East West University

Aquametry
PHRM 309

Tareq Hasan 8/11/2011

Table of Contents
Aquametry..............................................................2 Introduction ..........................................................2 Importance of Water Determination ................2 Methods of Water Determination ....................2 Thermal Methods of Water Determination ..........2 Loss on Drying ...................................................2
Theory ..................................................................2 Factors Affecting Weight Loss of Sample in Loss on Drying ..............................................................3 Modification of Loss on Drying Technique for Specific Water Determination ..............................3 Overcoming the problem of Degradation of Sample due to High Heat .................................3 Overcoming the problem of Interference by Volatile Materials present in Sample ...............3 Limitation of Loss on Drying .................................3 Justification for Using Pyridine and Methanol in Karl Fischer Reagent ............................................ 8 Justification for Using Anhydrous Pyridine ..... 8 Justification for Using Anhydrous Methanol ... 9

Classification of Karl Fischer Titration ...............9

Volumetric Karl Fischer Titration ......................... 9 Purpose of Performing Volumetric Karl Fischer Titration .......................................................... 9 Volumetric Karl Fischer Direct Titration .......... 9 End Point Detection (Volumetric Karl Fischer Direct Titration)............................ 10 Volumetric Karl Fischer Back Titration .......... 10 End Point Detection (Volumetric Karl Fischer Back Titration) .............................. 11 Purpose of Performing Volumetric Karl Fischer Back Titration ............................... 11 Coulometric Karl Fischer Titration ..................... 11 Purpose of Performing Coulometric Karl Fischer Titration ............................................ 12 Instruments for Coulometric Karl Fischer Titration ........................................................ 12 Coulometric Karl Fischer Direct Titration ...... 13 End Point Detection (Coulometric Karl Fischer Direct Titration)............................ 13 Coulometric Karl Fischer Back Titration ........ 14 End Point Detection (Coulometric Karl Fischer Back Titration) .............................. 14

Azeotropic Distillation .......................................4

Azeotrope / Azeotropic Mixture...........................4 Principle of Azeotropic Distillation Method .........4 Criteria of Organic Solvents in Azeotropic Distillation Method ..............................................4 Procedure of Azeotropic Distillation Method (Dean Stark Trap) ...............................................5 Advantage of Azeotropic Distillation Method ......6 Limitations of Azeotropic Distillation Method .....6

Chemical Method of Water Determination ..........6 Karl Fischer Titration .........................................6

Theory ..................................................................6 Karl Fischer Reagent .............................................6 Composition of Karl Fischer Reagent ...............6 Problem with Karl Fischer Reagent and the Solution.......................................................7 Chemistry of Karl Fischer Titration .......................7 Primary Condition of Titration .........................7

Comparison between Different Karl Fischer Titration (KFT) Method ...................................15 Advantages of Karl Fischer Titration ...............16 Limitations of Karl Fischer Titration ................16

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Aquametry
In tro d u cti o n

Methods of Water Determination

Methods of Water determination can be of 2 types 1. Thermal Methods Loss on Drying Azeotropic Distillation Method 2. Chemical Methods Karl Fischer Titration
Ther m al Method s of W ate r Det e rm ina tion

Aquametry is a quantitative method of determining water content present in a sample. Determination of water is one of the most important and widely practiced analytical methods in pharmaceutical industry.

Importance of Water Determination

Quantitative Determination of Water in pharmaceutical products is important

Loss on Drying Theory

Loss on Drying involves the loss of weight of a material upon drying. The lost weight is usually the weight of water and Volatile materials present in the sample. So,

because H2O may be present in the products as Solvent (E.g. H2O in Syrup, Suspension or Emulsion) Absorbed water (E.g. Absorbed H2O by Powder for Suspension) Water of Crystallization (E.g. Crystals of salts containing H2O) Adulterant (E.g. Excess Water in

Digitalis Leaves) This is important, because Physical of a drug or raw material is modified by the presence of H2O. Pharmaceutical Granulation, Procedures formation of and

Both BP and USP have specified the temperature and time for drying for a fixed amount of Material / Sample and also the amount of lost weight in percentage. E.g. BP has specified that At least 5% weight will be lost for 1 gm

Tablet

coating operations are affected by H2O Content.

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Paracetamol

dried

at

120 0C

sample at a lower temperature and reduced pressure environment. Overcoming the problem of Interference by Volatile Materials present in Sample Interference by volatile material can be controlled by using water selective absorbents such as Anhydrous (Dehydrite) CaSO4 Phosphorous pentaoxide (PO5) Barium oxide CaCl2 Anhydrous Silica Gel In this case, an inert gas is used to carry the formed H2O vapor to the absorbents. So, Mg perchlorate

temperature for 2 hours.

Factors Affecting Weight Loss of Sample in Loss on Drying

Loss on Drying is not specific for H2O determination, since weight loss of material upon drying depends on the following factor Evaporation of Moisture / H2O Evaporation of Volatile material present in the sample Degradation of the Material / Sample due to High Heat

Modification of Loss on Drying Technique for Specific Water Determination

The usual loss on drying technique can be modified for more specific and accurate measurement of H2O by overcoming the factors which affect Loss of weight in Loss on Drying except H2O Evaporation. These are 1. Degradation of Sample due to High heat 2. Interference by Volatile Material Overcoming the problem of Degradation of Sample due to High Heat Degradation of Sample due to high heat can be overcome by drying the

Limitation of Loss on Drying

The Loss on drying process is not specific for water, because weight loss of sample occurs also due to Degradation of Sample due to High heat Interference by Volatile Material

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 Sample totally degrades due to high heat involved and cannot be reused. So, it is not good for expensive materials. The process is time consuming.

A known weight of sample is placed in flask with an organic solvent.

Azeotropic Distillation Azeotrope / Azeotropic Mixture

Azeotrope is a mixture of 2 liquids which boils at a constant Temperature regardless of the boiling points of the individual liquids in the mixture. Ratio of Composition as in their liquid forms and as in their formed vapor due to evaporation. E.g. Water (110.6 C)
0 0

Criteria of Organic Solvents in Azeotropic Distillation Method

Solvents used in Azeotropic Distillation Method must have

the following criteria:

Solvent must be water immiscible and insoluble Density of the solvent must be lesser than H2O. Boiling point of solvent must be higher than Water. The solvent must be safe to use.

(100 C) mixture

Toluene forms an The flask containing the sample and the solvent is attached to a condenser and the mixture is heated. Upon heating, the water in the sample evaporates and moves up into the condenser where it is cooled and condensed back into water which is then deposited into the graduated tube. When all of the water in sample is collected into the tube, the distillation process is stopped and the volume of

Azeotrope with a boiling point 84.10C and the Azeotrope contains 19.6% H2O. Water (100 0C) Xylene (139.10C) mixture forms an Azeotrope with a boiling point 94.50C and the

Azeotrope contains 40% H2O. Water (100 0C) CCl4 (76.80C) mixture forms an Azeotrope with a boiling point 66.80C and the

Azeotrope contains 4.1% H2O.

Principle of Azeotropic Distillation Method

the water is read directly from tube.

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Procedure of Azeotropic Distillation Method (Dean Stark Trap)

Azeotropic Distillation method follows the addition of a water insoluble and immiscible organic solvent to the Sample containing water / moisture and in this manner to co distill any water present. Upon heating, the water present in sample will evaporate at first, since Boiling point of organic solvent is higher than Water. It continues until the ratio of

temperature and evaporate at the same Azeotropic composition.

composition for Azeotropic mixture is achieved.

Figure 2: Apparatus used in Azeotropic Distillation

Evaporated solvents will be condensed in the condenser and go downwards to deposit in the graduated tube of the Dean Stark Trap.
Figure 1: Dean - Stark Trap

Since, organic solvent is less dense than water it will deposit above the water composition is

When

Azeotropic

layer and volume of the water can be measured directly.

achieved, both water and the organic solvent will start to boil at a constant

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Advantage of Azeotropic Distillation Method

Azeotropic Distillation method for

of determination of water in the field of pharmaceutical analysis. The fundamental principle behind it is based on the Bunsen reaction

water determination is popular because of being Accurate An Easy Method for Water

between I2 and SO2 in an aqueous medium.

Determination An Efficient method Economic This method moisture is advantageous for in bulk

Karl Fischer Reagent

Karl Fischer Reagent is the titrant used in the Karl Fisher Reagent to determine the amount of H2O in the Sample / Analyte. Composition of Karl Fischer Reagent Karl Fischer Reagent is a mixture containing components
Table 1: Composition of Karl Fischer Reagent

determination

materials such as plant parts, soap solution etc.

Limitations of Azeotropic Distillation Method

Large amount of samples are needed. Applicable amount of for sample with (E.g. large Syrup,

the

following

moisture

Suspension, Plant Materials). Trace amount detected. Sample totally degrades due to high heat and cannot be reused.
Chem ica l Me th o d o f W ate r Det e rm in a tion

of

moisture

cannot

be Component Iodine Anhydrous Pyridine Amount 125 gm 170 ml

Anhydrous Methanol 670 ml Liquid Sulfur Dioxide 100 ml

Karl Fischer Titration Theory

Karl Fischer Titration method is the most widely used and accepted method

Recently, the pyridine has been replaced due to its objectionable odor by amine compounds such as Imidazole. 6|Page

containing Pyridine Methanol

I2, and

Anhydrous Anhydrous before

promptly

performing the titration.

Figure 3: Pyridine & Imidazole

Chemistry of Karl Fischer Titration

In presence of water from the sample I2 and SO2 from Karl Fischer Reagent will be reduced and oxidized respectively in the following manner

1 ml of Karl Fischer Reagent is equivalent to 5 mg (or 3 6 mg) water. Problem with Karl Fischer Reagent and the Solution Problem: If all of the

components of Karl Fischer Reagent are premixed way long before titration, then

Primary Condition of Titration

The primary condition of

performing titration is that all reactions must be Irreversible or Unidirectory Rapid

components reacts with each other resulting in numerous side reactions and as a

consequence, the effectiveness of the reagent reduces. This reversible and slow reaction becomes unidirectory and rapid by the
A Freshly prepared Reagent has strength about 80% of theoretical value. A 1 month old reagent has 50% strength. A 3 months old reagent has 40% strength.

large quantity of pyridine present in the Karl Fischer Reagent which can form complexes with I2 and SO2 as C5H5N.I2 and C5H5N.SO2 respectively and the reaction will occur in the following manner

Solution: This problem can be overcome by adding the Liquid SO2 to the stock of solution 7|Page

Justification for Using Pyridine and Methanol in Karl Fischer Reagent

Justification for Using Anhydrous Pyridine
In presence of water from the sample, I2 and SO2 from Karl Fischer

The formed Pyridinium Sulfite, an inner salt reacts with water from sample giving Pyridinium Salt of Hydrogen Sulfate. As a result, H2O does not react with iodine in the main Bunsen reaction.

Reagent will be reduced and oxidized respectively in the following manner

Large quantity of pyridine present in the Karl Fischer Reagent forms complexes with I2 and SO2 as C5H 5N.I2 and C5H5N.SO2 respectively. As a result, the reaction becomes Irreversible or Unidirectory Rapid

Presence

of

large

amount

of

the reaction

will occur

in the

Anhydrous Methanol in the Karl Fischer Reagent prevents the reaction of water with Pyridinium sulfite by forming Pyridinium salt of methyl sulfate.

following manner

For this, Anhydrous Pyridine is used in the Karl Fischer Reagent.

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Justification for Using Anhydrous Methanol

Due to the presence of Pyridine in the Karl Fischer Reagent, an inner salt Pyridinium sulfite is formed.

Classification of Karl Fischer Titration

Karl Fischer Titration can be of 2 type 1. Volumetric Karl Fischer Titration 2. Coulometric Karl Fischer Titration

Volumetric Karl Fischer Titration

Volumetric Karl Fischer Titration can be defined as a method of titration where an exact volume of Karl Fischer Reagent is consumed during the course of
Pyridinium sulfite reacts with water from sample giving pyridinium salt of hydrogen sulfate. As a result, H2O does not react with iodine in the main Bunsen reaction.

titration from which equivalent amount of Water present in the sample can be detected; i.e. 1 ml of Karl Fischer Reagent is equivalent to 5 mg (or 3 6 mg) water. End point in this titration is detected by color change in the solution. Volumetric Karl Fischer Titration can be of 2 type

Presence

of

large

amount

of

1. Volumetric Titration 2. Volumetric Titration

Karl

Fischer

Direct

Anhydrous Methanol in the Karl Fischer Reagent prevents the

reaction of water with Pyridinium sulfite by forming Pyridinium salt of methyl sulfate.

Karl

Fischer

Back

Purpose of Performing Volumetric Karl Fischer Titration Volumetric Karl Fischer Titration is performed when the sample is not

For this, Anhydrous Methanol is used in the Karl Fischer Reagent.

colored. Volumetric Karl Fischer Direct Titration

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 In Volumetric Karl Fischer Direct Titration, sample is dissolved in excess anhydrous methanol and the solution is then filtered to remove impurities. Then, Karl Fischer Reagent is added to the solution drop by drop with the help of a burette; thus, forming a pale yellow solution. When all of the H2O present in the solution will react with Karl Fischer Reagent, the color of the solution will suddenly change into Dark Brown from pale yellow; thus indicating the End Point of the Titration. End Point Detection (Volumetric Karl Fischer Direct Titration) An End Point in Volumetric Karl Fischer Direct Titration can be observed visually based on the color change from Pale Yellow to Dark Brown color of the excess Karl Fischer Reagent.

Colorless Solution of Sample in Anhydrous Methanol Addition of Karl Fischer Reagent drop by drop

No Color

Formation of Pale Yellow Liquid

End - Point

Change of Clolor from Pale Yellow to Dark Brown

Figure

4:

Diagrammatic

Representation

of

Detection of End Point in Volumetric Karl Fischer Direct Titration

Volumetric Karl Fischer Back Titration In Volumetric Karl Fischer Back Titration, at first the sample is mixed with excess Karl Fischer Reagent giving the solution a Dark Brown color of excess Karl Fischer Reagent, since all of the water in the sample has already reacted with the Karl Fischer Reagent. Then, A Standard Water in Methanol is added drop by drop to the solution with the help of burette. When all of the H2O in Methanol reacts with Excess Karl Fischer Reagent, the color of the solution

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will suddenly change into Pale Yellow from Dark Brown; thus indicating the End Point of the Titration. End Point Detection (Volumetric Karl Fischer Back Titration) An End Point in Volumetric Karl Fischer Back Titration can be observed visually based on the color change from Dark Brown of excess Karl Fischer Reagent into Pale Yellow.

Purpose of Performing Volumetric Karl Fischer Back Titration Purpose of Performing

Volumetric Karl Fischer Back Titration is to determine the accuracy of the Volumetric Karl Fischer Direct Titration. After the reaction in Volumetric Karl Fischer Direct Titration is complete, the excess amount of Karl Fischer Reagent is

determined by titration with a standard H2O in Methanol Solution. The Actual Amount of Karl Fischer Reagent reacting with desired amount of Water is

Solid Sample

No Color

Addition of Karl Fischer Reagent in Excess Amount

calculated by subtracting the

Dark Brown Color of Excess Karl Fischer Reagent

volume consumed in the Back Titration from the volume

added in the Direct Titration.

Addition of Standard Water - in Methanol Reaction of Water with Excess Karl Fischer Reagent

End - point

Sudden Color Change into Pale Yellow from Dark Brown

Coulometric Karl Fischer Titration

Figure 5: Diagrammatic Representation of Detection of End Point in Volumetric Karl Fischer Back Titration

Coulometric Karl Fischer Titration can be defined as a method of titration where an exact volume of Karl Fischer 11 | P a g e

Reagent is consumed during the course of titration from which equivalent amount of Water present in the sample can be detected; i.e. 1 ml of Karl Fischer Reagent is equivalent to 5 mg (or 3 6 mg) water. End point in this titration is detected by sudden change in the electricity current flow. Coulometric Karl Fischer Titration can be of 2 type 1. Coulometric Karl Fischer Direct Titration 2. Coulometric Titration Karl Fischer Back

Purpose of Performing Coulometric Karl Fischer Titration Coulometric Karl Fischer Titration is performed when the sample is colored. As a result the End Point cannot be detected by color change. Instruments for Coulometric Karl Fischer Titration The Coulometric Karl Fischer

Titration vessel is fitted with 1.5 2.9 V Dry cell across a variable resistance of about 2000 which is in series with two platinum

electrodes and a microammeter, mechanical stirrer and a burette.

Figure 6: Instruments for Coulometric Karl Fischer Reaction

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Coulometric Karl Fischer Direct Titration In Coulometric Karl Fischer Direct Titration, at first sample is dissolved in excess anhydrous methanol and the colored solution is then filtered to remove impurities. Then, Karl Fischer Reagent is added to the solution drop by drop with the help of a burette. As a result the following reaction occurs, assuming that pyridine is present in the Karl Fischer Reagent making the reaction rapid and unidirectory.
Colored Solution of Sample in Anhydrous Methanol

which indicates the End point of the Titration. End Point Detection (Coulometric Karl Fischer Direct Titration)

Colored Solution

Addition of Karl Fischer Reagent drop by drop

Flow of low voltage of current within the circuit observed through the microammeter due to the presence of only Iodide Ion

Under these conditions, a constant low voltage of current flows within the circuit observed through the microammeter due to the presence of only Iodide ion (I) from HI in the reaction medium.

End - Point / Kick-off point

Maximum Increase in flow of Current from Very Low Current due to the presence of I2/ICouple

Figure 7: Diagrammatic Representation of End point Detection in Coulometric Karl Fischer Direct Titration

An End Point in Coulometric When all of the H2O water reacts with reagent, free I2 enters the system and reversible Iodine / Iodide couple / (I2/I ) completes the circuit resulting in a sudden increase in the flow of current,

Karl Fischer Direct Titration can be observed visually on the Microammeter when the flow of current suddenly increases to maximum current flow. from very low

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 This point is also called Kick off Point. Coulometric Karl Fischer Back Titration In Coulometric Karl Fischer Back Titration, at first the sample is mixed with excess Karl Fischer Reagent. As a result All of the water in the sample reacts with the Karl Fischer Reagent. The reaction medium contains I2/I Couple due to the

End Point Detection (Coulometric Karl Fischer Back Titration)

Solid Sample

No Color

Addition of Karl Fischer Reagent in Excess Amount

Reaction of Water and Karl Fischer Reagent

presence of free excess Iodine and the Iodide ion as a result of reaction between iodine and water. The Microammeter shows

Presence of I2/I Couple due to free excess Iodine and the Iodide ion in Reaction Medium

Maximum Flow of Current

Maximum Flow of Current. Then, A Standard Water in Methanol is added drop by drop to the solution with the help of burette. When all of the H2O in Methanol reacts with Excess Karl Fischer Reagent, the excess Iodine is reduced into Iodide ion. As a result, there will be a sudden drop of flow of current from the maximum; thus indicating the End Point of the Titration.

Addition of Standard Water - in Methanol

Reaction of Water with Excess Karl Fischer Reagent

End - point / Dead Stop Point

Sudden drop of Flow of Current

Figure 8: Diagrammatic Representation of End Point Detection in Coulometric Karl Fischer Back Titration

An End Point in Coulometric Karl Fischer Back Titration can 14 | P a g e

be observed visually on the Microammeter when the flow of current suddenly drops down from maximum current flow.

This point is also called Dead Stop Point.

Comparison between Different Karl Fischer Titration (KFT) Method

Table 2: Comparison between Different Karl Fischer Titration (KFT) Method

Types of Volumetric KFT KFT General

Coulometric KFT End point Detection is depended on Change of flow of current Performed for both colored and colorless samples Process is automated Instruments used is complex Initial Solution of Sample and Anhydrous Methanol is colored Addition of Karl Fischer Reagent Drop by Drop results in a constant flow of current at low voltage At End point, Flow of current suddenly changes into maximum flow of current. End point appears when all of the Water reacts with Karl Fischer reagent, so that excess Free Iodine and Iodide Ion forms (I2/I) Couple. End point is called Kick off Point. Use of Methanol to dissolve the sample is not necessary Titrant is the Standard Water in methanol. Excess Karl Fischer Reagent is used to dissolve the sample Solution shows a max flow of current at initial stage, because excess Free Iodine and Iodide Ion forms (I2/I) Couple and completes the circuit At End point, Flow of current suddenly changes from maximum flow of current into minimum. End point is called Dead Stop Point. 15 | P a g e

End point Detection is depended on Color Change Performed only for colorless samples Process is manual Instruments used is simple Initial Solution of Sample and Anhydrous Methanol is colorless Addition of Karl Fischer Reagent Drop by Drop forms a pale yellow liquid in the solution At End point, Pale Yellow Color suddenly changes into Dark Brown Color of Karl Fischer Reagent. End point appears when all of the Water reacts with Karl Fischer Reagent to show the Dark Brown Color of the Reagent. End point has no other synonym. Use of Methanol to dissolve the sample is not necessary Titrant is the Standard Water in methanol Excess Karl Fischer Reagent is used to dissolve the sample Solution forms a dark brown color liquid at initial stage, because all of the H2O reacts with the Karl Fischer Reagent. At End point, Dark Brown Color of Karl Fischer Reagent changes into Pale Yellow Color suddenly. End point has no other synonym.

Direct Titration

Back Titration

Advantages of Karl Fischer Titration

Karl Fischer Titration Method is very popular for a large practical advantage which it holds over other moisture

2. Carbonyl

Compounds

can

react

with

Methanol to form Acetals of Ketals and the liberation of water.

determination technique, These includes Easy Sample Preparation High Accuracy and Precision Independence of presence any volatile materials Nearly Unlimited Measuring Range Selectivity for Water Short Analysis Duration Small Sample Quantities required Suitability for analyzing Solids Liquids Gases Suitability for Automation 3. The optimal pH range for the Karl Fischer Reaction is 5 8. So, highly acidic / basic samples need to be buffered to bring the overall pH into this range.

Limitations of Karl Fischer Titration

1. Presence of Compound which reacts with Iodine or Iodide will interfere in the process. E.g. Ascorbic Acid will be oxidized by Iodine present in the reagent. Quinone will be reduced by Iodine formed during the reaction.

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