Post-mortem Toxicology
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Transcript Post-mortem Toxicology
Post-mortem Toxicology
DR A J JEFFERY
MBChB MD FRCPath (Forensic) MFFLM
HOME OFFICE REGISTERED FORENSIC PATHOLOGIST
Areas to Cover
Why take toxicology
What samples
How to take them
What does the toxicologist do with the samples?
How to interpret the results – general considerations
Alcohol
Drugs of abuse
Toxicology in other causes of death
Other specimens
Case examples
WHY TAKE
TOXICOLOGY
?
Why take toxicology ?
To ascertain if the deceased was under the influence
of alcohol or drugs of abuse at the time of their
death.
RTAs / Accidental deaths / suicides
To confirm or refute overdose / poisoning
To confirm presence / levels of therapeutic drugs.
Eg epilepsy / antidepressants
WHAT SAMPLES
ARE
APPROPRIATE
?
Samples
Blood
Blood
Urine
Urine
(plain)
(preserved)
(plain)
(preserved)
(peripheral)
(peripheral)
Fluoride – inhibits further alcohol production but won’t undo the damage
already done.
Vitreous
Stomach Contents
Tissues
Liver
Skeletal muscle
(mid R lobe)
(eg psoas) (if embalmed buttock)
OBTAINING
THE
BLOOD SAMPLE
Femoral Vein Sampling
Vein NOT artery
Before evisceration
Before urine sampling
Ideal = tie off / clamp, then sample by wide-bore
needle below
Routine = clean catch
Ideal = don’t milk the leg
Routine = required to gain sufficient sample
Problem:
MINIMAL FEMORAL
BLOOD
Insufficient Femoral Blood
Take what ever you can in preserved tubes
Subclavian = reasonable alternative
Could take free-lying chest blood etc for screening
for the general presence of drugs
Make sure you say where each sample has come from
Obtain an alternative specimen
OBTAINING
THE
URINE SAMPLE
Urine Sampling
Needle and syringe
or
Open dome of bladder and aspirate with syringe
alone
Presence of a catheter may be important
toxicologically as the urine may contain artefactually
high lignocaine due to catheter lubricant gel
Vitreous
WHAT DO THE
TOXICOLOGISTS
DO WITH THE SAMPLE
?
Analysis
Screening (GC / Immunoassay)
What classes of drug are present
Confirmation (GCMS)
Specific drugs found by breaking them down & looking at the
breakdown products.
Quantification
Technique may vary dependent on the nature of the drug being
analysed.
HOW TO
INTERPRET
THE RESULTS
General Considerations
Accuracy of reference ranges
Re-distribution – site matters!
Individual variation (e.g. renal disease)
Decomposition
Tolerance
Accuracy of reference ranges
Interpretation of absolute drug levels / Reference
ranges
Based on individual reports
Variable from lab to lab due to varying techniques
Need to consider the previous list
General Considerations
Re-distribution – site matters!
Individual variation (renal disease)
Decomposition
Tolerance
Redistribution
Discovered with digoxin
Most drugs that undergo
redistribution do so
because of their relative
lipid solubility.
Due to
Loss of cell integrity
Diffusion
GI tract – to adjacent
structures
Through conduits – lymph
Diffusion from bladder
Natural Disease
Depends or route of administration
1st pass / second pass metabolism
Absorption
With or without meal
GI surgery
Elimination / Clearance
Renal impairment
Liver impairment
Decomposition
Significant redistribution
Some drug levels increase
Alcohol production by bacterial action
Others degrade
If there is a degree of decomposition make sure you
write it on the tox request
Tolerance
Increasing doses required over time to achieve same
effects.
What is lethal to a naïve user may have no effect at
all in a chronic user.
First dose deaths
People walking around and working with enough drugs on
board to kill an elephant!
Prison release deaths
Alcohol
Deaths due to Alcohol
What alcohol related causes of death do you know?
How might you classify them?
Which specific toxicological causes do you know?
Alcohol
Acute alcohol toxicity
Ketoacidosis
Alcohol in combination with
other drugs
Problems with Interpretation of Alcohol
Redistribution
Dealing with decomposition
Back calculations
Acute alcohol toxicity
How does it cause death?
>30 = higher skills
Death – respiratory
30 – 50 = deterioration in
depression due to action
on brainstem
UK legal driving limit?
Driving limit 80
mg/100ml
Less than 20 mg/100ml
generally considered
insignificant.
driving
50 – 100 = inhibitions /
laughter
100 – 150 = slurring,
insteadiness, poss nausea
150 – 200 = obvious
drunkenness, nausea
staggering
200 – 300 = stupor, vomiting,
coma
300 + = stupor, coma,
aspiration &
Alcohol – fatal level or not?
LD 50 = 400 mg/100ml
Alcohol has symbiotic relationship with other drugs.
e.g.
< 200mg/100ml can be fatal if opioids are taken.
> 200mg/100ml can ½ the fatal dose of opioids
> 100mg/100ml may enhance heroin toxicity
Ethyl glucuronide (minor breakdown product) in
urine if imbibed within 5 days of death.
What is ketoacidosis?
Can you explain why this happens?
Ketoacidosis
Brain can utilise ketone bodies when glucose is unavailable – fasting /
starvation
Ketone bodies, formed by the breakdown of fatty acids and the de-amination of
amino acids.
Ketoacidosis is an extreme and uncontrolled form of ketosis, which is a normal
response to prolonged fasting. In ketoacidosis, the body fails to adequately
regulate ketone production causing such a severe accumulation of keto acids
that the pH of the blood is substantially decreased.
Alcoholic ketoacidosis
Metabolic acidosis
Malnutrition
Binge drinking superimposed on chronic alcohol abuse
Ketoacidosis
Ketones:
Acetone (can be produced pm)
<0.5 mg/100ml
Beta hydroxybutyrate (less likely to be raised artefactually)
<0.5 mmol/L
1.26 – 47.2 mmol/L (assoc with fatalities)
Causes
Alcoholic ketoacidosis
Diabetic ketoacidosis
Alcoholic vs Diabetic
How might you differentiate?
Urine glucose
HbA1c
4 - 6.1%
Calculations
AVOID !
Clearance
10 – 25 mg/dl/hr ( about a unit an hour)
In 10 hours you can clear ~ 100-200 mg/dl
Alcoholics can clear 30 – 40 mg/dl/hr (due to training!)
Widmark equation
Used by some to predict amount of alcohol consumed
Decomposition
70 – 190 mg/100ml reported as artefact
Consider pm findings
Look for other substances produced pm
Use vitreous and urine as supportive evidence
These are relatively protected from redistribution
Normal ratios (if in equilibrium)
Urine : Blood
1.23 : 1
Vitreous : Blood
1 : 0.81
Drugs of Abuse
OPIOID AGONISTS
SYMPATHOMIMETICS
Opioid agonists
Opioid agonists
Analgesia / euphoria / dysphoria
Respiratory depression
miosis
Morphine and other opioids
Morphine
Heroin (diamorphine) – IV, smoked, sniffed
Methadone (green liquid – oral or IV)
Pethidine, buprenorphine
** CNS depression **
Findings
History / scene / paraphernalia
External
iv sites
Foam at nose / mouth
Limited & non specific
Pulmonary congestion and oedema
Stomach contents – methadone is usu green!
Morphine / Heroin
Heroin / diamorphine – synthetic morphine
derivative
Powerful opioid analgesic
Metabolised almost immediately (10 – 15 mins) to 6
monoacetyl morphine 6MAM and then within 24 hours to
morphine.
Presence of 6MAM is consistent with use within 12-24 hours
Ie recent intake / top up injections
Acute alcohol intoxication potentiates the effects
Total morphine : Free Morphine
Gives some idea of time since administration
Eg in IV admin
15 mins post admin
60 mins
4
9
Therapeutic
Free morphine 10 – 100ng/ml
:
:
1
1
Lethal_______
50 – 4000 ng/ml
Heroin
A contaminate of street heroin is acetylcodeine
Hence may have +ve codeine levels
Most heroin deaths occur several hours after taking
the drug
Sleepy / snoring
May have time to metabolise drug despite irreversible
respiratory depression
Methadone
Therapeutic
75 – 1100 ng/ml
Toxic
200 – 2000 ng/ml
Lethal
400 – 2000 ng/ml
Significant overlap
Tolerance becomes very important
Interpretation requires knowledge of drug history
Long & variable T ½
Methadone
Breakdown product – EDDP
This is inactive
Titration is important
Many deaths occur during first few weeks of treatment
Can cause respiratory depression at therapeutic doses
Lipophilic so undergoes significant redistribution
Even peripheral samples can be 2x in
and 3x in
Opioids
Tolerance = V V important consideration
Eg. Prison release
Palliative care
Nb worth remembering that 10% of codeine will
breakdown to become morphine.
Free Codeine
Therapeutic
Lethal_______
30 – 340ng/ml
>1600 ng/ml
Sympathomimetics
Sympathomimetics
Incr activity of adrenaline and serotonin
Adrenalin
Hypertension
Tachycardia
Mydriasis
Serotonin
Excitement
Hyperthermia
Stimulants
Cocaine
Amphetamine
Ecstasy
Other methamphetamines
Associated with subarchnoid haemorrhage
80% of these assoc with aneurysms
Intracerebral haemorrhage
Associated with AVMs & hypertension
Findings
Hearts of stimulant users tend to be heavier than
controls
Fibrosis
Contraction band necrosis
Accelerated atherosclerosis
Non specific pulmonary changes
Crack cocaine smokers – prominent anthracosis esp
in alveolar macrophages & emphysematous changes.
Cocaine
Naturally occurring plant alkaloid stimulant
Snorted, smoked, cutaneous, injected
Nb always consider in sudden death in the same way
that you might consider HOCM.
Breakdown Products
Benzoylecgonine
Inactive
Methylecgonine
Cocaethylene
As active as cocaine itself & indicative of alcohol
consumption at the same time
Cocaine Toxicity
Less than 50 ng/ml cocaine is considered not to
produce measurable physiological effects
T ½ can be as little as 40 mins
Benzoylecgonine – 1-4 days in urine
Toxic
>900 ng/ml
Lethal
>1000 ng/ml
Benzoylecgonine
Lethal - >1000 ng/ml *
Cocaine
But lethal nature not dose related
Long term effects:
Cardiovascular damage –
incr ischaemic event / Coronary Art thrombosis
coronary artery spasm
contraction band necrosis
fibrosis / sudden arrhytmia
myocarditis/cardiomyop/valvular/aortic
dissection/hypertensive crisis
Non cardiac -
Cerebral infarction / intracerebral haemorrhage
So death can be attributed to cocaine even if not found in blood.
Cocaine can decrease rapidly in unpreserved blood samples stored
at room temperature.
Cocaine, death & Excited delirium
Excited delirium
Hyperthermia
Mental & physiological arousal
Excited, erratic & sometimes bizarre, violent behaviour
May have florid psychosis
May exhibit extra-ordinary strength
Tends to result in sudden respiratory arrest
Blood cocaine and benzoylecgonine may be low but there is usually
concentration of benzoylecgonine within the brain indicating long
term use
Marked decease in D2 receptors in hypothalamus in psychotic
cocaine abusers.
D2 receptors play a role in temperature regulation
Amphetamine
Prevalence second only to cannabis
Synthetic stimulant
Effects similar to cocaine
Stimulate release of catecholamines, particularly
adrenaline
Tolerance & dependence develop
Absorbed by GIT, clinical effects commence within
20 minutes, last 4-6 hours.
Amphetamine
Toxic
>500 ng/ml amphetamine
>1800 ng/ml methamphetamine
Lethal
Usu > 1000 ng/ml amphetamine
But can be seen if >50ong/ml
Usu > 10 000 ng/ml methamphetamine
Different Forms
Amphetamine (Benzedrine, uppers, 'A', speed, whizz,
cranks, wake-up, sulph, hearts)
Dextroamphetamine (Dexedrine, dex, dexy, dexies)
Methamphetamine (ICE, crystal, glass, meth)
Methylenedioxyamphetamine (MDA, EVE)
3 ,4, Methylenedioxymethamphetamine (MDMA, ADAM,
Ecstasy, 'E', doves, Dennis)
Amphetamine
Alcohol can potentiate effects on the heart.
Rare toxic effects:
Coma
Cerebral vasculitis
Cerebral haemorrhage
Rhabdomyolysis
D.I.C.
Renal dysfunction
Cardiac – long term users
Accelerated coronary atherosclerosis
Microvascular disease
MDMA / Ecstasy
Amphetamine-like drugs
3,4-methylenedioxymethamphetamine
Serotinergic and noradrenergic effects
• Hyperthermic effects
Liquid ecstasy – GHB
Gamma hydroxybutyrate
Other
Benzodiazepines
Diazepam – 20 – 4000 ng/ml
Nordiazepam – 20 – 1800 ng/ml
Need in the order of thousands to consider fatal.
Cannabis
Cannabinoids
THC – tetrahydrocannabinol
Delta 9 THC carboxylic acid
Rapidly distributed into tissues
Blood levels drop >90% within 2 hours of intake
THC can only be found within 4-12 hours post intake
>2 ng/ml suggestive of recent intake
THC metabolites remain in
Blood – up to ~ 5 days
Urine – up to ~ 12-36 days
Volatile substance abuse - VSA
Adhesives, aerosols, petrols, paint stripper, nail varnish
remover ….. amongst others
Increased risk taking behaviour
Accidental suffocation
CNS depression
Deaths thought to be due to cardiac arrhythmias
Sensitisation of myocardium to effects of adrenaline
Deaths often seen in association with physical exertion
Blood sample must be in a glass tube with a foil top filled
to the top.
Tie off whole lung and place within a nylon bag.
Head space
‘New’ Drugs
Mephadrone
‘Cream’
Toxicology
IN
OTHER
CAUSES OF DEATH
Fire deaths
Carboxyhaemoglobin
Normal <10%
Toxic 15 – 35%
Lethal >48%
Cyanide
Normal < 0.1mg/L
Develops within the potted blood sample if not preserved
Carbon Monoxide
In an individual breathing air
T ½ = 4 hours
Breathing O2 in Hospital
T ½ = 60 minutes
Therefore always consider survival time.
Therapeutic Drugs
Anti-depressants
Anti-convulsants
Overdose
Aspirin
Therapeutic
Hyper ventilation
Resp
Alkalosis
Enhanced
BBB transfer
Met
Acidosis
Body
Compensates
Toxic
20 – 100mg/l
Stimulation
of Resp
Centres
>150 mg/l
Lethal
>500 mg/l
NB those on reg Rx (eg arthritis – 3g/day) – 44-
330mg/L
T ½ up to 36 hours in massive OD
Findings
Pm
Blood stained gastric content / frank haematemesis
Rarely skin petechiae
Mucosal gastric erosions
Malaena if survival sufficiently long
Petechiae through other organs due to anticoagulant effect
esp parietal pleura and epicardium
Paracetamol
Therapeutic
Toxic
10 – 20 mg/l
>150 mg/l
Lethal
>160 mg/l
Other samples of interest
IMMUNOLOGY – ANAPHYLAXIS
BIOCHEMISTRY – DIABETES
Immunology - Anaphylaxis
Secure ante-mortem samples
Needs to be peripheral as mast cell rich organs can
release tryptase after death.
Serum (plain tube – spun down)
Mast cell tryptase ( = more specific)
Specific IgE
Make sure you give details of any suspected cause
Available for venoms, foods, medicines, contrast agents, latex…….
Mast cell tryptase
T ½ during life is ~ 2 hours so may be unhelpful if they have been resuscitated
and have survived and die later ( usu from cerebral anoxia)
Antemortem!!!
Tryptase is a sensitive marker for mast cell activation
High levels will be found post severe anaphylaxis
Levels are not raised in local allergic reaction eg rhinitis
Can be raised in pure asthma deaths but not of the same order of magnitude
Slight increases can be seen in
non-anaphylactic mast cell degranulation – EG opioids for chest pain
Trauma – disruption of mast cell rich tissues
Unless grossly elevated – interpret with caution
In presence of suggestive history and absence of pm findings it may provide
confirmatory evidence.
Normal Levels:
IgE
MCT
MCT can be produced pm
0 – 122 kU/L
2-14 mg/L
Biochemistry - Diabetes
Vitreous is best for biochem as most blood is already haemolysed
Glucose drops significantly after death
Bacterial metabolism
Drops even in vitreous
So low glucose ≠ hypoglycaemia
It is not possible to diagnose hypoglycaemia accurately at pm.
A high vitreous glucose virtually rules out hypoglycaemia as one can assume it was
the same or higher in the antemortem period unless peri-mortem dextrose admin
HbA1c – heparinised sample
Insulin
If endogenous the body has to cleave C peptide from pro-insulin to make active
insulin.
If exogenous the insulin is already cleaved and so they will have no C peptide.
Case Examples
Case 1
60 yr old male, in house fire, extensive burns, no suspicious injuries, no
soot in airways or stomach.
Ranges
Norm
Tox
Leth
CO = 40%
<10%
15-35%
>50%
Cyanide = 0.5 mg/L
<0.1mg/L
Paracetamol
6mg/l
10-20mg/l
>150mg/l
>160mg/l
Codeine
56ng/ml
30-340ng/ml
>1600ng/ml
Morphine (free)
<5ng/ml
10-100ng/ml
50-4000ng/ml
Case 2
30 year old female found on floor with green fluid trailing
from corner of mouth
Ethanol
Blood
181 mg/100mls
Urine
244 mg/100ml
Ethylglucuronide present in urine
Acetone
negative
Methanol negative
Isopropanol negative
What effect might you expect?
What caveat would you include?
Is this likely to by PM alcohol production?
Ranges:
Normal
Toxic
Lethal
Methadone
75-1100ng/ml
413 ng/ml
EDDP = 276 ng/ml
200-2000ng/ml
400-2000ng/ml
Amphetamine
471 ng/ml
>500ng/ml
>1000ng/ml
Diazepam
21 ng/ml
Nordiaz = 73ng/ml
>5000ng/ml
>30 000ng/ml
Cause of Death??
20-4000ng/ml
20-1800ng/ml
Case 3
Chronic alcoholic found dead at home, head injury
consistent with fall to the ground.
Ethanol
16 mg/100mls
72 mg/100mls
25 mg/100mls
145 mg/100mls
Supports ante-mortem consumption
Acetone
Blood
Urine
Vitreous
Stom Cont
Blood
Urine
Vitreous
3mg/100mls
9mg/100mls
4mg/100mls
Methanol & Isopropanol
2mg/100mls of each in Blood, urine and vitreous
Ketone
But
Can be produced pm
What else do you want to know?
Urine Glucose – negative
Vitreous glucose – unrecordable
Urine Ketones – present
Blood Betahydroxybutyrate 2.3 mmol/L (<0.5 mmol/L)
more specific than acetone
HbA1c – 12.1% (4.0 – 6.1%)
Urine ethyl glucuronide - present
Diabetes – reconsider
urine alcohol
Case 4
Decomposed @ home on sofa with drug paraphenalia around
LIVER
Homogenate
Ethanol
Methanol
0.88 mg/g
Not detected
Not detected
Isopropanol
Not detected
Acetone
General drug screen by gas chromatography – mass spectrometry (GC-MS)
Liver homogenate:
Morphine, cocaine metabolites, flupenthixol and metabolites, paracetamol,
chlorpromazine metabolites and cotinine detected detected
Liver Tissue Homogenate Quantitative Analysis by Gas chromatography –
Mass spectrometry (GC-MS)
Cocaine
= <0.02 ug/g
Benzoylecgonine = 0.28 ug/g
Methylecgonine = 0.26 ug/g
Cocaethylene
= 0.42 ug/g
Morphine (free) = 1.05 ug/g
Morphine (total) = 1.50 ug/g
Was it worth doing if the concentrations mean
nothing?
COMMENTS
The external examination showed an advanced state of decomposition with no evidence
of injury to suggest involvement of another individual.
Within the limits imposed by the degradation of the tissues, the internal examination
showed no apparent pathology which could have caused or contributed to death.
Interpretation of toxicological results is complicated in cases where the individual has
been dead for some time. Drug levels are altered after death by diffusion of the
substances throughout the body (post-mortem redistribution) and certain substances are
produced or degraded in the post-mortem period. As such, it would be unreliable to draw
conclusions from the drug concentrations themselves. However cocaine and morphine
are not produced endogenously by the body and so their presence indicates that cocaine
and morphine (possibly heroin) were taken by the deceased. Alcohol can be produced by
the body after death but the presence of cocaethylene would suggest that alcohol and
cocaine were used concurrently.
The exact levels of the fore-mentioned drugs within the body at the time of death cannot
be determined with any accuracy. However, we feel that their presence, along with the
drug paraphernalia noted in the vicinity of the body and the absence of identifiable
natural disease is significant. Based on the above information, we are of the opinion that,
on the balance of probabilities, death was in keeping with polydrug toxicity.
The information given within this report represents our understanding of the views,
opinions and circumstances of this case based on the information that we have received
to date, either in writing (all forms) or by oral communication. We recognise that in
part this may reproduce or rely upon witness statements, oral communications or
hearsay evidence of second parties and that the information given to us by others may
or may not be factually correct at the time of our consideration.
We reserve the right to reconsider any aspect of this report should further factual
information arise that contradicts the information provided at the time of the postmortem examination, upon which we have based our interpretations.
Cause of Death
Ia. In keeping with polydrug toxicity
Confounding Factors
Considerations
Ask Yourself
Arterial vs venous variation
Where was it from?
Continuing gastric residue
absorption
Redistribution
Post-mortem production or
degradation
Tolerance
Limited reference range data
Vitreous / blood / urine
Site
Was it appropriately preserved
Is there decomposition?
Is it a drug prone to
redistribtution / pm production
Do you know about their drug
taking / drinking habits
In Practice
Seek a drug history from coroner’s officer
Always give the toxicologists as much info as you
have
If you are looking for a specific substance – tell them
as it might not be on their routine screen
Appropriate specimens / appropriate site
If tox is important talk to coroner’s officer about
accessing ante-mortem bloods.
Resources
http://www.dundee.ac.uk/forensicmedicine/
C. Baselt, Disposition of Toxic Drugs & Chemicals in
Man.