Aquametry

Aquametry can be defined as the quantitative

determination of water. The determination of water is one of
the most important and most widely practiced analysis in
pharmaceutical industry.
Importance of water determination
Quantitative determination of water is important because
many drugs contain water:
● As a solvent
● As absorbed water
● As water of crystallization
● As an adulterant
Physical properties of a drug or a raw material are
modified by its water content. Pharmaceutical procedures of
granulation, tablet formation & coating operations are affected
by water content.
Physical methods of water determination
a) Thermal method:
The technique includes the loss of weight by drying. Both
the BP and USP describe such measurement under the general
term “loss on drying”. The limitation of this method is that such
method also involve losses resulting from other volatile
materials or from decomposition.
These measurement can be made more specific by
limiting the decomposition effects at lower temperature, that is,
drying accomplished at reduced pressure.
Interference by other volatile materials can often be
controlled by a process of measuring the increase in weight of
an absorbent selective for water.
Absorption agents used for this purpose are:
• Dehydrite (anhydrous magnesium perchlorate)
• Drierite (calcium sulphate)
• Phosphorous pentaoxide
• Barium oxide
• Calcium chloride
• Anhydrous silica gel

An inert gas is allowed to carry the water lost from a

known quantity of sample to the absorbent whose gain in
weight is then determined.
Azeotrope

A mixture of two liquids that boils at constant

composition; i.e. the composition of the vapor is the same as
that of the liquid known as azeotrope.
Chloroform and acetone
Alcohol and water

Water (100o C) and toluene (110.6o C) form an

azeotrope with bp 84.1o C, the azeotrope contains 19.6%
water.
Azeotropic distillation method

1. A known weight of sample is placed in a flask with an

organic solvent such as xylene or toluene.

2. The flask containing the sample and the organic solvent is

attached to a condenser and the mixture is heated.

The organic solvent must be

- insoluble with water;
- have a higher boiling point than water;
- be less dense than water; and
- be safe to use.
Azeotrope

3. The water in the sample evaporates and moves up into the

condenser where it is cooled and converted back into liquid
water, which then trickles into the graduated tube.

4. When no more water is collected in the graduated tube,

distillation is stopped and the volume of water is read from the
tube.
b) Azeotropic distillation method (Dean and Stark trap):

The usual procedure is to add a water immiscible

solvent to the material containing moisture (water) and in this
manner to co-distill any water present.
Recondensation of the vapors results in separation of
water from the immiscible solvent making it available for
volumetric measurement.

The hydrocarbons benzene, toluene and xylene are the

solvents usually used in this determination.
 These solvents with a specific gravity less than 1 (or
density less than water), have the added advantage of allowing
the water to form a layer at the bottom of the Dean and Stark
trap, where it can be measured directly.

Fig: Dean - Stark apparatus

Reflux condenser

Dean-Stark apparatus

Glass flask Fig: Apparatus for

azeotropic distillation method
 Azeotropic distillation determination of moisture have
extensive applications because of their, simplicity, economy,
efficiency and accuracy.

The method is specially successful for moisture

determination in bulk materials, such as plant parts and for
medicinal soap solutions.

The main disadvantage to this procedure is that

relatively large samples are required, making the technique
unsuitable for trace amount of water in expensive
pharmaceutical materials.
Chemical methods of water determination
Karl Fischer titration
In 1935, German Chemist Karl Fischer described a specific
titrimetric method for the determination of water, remains the
most generally applicable procedure.
Karl Fischer titration is a widely used analytical method
for quantifying water content in a variety of products.
The fundamental principle behind it is based on the
Bunsen Reaction between iodine and sulfur dioxide in an
aqueous medium.

2H2O + SO2 + I2 → H2SO4 + 2HI

Composition of Karl Fischer reagent (USP)
Karl Fischer reagent is a mixture of-

Iodine 125 gm
Anhydrous pyridine 170 ml
Anhydrous methanol 670 ml
Liquid sulfur dioxide 100 ml

In recent years, pyridine, and its objectionable odor,

have been replaced in the Karl Fischer reagent by other amines,
particularly imidazole. These pyridine-free reagents are
available commercially for both volumetric and coulometric Karl
Fischer procedures.
 1 ml of Karl Fischer reagent when freshly prepared will
react with about 5 mg (or 3-6 mg) of water.

When prepared it is general practice to increase the

stability of the reagent by adding sulfur dioxide to a stock
solution of the other components the day before actual use.

Numerous side reactions may occur among the

constituent substances. Freshly prepared reagent therefore has
a strength about 80% of the theoretical value, but this rapidly
falls to about 50% in 1 month and 40% in 3 months.
Chemistry of the reaction

In the presence of water, iodine will be reduced and

sulfur dioxide oxidized in the following manner:

H2O + I2 + SO2 2HI + SO3

The reversibility of the reaction can be prevented by

using large quantity of pyridine. The concentration of pyridine
is sufficiently large so that I2 and SO2 are complexed with the
pyridine as C5H5N  I2 & C5H5N  SO2.

rapid
C5H5N  I2 + C5H5N  SO2 + H2O + C5H5N 

2 C5H5N  HI + C5H5N  SO3 ------------ (1)

 The pyridine sulfur trioxide (pyridinium sulfite)
compound, an inner salt, reacts, in turn, with the methanol
present to form the pyridine salt of methyl sulfate.

C5H5N  SO3 + CH3OH  C5H5N+(H)-SO4CH3

------------- (2)

Thus, methanol prevent the further reaction of

C5H5N  SO3 with water.

C5H5N  SO3 + H2O C5H5N+(H)-SO4H

The last reaction is undesirable because it is not specific

for water. It can be prevented by using excess amount of
methanol.
 The primary reaction (1) occurs rapidly and permits the
direct titration of any available water with the reagent.

Karl Fisher determination can be performed by direct

titrations or excess of the reagent can be added and the excess
can be back titrated with a standard water-in-methanol
solution.

For a direct titration, methanol is added to dissolve the

sample or to assist in the penetration of an insoluble sample. In
back titration procedure, the reagent alone often serves this
purpose.
Instrument
The titration vessel is fitted with 1.5- or 2- volt dry cell
across a variable resistance of about 2000 ohoms (2000 ),
which is in series with two platinum electrodes and a
microammeter (A), a mechanical stirrer and burette.

Burette
Dry cell

Resistance

Fig: Karl Fischer instrument Platinum electrode

End point detection
An end point in a Karl Fischer titration can be observed
visually based on the color change from pale yellow to dark
brown color of the excess reagent (volumetric titration).

An amerperometric or electrometric detection of the

Karl Fisher titration end point is employed and found useful
specially when the sample is colored (coulometric titration).

Under these conditions of constant low voltage with the

direct titration procedure, there is a small constant residual
flow of current until the end point is reached, accompanied by
a large increase in current.
 Thus, when water is titrated with the Karl Fisher reagent
there is a “kick-off” or major deflection in the microammeter to
indicate the end point when last drop of excess Karl Fisher
reagent enter the titration flask.

Conversely, when a back titration is employed, there is a

sudden drop in current or a “dead-stop” end point occurs.
 Up to the end point in the direct titration of water with
Karl Fisher reagent, there are iodide ions present but no free
iodine.

At the potential used, the system is irreversible and

the electrodes are polarized, that is, they have an impressed
potential with a little flow of current.

However, as the free iodine enters the system there is

a reversible iodine–iodide couple established with the
depolarization of electrodes and an increase in the flow of
current.
Limitations
The Karl Fischer reagent is highly specific for water but there are
some limitations-
 Compounds which react with either iodine or iodide
will interfere the process. For example, ascorbic acid will be
oxidized by the iodine present in the reagent.

 Carbonyl compounds under the conditions of the Karl

Fischer determination react with methanol with the formation of
acetals or ketals and the liberation of water.
R2CO + 2CH3OH R2C(OCH3)2 + H2O

 The optimal pH range for the Karl Fischer reaction is

from 5 to 8, and highly acidic or basic samples need to be
buffered to bring the overall pH into that range.
Advantage of analysis

The popularity of the Karl Fischer titration is due in large

part to several practical advantages that it holds over other
methods of moisture determination, including:

* High accuracy and precision

* Selectivity for water
* Small sample quantities required
* Easy sample preparation
* Short analysis duration
* Nearly unlimited measuring range (1ppm to 100%)
Advantage of analysis
* Suitability for analyzing:
o Solids
o Liquids
o Gases
* Independence of presence of other volatiles
* Suitability for automation

In contrast, loss on drying will detect the loss of any

volatile substance.

Reference: Karl Fischer titration. (n.d.) In Wikipedia, the free encyclopedia online. Retrieved from

http://en.wikipedia.org/wiki/