COMPLEXOMETRIC TITRATIONS

Introduction to Complexometric Titration

Complexometric titrations are an essential tool in the pharmaceutical industry for ensuring the

safety, quality, and efficacy of drug formulations. They are used to quantify metal ions in a wide

range of pharmaceutical products, including supplements, parenteral solutions, and raw materials.

By accurately measuring metal ion content, complexometric titrations help ensure that

pharmaceutical products meet regulatory standards and provide the intended therapeutic effects.

Complexometric titration is a type of volumetric analysis used to determine the concentration of

metal ions in a solution by forming a complex between the metal ion and a suitable ligand. A

ligand is a molecule or ion that can donate electron pairs to form a coordinate bond with a metal

ion. The titration involves the use of a chelating agent, often a disodium

ethylenediaminetetraacetate (EDTA) solution, which can form stable complexes with metal

ions.

Complexometric titration revolves around the formation of a stable, soluble complex between

a metal ion (the analyte) and a chelating agent (the titrant), During the titration, the ligand binds to

the metal ion, and the endpoint is typically determined using an indicator that changes color when

the metal ion is fully complexed. The formation of a simple complex rather than a series of

complexes will simplify complexometric titration and facilitate the detection of endpoints.

Aminopolycarboxylic acids are excellent complexing agents, the most used being

ethylenediaminetetraacetic acid (EDTA) which chelates with many metals. This analytical method

is based on the donation of electrons from the chelating agent to the metal ion to form a ring

structure (water soluble chelates). The electron donor atom within the chelating agents are almost

always nitrogen, oxygen and sulphur. The formation of the complex is governed by the equilibrium
between the free metal ion and the metal-ligand complex, and the stability of the complex is

described by the formation constant (Kf), a measure of how strongly the ligand binds to the metal

ion. Chelating agents that are able to form water soluble chelates are referred to as sequestering

agents. Types of EDTA titration include direct EDTA titration (assay of magnesium sulphate), back

EDTA titration (assay of aluminum potassium sulphate), and replacement EDTA titration (assay

of calcium lactate

EDTA Titration Procedure

The typical steps involved in a complexometric titration using EDTA are as follows:

Step 1: Preparation of the Sample

• A known volume of the solution containing the metal ion is measured and transferred into

a clean Erlenmeyer flask.

Step 2: Addition of Buffer

• A buffer solution is added to maintain the pH at an optimal value for the metal-ligand

complex formation. For most metal ions titrated with EDTA, the pH is maintained between

4.5 and 7.5. The buffer ensures that the metal ions stay in the appropriate ionic form for

complexation.

Step 3: Addition of the Indicator

• A suitable indicator is added to the solution. The indicator is chosen such that it forms a

colored complex with the metal ion but changes color once the metal ion is complexed with

the EDTA.

Step 4: Titration with EDTA

 • The EDTA solution is added to the sample solution from a burette. As the EDTA is added,

it forms a complex with the free metal ions in the solution, displacing the indicator from

the metal, causing a color change.

Step 5: End Point Detection

• Endpoints in complexometric titration are often detected with the use of indicators. In some

cases, a visual or instrumental method may be used.

Step 6: Calculation of Metal Ion Concentration

• The amount of EDTA used to reach the endpoint is noted, and from this, the concentration

of the metal ion in the solution can be calculated using stoichiometry.

Factors Affecting Complexometric Titrations

Several factors influence the outcome of a complexometric titration:

a. pH: The pH of the solution is crucial because it affects the charge on the metal ion and the

ligand. EDTA forms stable complexes with metal ions in a specific pH range, and the

complexation efficiency can decrease outside this range.

b. Concentration of EDTA: The concentration of the EDTA solution must be known

precisely since it directly affects the accuracy of the titration.

c. Nature of the Indicator: The choice of indicator is essential for detecting the endpoint

accurately. The indicator should have a clear color change when the metal ion is displaced

by EDTA.

d. Temperature: The temperature can affect the complexation reaction. For instance, higher

temperatures may increase the rate of the reaction but can also influence the stability

constant of the metal-ligand complex.

 e. Interferences from Other Ions: Other metal ions or substances in the solution can

interfere with the titration, requiring additional steps such as masking agents to prevent

such interferences.

Indicators in Complexometric Titrations

Disodium edetate and its metal complexes are colourless. Therefore, sensitive indicators are

necessary to detect the endpoint in the titration of metallic salt solutions with edetate. Such

indicators are dyes which form weak complexes with metal ion, the complexes having a different

colour from a free indicator. A metal-indicator complex must be of lower stability than the

corresponding metal-edetate complex, so that the metal-indicator complex is formed only hen free

edetate is absent. Each indicator has a certain pH range over which it functions best. Hence, buffers

are often used in complexometric titration to maintain the pH at an optimum.

a. Eriochrome Black T (EBT): EBT is commonly used for the titration of calcium and

magnesium ions in water hardness determination. It forms a red complex with the metal

ions, which changes to blue when the metal is fully complexed by EDTA.

b. Calmagite: Calmagite is used for titrations of calcium and magnesium ions. It forms a pink

complex with calcium or magnesium ions, which changes to blue when the metal ions are

complexed by EDTA.

c. Murexide: Murexide is used for the titration of calcium and other metal ions. It forms a

purple complex with calcium, which changes to yellow when displaced by EDTA.

d. Sulfonphthalein Indicators: These are synthetic indicators that can be used in certain

complexometric titrations, where the color change is a result of the displacement of the

indicator from the metal ion by EDTA.

Applications of Complexometric Titration

a. Water Hardness Testing in Pharmaceutical Preparations: Many pharmaceutical

formulations (e.g., liquid syrups, injectable solutions, or oral solutions) are sensitive to

water quality. The hardness of water, due to dissolved calcium and magnesium salts, can

affect the stability and efficacy of these products. EDTA is used to titrate these ions, and

the amount of EDTA used is proportional to the hardness of the water

b. Determination of Metal Ions in Biological Samples: Complexometric titration is used to

determine essential metal ions such as calcium, magnesium, iron, and zinc in biological

fluids (e.g., blood serum, urine, or tissue samples)

c. Determination of Trace Metal Contaminants in Pharmaceutical Products: Certain

pharmaceutical products, especially those intended for intravenous use or sterile

formulations, need to be free from metal contaminants that can affect patient safety.

d. Determination of the Active Ingredient in Chelated Metal Complexes: Pharmaceutical

formulations containing chelated metal complexes (e.g., chelated iron, cobalt, or zinc)

require analysis to ensure the correct metal concentration.

e. Assay of Metal-based Drugs and Injectables: Some drugs, such as those containing

metals (e.g., copper sulfate, zinc sulfate, cobalt chloride, or manganese sulfate), require

precise measurement of the metal content for correct dosage.

f. Quality Control in Parenteral Preparations: Parenteral preparations (e.g., intravenous

(IV) fluids and injections) are particularly sensitive to contamination, including metals.

Accurate determination of metals such as zinc, copper, and iron is crucial for the safety and

efficacy of these preparations.

g. Analysis of Antacids Containing Magnesium and Calcium Carbonates

 h. Analysis of Soils and Fertilizers: The concentration of essential metal ions like potassium,

magnesium, and calcium in soils or fertilizers can be determined using complexometric

titration.

i. Environmental Monitoring: Complexometric titration is used to assess the concentrations

of toxic metals (such as lead, copper, and mercury) in environmental samples like water

and soil.

j. Determination of Metal Ions in Pharmaceutical Excipients: Excipients used in

pharmaceutical formulations (e.g., binders, stabilizers, fillers) may contain metal ions that

influence the drug's performance, especially in sensitive formulations.

Advantages of Complexometric Titrations

a. High Sensitivity: Complexometric titrations are highly sensitive and can detect low

concentrations of metal ions in a sample.

b. Versatility: A wide variety of metal ions can be titrated using complexometric methods,

including both transition metals and alkaline earth metals.

c. Accuracy: With appropriate indicators and buffer systems, complexometric titrations can

provide very accurate results.

d. Specificity: Complexometric titrations can be selective for specific metal ions, especially

when using masking agents to prevent interference from other ions.

Disadvantages of Complexometric Titrations

a. Interferences: The presence of other metal ions may interfere with the titration. In such

cases, masking agents are required to selectively block the interference of certain ions.
b. Complex Procedures: The process of selecting the appropriate indicator, buffer, and pH

conditions may be complex and require careful optimization for each metal ion being

analyzed.

c. Need for Proper Calibration: Precise preparation of the EDTA solution and calibration

against a standard is necessary to ensure accurate results.

FDA Approved Clinically Relevant Chelating Agents

1. Ethylenediaminetetraacetic acid (EDTA)

There are two different types of EDTA that have been formulated for pharmaceutical use:

EDTA complexed with calcium and EDTA without calcium. The formulation with calcium is

known as edetate calcium disodium, also known as disodium versenate. The formulation

without calcium is edetate disodium. One can easily be confused between these types of EDTA,

but the difference is critically important, as the formulation with calcium will not bind calcium,

while the one without calcium will bind calcium. Edetate calcium disodium was first approved

in 1953. It is available by prescription and is approved for the treatment of lead poisoning.

Adverse reactions associated with the use of edetate calcium disodium include acute renal

failure, mild increase in hepatic transaminases, hypotension, cardiac arrhythmias, and allergic

reactions.
 Disodium EDTA
EDTA metal complex
2. Penicillamine Penicillamine is a unithiol possessing only one thiol group. It is an oral

chelator and is available by prescription only. It is approved for the treatment of Wilson's

disease, which is chronic copper storage disease, cystinuria, and refractory rheumatoid

arthritis. It has also been used to treat lead poisoning, but that use has been off-label.

Penicillamine promote urinary excretion of excess copper , lead and mercury. Serious

hematological and renal adverse reactions have been associated with penicillamine

including leukopenia, thrombocytopenia, aplastic anemia, proteinuria, hematuria, and

nephrotic syndrome.

A. Penicillamine

B. Penicillamine metal complex

3. Deferoxamine, originally approved in the 1960s, is an intravenous chelator that is

approved for the treatment of acute iron poisoning and chronic iron overload due to

transfusion-dependent anemia (such as thalassemia) in the setting of numerous blood

transfusions. Adverse reactions associated with deferoxamine use include hypotension,

hypersensitivity reactions, ARDS, renal failure, and susceptibility to Yersinia infections.

Deferasirox and deferiprone are two oral iron chelators that are also approved for chronic

iron overload.

Deferoxamine

Deferoxamine metal complex