BIOMONITORING

Nendyah Roestijawati
4 Juni 2013
Biomonitoring
In human : direct measurement of levels of
chemical substances in blood, urine,
breast-milk or saliva; and such other
tissues as bone, teeth, skin, hair and nails
Wildlife and other species may include
additional tissues such as muscle (for
example fish fillets), liver, fat, eggs, and
reproductive tissues
Contd
biomonitoring has become the standard
for assessing people's exposure to toxic
substances
for responding to serious environmental
public health problems
Biotransformation
is the process whereby a substance is
changed from one chemical to another
(transformed) by a chemical reaction
within the body
vital to survival in that it transforms
absorbed nutrients (food, oxygen, etc.)
into substances required for normal body
functions
it is a metabolite that is therapeutic and
not the absorbed drug
an important defense mechanism in that
toxic xenobiotics and body wastes are
converted into less harmful substances
and substances that can be excreted from
the body.
Contd
Contd
Lipophilic toxicant, non-polar, and low
molecular weight are readily absorbed
through the cell membranes of the skin,
gastrointestinal (GI) tract, and lung.
Lipophilic toxicants are hard for the body
to eliminate and can accumulate to
hazardous levels.
Most lipophilic toxicants can be
transformed into hydrophilic that are easier
for the body to eliminate than lipophilic
substances
Contd
The human body has a well-developed capacity
to biotransform most xenobiotics as well as body
wastes
Hemoglobin the normal destruction of red
blood cells biotransformed to bilirubin
toxic to the brain of newborns irreversible
brain injury
Biotransformation of the lipophilic bilirubin
molecule in the liver results in the production of
water-soluble (hydrophilic) metabolites excreted
into bile and eliminated via the feces.
Contd
The biotransformation process is not perfect
Biotransformation results in metabolites of lower
toxicity detoxification
Metabolites are more toxic than the parent
substance bioactivation.
An unusually reactive metabolite that may
interact with cellular macromolecules (e.g., DNA)
cancer or birth defects
Biotransformation of vinyl chloride to vinyl
chloride epoxide, which covalently binds to DNA
and RNA, a step leading to cancer of the liver.
Chemical Reaction
Most of these chemical reactions occur at
significant rates only because specific
proteins, known as enzymes, are present
to catalyze them, that is, accelerate the
reaction.
A catalyst a substance that can
accelerate a chemical reaction of another
substance without itself undergoing a
permanent chemical change.
Contd
Most biotransforming enzymes are high
molecular weight proteins, composed of
chains of amino acids linked together by
peptide bonds
Most enzymes will catalyze the reaction of
only a few substrates, meaning that they
have high "specificity
Specificity is a function of the enzyme's
structure and its catalytic sites, referred to
as the "lock and key" relationship
Three main type of specificity
The biotransformation of ethyl alcohol to acetaldehyde

ADH = alcohol dehydrogenase
Phase I Reaction
Phase I reactions are
1. Oxidation,
2. Reduction, and
3. Hydrolysis
Oxidation
Chemical reaction in which a substrate
loses electrons
Addition of oxygen was the first of these
reactions discovered
The simplest type of oxidation reaction is
dehydrogenation, that is the removal of
hydrogen from the molecule
Another example of oxidation is electron
transfer

Types of oxidizing reactions
Oxidizing Reactions
alcohol dehydrogenation
aldehyde dehydrogenation
alkyl/acyclic hydroxylation
aromatic hydroxylation
deamination
desulfuration
N-dealkylation
N-hydroxylation
N-oxidation
O-dealkylation
sulphoxidation

Reduction
Chemical reaction in which the substrate gains
electron
Most likely to occur with xenobiotics in which
oxygen content is low
Can occur across nitrogen-nitrogen double
bonds (azo reduction) or on nitro groups (NO2).
Some chemicals such as carbon tetrachloride
can be reduced to free radicals, which are quite
reactive with biological tissues
Frequently result in activation of a xenobiotic
rather than detoxification
Reduction reaction in the nitro group
Reducing Reactions
azo reduction
dehalogenation
disulfide reduction
nitro reduction
N-oxide reduction
sulfoxide reduction

Hydrolysis
Chemical reaction in which the addition of
water splits the toxicant into two fragments
or smaller molecules
The hydroxyl group (OH-) is incorporated
into one fragment and the hydrogen atom
is incorporated into the other
Larger chemicals such as esters, amines,
hydrazines, and carbamates are generally
biotransformed by hydrolysis
Procaine (local anesthetic) hydrolysis
Phase II Reactions

Are conjugation reactions, that is, a
molecule normally present in the body is
added to the reactive site of the Phase I
metabolite
The result is a conjugated metabolite that
is more water-soluble than the original
xenobiotic or Phase I metabolite
Usually the Phase II metabolite is quite
hydrophilic and can be readily eliminated
from the body.
Contd
The primary Phase II reactions are:
glucuronide conjugation - most important
reaction
sulfate conjugation - important reaction
acetylation
amino acid conjugation
glutathione conjugation
methylation

Glucuronide conjugation
One of the most important and common Phase II
reactions
One of the most popular molecules added
directly to the toxicant or its phase I metabolite is
glucuronic acid, a molecule derived from
glucose, a common carbohydrate (sugar) that is
the primary source of energy for cells
The sites of glucuronidation reactions are
substrates having an oxygen, nitrogen or sulfur
bond
Includes a wide array of xenobiotics as well as
endogenous substances, such as bilirubin,
steroid hormones and thyroid hormones
Glucuronide conjugation
Glucuronidation is a high-capacity pathway for
xenobiotic conjugation
Glucuronide conjugation usually decreases
toxicity, although there are some notable
exceptions, for example, the production of
carcinogenic substances
The glucuronide conjugates are generally quite
hydrophilic and are excreted by the kidney or
bile, depending on the size of the conjugate

Glucuronide conjugation of aniline
Sulfate conjugation
another important Phase II reaction that occurs
with many xenobiotics
In general, sulfation decreases the toxicity of
xenobiotics
Unlike glucuronic acid conjugates that are often
eliminated in the bile, the highly polar sulfate
conjugates are readily secreted in the urine
In general, sulfation is a low-capacity pathway
for xenobiotic conjugation
Often glucuronidation or sulfation can conjugate
the same xenobiotics.
The major transformation reactions for xenobiotics
Biotransformation Sites

Biotransforming enzymes are widely distributed
throughout the body
The liver is the primary biotransforming organ
due to its large size and high concentration of
biotransforming enzymes, it is also potentially
quite vulnerable to the toxic action of a
xenobiotic that is activated to a more toxic
compound
The kidneys and lungs are next with 10-30% of
the liver's capacity
Low capacity exists in the skin, intestines,
testes, and placenta.
Contd
The liver receives blood directly from the
gastrointestinal tract via the portal vein
"first pass" phenomena
Blood leaving the liver is eventually
distributed to all other areas of the body
the liver may have removed most of the
potentially toxic chemical, or some toxic
metabolites are in high concentration in
the liver.

Contd
Within the liver cell, the primary subcellular
components that contain the transforming
enzymes are the microsomes (small vesicles) of
the endoplasmic reticulum and the soluble
fraction of the cytoplasm (cytosol)
The mitochondria, nuclei, and lysosomes
contain a small level of transforming activity
The most important enzyme system involved in
Phase I reactions is the cytochrome P-450
enzyme system "mixed function oxidase
(MFO) " system. It is found in microsomes and is
responsible for oxidation reactions of a wide
array of chemicals.

Modifiers of Biotransformation
The relative effectiveness of biotransformation
depends on several factors, including species,
age, gender, genetic variability, nutrition,
disease, exposure to other chemicals that can
inhibit or induce enzymes, and dose levels
Differences are normally the basis for selective
toxicity, used to develop chemicals effective as
pesticides but relatively safe in humans
Malathion in mammals is biotransformed by
hydrolysis to relatively safe metabolites, but in
insects, it is oxidized to malaoxon, which is lethal
to insects.
Contd
Age may affect the efficiency of
biotransformation
Human fetuses and neonates (newborns) have
limited abilities for xenobiotic biotransformations
inherent deficiencies in many, but not all, of
the enzymes responsible for catalyzing Phase I
and Phase II biotransformations
Biotransformation capability is also decreased in
the aged
Gender may influence the efficiency of
biotransformation for specific xenobiotics
limited to hormone-related differences in the
oxidizing cytochrome P-450 enzymes.
Contd
Genetic variability in biotransforming capability
accounts for most of the large variation among
humans
The Phase II acetylation reaction in particular is
influenced by genetic differences in humans
The most serious drug-related toxicity occurs in
the slow acetylators "slow metabolizers
acetylation is so slow that blood or tissue levels
of certain drugs (or Phase I metabolites)
exceeds their toxic threshold
Contd
Poor nutrition can have a detrimental effect on
biotransforming ability inadequate levels of protein,
vitamins, and essential metals decrease the ability to
synthesize biotransforming enzymes
Enzyme inhibition exposure to a substance will inhibit
the biotransformation capacity for another chemical due
to inhibition of specific enzymes
Enzyme induction enhanced capability for
biotransforming a xenobiotic increase the production
of some enzymes increased level of enzyme activity
increased biotransformation of a chemical
subsequently absorbed.
Contd
Dose level can affect the nature of the
biotransformation
the biotransformation may be quite different at
high doses versus that seen at low dose levels
can be explained by the existence of different
biotransformation pathways.
At low doses, a xenobiotic may follow a
biotransformation pathway that detoxifies the
substance
If the amount of xenobiotic exceeds the specific
enzyme capacity, the biotransformation pathway
is "saturated
Contd
a dose-related difference in biotransformation occurs
with acetaminophen
At normal doses, approximately 96% of acetaminophen
is biotransformed to non-toxic metabolites by sulfate and
glucuronide conjugation, about 4% of the acetaminophen
is oxidized to a toxic metabolite toxic metabolite is
conjugated with glutathione and excreted
With 7-10 times the recommended therapeutic level, the
sulphate and glucuronide conjugation pathways become
saturated and more of the toxic metabolite is formed
the glutathione in the liver may also be depleted so that
the toxic metabolite is not detoxified and eliminated
react with liver proteins and cause fatal liver damage.