRUG ABUSE
SOLVENTS
Pesticides
ABBREVATION – IN POLISH FORENSIC STATISTIC ON THE 3RD PLACE THE CARBON MONOXIDE IS LOCATED
45%
55%
15%
5%
THE COLLECTION ROUTES AND TYPE OF POST MORTEM SAMPLES
• Blood ( 2 x 10 mL) – alcohols & drug abuse, drugs & other organics, • Urine (25-50 mL) – alcohols, drug metabolites, drug of abuse • Brain tissue (50g) – psychotropic substances• Kidney (1) – nophrotoxicity estimation and elimination • Liver (50-100 g) – hepatotoxicity and high conc. of metabolites• Intenstine with contents (50g) • Stomach with separated colected contents• Lungs (50 g) – excelent matherial for solvents and gasses int.
When the results of authopsy are unknown and we suspect neccessary of multi-direct toxicological examination
THE COLLECTION ROUTES AND TYPE OF POST MORTEM SAMPLES
• BLOOD COLLECTED FOR ETHANOL AND OTHER ALCOHOLS DETERMINATION SHOULD BE TAKEN FROM SURFACE VESSELS (VEINS)
• BLOOD COLLECTED FOR CARBOXYHEMOGLOBIN SHOULD BE TAKEN FROM DEEP LOCATED
VESSELS• VIALS WITH SAMPLES COLLECTED FOR
VOLAITABLE SUBSTANCES DETERMINATION SHOULD BE
PERFECT CLOSED
ALL BIOLOGICAL SAMPLES SHOULD BE PRESERVED IN TO ABSOLUTLY CHEMICALY CLEAN VIALS,
BOTLES AND JARS
IMPORTANT TO REMEMBER
DIFFERENTIATE OF DEATH CASE
Observation during authopsy: pink-redish colour of body surface, internal organ surface and blood
That can be resulted from:
• death in the course of carbon monooxide fatal intoxication (COHb > 40% detected in the blood)
• death in the course of cianides intoxication
(bitter almond smell)
• death in the course of low temperatures action (freezing)
Differentiate course of death
Observation durring authopsy: gray-blue colour of corps and organs surfaces, chocolade colour of blood
Result from:
• Death in course of methemoglobinemic substances intoxication (nitrates, nitrites and anilin e.g. – MtHb > 30%)
• deep anoxemia, metabolic acidosis
• putrification of body
OTHER CHARACTERISTIC CHANGES ILLUSTRATED EFFECTS OF POISON
INFLUENCE
TYPE OF INJURY EXAMPLES
• CAUSTIC POISONS (CORROSIVE – ACUTE LYE BURNS, CORROSION OF GASTRIC MUCOSA, HYPEREMIA, HAEMORAGIAE, NECROSIS) - (STRONG ACIDS, ALKALIAS, HOME SANITARY-CLEANING AGENTS)
• SKIN BLISTERS (BARBITURATES) • RENAL DAMAGES & NECROSIS – METALS & METALOIDS TETRACYCLINES, BABRBITURATES, METHANOL,
GLYCOLS, • PULMONARY & GI CHANGES (OEDEMA,HYPEREMIA,
HEMORRAGIAE, IRRITANTS,) – GASES, ARSEN OXIDE, INSECTICIDES, PETROLEUM DISTILATE PRODUCTS (GASOLINE, KEROSENE....)
• HYPEREMIA – PINK-RED BLOOD AND ORGAN SURFACES – C=O
• SKIN AND BLOOD CYANOSIS – METHEMOGLOBINEMIC AGENTS
MECHANISMS OF CELL DEATH FOLLOWING DRUG INDUCED LIVER INJURY (By J.G.O’Grady)
DRUG
COVALENT BINDING TO CELLULAR STRUCTURES
Covalently modified P-450
Covalently modified proteins
Haptenic epitopes
Auto-antigen epitopes
Reactive metabolites Covalently modified
DNA
Mutations,
cancerogenesis
DEATH OF CELLNeoantigens Immune system
MORPHOLOGIC CLASSIFICATION OF DRUG-RELATED CHRONIC LIVER INJURY
TYPE OF INJURY EXAMPLES • CHRONIC HEPATITIS HALOTANE, ISONIAZIDE,
DANTROLENE, SULPHONAMIDES,• CHRONIC CHOLESTASIS TCA, FENOTHIAZINES,
BARBITURATES, PHENYTOIN, TOLBUTAMIDE,
• CHRONIC STEATOSIS ETHANOL, METHOTREXATE, ANTINEOPLASTIC AGENTS,
• PHOSPHOLIPIDOSIS AMIODARONE, THIORIDAZINE,• VASCULAR (occlusion, ESTROGENS, ANABOLICS, hypertension, peliosis)• NEOPLASM ESTROGENS, ANABOLICS, VINYL
MORPHOLOGIC CLASSIFICATION OF DRUG-RELATED ACUTE LIVER INJURY
(HEPATITIS)
TYPE OF INJURY EXAMPLES___________________________________________
• HEPATOCELLULAR (cytolytic) • zonal necrosis - CCL-4, halothane, Acetaminophen
• steatosis (makro and mikrovesicular) – ethanol,
methotrexate, tetracyclines, valproic acid, • CHOLESTATIC • hepatocanalicular -Amoxiciline, chlorpromazine,• Canalicular – estrogens, steroids, anabolics
• VASCULAR (vein occlusion) estrogens, anabolics.
MEDICO-LEGAL ASPECTS OF ETHANOL
TYPE OF CRIMES AND ACCIDENTS STRONGLY ASSOCIATED WITH USE OF ETHANOL :
• MAJOR TRAUMA• MOTOR VEHICLE COLLISIONS• FIRES, BURNS AND FALLS• DOMESTIC VIOLENCE, CHILD ABUSE• SUICIDES ATTEMPT• TRAUMA AND INJURES OUTCOM COMPLICATIONS
• Ethyl alcohol is most important constituent of different „alcoholic” beverages (Long-drinks), and basic solvent used
in several medicinal preparations.
• Ethanol is most frequent abused intoxicating substance companioned of human from several hundred years.• It is most commonly encountered toxic substance in the forensic toxicology also.
CLINICAL AND MEDICO-LEGAL CLASSIFICATIONOF ETHANOL
ACTION
> 4,0 - 4,5 ‰ PARALYTIC PHASE (V)3,0 - 4,0 ‰ NARCOTIC PHASE (IV)
2,0-3,0 ‰ EXCITATION PHASE (III)
1,0-2,0 ‰ EUPHORIC PHASE (II)
0,2-1,0 ‰ DYSPHORIC ACTION (I)
< 0,2 - 0,5 > ‰ – STATUS AFTER USAGE OF ALCOHOL - OFFENSE § 87 KW
> 0,5 ‰ DRUNKNESS STATUS - CRIME ACCORDING TO § 178 CRIME CODE
2000-2004 YEARS
• 240 from 760 clinical examination (alcohols)
• 975 drivers examined
• 343 under alcohol influence
• near 110 under drug abuse (THC, AMPH.)
• 84 - both substances W 2005 - 2007
• Ca. 2800 drivers and victims of accidents were examined (alcohol + drug abuse): 800 -1200 per year
• 2007 - under influence of drug abuse 221 examined (from 595)
• THC, AMPH, MDMA, Cocaine
STATISTIC DATA OF PAM FOR. DEPARTM.
AMOUNT OF ETHANOL IN THE DIFFRERENT ALCOHOLIC BEVERAGES
• GLASS OF VODKA 100g = 32g PURE SPIRIT
• 1-2 GLAS OF 12-14% VINE 500 mL = 32g PURE SPIRIT
• 4 GLAS OF PORTER BEER (ALE, GUINESS – 8-9%) = 32g PURE SPIRIT
INTAKE ANY OF ABOVE LONG DRINKS CAUSED OVERCROSS THE LEGAL LIMIT OF ETHANOL IN
BLOOD
OVER 0.5 PROMILLE (0.5 mg/mL)
ETHANOL CONCENTRATION IN SOME COMMERCIAL PRODUCTS
PRODUCT ETHANOL
C [%]
„PROOF” AMOUNT
in [g]BEER 2-6 4-12 1,6-4,8
WINES 10-20 20-40 8-16
DESTILATED SPIRIT (vodka,
brandy, cognac)40-60 80-120 32-48
COUGH MEDICINES
3-25 6-50 2,4-20
MOUTHWASH 16-27 32-54 12,8-21,6
COLOGNES 40-60 80-120 32-48
Drinking and driving-the limits in Europe
BULGARIA, HUNGARY, CZECHY, SLOVAKIA, FSRR
PORTUGAL, YUGOSLAVIA, NEDERLAND, BELGIUM, FINLAND, GERMANY, UK
ENGLAND,SPAIN, FRANCE, SWITZERLAND, AUSTRIA
IRELAND
POLAND *, SWEDEN, NORWAY
1,0
0,8
0,5
0,2
0,0
‰ (PROMILLE 1/1000) = MG/ML
‰ x 10 = MG/L
INFLUENCE OF DIFFERENT FACTORS ON THE BODY-KINETIC OF ETHANOL
EXTERNAL AGENTS
• TIME-DEPEND AND FREQUENTNES OF DRINKING
• KIND OF BEVERAGES
• DIET
• ENVIROMENTAL TEMPERATURE
• BIOMETOROLOGY STATUS
CONSTITUAL AGENTS
• RASE
• SEX
• AGE
• HEALTH STATUS
• GENETIC PREDISPOSITION
THEREFORE EFFECTS OF ETHANOL INFLUENCE ARE PERSONAL, INDIVIDUAL AND CHANGEBLE IN TIME
HEPATIC CELLULAR OF ETHANOL BIOTRANSFORMATION
ETHANOL
ALCOHOLDEHYDROGENASE
ACETYTALDEHYDE
ACETALDEHYDE
DEHYDROGENASE
ACETATE
CYTOCHROMEP450 2E1
MEOS
ENDOPLASMIC RETICUKUM
CYTOPLASME OF
HEPATOCYTE
MITOCHONDRIUM
XANTHINE OXIDASEACETALDEHYDE OXYIDASE
OXIDANT STRESS
INACTIVE METABOLITES
CURVES ILLUSTRATED DIFFERENT BODY-KINETIC OF ETHANOL
30 60 90 120 (t) min.
0,0 5 %
0,1 %
0,15 % ETHANOL DEFFICIT
AVERAGE EMPTY FULL CUMULATION
β
β
β
/ R / - DISTRIBUTION COEFFICIENT
ELIMINATION COEFFICIENT
ABSORPTION
EFFECTS OF THE EXCESSIVE ALCOHOLS INTAKE ON THE LIVER TISSUE
ETHANOL INTAKE,MORE THAN 40 UNITS A WEEK
METABOLISM ABNORMALITIES
TRIGLICERIDES, SUGARS, GLIKANES
FATTY LIVER PROCESS
ALCOHOLIC HEPATITIS
ALCOHOLIC CIRRORIS
FIBROTIC INJURYFATTY LIVER
70%
30%
1 UNIT = 10 g PURE (spirit) ALCOHOL (100%)
100 ml of vodka (40%) included 32 g of pure alcohol
THE RETROSPECTIVE AND PROSPECTIVE ALCOHOLOMETRIC CALCULATION
EXPERT’S OPPINION FOR COURTS, PROSECUTORS AND INSURANCE
COMPANIES
WHEN SHOULD BE PERFORMED ?
• WHEN DRIVER-PERPETRATOR OF ROAD FATAL ACCIDENT WAS ESCAPED
• WHEN ETHANOL ESTIMATION WAS PERFORMED SOME TIME AFTER ACCIDENT
• WHEN WHITNESES HAVE DIFFERENT OPPINIONS ON THE SOBRIETY OF DRIVER
• WHEN THERE IS POSSIBILITY OF OVERDRINKING OF ETHANOL POST FACTUM
• WHEN POSSIBLE ADVERSE OR PATHOLOGIC AFTER ALCOHOL REACTIONS MUST BE RESPECT
RETROSPECTIVE ALCOHOLOMETRIC CALCULATION
THEORETIC, CALCULATED CONCENTRATION OF BLOOD ETHANOL, BASED ON THE RESULTS OF ALCOHOLOMETRIC EXAMINATION
C(t) = C(a) + B (60) x T
When: C(t) – ethanol concentration in blood in the road accident moment (time)
C(a) – ethanol concentration estimated in blood in the alcoholometric examination
B (60)- elimination valuue factor (0,07-0,28 mg/h) for expert apply limited (0,1-0,2 mg/h)
T - time difference from accident to examination (in hours) for example 3 hours
RETROSPECTIVE ALCOHOLOMETRIC CALCULATION continued
THE OBLIGATORY CONDITIONS LEAD TO ATTEMPT OF RETROSPECTIVE CALCULATION
• THE ETHANOL ELIMINATION PHASE SHOULD BE NOTED (DECREASING RESULT IN THE FOLLOWING EXAMINATIONS e.g. 1.0 ; 0.8 ; 0.7 mg/mL)
• TIME PERIOD FROM ACCIDENT TO EXAMINATION NOT OVER 5-7 HOURS
• THE ESTIMATED IN BLOOD CONCENTRATION - NOT LESS THAN 0.5 mg/mL
PROSPECTIVE ALCOHOLOMETRIC CALCULATION
APPLIED FOR VERYFICATION OF PENETRATOR WITNESS OR OTHER PARTICIPANTS OF ROAD
ACCIDENT BASED ON THE DECLARED AMOUNT OF INTAKE ETHANOL
C(o) = A x P x R• C(o) – [mg/ml] theoretical, calculated concentration of
blood ethanol based on the known amount and kind of drinken ethanol (declared by driver or whitnes)
• A - amount of intake ethanol expressed in the grams of pure (100%) ethanol (e.g. 100 ml 40% vodka = 32 g pure ethanol)
• P - body weight (kg) standard range 65-85 kg, at present more frequently BMC (body mas index) is used
• R - body distribution coefficient (male - 0.7 ; female – 0.6)
ANALYTICAL METHODS OF ETHANOL DETERMINATION
THE PHYSICO-CHEMICAL METHODS DETERMINATION OF ETHANOL IN THE BREATH AIR (used mainly by Police,
industry security...)
Oxygen Ethanol, carbon dioxide
Vessel-pulmonary membrane
HENRY LAW (1803) - gases diffusion balance; CUSHING R. (1910) non-active diffussion
INFRARED SPECYFIC DETECTORS FOR ETHANOL
ANALYTICAL METHODS OF ETHANOL DETERMINATION cont.
Oxygen Ethanol, carbon dioxide
Vessel-pulmonary membrane
•PERSONAL (ENSURE) – No specyfic, only for drivers autocontrol, may be used also as screening test
•PROFFESSIONAL (Police) – different type of detectors
ALCOMAT (Drager), ALCOTEST 7110, ALCOMETR A.2.0
(estimation of infra-red abssorbance)
ALCOTEST 7310,7410,ALERT, AVAT IV (electrochemical)
ANALYTICAL METHODS OF ETHANOL DETERMINATION cont.
Aldehyde and acid acetic, carbon dioxide
•GAS CHROMATOGRAPHY (GC) the best procedure: specyfic, fast, multi-detectable, sensitive
•GC-MS, GC headspace – mostly use at present
•SPECTROMETRIC/ENZYMATIC ( applied ADH for estimation of ethanol) measurement with UV/VIS spectrometers
•IMMUNOENZYMATIC (FPIA, EMIT) ABBOT, VIVA
Laboratory tube with blood, urine, stomach contents...
ETHANOL +Oxidation agentor enzymes