AQUAMETRY

Course Instructor: Marjana Khalil

 What is Aquametry?
Aquametry can be defined as the quantitative determination of water
present in any samples or products.
Methods/Processes include Karl Fischer titration, Reactions with acid
chlorides and anhydrides, Oven drying, Distillation and Chromatography.
Samples can contain water:
 as solvent
 as an absorbed water
 as water of crystallization
 Importance of Moisture Determination
(Aquametry)
1. When dealing with expensive materials and even with large quantities
of less costly materials, water can be considered an adulterant.
2. Physical properties of a material are modified by its water content.
3. Physical stability of the product may be hampered in presence of
water.
4. Many pharmaceutical procedures such as granulation, tablet
formation and coating operation are also affected by water content. 3
major problems in tablet manufacturing: capping, picking and sticking
may arise due to moisture content.
5. Many products are hydrolyzed in presence of moisture.
6. Microbial growth is also potentiated by the presence of moisture.
7. Many chemical reactions involve water either as a reactant or product.
Such process can be followed or a reactant can be assayed by the
quantitative determination of water. Such as:
 Methods of Measuring of Moisture Content
A. Gravimetric methods  Spectroscopic method
B. Azeotropic method E. Other chemical methods:
C. Chemical methods: Karl Fischer Titration  By using inorganic reagents
methods
By using organic reagents
D. Other physical methods:
 Gas chromatographic method  By colorimetric procedures
 Electrogravimetric method etc
 Gravimetric method
• Gravimetric method can be of 2
types:
1. Thermal method:
• This technique is based on loss of
weight upon drying.
• Procedure:
• Substance (sample) → Heat →
Loss of weight → Lost weight is
the amount of moisture.
2. By using absorbent:
In this method, water absorbents are used
to absorb moisture present in the sample.
Then the gained weight of absorbent is
determined which is actually the amount
of moisture of that sample.
Absorbents that have been classically used
for this purpose include:
Anhydrous Mg perchlorate (dehydrate)
,BaO2 , P2O5 + H2SO4 mixture
CaCl2,CaSO4
 Azeotropic method
• In this method, water-immiscible solvent
(Benzene, Toluene or Xylene) is added to the
material containing moisture and then co-
distilled any water present. Recondensation of
the vapors results in the separation of water
from the immiscible solvent, making it
available for the volumetric measurement. It is
also known as co-distillation method.
• These method is specified for the
determination of water content more than 2%.
• The solvents with a specific gravity less than
one have the added advantage of allowing the
water to form a layer at the bottom of the trap,
where it can be measured directly.
 Azeotropic method
• Advantage:
Recovery of solvent is possible.
Useful for moisture determinations in bulk material such
as plant parts and for aqueous solutions such as medicinal
soap solutions.

• Disadvantage:
Not applicable for the heat sensitive materials.
Not applicable for small amount of samples.
 What is Karl Fischer Titration?
The fundamental
The principle behind
fundamental principle behind this
this method
method isis based
based onon the
the reaction
reaction between
between iodine
iodine and
and sulfur
sulfur
dioxide in
dioxide in an
an aqueous
aqueousmedium.
medium.
H2O + I2 + SO2 → 2HI + SO3
H2O + I2 + SO2 → 2HI + SO3
It was the German chemist Karl Fischer who discovered in 1935 that this reaction could be modified to
It
bewas thefor
used German chemist Karl Fischer
the determination whoindiscovered
of water in 1935system
a non-aqueous that thiscontaining
reaction could be modified
an excess to
of sulfur
be used for
dioxide. He the
useddetermination of water
a primary alcohol in a non-aqueous
(methanol) systemand
as the solvent, containing
a base an excess of
(pyridine) assulfur dioxide.
the buffering
agent.
He used a primary alcohol (methanol) as the solvent, and a base (pyridine) as the buffering agent.
ROH ++ SO
ROH SO22 ++ RʹN
RʹN → → [RʹNH]SO
[RʹNH]SO33RR
(alcohol) (base)
(alcohol) (base)(alkylsulfite
(alkylsulfitesalt)
salt)
[RʹNH]SO33RR ++ H
[RʹNH]SO H2OO ++II22++22RʹN
RʹN→→[RʹNH]SO
[RʹNH]SO44RR++22[RʹNH]I
[RʹNH]I
2

(water)(water)
(iodine) (iodine) (alkylsulfate
(alkylsulfate salt) (hydroiodic
salt) (hydroiodic acid salt)
acid salt)
The alcohol (typically methanol) reacts with sulfur dioxide (SO2) and base to form an intermediate
The alcohol salt,
alkylsulfite (typically
whichmethanol) reacts with
is then oxidized sulfur dioxide
by iodine (SO 2) and base
to an alkylsulfate salt. to form
This an intermediate
oxidation reaction
consumes water.
alkylsulfite salt, which is then oxidized by iodine to an alkylsulfate salt. This oxidation reaction consumes
water.
Classic Karl Fisher reagents contained pyridine, a noxious carcinogen, as the base. The reagents most
frequently
Classic Karlused
Fishertoday are pyridine-free
reagents and contain
contained pyridine, imidazole
a noxious or primary
carcinogen, amines
as the base.instead.
The reagents most
frequently used today are pyridine-free and contain imidazole or primary amines instead.
 What is Karl Fischer Reagent?
• A typical Karl Fischer reagent is a mixture of –
1. 125 gm Iodine
2. 170 mL Pyridine
3. 670 mL methanol
4. 100 mL of liquid SO2

Mechanism of action of a Karl Fischer Titrator:

• Water and iodine are consumed in a 1:1 ratio in the above reaction. Once all of the
water present is consumed, the presence of excess iodine is detected voltametrically
by the titrator’s indicator electrode. That signals the end-point of the titration.
• The amount of water present in the sample is calculated based on the concentration
of iodine in the Karl Fisher titrating reagent (i.e., titer) and the amount of Karl Fisher
reagent consumed in the titration.
Effect of pH in Karl Fischer Titration:
• 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.
• When the pH is lower than 5, the titration speed is very
slow. On the other hand, when pH is higher than 8,
titration rate is fast, due to an interfering esterification
side reaction which produces water, resulting in a
vanishing endpoint.
 Types of Karl Fischer titration

There are 2 types of Karl Fischer titration

1. Volumetric KFT and 2. Coulometric KFT

Volumetric KFT
• In this method, iodine is added mechanically to a solvent containing the sample by
the titrator’s burette during the titration. It detects the endpoint of the titration
using the double platinum pin indicator electrode. Water is quantified based on the
volume of Karl Fischer reagent consumed. Volumetry is best suited for determination
of water content in the range of 100 ppm to 100%.
Coulometric KFT
• In coulometric Karl Fischer, iodine is generated
electrochemically (in situ) just in time from iodide during the
titration.
• In Cathode, 2H+ + 2e → H2
• In Anode, 2I– → I2 + 2e
• Water is quantified based on the total charge passed (Q), as
measured by current (amperes) and time (seconds), according
to the following relationship:
• Q = 1 C (Coulomb) = 1 A x 1 s [where, 1 mg H2O = 10.72 C]
• Coulometry is best suited for determination of water content
in the range of 1 ppm to 5%.
The Coulometric System

• Micro Detection System

• “Absolute Method”
• High Water Capacity Reagents
• Designed for Titration of Liquids & Gases Only
• Less System Flexibility
• Co-Solvents are limited
• No Option for Temperature Modification
• No Option for Homogenizer
There are two main types of coulometric KFT reagent systems:
a. Fritted-cell, coulometric KF:
In fritted-cell, a diaphragm – or frit – separates the anode from
the cathode that form the electrolytic cell known as the
generator electrode. The purpose of the frit is to prevent the
iodine generated at the anode from being reduced back to
iodide at the cathode instead of reacting with water.  
b. Fritless-cell, coulometric KF:
In fritless-cell coulometric KF, an innovative cell design is used
that through a combination of factors, but without a frit, makes
it nearly impossible for iodine to reach the cathode and get
reduced to iodide instead of reacting with water.
Fritted-cell, coulometric KF

Anode compartment filled Titration begins. Note H2

Cathode compartment with anolyte solution formation (bubbles) at
filled with catholyte I2 formation occurs at the the cathode:
solution
anode: 2I- I2 +2e- 2H+ + 2e-  H2(g)
Fritless-cell, coulometric KF
Anolyte and Catholyte
• An Anolyte is the electrolyte on the anode side of an
electrochemical cell that is divided into compartments.
• A catholyte is the part of an electrolyte which is on the cathode
side of an electrochemical cell that is effectively divided into
two compartments.
• Catholytes have various uses, including:
• Reducing surface tension of water, and hence conditioning
of water to improve oil production in wells.
• Reducing microbe contamination along with anolytes.
• Used as detergents or cleaning agents in the food and
beverage industry.
The Volumetric System
In volumetric Karl Fischer, iodine is added mechanically to a
solvent containing the sample by the titrator’s burette during the
titration. Water is quantified on the basis of the volume of Karl
Fischer reagent consumed.

Designed for Higher Water Concentrations

• Fast Titration (30 mg Water per minute)
• Designed for Titration of Solids, Liquids & Gases
• System Flexibility
• Modified Solvent Systems
• Temperature Adjustment
• Peripheral Equipment (Homogenizer, Oven)
One Component System
• The one component systems are the most popular. The benefit of a one
component volumetric titration is that it has almost unlimited water
capacity. One-component volumetric reagents are easier to handle, and are
usually less expensive than two-component reagent.

Titrant
Contains all reactants (SO2, I2
and base) except the water

Sample solvent
Methanol or other solvent
to which sample is added
Two Component System
The benefits of a two component volumetric titration are:
• Higher titration speed
• Greater accuracy for small amounts of water
• Higher buffer capacity
• Usually more costly, and have lower solvent capacity .

Titrant
• Exact and stable titer
Contains I2 in methanol or
ethanol

Sample solvent
Contains SO2 and base in
methanol or ethanol
Bipotentiometric titration:
Measuring I2
• The second reaction in the K-F titration – the one that actually
consumes the water – is a redox reaction:
[RNH]SO3CH3 + I2 + H2O +2RN [RNH]SO4CH3 + 2[RNH]I
• Sulfur is oxidized from alkylsulfite (oxidation number +4) to
alkylsulfate (oxidation number +6):
SO3CH3-2  SO4CH3-2 + 2e-
• Iodine is reduced:
I2 + 2e-  2I- (E0 = 0.54 V)
• The reduction of iodine consumes electrons generated by the
oxidation of sulfur, which changes the electrical potential of the system.
• The change in potential is detected by an electrode (a double platinum
electrode).
 End Point Detection
End point of a Karl Fischer titration can be detected in 3 ways:
A. Visible method: The color change accompanying the KF reaction is
suitable for the detection of the end point in many titrations.
CH3OH + H2O → Yellow → Dark brown
• But some samples are colored. In that case automatic titrations are
employed.
B. Photometric method: In this case absorbance of the titrating
solution is measured at 525 nm. A plot of absorbance versus titrant
volume is made.
• Before the end point the absorbance remains close to zero but after
the end point it increases linearly with the Volume
C. Potentiometric method: In this process, 2 Pt electrodes are
polarized at a constant current.
• Before the end point there is a small residual potential established
between the two electrodes, whereas in end point there is a large
increase in potential with the introduction of the free iodine.
 Acceptable Sample Size
The amount of sample used depends on the anticipated water content and
the desired degree of accuracy. The following convenient reference table
shows the relation between water content and proper sample size:
 Standardization of Karl Fischer
reagent
• KF reagent is not a primary standard reagent. So, standardization is required before use. The USP
lists Na-tartrate (Na2C4H4O6. 2H2O) as its primary standard.
• Molecular weight (M.W.) of Na-tartrate = 230.06 gm. [Where 2 H2O = 2 × 18 = 36 gm.]
• So, 1 gm. of Na-tartrate contains = 36/230.06 gm. of water = 0.1566 gm. of water
• If ‘w’ mg (100 – 350 mg) of Na-tartrate is titrated by KF reagent (v ml) from the burette, then
• 1 mg of Na-tartrate contains = 0.1566 mg of water
• So ‘w’ mg of Na-tartrate contains = (w × 0.1566) mg of water
• Now,
• v ml of KF reagent = (w × 0.1566) mg of water
• So, 1 ml or each ml of KF reagent = (w x 0.1566)/v mg of water = F
• Here, F is known as the water equivalence factor that indicates the amount of water reacting
with each ml of KFR.
 Moisture Determination:

• After determination of F value (standardized), the

moisture of the sample is determined.
• If ‘S’ ml of KFR is required for W1 mg of sample then,
• W1 mg sample contains = (F × S) mg of moisture
• So, 100 mg contains = (F×S)/W1×100 mg of moisture
• Therefore, % moisture = (𝐅×𝐒)/𝐖𝟏×𝟏𝟎𝟎
 Problems
1. Calculate the % of moisture in amino salicylic acid if 9 mL KFR having a
water equivalence factor 4.1, is consumed by 5.1 gm. sample.

Answer
•Total moisture content = 9 × 4.1 = 36.9 mg
•5.1 gm or 5100 mg of sample contain = 36.9 mg of water
•So, 100 mg of sample contain = (36.9 × 100)/5100 = 0.724%
•2. Calculate the % of moisture in 6.05 gm aspirin, if 7.5 ml of KFR
having a water equivalence factor 4.21, is consumed by the sample.
[Answer = 0.522 %]
•3. Calculate the % of moisture in amino salicylic acid if 9.3 ml of KFR
having a water equivalence factor 3.99 was consumed by 5.5 gm of
the sample. [Answer = 0.675 %]
•4. Calculate the water equivalence factor of KFR if 180 mg of Na-
tartrate required 15 ml for titration.
Answer:

F = (0.1566 × w)/v = (0.1566 × 180)/15 =1.88

 Advantages of the KFT method over other
methods

1. Azeotropic method can be effective in case of high moisture containing

substance. But KF method can detect trace amount of moisture.
2. Heat is applied in case of Azeotropic method. So heat sensitive products
cannot be analyzed. But in KF method no heat is applied, so the heat
sensitive material can also be used.
3. In case of gravimetric method volatile substance can cause error in the
result but in KF method there is no such kind of problems.
4. KF method is accurate than other methods. Only 0.1-0.2% error may occur.
5. The end point of KF titration can be detected visually, photometrically or
electrometrically.
6. Water can be detected from most classes of organic sample like acids,
esters, alcohols
• Limitations of KFT method:
1. The method is only applicable for those
substances who neither react with any
components of the reagent nor with the hydrogen
iodide, which is formed during the reaction with
water. E.g.:
• 1. Ascorbic acid will be oxidized by the iodine
present in the reagent.
• 2. Carbonyl compounds react with methanol
forming acetals or ketals.
• 3. Oxidizing agents, for instance: chromates, Cu
(II), Fe (III), Cr2O72–, peroxides, salts, higher
oxides etc.
• MnO2 + 4C5H5NH+ + 2I– → Mn2+ + 4C5H5N + I2 +
H2O
• 4. Reducing agents, such as: Sn (II) salts,
sulphides and S2O32–
2. Compounds that have a tendency to form water
with the ingredients of the Karl Fischer reagent,
for instance :
• a) basic oxides: e.g. ZnO;
• ZnO + 2C5H5NH+ → Zn2+ + C5H5N +
H2 O
• b) salts of weak oxy-acids e.g.
NaHCO3;
• NaHCO3 + C5H5NH+ → Na+ + H2O +
CO2 + C5H5N
3. Carbonic acid (H2CO3) is very
unstable; hence it splits up to yield a
mole each of water and CO2.
• H2CO3 → H2O + CO2