Principles of Environmental Toxicology

Learning Objectives
Explain the role of biotransformation in toxicokinetics.
Describe how biotransformation facilitates elimination
of toxicants.
Biotransformation and Elimination Distinguish between Phase I
of Toxicants and Phase II reactions.
Define bioactivation
Principles of Environmental Toxicology or toxication.
Instructor: Gregory Mller, Ph.D.
University of Idaho

Principles of Environmental Toxicology Principles of Environmental Toxicology

Learning Objectives, 2 Metabolism

Identify tissues and factors Sum of biochemical rxns occurring to a
involved in biotransformation. molecule within the body.
Anabolism - build-up
Summarize the role of elimination in
Catabolism - break-down
toxicokinetics.
Occurs in the cytoplasm or
Describe processes occurring in at specific organelles within
the kidney, liver and lung the cell.
related to the elimination Storage affects the bodys
of toxicants. ability to biotransform and
eliminate.
Bone, lipid.

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Principles of Environmental Toxicology Principles of Environmental Toxicology

Biotransformation Biotransformation Reactions

Process that changes substances from hydrophobic Grouped as Phase I (functional group modification)
to hydrophilic to aid in elimination (grease to salt). and Phase II (conjugation).
Hydrophilic molecules are less able to cross cellular Goals
membranes, hence fluid filterable (kidneys). Produce water soluble metabolites.
Major elimination routes are Activate natural/endogenous compounds
feces (biliary) and urine. for normal function.
Biological half-life, T
allows comparison of
Some compounds undergo
rates of removal. bioactivation.
The biotransformed metabolite
is more toxic than the original
compound.

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 Principles of Environmental Toxicology Principles of Environmental Toxicology

Results of Biotransformation Major Categories/Reactions

Increase toxicity via a toxic metabolite.
Phase I
Decrease toxicity via metabolism of a toxic
parent compound. oxidation
reduction
No effect on toxicity. hydrolysis
Present to metabolize
endogenous compounds.
polar Phase II
conjugation
synthesis
very
polar
Elimination
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Principles of Environmental Toxicology Principles of Environmental Toxicology

Enzymes of Biotransformation Phase I Reactions

R NHOH
Phase I Enzymes N-oxidation
R NH2
Hughes
Oxidation (most important). R1 R1
Add O, remove H, increase valence. S S O
S-oxidation
Cytochrome P-450, MFO, alcohol dehydrogenase, R2 R2
oxidases, others. O OH
Carbonyl reduction R1 C R1 CH
Reduction (less important).
R2 R2
Remove O, add H, decrease valence. O
R2
Reductases. Ester Hydrolysis R1 C-O R2 R1 CH2OOH + HO

Hydrolysis. R1 R1
Add water. Desulfuration C S C O
Esterases, phosphtases, others. R2 R2

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Dehydrogenation RCH2OH RCHO

Principles of Environmental Toxicology Principles of Environmental Toxicology

Enzymes of Biotransformation, 2 PII Cofactors: GSH

Phase II Enzymes NH2 O
Conjugation reactions. H
HO N O
Enzymes (tranferases) + cofactor.
Enzyme catalyzes. N
Cofactor donates group. O O H OH
Glucuronic acid, glutathione, sulfate, HS
acetyl group, methyl group.
Tends to increase
molecular size and
Glutathione
polarity for excretion.

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 Principles of Environmental Toxicology Principles of Environmental Toxicology

PII Cofactors: Acetyl-CoA PII Cofactors: PAPS

O O NH2
HO OH
P P
N 3-Phosphoadenosine
O
O O Acetyl Coenzyme A N
O O N
5-phosphosulfate
O N N
HO NH O N O O N
P
OHHO
HO
N
NH2 O
O S O P O
O OH OH
NH
O OH
O P OH
S
OH
13 O 14

Principles of Environmental Toxicology Principles of Environmental Toxicology

PII Cofactors: UDPGA Benzene Metabolism

H O
HO H UDP-GT Glucuronide
HO OH
Uridine-5-
Phenol
diphosphoglucuronic acid H N ST
O OSO3
H PAPS
O N O
H2C H
OH
O P OH P450 Epoxide
HO O
O Epoxidation Hydratase
P OH
H O Toxic Epoxide Dihydrodiol
HO H OH O
H GSH
HO GST
O
H OH
H O
-
O
15 16 Glutathione

Principles of Environmental Toxicology Principles of Environmental Toxicology

Aniline De-Alkylation
Phase II
H
NH2 N
P450 OH Phase II O
P450 H
N N + HC
Amine N-hydroxylation
Dimethyl-propyl-amine Methyl-propyl-amine Acetaldehyde

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 Principles of Environmental Toxicology Principles of Environmental Toxicology

Free Radical Generation Case Study: Fluorocitrate and Kangaroos

Fluorocitrate found in legume
Cl NADH
P450 Cl pasture plants
Cl C Cl Cl C GSH of Western Australia.
Reducatase Gastrolobium and Oxylobium.
Cl Cl
Highly lethal (TD 1 mg/1080 kg).
Tetrachloro-methane Toxic Free Radical WACALM
Leaf concentrations can be 2.6 g/kg.
The rat and gray kangaroo
of Western Australia have
evolved resistance.
In vivo defluorination w/ glutathione.
Other kangaroos from areas
w/o these plants are not tolerant.

Harborne
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Rodenticide: Fluoroacetic Acid Fluorocitrate Metabolite

CoASH
O H O AcCoA O OH
O

F C COSCoA O OH
F HO HO O
OH H O OH FAcCoA F
HO
Fluoroacetate Fluoroacetyl CoA

Sodium Fluoroacetate
Compound 1080
rodenticide
predator control
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Krebs Cycle Deoxynivalenol, Vomitoxin

(Fluoro)Citrate

O O
AcCoA O OH
OH OH
O OH O O O O
HO HO O
O OH FAcCoA F
HO Mitochondrial
Oxaloacetate energy production HO HO
HO
HO O CH2
Aconitase H2 O

Fusarium trichothecene
O
O mycotoxin found on
OH corn and barley
HO O

23 Cis-aconitate HO 24

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 Principles of Environmental Toxicology Principles of Environmental Toxicology

Aflatoxin B1 Benzo[a]pyrene
H O O H O O

O O OH
OR
H H
O O

O O R = sulphate or glucuronic acid

O O
Q1 = hepatic metabolite
B1 Polycyclic aromatic
hydrocarbon.
Aspergillus mycotoxin Environmental carcinogen.
found on corn, peanuts Cell cultures from rodents,
and cottonseed fish and humans
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Heavy Metal Toxicity - Pb Heavy Metal Toxicity - Pb, 2

Absorbed via Ca channels as divalent ion. Sensitivity of a system and degree of
Capable of reacting with a variety of binding sites. interference determines clinical effects.
Protein precipitation. Digestion/respiration absorption.
Liver detoxication.
Specific toxic effect depends on rxns with ligands
Kidney excretion.
that are essential for the living system.
Antidotes are competing ligands.
Metal ligands are formed with
sulfhydryl groups, as well as O O
OH
amino, phosphate, imidazole, HO N
and hydroxyl groups of enzymes EDTA
and essential proteins. N O
HO
O OH
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Heavy Metal Toxicity - Pb, 3 Heavy Metal Toxicity - Pb, 4

Metallic lead absorbed most efficiently Some endpoints. SH

by the respiratory tract. Sulfhydral enzyme inhibition. HO CH SH

C C
10% of ingested lead is absorbed. K transport in RBC inhibited H2 H2
Anemia.
Small intestine.
Porphyrinuria. 2,3-Dimercapto-propan-1-ol
Lead salts are soluble in gastric juices; absorbed.
Excreted chiefly in
Plasma to blood cells erythrocytes.
feces and urine.
After oral ingestion:
Chelating agents:
60% bone (also hair, teeth).
Ca - EDTA.
25% liver (hepatocytes).
Penicillamine.
4% kidney (renal tubules).
Dimercaptrol (BAL).
3% intestinal wall.
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Case Study: Elevated PbB Associated with
Illicitly Distilled Alcohol, Alabama 1991 Case Study: Moonshine Lead Toxicity
The use of automobile radiators containing Seven patients required hospitalization for
48 hours or longer (range: 2-18 days). Three
lead-soldered parts in the illicit distillation of of these received chelation therapy; initial
alcohol (i.e., "moonshine") is an important BLLs were 67, 228, and 259 ug/dL. One
source of lead poisoning among persons in patient, whose BLL was 67 ug/dL, died
some rural Alabama counties. during hospitalization from alcohol-
In 1991, eight persons were diagnosed with withdrawal syndrome complicated by
elevated blood lead levels (BLLs) at a local aspiration pneumonia.
hospital. Patients reported moonshine ingestion
ranging from 0.2 L per day to 1.5 L per day.
9 patients had been evaluated for alcohol-
The lead contents of specimens of
related medical conditions at the hospital. moonshine confiscated from two radiator-
Manifestations included generalized tonic-clonic containing stills in the county in 1991 were
seizures (six), microcytic anemia (five) 7400 ug/L and 9700 ug/L, compared with
(hematocrit mean: 32.1%), encephalopathy nondetectable amounts (less than 1.0 ug/L)
(two), upper extremity weakness (one), and in municipal water from the county.
abdominal colic (one). BLLs ranged from 16 Consumption of 0.5 L per day of moonshine
ug/dL to 259 ug/dL (median: 67 ug/dL). containing 9700 ug/L lead would result in a
steady state BLL of approximately 190
ug/dL.

31 MMWR (1992) 41(17);294-295 32

Principles of Environmental Toxicology Principles of Environmental Toxicology

Elimination of Toxicants Kidney

Urinary.
Fecal.
Respiratory.
Other:
Saliva.
Sweat.
Milk (transfer to child).
Nails, Hair, Skin.
Cerebrospinal fluid.

Hughes
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Renal Macrostructure Renal Filtration Microstructure

Renal cortex Renal medula

Ureter

Bovine

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Renal Histology Urinary Excretion

Glomerular filtration
Tubules Tubular secretion
Tubular reabsorption

Glomerulus
Microscopic
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Fecal Excretion Exhaled Air

NLM
Excretion in bile to intestine. Gas phase xenobiotics.
Active transport of toxicant parent and metabolites. Passive diffusion from blood
Highly soluble Phase II metabolites (large, ionized) to alveolus via concentration
Excretion into the lumen of the GI tract. gradient.
Direct diffusion from capillaries. The total alveolar epithelial Gray's Anatomy 1918

surface area within an average

adult human lung has been
estimated to be as large as
100-140 m2.

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