TOXICOLOGY
Rmt2023
CLINICAL CHEMISTRY 2 LECTURE: FINALS
TOXICOLOGY
study of the mechanism on how a toxin causes damage or
disease to the body; talks about toxic substances or poisons
⮚ study of the adverse effects of xenobiotics
(substances or drugs not produced in the body) in
humans.
⮚ Five Major Disciplines of Toxicology
1. Mechanistic Toxicology
2. Descriptive Toxicology
3. Regulatory Toxicology
4. Forensic Toxicology
5. Clinical Toxicology
MECHANISTIC TOXICOLOGY
⮚ cellular and biochemical effects of toxins
⮚ provide a basis for rational therapy design and the
development of tests to assess the degree of
exposure of poisoned individuals.
DESCRIPTIVE TOXICOLOGY
introduces toxin to an experimental animal to determine the
level of toxicity that can damage the animal → predict level
of toxicity that can harm to humans → risk assessment
⮚ uses the results from animal experiments to predict
what level of exposure will cause harm in humans
REGULATORY TOXICOLOGY
⮚ Involves the interpretation of the combined data
from mechanistic and descriptive studies
⮚ is used to establish standards that define the
level of exposure that will NOT POSE A RISK to
public health or safety.
Example: Food and Drug Administration (FDA) - tests
products in the market if it contain high amount of toxins (e.g.
mercury, lead) or chemicals that can cause risk to the public
health
FORENSIC TOXICOLOGY
performed for medico-legal purposes to know the cause of
death (specific toxin)
⮚ primarily concerned with the harmful consequences
of toxin exposure.
⮚ Focuses on establishing and validating the analytic
performance of the methods used to generate
evidence in legal situations, including the
___________________.
Example: Scene of the Crime and Operatives (SOCO)
CLINICAL TOXICOLOGY
performed in the laboratory (CC department)
⮚ the study of interrelationships between toxin
exposure (xenobiotics) and diseases
determine the cause of the signs and symptoms → identify
the toxin causing it
⮚ emphasizes not only diagnostic testing but also
therapeutic intervention.
once the cause is identified, therapy can also be determined
(antidotes binding to toxins to remove its toxic effects and will
be eliminated from the body)
ENVIRONMENTAL TOXICOLOGY
OUR LADY OF FATIMA UNIVERSITY I PAMPANGA CAMPUS I COLLEGE OF MEDICAL LABORATORY SCIENCE I CLINCHEMISTRY
studies pollutants to the water, soil and air
⮚ includes evaluation of environmental chemical
pollutants and their impact on human health
Example: Department of Environment and Natural
Resources (DENR)
EXPOSURE TO TOXINS
⮚ 50% - intentional ex: suicide attempt (intake of
poisonous substances)
⮚ 30% of cases - accidental exposure ex: negligence
(cleaning agent used for silver which is colorless and
odorless)
⮚ 20% - homicide or occupational exposure
(industrial worker in manufacturing paints or uses
heavy metals; farmers using chemical pesticides
and fertilizers)
ROUTES OF EXPOSURE
⮚ Toxins can enter the body via several routes:
● ingestion
● inhalation esp. if the toxin is in a gaseous or vapor
form
● transdermal meaning absorbed thru the skin
⮚ Mechanism of Absorption of toxins from the
gastrointestinal tract
✔ taken up by processes intended for dietary
nutrients passive diffusion - toxins, along
with the food is absorbed at the same time
bec. intestines cannot distinguish the
harmful substances from non harmful →
liver which is a metabolic organ that filters
substances absorbed from the intestine
before it reaches the general circulation →
first pass (everything absorbed in the
intestine must first pass through the kidney
for detoxification) so it is the first organ to be
affected by poisoning
✔ absorbed by intestine
TERMINOLOGIES
1. acute toxicity – single, short-term exposure ex.
accidental exposure
2. chronic toxicity – repeated exposure for extended
periods of time ex. occupational exposure (paint or
battery manufacturing company)
3. toxic dose 50 (TD50) – the dose that would be
predicted to produce a toxic response in 50% of the
population
4. lethal dose 50 (LD50) – the dose that would predict
death in 50% of the population
5. effectivity dose 50 (ED50) – the dose that would be
predicted to be effective or have therapeutic benefit
in 50% of the population
DOSE-RESPONSE RELATIONSHIP
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 ⮚ full stomach will have a 3 times slower
absorption of alcohol than an empty stomach

a. Ethanol (Grain Alcohol)

b. Methanol (Wood Alcohol)
c. Isopropanol (Rubbing Alcohol)
d. Ethylene glycol (1,2-ethabediol)

Among these substances, ethanol is the most abused,it is

used to make alcoholic beverages

a. ETHANOL (GRAIN ALCOHOL)

COMMON INDICATORS OF ETHANOL ABUSE
TEST COMMENTS

GGT gamma- -increases can be seen before

Categorize diff. chemicals using the toxicity rating → glutamyl transferase the onset of pathologic
depends on the amount that will show toxicity effects to an (GGT) → an indicator consequences
individual for chronic alcoholism → sensitive marker for
Dose is per kilogram not per individual or for occult alcoholism but the problem is
alcoholism increases in serum activity can
ANALYSIS OF TOXIC AGENTS occur in many non-ethanol-
⮚ Select the best specimen for selected test saliva, related conditions → meaning it
blood, urine, hair or nails (both for chronic exposure is non specific marker
since toxins can be deposited in these areas -
keratin, a protein that has an affinity for metals) so AST -increases in serum activity can
it is important to know what type of spx should be → a liver enzyme occur in many non-ethanol-
used to a specific type of toxin related-conditions
mercury - DO NOT use urine, due to a possibility → non specific marker
that this metal cannot be found in the urine
arsenic - could be missed if blood is used AST/ALT ratio -a ratio of greater than 2.0 is
⮚ specimen collection, handling, and storage highly specific for ethanol-
De Ritis Ratio→ ratio related liver disease →
TWO-STEP ANALYSIS OF TOXIC AGENTS ANALYSIS: of the AST/ALT alcoholic hepatitis
1. SCREENING TEST
highly sensitive test that can detect even in low HDL -high serum HDL is specific for
concentrations but lacks specificity and has cross reactivity ethanol consumption
so it is validated using confirmatory testing
rapid, simple, qualitative test (presence or absence) for toxic MCV -increased erythrocyte MCV is
substances commonly seen with excessive
⮚ Immunoassays, thin layer chromatography (TLC) ethanol consumption
-increases are not related to
2. CONFIRMATORY METHOD folate or vitamin B12 deficiency
⮚ GC-MS gas chromatography and mass
spectrometry ➢ also called as ethyl alcohol, beer alcohol
⮚ ICP-MS Inductively coupled plasma mass ➢ called grain alcohol since it is prepared thru
spectrometry fermentation of grains like wheat, rice (rice wine)
⮚ AAS Atomic absorption spectrometry ⮚ Ethanol abuse causes acidosis due to the
production of acetic acid – accumulation of ketones
TOXIC AGENTS and lactate
1. ALCOHOL ⮚ Causes diuresis → frequent urination/polyuria
⮚ commonly abused substance (ethanol suppress the action of the ADH)
⮚ CNS depressant ⮚ Hangover symptoms are due to the effect of the
⮚ disorientation, confusion, and euphoria, which by product of ethanol metabolism called
can progress to unconsciousness, paralysis, and, “acetaldehyde”
with high-level exposure,even death ⮚ 80 mg/dl of alcohol is the statutory limit for the
⮚ some of the alcohols can even cause blindness, operation of motor vehicle
and acidosis ⮚ 50 g/day of ethanol can lead to the accumulation
⮚ Symptoms of intoxication begin when the of lipids into the hepatocytes can lead to fatty liver
concentration of the alcohol to the blood is → alcoholic hepatitis → can progress to liver
> 0.05% w/v cancer
⮚ proof: pertains to the amount of alcohol that is
present in a solution HOW DOES THE ETHANOL CONVERTED INTO OUR
⮚ 80 proof alcohol: contains 40% of alcohol (divided BODY?
by 2) BIOTRANSFORMATION:
ETHANOL → converted to ACETALDEHYDE thru the
action of the enzyme alcohol dehydrogenase → converted

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 to ACETIC ACID thru the enzyme aldehyde ➔ more NADH produced → more
dehydrogenase alcohol is present to the sample
✔ GC/ gas chromatograpy: when combined
b. METHANOL (WOOD ALCOHOL) to the mass spectrometery = GOLD
⮚ extracted from the body of the tree by burning STANDARD
⮚ Commonly used solvent and a contaminant of ✔ Osmometry- osmolality inc. by about 10
homemade liquors specially for native beer mOsm/kg for each 60 mg/dl inc in serum
“lambanog” ethanol.
⮚ a very toxic substance ➔ when there’s an increase in the
⮚ some of symptoms is stomach pain → blindness ethanol level in the blood it will also
⮚ Fatal dose: 60-250 ml increase the osmolality in the blood

BIOTRANSFORMATION: ⮚ Detection Limit: 12 hours

METHANOL → converted to FORMALDEHYDE → ⮚ Fatal Dose: 300-400 ml of pure alcohol equivalent
converted to FORMIC ACID (can cause acidosis and it’s very to 1 and ½- 2 glasses of alcohol
poisonous; blindness) ⮚ Toxic Blood Level: 250-400 mg/dL

c. ISOPROPANOL (RUBBING ALCOHOL) CARBON MONOXIDE

⮚ usually masseur used isopropanol in massaging ⮚ odorless, colorless and tasteless gas
due to its high volatility (vaporizes quickly) → ⮚ Produced from incomplete combustion of
causes cooling effect carbon-containing substances
⮚ metabolized by hepatic ADH to acetone, which is its (cigarette,gasoline)
primary metabolic end product ⮚ also detected when a person slept in a car without
⮚ Intoxication with isopropanol, may result in an opening → filled with carbon monoxide
severe acute-phase ethanol-like symptoms that ⮚ Toxic effects are manifested by binding with
may persist for an extended period → meaning, heme proteins (has a high affinity for hemoglobin;
when ingested, it will have a longer half-life 200-225 times stronger compare to the affinity of the
⮚ Isopropyl alochol toxication is a rare occurrence oxygen to the hemoglobin)
because it promotes vomiting → will eliminate ⮚ inhibits cellular respiration and electron
alcohol transport
⮚ Fatal dose: 250ml or equivalent to 1 cup ⮚ Treatment: 100% oxygen therapy
⮚ Major Organ Affected: organs that have high
d. ETHYLENE GLYCOL (1,2-ethabediol)
demand for oxygen (heart, brain)
⮚ common component of hydraulic fluid and
⮚ Toxic Levels: 20%
antifreeze used for automobile cooling system
⮚ Indicator of Toxicity: Cherry Red Color of the
⮚ sweet taste
Face
⮚ Final product leads to deposition/accumulate of
⮚ Sample for Testing: EDTA whole blood
CaOx in renal tubules
⮚ Method for Testing:
➔ under the microscope CaOx has the
appearance of an “envelope” ✔ Spot Test→ 5ml of the 40% sodium
hydroxide is used and is added into
⮚ Fatal Dose: 100 grams
aqueous diluted whole blood
BIOTRANSFORMATION: ➔ (+): pink color (level of
ETHYLENE GLYCOL → converted to OXALIC ACID and carboxyhemoglobin is 20% or
GLYCOLIC ACID → greater)
✔ Differential Spectrophotometry
ETHANOL DETERMINATION ✔ Gas Chromatography
LABORATORY ANALYSIS ✔ Co-oximetry→ also measures
⮚ Specimen- serum, plasma and whole blood carboxyhemoglobin
⮚ container must be properly sealed or covered with
cap because the alcohol might evaporate → falsely CAUSTIC AGENTS
decreased level of alcohol corrosive substances can be acquired thru inhalation
⮚ Sealed specimens can be refrigerated or stored ⮚ Cyanide
at room temperature for up to 14 days without ⮚ Metals and Metalloids
loss of ethanol. ✔ Arsenic
⮚ Nonsterile specimens- should be preserved with ✔ Cadmium
sodium fluoride (best anticoagulant) ✔ Lead
⮚ Lab test – elevated GGT, AST, AST/ALT ratio, ✔ Mercury
HDL and MCV ⮚ Pesticides
⮚ Method: ✔ organic phosphates
✔ Enzymatic(uses non-human ADH which ✔ carbamates
oxidizes ethanol to acetaldehyde with
reduction of NAD to NADH @ 340 nm) CYANIDE
➔ this method is dependent to the ⮚ component of some insecticides
conversion of NAD to NADH ⮚ Expresses toxicity by binding to heme iron

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 ⮚ Inhibits cellular respiration, electron transport
and ATP formation
⮚ At low levels of exposure, patients experience
headaches, dizziness, and respiratory
depression, which can rapidly progress to
seizure, coma, and death at slightly greater
doses
⮚ has a bitter almond odor
⮚ Antidote: sodium thiosulfate
⮚ Toxic levels: more than 2 ug/ml
⮚ Laboratory Analysis:
✔ Ionspecific electrode methods and
photometric analysis following two-well
microdiffusion separation
✔ urinary thiocyanate concentration.
ARSENIC
⮚ odor of garlic and has a metal base
⮚ Component of ant poisons,rodenticides, paints
and metal alloys
⮚ common homicide and suicide agent
⮚ heavy metal poisoning
⮚ It expresses its toxicity by high affinity binding
to thiol groups in protein
⮚ high affinity binding to keratin → can be used in
chronic exposure
⮚ salvarsan→ used to treat syphilis; made up of
arenic
⮚ fish arsenic→ found among predatory fish (salmon,
mackerel)
⮚ Evaluating long-term exposure
✔ Specimen: hair and nail especially for
chronic exposure
-one of the clue for arsenic exposure is the
presence of the mees lines in nails (might
also indicate ascorbic acid def.)
✔ Ion Emission Spectroscopy
⮚ Evaluating short-term exposure
✔ Specimen:
● Blood (arsenic will only last in the
blood for few hours)
● urine (specimen of choice since
arsenic can persist up to 6 days)
✔ AAS
⮚ Acute Fatal Dosage: 120 mg
⮚ Other Method for Testing:
✔ Reinsch test → not only specific for arsenic
CADMIUM
⮚ use in electroplating and galvanizing
⮚ also present in tobacco
⮚ pigment found in paints and plastics and is the
cathodal material of nickelcadmium batteries
⮚ expresses its toxicity primarily by binding to
proteins
⮚ Toxicity may also result to destruction of type 1
epithelial cells in the lungs (collapse of the lungs)
and decreased resistance to bacterial infection
⮚ also targets the kidney→ renal tubular defect
(RTD), glucosuria, aminoaciduria
⮚ has a half-life of 10-30 years in our body
⮚ Laboratory Analysis:
✔ Whole blood or urine- AAS
LEAD
OUR LADY OF FATIMA UNIVERSITY I PAMPANGA CAMPUS I COLLEGE OF MEDICAL LABORATORY SCIENCE I CLINCHEMISTRY
⮚ common environmental contaminant
⮚ A potent enzyme inhibitor
⮚ most distinctive feature: can decrease the IQ
points especially in the children
⮚ Low level exposure may cause behavioral
changes- hyperactivity and attention deficit
disorder/ADHD/PDD and also affects IQ because
the toxic effect of the lead may cause ??? of the
peripheral nerves → causes decreased in nerve
conduction demyelination
⮚ lead is also used in paints and also in children toys
⮚ Vitamin D and heme synthesis are affected – lead
blocks D-ALA synthetase, producing anemia →
lead poisoning can cause microcytic hypochromic
anemia
⮚ to the peripheral blood (PBS), basophilic stippling
can be seen
⮚ It has characteristic “wrist drop or foot drop”
manifestation → lead affects nerve conduction
⮚ Treatment: EDTA and Dimercaptosuccinic acid
⮚ Toxicity Dose: > 0.5 mg/day
⮚ Fatal Dose: 0.5 g
⮚ Toxic Blood Levels: >70 ug/dl (definitive)
⮚ Samples: whole blood (used for quantitative
testing exact value of the lead), urine (used for
recent/acute lead exposure) and hair (used for
chronic exposure)
⮚ half-life of > 20 years in the hard tissue
⮚ half-life of 120 days in the soft tissue
EFFECTS OF LEAD
⮚ Acute Exposure- Abdominal or neurological
symptoms manifest.
⮚ Neurologic symptoms- encephalopathy
characterized by a cerebral edema and ischemia.
⮚ Severe lead poisoning can result in stupor,
convulsions, and coma.
⮚ CDC cut-off for normal level of lead in children:
less than 10 ug/dl
⮚ FDA is very important in checking different products
for their lead conten
INDICATORS OF LEAD
⮚ Urinary D-ALA
⮚ Free RBC porphyrin
⮚ Presence of Basophilic Stippling in RBC
TESTS
⮚ Graphite Furnace AAS
⮚ Inductively Coupled Plasma Emission
Spectrophotometry (ICPS)
⮚ Anodic Stripping Voltammetry
⮚ Zinc protoporphyrin or free RBC protoporphyrin test
MERCURY
⮚ can be seen in sphygmomanometer,
thermometer
⮚ single drop of mercury can poison the whole room
⮚ Binds with protein and also an enzyme inhibitor
⮚ Exists in three forms:
✔ Elemental (liquid)
✔ Inorganic salts
✔ Component of organic compounds
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 ⮚ Inorganic mercury- tachycardia, tremors,
Thyroiditis, and, most significant, a disruption of
renal function
⮚ Organic mercury- neurologic symptoms
✔ Low levels of exposure cause tremors,
behavioral changes, mumbling speech,
and loss of balance. Higher levels of
exposure result in hyporeflexia,
hypotension, bradycardia, renal
dysfunction, and death
⮚ Sample for Testing:
✔ Whole blood (organic mercury) – AAS
✔ Urine (inorganic mercury) - anodal
stripping voltammetry
⮚ Method for Testing:
✔ Reinsch Test
⮚ Significant Exposure: > 50 ug/dl (whole blood)

PESTICIDES
⮚ Organophosphates and carbamates function by
inhibition of acetylcholinesterase
(neurotransmitters which are important in signaling
between nerve endings)
⮚ Low levels of exposure are associated with
salivation, lacrimation, and involuntary urination
and defecation
⮚ Higher levels of exposure result in bradycardia,
muscular twitching, cramps, apathy, slurred
speech, and behavioral changes.
⮚ Death due to respiratory failure may also occur.
⮚ Pesticide exposure usually happen to farmers called
occupational exposure)

METHODS
⮚ Evaluation of erythrocytic acetylcholinesterase
activity
⮚ measurement of serum pseudocholinesterase
⮚ cholinesterase and pseudocholinesterase
are markers pesticide and insecticide poisoning
where their concentrations decreases

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