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Post Mortem Forensic Toxicology. Jeffery Hackett MSc PhD CSci CChem MRSC jhackett@unitedchem.com. What is it?.

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Post mortem forensic toxicology l.jpg

Post Mortem Forensic Toxicology

Jeffery Hackett

MSc PhD CSci CChem MRSC

jhackett@unitedchem.com

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What is it l.jpg
What is it?

Post-Mortem Forensic Toxicology: determines the absence or presence of drugs and their metabolites, chemicals such as ethanol and other volatile substances, carbon monoxide and other gases, metals, and other toxic chemicals in human fluids and tissues, and evaluates their role as a determinant or contributory factor in the cause and manner of death

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Where do I find out about it?

Journals:

Forensic Science International

J.Forensic Science

J. Anal. Toxicology

J.Chromatogr. B (and A)

Books:

R.C. Baselt: Disposition of Toxic Drugs and Chemicals in Man

Clarke’s Analysis of Drugs and Poisons

J.Garriot: Medico-legal Aspects of Alcohol

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Interesting cases

  • Dr. Crippen: Hyocine

  • Georgi Markov: Ricin

  • Justin Devilliers: Fentanyl

  • Ivan Litvanenko: Polonium

  • Mrs Cahill: Cyanide

  • Stacy Castor: Ethylene Glycol

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What’s it all really about

1. How drugs get into people

2. How we get the drugs out

3. How drugs are tested for

4. What does it all mean??

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1 how do drugs get into people l.jpg
1. How do drugs get into people??

Intoxicants: Ethanol (Other Alcohols)

Oral administration

Butane (Other gases)

Toluene(also Solvents)

Via Inhalation

Marilyn Monroe?

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Illicit Drugs:

Cannabinoids: Smoked

Amphetamines:Oral

Cocaine/ Heroin :Smoked, Snorted, Injected

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Prescription Drugs:

Acetaminophen

Propoxyphene

Fentanyl

Oxycodone

Zolpidem

Temazepam

Methadone

Mainly oral administration, some have been abused in other ways

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Weird and Wonderful

Mephedrone

MCat

Spice (JWH-018)

Salvia Divinorum

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Route of Drugs:Oral administration

A.D.M.E.

Drugs will leave from the stomach

Pass through the Liver

Enter into Blood stream/ CSF

Leave through Liver/Kidneys

Pass into Urine

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What does this mean for sampling?

Bloodis always important (Site?)

Liverwill sequester higher levels of drugs

Lungs are excellent for solvents

Urine is the last point for drugs

Hair has problems all of its own

Vitreous Humor: Alcohol Data

Brain??: Oral Fluid

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THC in Blood (LC-MSMS)1 ng per mL

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2. Getting the drugs out:

Volatile/Semivolatile: Headspace

Acid drugs : Solvent extraction

+

Basic drugs : Solid phase (micro) extraction

Metals/Metalloids: Digestion

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Headspace analysis l.jpg
Headspace Analysis:

  • Samples are warmed to create an atmosphere

  • Aliquots are taken of the vapour

  • This is free of contamination/ interferences

    Low boiling:Propane

    Medium : Ethanol

    High : Toluene

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Solvent extraction l.jpg
Solvent extraction:

pH Modification

Addition of acid/ base

COOH COO- + H+

H+ + R-COO- COOH

Neutral Species are more soluble in organic solvents

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Solid phase extraction:

Samples are pH modified

Filtered through porous sorbents

Drugs are collected onto modified surfaces

Interferences are washed off

Compounds of interest are eluted off

**pH= pKa +2**

pH=pKa +log [ ionised/unionised]

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Digestion l.jpg
Digestion:

Dry digestion

Samples are: heated, ignited and ashed

Residue is dissolved in dilute acid

Good for Copper, Lead, Zinc etc

Wet digestion

Samples are heated in strong acid solution

Solution is reduced in volume, neutralized and diluted

Good for volatile metals: Mercury

Thallium

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Drug Testing:

Immunoassay:

ELISA/EMIT/FPIA/RIA

Chromatography:

Gas Chromatography (FID, ECD, MS)

Liquid Chromatography (PDA, FLD, MS/MS)

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Method:

*Immunoassays are Presumptive Tests

*All operators are competency trained.

Calibrators are matrix matched

Set at Negative, Cut off and Positive values

Good for Blood, Urine and Tissue

Wide range of analytes available

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Analyte

Negative

Cut-Off

Positive

Amphetamine

2.816

1.023

0.181

0.992

2.562

Barbiturate

1.609

0.504

0.196

1.266

1.555

Benzodiazepine

2.151

0.413

0.113

2.136

0.155

Cannabinoids

1.238

0.658

0.398

1.326

0.497

Cocaine

1.070

0.739

0.430

1.045

0.114

Methadone

1.240

0.668

0.156

1.426

1.529

Methamphetamine

2.170

0.799

0.310

1.658

2.069

Opiate

1.393

0.331

0.145

1.504

0.104

Oxycodone

1.704

1.266

0.488

1.612

1.039

Phencylcidine

2.195

1.210

0.793

2.237

2.332

ELISA Analysis:

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Gas-ChromatographyInstrumentation

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Gas-Chromatography Mass Spectrometry

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Liquid ChromatographyInstrumentation

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Example of liquid chromatography xanthines theophylline theobrmine caffeine 8 chlorotheophylline l.jpg
Example of Liquid Chromatography: (xanthines: Theophylline/ Theobrmine/ Caffeine/ 8-Chlorotheophylline)

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Different Detectors (LC) Theobrmine/ Caffeine/ 8-Chlorotheophylline)UV and Fluorescence

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LC-MSMS Theobrmine/ Caffeine/ 8-Chlorotheophylline)

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Chromatographic Methods Theobrmine/ Caffeine/ 8-Chlorotheophylline)

  • Use Validated/Peer Reviewed ones

  • Internal Standards (IS) must match analytes

  • Best IS are: deuterated (MS)

    : Substituted Analogues (FID,NPD)

    : Non-drug materials

    extraction involves : Volatile Solvents (GC)

    : Miscible Solvents (LC)

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Chromatographic Analysis Theobrmine/ Caffeine/ 8-Chlorotheophylline)

Involves:

Calibration curves with known standards and controls

Derivitization with recognized modifiers (BSTFA, PFPA)

Recognized detection methods

(SIM, MRM, PDA)

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Calibration Curve: Theobrmine/ Caffeine/ 8-Chlorotheophylline)THC and Metabolite

THC (0.25 to 50 ng/ mL)

Carboxy-THC (0.25 to 50 ng/ mL)

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4. What does it all mean: Theobrmine/ Caffeine/ 8-Chlorotheophylline) Toxicology Report

RESULTS:

Blood (Iliac):

Ethyl Alcohol – Not Detected5

Benzoylecgonine – 0.05 mg/L1,2,4

Sertraline – 0.11 mg/L2,4

Desmethylsertraline – 0.01 mg/L2,4

Quetiapine – 0.50 mg/L

Negative for Acetaminophen, Amitriptyline, Barbiturates, Chlordiazepoxide, Citalopram, Clomipramine, Cocaethylene, Cocaine, Desipramine, Diazepam, Doxylamine, Ibuprofen, Imipramine, Meperidine, Methadone, Methamphetamine, N-desmethyltramadol, Norpropoxyphene, Pentazocine, Phenytoin, Procainamide, Procaine, Strychnine, Thioridazine, Tramadol, Trazodone, Trimipramine, Venlafaxine, and Verapamil.2,4

For example use only………Not a real report.

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Interpretation of Drugs: Theobrmine/ Caffeine/ 8-Chlorotheophylline)

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Psilocybin/ Psilocin Question Theobrmine/ Caffeine/ 8-Chlorotheophylline)

PB IS PC

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Interpretation of Alcohol: Theobrmine/ Caffeine/ 8-Chlorotheophylline)Clinical Values

BAC/g% Effect

0.02 Relaxed

0.05 Tranquil

0.10 Coordination problems 0.20 Intoxication Obvious

0.30 Passing Out

0.40 Comatose

0.50 Death

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Post Mortem Alcohol: Theobrmine/ Caffeine/ 8-Chlorotheophylline)A Point of View

Widmark:

BAC= 100* A

Wt*Wf

Back-Extrapolation:

*Post Absorptive Period

Linear Elimination

No Alcohol Consumption

**Should not be applied because of the number variables**

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New Issue (EtG/ EtS) Theobrmine/ Caffeine/ 8-Chlorotheophylline)

Ethylglucuronide molecule

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My Thanks: Theobrmine/ Caffeine/ 8-Chlorotheophylline)

Albert Elian

Bob Osiwiecz (Erie Co. Med Examiner Lab)

J. Spencer

Mike Telepchak

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