Phr.

Rautbhola, Quality control officer

COMPLEXING & CHELATING AGENTS USED IN THERAPY

COMPLEXING & CHELATING AGENTS are those agents which form complex (via coordinate
covalent bond) with simple metal ion and transformed into complex ion (coordinate compound)
by addition of reagent which is known as ligand.
Metal ion + Electron donor = Complex (Coordinate compound) e.g. Cd2+ + CN- = CdCN+

The complex formed is stable and water soluble. In complex formation metal ions accepts
electron and ligand donate it.
Thus in a ligand molecules there is presence of at least one lone pair of electrons through which
co-ordinate linkage with metal ion take place.
In chemistry, a coordination complex or metal complex consists of a central atom or ion, which
is usually metallic and is called the coordination centre, and a surrounding array
of bound molecules or ions, that are in turn known as ligands or complexing agents.

Many metal-containing compounds, especially those of transition metals, are coordination

complexes.
Complexation is the combination of individual atom groups, ions or molecules to create one
large ion or molecule. One atom or ion is the focal point of the complex. This central atom
contains empty electron orbitals that enable bonding with other atoms as well as unshared
electrons.
The last stage in complexation involves the sum of individual components' charges. Therefore,
there can be zero, negative and positive charges in a complex within a solution.
Complexing and chelating agents are the compounds mainly used in heavy metal poisoning

Combines (complex) with metallic ions, forming ring structures within their molecule
(Chele crab)
Form stable, non-toxic and easily excreatable complexes with toxic metals
Contains 2 or more reactive groups (ligand) such that can hold metal from two sides

A chelating agent is a substance whose molecules can form several coordinate bonds to a single
metal ion. That is, a chelating agent is a polydentate ligand. The most common and most widely
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used chelating agents are those that coordinate to metal ions through oxygen or nitrogen donor
atoms, or through both.
A chelate is a chemical compound composed of a metal ion and a chelating agent. Chelating
agent + metal complex is known as Chelate.

A chelate is a chemical compound composed of a metal ion and a chelating agent. A chelating
agent is a substance whose molecules can form several bonds to a single metal ion. In other
words, a chelating agent is a multidentate ligand. An example of a simple chelating agent is
ethylenediamine.
Chelation means "to grab" or "to bind." Chelation is a type of bonding of ions and molecules to
metal ions. It involves the formation or presence of two or more separate coordinate
bonds between a polydentate (multiple bonded) ligand and a single central atom.
Three widely used chelating agents are ethylenediamine, ethylenediaminetetraacetic acid, and
dimercaprol.
A ligand is a molecule or ion that is directly bonded to metal ion in a coordination complex.
Molecules or ions having lone pair of electron can be used to form bond with a metal ion. A
ligand is an ion or molecule (functional group) that binds to a central metal atom to form
a coordination complex. The bonding between metal and ligand generally involves formal
donation of one or more of the ligand's electron pairs. The nature of metal-ligand bonding can
range from covalent to ionic.
Ligand molecules may have no. of site present in them like

Unidentate: Cyanide ion

Bidentate: Glycine and oxalic acid
Multidentate: EDTA

Usually ligands are organic compounds, and are called chelants, chelators, chelating agents, or
sequestering agents.
Chelation therapy is a medical procedure that involves the administration of chelating agents to
remove heavy metals from the body. For the most common forms of heavy metal intoxication
those involving lead, arsenic or mercury. Chelation therapy is the use of chelating agents
to detoxify a patient's body of poisonous metal agents, such as mercury, arsenic, and lead, by
converting them to a chemically inert form that can be excreted without further interaction with
the body.

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Co-ordinate linkage: Particular no. of ligand molecules associated with metal ion is called as
co-ordinate linkage with metal ion take place.
Chelating agents have high affinity for such metals and combine with them to form non toxic and
water soluble complexes for elimination. Possesses: ve charged groups to attract + ve
charged toxic metals.
Chelating agents are chemical compounds (usually organic compounds) that form complexes
with metal ions or other substrates. They are also called chelators or sequestering agents. The
chelating agent has a ring like center, which forms a complex with the metal ion/substrate by two
or more bindings and the metal ion is bound and excreted.
e.g. Ethylenediamine ligand chelating to a metal with two bonds.

Chelation is a type of bonding of ions and molecules to metal ions. It involves the formation or
presence of two or more separate coordinate bonds between a polydentate (multiple
bonded) ligand and a single central atom. Usually these ligands are organic compounds, and are
called chelants, chelators, chelating agents, or sequestering agents.
Chelation therapy is used as a treatment for acute mercury, iron (including in cases of
thalassemia), arsenic, lead, uranium, plutonium and other forms of toxic metal poisoning.
The chelating agent may be administered intravenously, intramuscularly, or orally, depending on
the agent and the type of poisoning.
Mechanism of action heavy metals poisoning:
Heavy metals combine with
one or more reactive groups(Ligands)

Oxygen (-OH, -COO, -OPO)

Nitrogen (-NH2, -NH)
Sulphur (-SH, -S-S)
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Hamper physiological function

Enzyme inhibition, Oxidative stress

Mechanism of action of chelating agents

Chelating agents forms
complexes with heavy metals

Stable Chelates

Nontoxic, easily excreted

Heavy metals exert their toxic effects by combining with and inactivating functional groups
(ligands) of enzymes and important biomolecules - sulfhydril, hydroxyl, carboxyl etc. leading to
inactivation. Chelating agents compete with body ligands for the heavy metal also differ in
affinity for different metals. Chelating agents have high affinity for such metals and combine
with them to form nontoxic and water soluble complexes for elimination.
Factors affecting actions of chelating agents

Relative affinity of chelator to heavy metal

Distribution of chelator in body
Capacity to mobilize the complex
Half-life of heavy metal
Time after exposure

Ideal chelating agent

Resistant to biotransformation
Ability to reach sites of metal storage
Form nontoxic complexes with heavy metal

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Retain activity at pH of body fluids

Ideal chelating agents have higher affinity for toxic metals than for body Ca++ (readily
available in plasma and ECF)
Should also have higher affinity for toxic metals than body ligands (Greater affinity for
heavy metal than endogenous ligand)
Ideally should be water soluble (Highly water soluble) and distribution should correspond to
that of the metal intended to
Interval of administration between exposure to metals and chelating agents should be less
Ready excretion of chelate

Uses of chelating or complexation agents

Softening of water
Detection of some metals in qualitative analysis e.g. estimation of magnesium, calcium,
aluminium and zinc etc.
For sequestration of metals i.e. removal or separation and prevents from absorption
Stabilization of drugs vulnerable to oxidation in presence of trace elements
Treatment of heavy metal poisoning.
Treatment of certain metabolic disorders where metals like iron and copper are accumulated
in abnormal amount in various tissues.
Formation of complex can result in the alteration of various properties of the drug like
solubility, light absorption, conductance, partitioning behaviours and chemical reactivity.
Complexation is used to stabilize the drugs e.g. boric acid chelates with epinephrine so
stabilized epinephrine act against attack by bisulphate and sulphate.
Uses as antimicrobial drugs e.g. silver sulphadizine, anticancer drug e.g. cisplatin, as antiviral
drug e.g. zinc Gluconate etc.

Chelating Agent Classification

1.
2.
3.
4.
5.
6.

Dimercaprol (British antilewisite) or BAL As, Au, Bi, Ni, Sb and Hg poisoning
Dimercaptosuccinic acid (succimer) - Pb
Calcium disodium edetate (EDTA) lead poisoning
Disodium edetate
Penicillamine Cu, Pb, Hg, Zn
Desferrioxamine Iron overload

Poisoning and their antagonist

Poisoning of the body can be happened by various reasons. The most common poisoning occurs
because of followings:
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Heavy metals or metallic contamination of food

Due to insecticides or pesticides
Due to excessive use of drugs/overdose

Antidotes are the substances which react specifically with an ingested poison or toxic substances
or an overdose of a potent drug. They act by neutralizing the poison (neutralizing its toxic
effects), act as antagonist (chemically convert into non-toxic or less toxic compounds)
Cyanide poisoning

Cyanide poisoning normally takes place accidently or when cyanide poison is taken
intentionally to commit suicide. In cyanide poisoning cyanide ion combines with ferric ion of
cytochrome oxidase, an enzyme which is responsible for electron transfer reactions. This
causes stoppage of cellular respiration and metabolic reaction. Cyanide poisoning is usually
fatal, if it is not treated immediately and instantaneously.
For cyanide poisoning two inorganic antidotes such as sodium nitrite and sodium
thiosulphate are used. Both are used in conjunction with each other.

An antidote is a substance which can counteract a form of poisoning. Antidotes may be defined
as those substances which react specifically with ingested poisons or toxic substances or an
overdose of a potent drug. They act either by neutralising the poison or its toxic effect or
pharmaceutically (antagonistic action) or chemically by converting them to non-toxic or less
toxic forms (e.g. chelates, acids, insoluble derivatives)
Mechanism of Action of Antidotes
Antidotes act by different mechanism. The mechanisms of action of antidotes are given below:

Complex formation.
Metabolic conversion.
Prevention of toxic metabolite formation.
By changing the physio-chemical nature of toxicant.
Promotes return to normal function by repairing a defect or enhancing a function that corrects
the effects of poison.

It is possible to classify antidotes on the basis of their mechanism of action as follows:

1) Physiological antidotes: They act by producing the effect opposite to that of poison or to
counteract the effect of poison physiologically. For example, sodium nitrite converts hemoglobin
into methemoglobin in order to bind cyanide.
2) Chemical antidotes: They act usually by combining with the poison and thus change its
nature and poison cannot act any more. For example, sodium thiosulphate which changes toxic
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cyanide to the non-toxic thiocyanate; sodium calcium edetate chelates agents used for heavy
metal poison.
3) Mechanical antidotes: They usually act by preventing the absorption of poisons in the body
or expelling out the poison by emesis or elimination through urine. For example, activated
charcoal, Light Kaolin etc. absorb the poison prior to absorption across intestinal wall. Copper
sulphate, magnesium sulphate and sodium monohydrogen phosphate inactivate and precipitate
the toxic material as insoluble salts by chelation.
Therapeutic Uses of antidotes
Antidotes are used in treatment of poisoning as well as in case of over dose of drugs. Following
are the examples of antidotes which are used for treating poisoning:
Examples of Poisons and Antidotes
Poison/Drug

(Antidote)

Paracetamol (acetaminophen)

(N-acetylcysteine)

Anticoagulants, Warfarin

(Vitamin K)

Opioids

(Naloxone)

Cyanide

(Amyl Nitrite, Sodium Nitrite)

Organophosphates

(Atropine and Pralidoxime)

Calcium Channel Blockers (Verapamil, Diltiazem)

(Calcium Gluconate)

Isoniazid

(Pyridoxine)

Atropine

(Physostigmine)

ACTIVATED CHARCOAL

Preparation: Commercially, it is obtained as a residue during destructive distillation of various

organic matters or from burning of organic materials in a special manner. The coarse material is
crushed and powdered.
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Properties:

It is fine black, odourless, tasteless powder having smooth touch and free from gritty
particles.
It is almost insoluble in usual solvents.
The fine powder provides more surface area for adsorbent properties.

Action and uses of Activated Charcoal

It is a general type of adsorbent which finds use in poisoning.

It not only adsorbs heavy metals but also adsorbs drugs such as hypnotics, sedatives,
alkaloids etc. and also gases like carbon monoxide, carbondioxide, nitrous oxide etc.
It is normally employed in the ratio of 5:1 or 10:1 (charcoal to poison).
It is administered in the form of tablets. It also finds use in diarrhoea to adsorb toxins.
Alleviates Gas & Bloating, Whitens Teeth, Treats Alcohol Poisoning & Helps Prevent
Hangovers, Water Filtration
Digestive Cleanse: Activated charcoal uses help promote a healthy digestive tract by
removing toxins that cause allergic reactions, oxidative damage and poor immune system
function.
It doesnt absorb the toxins, however. Instead it works through the chemical process
of adsorption.

Activated charcoal works by trapping toxins and chemicals in its millions of tiny pores. The
porous surface of activated charcoal has a negative electric charge that causes positive charged
toxins and gas to bond with it.

Ethylenediaminetetraacetic acid (EDTA)

Ethylenediaminetetraacetic acid which is commonly shortened to EDTA, is the most widely used
complexometric titrant. Fully protonated EDTA has the structure
The EDTA molecule has six potential sites for bonding a metal ion: the four carboxyl groups and
the two amino groups, each of the latter with an unshared pair of electrons. Thus, EDTA is a
hexadentate ligand.

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Industrial synthesis of EDTA

The commercial preparation involves the reaction of ethylenediamine, formaldehyde

(HCHO), and sodium cyanide.

EDTA is also prepared commercially by the reaction of ethylenediamine and chloroacetic

acid.

Uses:

EDTA deactivates metallo-enzymes by removing the metal ion from the enzyme. In food,
some of these enzymes catalyze the reactions that produce spoilage.
EDTA dissolves the CaCO3 scale deposited from hard water without the use of corrosive
acid.
EDTA is also used as an anticoagulant for stored blood in blood banks. It prevents
coagulation by sequestering the calcium ions required for clotting.
As an antidote for lead poisoning, calcium disodium EDTA exchanges its chelated calcium
for lead, and the resulting lead chelate is rapidly excreted in the urine.
The calcium salt of EDTA, administered intravenously, is also used in the treatment of acute
cadmium and iron poisoning

Disodium Edetate

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Molecular formula: C10H14N2Na2O8.2H2O

Molecular weight: 372.2

Disodium Edetate contains not less than 98.5 per cent and not more than 101.0 per cent of
C10H14N2Na2O8.2H2O
Description: A white, crystalline powder; odourless. Soluble in water; practically insoluble in
ethanol (95 per cent).However, its sodium salts are quite soluble in water.
Category: Pharmaceutical aid; chelating agent in metal poisoning.
Dose: By intravenous injection, 50 mg per kg of body weight, up to a maximum of 3 g per day.
Identification:

Determine by infrared absorption spectrophotometry. Compare the spectrum with that

obtained with disodiumedetate RS.
Dissolve 2 g in 25 ml of water, add 6 ml of lead nitrate solution, shake and add 3 ml of
potassium iodide solution; no yellow precipitate is produced. Make alkaline to red litmus
paper with 2 M ammonia and add 5 ml of ammonium oxalate solution; no precipitate is
produced.
Dissolve 0.5 gin 10 ml of water, add 0.5 ml of a 10 per cent w/v solution of calcium chloride,
make alkaline to red litmus paper with 2 M ammonia and add 3 ml of ammonium oxalate
solution; no precipitate is produced.
Gives the reactions of sodium salts.

Preparation: It is prepared by adding hot solution of sodium hydroxide to solution of edetic

acid.
Assay: Weigh accurately about 0.5 g, dissolve in sufficient water to produce 300 ml and ad42 g
of hexamine and 2 ml of 2 M hydrochloric acid. Titrate with 0.1M lead nitrate using about 50 mg
of xylenol orange triturate as indicator.
1 ml of 0.1 M lead nitrate is equivalent to 0.03722 g of C10H14N2Na2O8.2H2O
Uses:

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Chelates calcium and magnesium in addition to various heavy metals.

Widely used in therapy to lower plasma calcium levels.
It is also used as anticoagulant. It is used as chelating agent in metal poisoning. It is most
widely used for lead poisoning.

DIMERCAPROL or BAL (British Anti -Lewisite)

Molecular Formula: C3H8OS2

Molecular Weight: 124.2

This chemical was originally employed to treat the toxic effects of an arsenic-containing mustard
gas called Lewisite [It is an organoarsenic compound], which was used in World War II.
Standard: Dimercaprol contains not less than 98.5 per cent w/w and not more than 101.5
percent w/w of C3H8OS2.
Category: Antidote in heavy metal poisoning; metal complexing agent.
Description: A clear, colourless or slightly yellow liquid; odour, strong, characteristic and
alliaceous.

Identification:
A. Dissolve 0.1 ml in 4 mI of water and to 2 mI of the solution add lead acetate solution; a
yellow precipitate is obtained.
B. To 2 mI of the solution prepared for test A add 1 mI of 0.05M iodine; the colour of iodine is
immediately discharged.

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Preparation: A methanolic solution of sodium hydroxide is saturated with hydrogen sulphide to
get sodium hydrogen sulphide. To it 2, 3-dibromopropanol prepared from allyl alcohol is added
and the mixture is heated at 40C under pressure to get dimercaprol.

Assay: Weigh accurately about 0.1 g, dissolve in 40 mI of methanol and add 20 mI of 0.1 M
hydrochloric acid and 50.0 mI of 0.05 M iodine. Allow to stand for 10 minutes and titrate with
0.1 M sodium thiosulphate. Repeat the operation without the substance under examination. The
difference between the titrations represents the amount of iodine required.
1ml of 0.05M iodine is equivalent to 0.00621 g of C3H8OS2.
Storage: Store protected from light in well-filled containers in a refrigerator (2 to 8).
Action and uses:
Dimercaprol is the most widely used antidote for arsenic, mercury, antimony, and gold
poisoning.
The chelated metal cannot enter living cells and is rapidly excreted from the body. Since
dimercaprol is water insoluble, it is dissolved in an oil base (often peanut oil) and injected
intramuscularly.

Dose: By intramuscular injection, 2 to 3 mg per kg of body weight every 4 hours during the first
day and subsequently, in accordance with the needs of the patient.

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Standard: Desferrioxamine Mesylate contains not less than 98.0 per cent and not more than
102.0 per cent of C25H48N608,CH4SO3, calculated on the anhydrous basis.
Description: A white or almost white powder. Freely soluble in water; slightly soluble in
methanol; very slightly soluble in ethanol (95 percent); practically insoluble in chloroform and in
ether.
Identification

Determine by infrared absorption spectrophotometry. Compare the spectrum with that

obtained with desferrioxamine mesylate RS or with the reference spectrum of
desferrioxamine mesylate.
Dissolve 5 mg in 5 ml of water, add 2 ml of a 0.5 per cent w/v solution of tribasic sodium
phosphate, mix and then add0.5 ml of a 2.5 per cent w/v solution of sodium1,2naphthoquinone-4-sulphonate; a blackish brown colour is produced.

Preparation: It is isolated from cultures of Streptomyces pilosus as ferric complex. After

removal of iron, the chelating agent is purified as the methane sulphonate (mesylate) salt.
Test for purity: It is tested for Appearance of solution, pH(3.7 to 5.5, determined in a freshly
prepared 10.0 percent w/v solution),Related substances, Heavy metals, Chlorides, Sulphates,
Sulphated ash, Water (NMT 2%,determine on 1.0 gram)
Assay: Weigh accurately about 50 mg of the substance under examination in 50.0 ml of water.
To 2 ml of this solution, add 3 ml of ferric chloride solution prepared by dissolving 6.7 g of ferric
chloride in 100 ml of 1 per cent v/v solution of hydrochloric acid and dilute to 25 ml with water.
Measure the absorbance of the resulting solution at the maximum at about 485 nm. Calculate the
content of C25H48N608,CH4SO3 from the absorbance obtained from a solution of known
concentration of deferoxamine mesylate RS.

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Storage: Store protected from light in a refrigerator (2 to 8).Do not freeze. If the substance is
sterile, store in sterile, airtight, tamper-evident containers sealed so as to exclude
microorganisms.
Labelling: The label states where applicable, that the substance is sterile.
Uses: It possesses high affinity for both ferrous and ferric iron; so it is used especially in acute iron
poisoning in small children. Removes iron from hemosiderin, ferritin but not from hemoglobin and
cytochromes. It is also used to chelate aluminium. It is given parenterally.

D-Penicillamine
Molecular Formula: C5H11NO2S

Mol. Wt.: 149.2

Penicillamine is 3-mercapto-o-valine. The thiol group is where metal ion interacts to form a
chelate.
Standard: Penicillamine contains not less than 98.0 per cent and not more than 101.0 per cent of
C5H11NO2S, calculated on the dried basis.
Category: Chelating agent in copper and lead poisoning; antirheumatoid arthritic.
Description: A white or almost white, crystalline powder. Freely soluble in water; slightly
soluble in ethanol (95 per cent); practically insoluble in chloroform and in ether.
Identification:

Determine by thin layer chromatography, coating the plate with silica gel G.
To 4 ml of a 1 per cent w/v solution add 2 ml of phosphotungstic acid solution and heat
nearly to boiling; a blue colour is produced.

Preparation: It is prepared by hydrolysis of Penicillin. It is precipitated from the hydrolysis

mixture as mercury salt which is then collected, suspended in water and treated with hydrogen
sulphide to liberate free acid. Purification involves only recrystallization from water.

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Test for purity: It is tested for Appearance of solution., pH: 4.5 to 5.5, determined in a 1.0 percent w/v
solution, Specific optical rotation, Heavy metals, Mercuric salts, Penicillamine disulphide, Sulphated ash,
Loss on drying etc.
Assay: Dissolve 0.1 gin 30 ml of anhydrous glacial acetic acid. Titrate with 0.1 M perchloric acid,
determining the endpoint potentiometrically. Carry out a blank titration.
1 ml of 0.1 M perchloric acid is equivalent to 0.01492 g of C5H11NO2S.
Storage: Store protected from moisture.

Uses:

It is also used as chelating agent in the treatment of Wilsons disease and biliary cirrhosis,
lead, gold or mercury poisoning.
It is also used for zinc intoxification.
This chelator is well absorbed from the GIT after oral administration. Penicillamine is often
given for long term treatment of chronic metal poisoning after the patient has been removed
from immediate danger. It is not considered as first choice of antidote.

Dose: In poisoning, 500 mg to 2 g daily, in divided doses or in accordance with the needs of the
patient.

Ethylenediamine

Ethylenediamine (molar mass 60.10 g) is a colorless, clear liquid with a boiling point of 116C
and a melting point of about 8 C. It has an ammonia-like odor, and it dissolves in water to form
an alkaline solution. Ethylenediamine is prepared commercially by heating 1,2-dichloroethane
with aqueous ammonia under pressure at 100-180 C.
Cl-CH2-CH2-Cl + 2 NH3

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H2N-CH2-CH2-NH2 + 2 HCl