Chapter 5
Absorption, Distribution,
Metabolism, and Elimination of
Toxics
Biotransformation: Basic Concepts (1)
• Renal excretion of chemicals
Biotransformation: Basic Concepts (2)
• Biological basis for xenobiotic metabolism:
– To convert lipid-soluble, non-polar, non-excretable forms
of chemicals to water-soluble, polar forms that are
excretable in bile and urine.
– The transformation process may take place as a result of
the interaction of the toxic substance with enzymes found
primarily in the cell endoplasmic reticulum, cytoplasm,
and mitochondria.
– The liver is the primary organ where biotransformation
occurs.
Biotransformation: Basic Concepts (3)
Biotransformation: Basic Concepts (4)
• Interaction with these enzymes may change the toxicant to
either a less or a more toxic form.
• Generally, biotransformation occurs in two phases.
– Phase I involves catabolic reactions that break down the toxicant
into various components.
• Catabolic reactions include oxidation, reduction, and hydrolysis.
– Oxidation occurs when a molecule combines with oxygen, loses hydrogen,
or loses one or more electrons.
– Reduction occurs when a molecule combines with hydrogen, loses oxygen,
or gains one or more electrons.
– Hydrolysis is the process in which a chemical compound is split into
smaller molecules by reacting with water.
• In most cases these reactions make the chemical less toxic, more
water soluble, and easier to excrete.
Biotransformation: Basic Concepts (5)
– Phase II reactions involves the binding of molecules to
either the original toxic molecule or the toxic molecule
metabolite derived from the Phase I reactions. The final
product is usually water soluble and, therefore, easier to
excrete from the body.
• Phase II reactions include glucuronidation, sulfation, acetylation,
methylation, conjugation with glutathione, and conjugation with
amino acids (such as glycine, taurine, and glutamic acid).
– It is important to understand that these Phase I and II
reactions may occur simultaneously or sequentially.
Biotransformation: Basic Concepts (6)
• Whether the more toxic intermediate produces
an effect is dependent on
– how rapidly the intermediate undergoes further
metabolism to less toxic substances,
– how much of it is produced and accumulated in
cells,
– what type of cellular damage is caused by the toxic
intermediate, and
– what factors may affect excretion of the toxic
material.
Example : Result of Oxidation of Benzene
Phase I Reaction (1)
• Exposing a functional group on the starting
compound
Phase I Reaction (2)
• Sequential metabolism of benzene
– Adding a functional group
Phase I Phase II
Phase I Reaction (3)
• Oxidation
– Many toxicants are metabolized by the enzymes
cytochrome P-450 reductase and cytochrome P-450 in
association with NADPH (nicotinamide adenine
dinucleotide phosphate).
• NADPH is a co-enzyme present in most cells; it interacts with
various substances during normal cell metabolism.
– These two enzymes are found in abundance in the
endoplasmic reticulum of liver cells.
– When these enzymes interact with the toxic molecule, one
atom of oxygen is attached to the toxic molecule and
another oxygen atom interacts with hydrogen to form water.
Phase I Reaction (4)
• Cytochrome P450 Characteristics
– can metabolize many xenobiotics (broad substrate
specificity).
– can catalyze many types of reactions
– is widely distributed among tissues, and tissue
distribution can be quite varied.
– exists in multiple forms (determined by different
genes)
– levels can be increased by exposure to chemicals in
the food, water, or air (induction)
Phase II Reaction (1)
• During Phase II reactions the toxic substance or its
metabolite is bound with a compound that generally
renders the newly synthesized molecule less toxic. This type
of reaction is referred to as conjugation.
• Conjugation is the joining of two substances to form s single
molecule, which can increase water solubility and excretion.
Phase II Reaction (2)
• There are several types of Phase II reactions, of which
glucuronidation is probably the most important.
• Conjugation with glucuronidation
– Glucuronidation occurs as a result of the conjugation of
glucuronidation acid with either a metabolite from Phase I reactions
or with the parent compound.
– There are several chemical compounds that interact with glucuronic
acid:
• alcohols (R-OH)
• carboxylic acids (R-COOH)
• sulfhydryl compounds (R-SH)
• amines (R-NH)
Phase II Reaction (3)
• Conjugation with glutathione
– Conjugation with glutathione is an important Phase II reaction
that renders highly toxic metabolites harmless.
– Conjugation of these intermediates with glutathione prevents
binding with the nucleic acids, therefore preventing the
occurrence of mutations.
• Metabolites of organic solvents such as benzene, chloroform, and
carbon tetrachloride are conjugated with glutathione resulting in
decreased toxicity.
Phase II Reaction (4)
• Illustation of glucuronidation
– I
Phase II Reaction (5)
• Sulfation
Phase II Reaction (6)
• Acetylation and methylation
– Acetylation and methylation may increase toxicity by
decreasing water solubility and, therefore, excretion.
– Methylation plays only a minor role in the transformation of toxic
substances.
• Methylation of nicotine. A methyl group is added from SAM (S-adenosyl
methione) to nicotine.
Deactivation versus Bioactivation (1)
• In general, Phase I and Phase II reactions are
designed to deactivate toxic substances by
increasing water solubility and excretion of toxic
substances. However, these reactions may also
bioactivate some toxicants by transforming
inactive non-toxic molecules to active toxic forms.
Deactivation versus Bioactivation (2)
• Four potential end results of any chemical undergoing
metabolism.
Deactivation versus Bioactivation (3)
Deactivation versus Bioactivation (4)
• Metabolism of benzo(a)pyrene
(BP)
Cancer-causing
agents
Deactivation versus Bioactivation (5)
• Metabolism of benzene in the liver by cytochrome P450
enzymes.
Deactivation versus Bioactivation (6)
• Metabolism of dimethylnitrosamine.
carcinogenic
Deactivation versus Bioactivation (7)
• Metabolic pathway for the bioactivation of bromobenzene
Factors Affecting Metabolism (1)
• The rate at which metabolism of toxic substances occurs is
dependent on a variety of factors that can be categorized
into two groups:
– those factors that affect the metabolic processes directly, and
– Those factors that affect the transport of toxic substance to tissues
where metabolism occurs.
• Biotransformation is affected by the species of the test
animal, age, sex, nutritional status, disease, enzyme
induction or inhibition, and genetics.
– Newborn babies and young infants are more susceptible to a
variety of chemicals such as pesticides because the cytochrome
p450 enzymes --- important in pesticide detoxification reaction ---
are not well developed.
Factors Affecting Metabolism (2)
• A balanced diet will provide the necessary protein as well
as essential metals and minerals such as copper, zinc, and
calcium to assist normal cellular enzymatic activities
associated with biotransformation.
• Protein-deficient diets can result in a decrease in protein
synthesis, thus affecting the synthesis of enzymes involved
in the metabolic reactions used in detoxification.
Factors Affecting Metabolism (3)
• Cirrhosis of the liver is often caused by excessive drinking
of alcohol. During the disease process the liver cells are
damaged and replaced by connective tissue. If enough
cells are killed, the ability of the liver to metabolize toxic
substances is dramatically reduced.
• Cirrhosis cal also be caused by repeated exposure to
arsenic or to high levels of vitamin A. Exposure to
chemicals such as carbon tetrachloride and vinyl chloride
may result in liver cell damage and decrease metabolism of
toxic substances. These two substances are also associated
with the development of liver cancer.
Factors Affecting Metabolism (4)
• Kidneys are damaged by absorption and concentration of
heavy metals (e.g., Hg, Cd, etc.) in the cells of the proximal
convoluted tubules of the nephron.
Factors Affecting Transport and
Absorption (1)
• Absorption rate, perfusion rate, plasma protein
binding, and storage will affect the rate at which a
toxic substance is delivered to the tissue where
metabolism occurs.
• The perfusion rate of a given tissue is important in
determining how quickly a toxic substance will be
transformed. Organs such as the liver and
kidneys have a high perfusion rate relative to
other tissue types. These organs have the potential
to extract and detoxify larger quantities of
toxicants from the blood.
Factors Affecting Transport and
Absorption (2)
• Toxic substances bound to proteins in the blood do not
easily move across cell membranes. In many cases this
slows the rate at which the toxicant is metabolized because
the substance may not be readily absorbed by the tissue
where detoxification occurs.
Excretion (1)
• The faster a substance is eliminated from the body, the
more unlikely a biological effect will be.
• The primary organs involved in excretion are the kidneys,
liver, and lung.
Excretion (2)
• Excretion by the liver
– Many toxic substances are stored and detoxified in the
liver. The toxic substances are then excreted into the
bile. Bile is produced in the liver by the hepatic cells.
– This mechanism is important in removing large
protein-bound toxicants such as heavy metals.
– Excretion of the toxicant from the liver to the intestinal
tract will usually result in the substance being removed
in the feces.
– However, the intestinal bacteria are also capable of
producing enzymes that cause the detoxified substance
to become less water soluble.
Excretion (3)
• Excretion by the liver
– As a result, the toxic substance may be reabsorbed from
the digestive tract. The process of excreting toxic
substances from the liver and their subsequent
reabsorption from the digestive tract is referred to as
enterohepatic circulation.
• Gluconated polycyclic aromatic hydrocarbons and glutathione
conjugates of trichloroethylene are reabsorbed by this mechanism and
therefore retained.
– Detoxification of toxic substances before they reach the other
portions of the systemic circulation is referred to as the “first-pass
effect.” This effect can decrease the systemic toxicity of those
substances absorbed from the digestive tract.
Excretion (4)
• Excretion by the kidneys
– The kidneys receive 25 percent of the cardiac output. The high
perfusion rate not only results in significant exposure to circulating
toxic substances, but also facilitate the excretion of the toxicants.
– Toxic substances enter the kidneys as a result of active and passive
transport mechanisms present in the glomerulus and the nephron
tubules.
– Several heavy metals --- such as cadmium, lead and mercury --- are
excreted by the kidneys. These metals are bound to plasma
protein. The protein-metal complex has a low molecular weight
and is able to pass through the glomerulus to the nephron.
However, this complex may be reabsorbed by active transport
mechanisms in the proximal convoluted tubules.
Excretion (5)
• Excretion by the lungs
– Excretion of volatile toxic gases, such as those
associated with organic compounds, occurs in the lungs.
The transfer of gases from the blood to the lungs is
influenced by concentration gradients and by their
solubility in water.
• Ethylene --- which is only slightly soluble in water --- will
readily diffuse from the blood into the lungs and will therefore
be easily removed.
• Chloroform however, is more water soluble and will not diffuse
as easily from the blood into the lungs.
Cytochrome P450
– Cytochromes P450 (CYPs) belong to the superfamily of
proteins containing a heme cofactor and, therefore, are
hemoproteins.
An Example of P450 Catalytic Cycle
