DETOXIFICATION AND METABOLISM OF XENOBIOTICS

Introduction
Humans are exposed to various xenobiotics (foreign
chemicals) from air (air pollutants), nutrition (food additives,
preservatives), therapy for disease (drugs), etc. Hence an
understanding of the ability of the body to cope with the
chemical onslaught at the cellular level is an important
aspect of metabolism.

Definition
Detoxification or detoxication is a series of biochemical
reactions to convert toxic compounds to less toxic and more
easily excretable forms.

Site of detoxification

Liver is the major site of detoxification. Kidney and intestines

are involved to a lesser extent.
Compounds that are detoxified include: Foreign chemicals
(xenobiotics)2
Drugs, food additives, insecticides, pollutants, etc.
Compounds produced in the body which are to be eliminated
Bilirubin, steroids, NH3, etc.
Compounds produced in the intestine by bacterial
putrefaction and
Fermentation.
Indole and skatole (from tryptophan), histamine (from
histidine), tyramine (from tyrosine), etc.

Medical Importance
Humans are increasingly exposed to various xenobiotics in
present times.
Hydrophobic (non-polar) xenobiotics would persist in adipose
tissue almost indefinitely if they were not converted to polar
forms. Eg: DDT
Knowledge of the metabolism of xenobiotics is basic to a
rational understanding of pharmacology and therapeutics,
pharmacy, toxicology, cancer research and drug addiction.
All these areas involve administration or exposure to
xenobiotics.

The term ‘detoxification’ is not always appropriate because

in some cases metabolism of toxins increases their biological
activity and toxicity (e.g. conversion of methanol to more
toxic formaldehyde). Metabolism of Xenobiotics is for foreign
bodies only.

Mechanism of Detoxification
The overall mechanism of detoxification is to increase the water
solubility (polarity) of toxic products and thus facilitate their excretion
from the body mostly in urine and also through bile and feces.

Toxic products are rendered water-soluble by adding polar functional

groups / conjugates.
Fig. 1. : Mechanism of detoxification of xenobiotics.
Reactions of Detoxification
It is convenient to consider metabolism of xenobiotics in 2 phases :
Phase I and Phase II. The overview of the metabolism of xenobiotics is
shown in fig. 2.
XENOBIOTIC EXCRETED
 Oxidation (hydroxylation),
PHASE I  Reduction or
Products
EXCRETED
 Hydrolysis
PHASE II Conjugation Products EXCRETED
Fig. 2 : Overview of the metabolism of xenobiotics.
Phase I :
Most of the reactions of phase 1 are either Oxidation (hydroxylation),
Reduction or Hydrolysis.
Minor reactions like deamination, desulfuration, dealkylation,
epoxidation, etc.. are also included under phase I.
2
However, in phase I, the chief reaction involved is hydroxylation.
Phase II
Reactions of phase II involve conjugation with glucuronic acid, sulfate,
glycine, cysteine, glutamine, acetate or methyl group.
Excretion of the detoxified xenobiotics : (Fig. 2)
 The products of phase I reactions (detoxification) are either
excreted directly or 
 undergo further metabolism by phase II reactions (entoxification)
and then excreted. 
 In some cases, a xenobiotic may undergo only phase II reaction
without undergoing phase I reaction. 

 Occasionally, a xenobiotic may be excreted unchanged. 
Reactions of Phase I :
a) Detoxification by Oxidation (Hydroxylation) Reactions
In general,
 Hydrocarbons are oxidised to their corresponding alcohols; 

 Alcohols to aldehydes and 

 Aldehydes to acids. 
Enzymes of oxidation reactions : Cytochrome P450 (hemeproteins) are
involved in majority of oxidation reactions 3.
Location of Cytochrome P450 : Liver microsomes (ER).
Mechanism :
 Cytochrome P450 needs a molecule of oxygen (O2), and one of the
oxygen atoms is incorporated into the xenobiotic to produce a hydroxyl
group. 

 Hence, Cytochrome P450 is also called monooxygenase / mixed
function oxidase. 

 General reaction4 of Cytochrome P450 
Cytochrome P450
RH + O2 R - OH
NADPH + H+ NADP+ + H2O
Reference 3 : So named after the absorption maximum at a
wavelength of 450 nm.
Reference 4 : RH represents a wide variety of xenobiotics including
drugs, carcinogens, pesticides, petroleum products and pollutants
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CytochromeP450 has isoforms and most of them are inducible 5 :
I. 50% of drugs are detoxified through this system Phenobarbital,
Acetanilide, Morphins, Benzodiazpene, etc.
II. Ethanol and drugs such as Phenobarbital induce cytochrome P450.
Examples
Phase 1 Phase 2
1. Benzene Phenol Phenyl Sulphate
-OH
- O - SO4
NADPH + H+
NADP+ + H2O
2. Toluene
Benzyl alcohol
Benzoic acid
-CH3
- CH2OH
COOH
NADPH + H+
NADP+
Example :
Ethyl alcohol / Ethanol is also acted upon by Cytochrome P450 system
in liver named as Microsomal Ethanol Oxidising System (MEOS) with
NADPH and O2. Toxic effects of alcohol are due to its oxidation to
acetaldehyde which forms adducts with Nucleic acids, proteins and
other molecules.
Detoxification by Oxidation Reactions without using cytochrome P450
 Oxidation is a process of detoxification of alcohol metabolism.
Alcohol dehydrogenase oxidises alcohol to aldehyde which is further
oxidised to acid. 
Alcohol dehydrogenase Aldehyde dehydrogenase
Alcohol Aldehyde Acid
Reference 5 : Some of the isoforms exhibit low activity which may be
the reason for variation in drug response in patients.
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However, Oxidation of some compounds may lead to further toxic
compound production.
Examples :
Alcohol dehydrogenase
Aldehyde dehydrogenase
 Methanol
Formaldehyde
Formic acid
NAD+
NADH + H+
(More toxic than methanol).
Alcohol dehydrogenase
Aldehyde dehydrogenase
 Ethyl alcohol
Acetaldehyde
Acetic acid
NAD
NADH + H+
b) Detoxification by Reduction Examples
 Nitrobenzene Amino benzene (Aniline)
NO2 NH2

Picric acid
Picramic acid

Chloral
Trichloroethanol
(Chloral is a sedative / hypnotic drug).
c) Detoxification by Hydrolysis
These include, hydrolysis of ester, amide, glycosidic bond, ether bond,
etc.
Examples H2O
 Aspirin Salicylic acid + Acetic acid
(Acetyl salicylic acid)
COOH H2O COOH
+ CH3COOH
O- CO- CH3 OH
5
H2O

Acetanilide
Aniline
+
acetic acid
H2O

Atropine
Tropic acid
+
tropine
Glycosidase
Reactions of Phase 2 :
a) Detoxification by Conjugation (Fig. 3)
Conjugation is a process in which a toxic compound (foreign or
endogenous) combines with a polar compound produced in the body.
This makes the toxic substance more water soluble and hence more
easily excretable.
Polar compound (produced in the body)
Toxic compound
Polar compound
(foreign or endogenous)
(Less toxic, water soluble)
easily excreted.
Fig. 3 : Detoxification by Conjugation.
Conjugation may occur either directly or after the phase I reaction.
The more important conjugating agents identified in the body are
glucuronic acid, glycine, cysteine, sulfate, thiosulfate, acetic acid and
methyl group.
 Glucuronic acid
Conjugation with glucuronic acid is the most common conjugation
reaction. UDP-Glucuronic , acid is the glucuronyl doner, UDP-
glucuronyl transferases are the enzymes 7.
General Reaction of Glucuronide Conjugation :
Reference 6 : Atropine is used as a therapeutic for organophosphorus
poisoning. It is an anticholinergic agent and is useful in dilatation of
pupil.
Reference 7 : UDP-glucuronyl transferase, present in the endoplasmic
reticulum of hepatocytes, and is inducible by drugs, such as
barbiturates.
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UDP-glucuronyl transferase
X – OH
+ UDP-glucuronic acid
X-O-glucuronide
+ UDP
‘Active’ Glucuronide
(Produced in the uronic acid pathway)
Examples:

Bilirubin
Bilirubin diglucuronide
(Endogenous)

Benzoic acid
Benzoyl glucuronide
(Benzoic acid is used as food preservative)

Phenol
Phenyl glucuronide
+
UDP
 Glycine 
Many aromatic carboxylic acids are conjugated with glycine.
Examples:
Glycine
 Benzoic acid Benzoyl CoA Benzoyl glycine 
(Hippuric acid)
Glycine
 Phenyl acetic acid 8 Phenylaceturic acid 

 Cysteine 
Cysteine, required for conjugation, is derived from the tripeptide,
glutathione, (GSH).
General reaction is as follows :
R + GSH R –cysteine A Mercapturic acid
Glutamate Glycine Acetyl CoA HSCoA
Reference 8 : Phenylacetic acid is derived from metabolism of
Phenylalanine in case of the enzyme defect in phenylketonuria.
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 Glutamine 

Example : 

 Phenyl acetic acid + glutamine Phenyl acetylglutamine9 

 Acetic acid 
Acetyl CoA (‘active’ acetate) is the acetate donor.
Example :
 Sulfanilamide + Acetyl CoA Acetyl sulfanilamide
(a sulfa drug)
 Sulfate : 
Doner of sulfate group : Phospho adenosine phospho sulfate (PAPS /
‘active’ sulfate).
Examples

Indole
+
PAPS
Indoxyl sulfate (Indican)
Sulfotransferase

Phenol
+
PAPS
Phenyl sulfate
 Thiosulfate :
Example
Rhodanase
 Cyanide + Sodium thiosulfate Thiocyanate + Sodium sulfate
 Methyl group10 
Methyl group donor : S-adenosyl methionine is the most common
(SAM / ‘active’ methyl group)
Example
 Pyridine + SAM N-methyl pyridine + SAH
Reference 9 : Phenylacetyl glutamine is derived from metabolism of
phenylalanine in case of the enzyme defect in phenylketonuria.
Reference 10 : Methylation may help in catabolising the compounds to
excrete through Mono Amino Oxidase (MAO) system. Metabolism of
catecholamines takes place through this pathway.
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