Titration

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Karl Fischer

KF Guide 1
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Introduction
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Titration
2
Editorial

Dear Reader,
The Karl Fischer method for the water content determination is one of
the most frequently used titration methods. Published by German petro-
chemist Karl Fischer in 1935, it has become well renowned for a wide
variety of applications and samples.
This brochure presents a short, historic overview of the development of
Karl Fischer’s method. It explains the chemical reaction and the two
major measuring techniques: Volumetry and coulometry.
Learn from METTLER TOLEDO’s extensive knowledge of moisture and
water content determination from 1 ppm to 100%.

3
Historic ­overview

Content 4

1. The Karl Fischer Titration 5
1.1. A historic overview 5
1.2. The Karl Fischer chemical reaction 7
1.3. Consequences for practical applications 9
2. Volumetric and Coulometric Karl Fischer Analyses 11
2.1. Volumetric KF analysis 12
2.1.1 One-component KF reagent 12
2.1.2 Two-component KF reagent 12
2.1.3 Pyridine-containing reagents 13
2.1.4 Special reagents for aldehydes and ketones 13
2.1.5 Karl Fischer reagents with ethanol 13
2.2 Coulometric KF analysis 14
2.2.1 KF coulometry 14
2.2.2 Stoichiometry of the coulometric KF rection 16
2.2.3 Iodine generation 16
2.2.4 Generator electrode without diaphragm 17
2.2.5 Limitations for the use of the cell without diaphragm 18
3. Further Information 19

4
 1. The Karl Fischer Titration

1.1. A historic 1935 Publication: “Neues Verfahren zur massanalytischen Bestim-

­overview mung des Wassergehaltes von Flüssigkeiten und festen
Körpern” by Karl Fischer.
1943 Publication: “The Dead-Stop End Point“, by G. Wernimont and
F.J. Hopkinson.
1950 Pyridine-containing two-component reagents and dead-stop
titration instruments are commercially available.
1952 Use of Karl Fischer method spreads, promoted by publications
by E. Eberius.
1955 Publication on stabilized single-component Karl Fischer rea-
gent by E. D. Peters and J. L. Jungnickel.
1956 First German DIN standard for the Karl Fischer titration (DIN
51777, April 1956, “Testing of mineral oil hydrocarbons and
solvents: Determination of water content according to Karl
Fischer - Direct method”).
1959 Publication: Coulometric Karl Fischer Titration by A. S. Meyer
and C. M.Boyd.
1960 KF titration instruments with piston motorized burettes.
Enormous spread of the use of KF titration.
1970 First coulometric KF titration instruments are commercially
available.
1980 Pyridine-free KF reagents are commercially available.
1984 First microprocessor controlled KF titrator (METTLER DL18)
with automatic drift compensation, and solvent dispensing
and removal.
1985 First fully automatic KF titration with laboratory robots
(METTLER DL18 and ZYMARK); DO185 Drying Oven for the
DL18 Karl Fischer Volumetric Titrator.
1989 First diaphragm-less cell for coulometric KF titration.
1990 DL37 KF Coulometer from METTLER TOLEDO.
1995 Water standards (10.0, 1.0, 0.1 mg/g) with test certificate
according to DIN 50049-2.3
First titrator (METTLER TOLEDO DL55) with online curves
E = f(t) and V = f(t) for Karl Fischer titration.

5
Karl Fischer chemical reaction

1997 New DV705 KF Titration Stand with very low drift value
(< 2μg/min) for the METTLER TOLEDO DL53/55/58, and
DL67/70ES/77 Titrators
1998 Introduction of the METTLER TOLEDO DL31/DL38 KF Titrators
with dedicated fuzzy logic control, titrant specific standard pa-
rameters and LEARN titration. They replaced the DL18/35 KF
Volumetric Titrators.
Introduction of less poisonous KF reagents based on ethanolic
solution.
2000 METTLER TOLEDO RONDO Sample Changer with Karl Fischer
Kit for automated KF volumetric determination.
2001 Improved METTLER TOLEDO DO307 KF Manual Drying Oven.
Solid KF Oven Standards with water contents of 5.5% and
1%, respectively.
2002 Introduction of the METTLER TOLEDO DL32/39 KF Coulometers
(generating cell with and without diaphragm).
Introduction of the METTLER TOLEDO STROMBOLI KF Oven
Sample Changer.
2008 Introduction of the METTLER TOLEDO Titration Compact Line
V20/V30 and C20/C30 Karl Fischer Instruments.

6
1.2 The Karl Fischer The water content determination is based on the reaction described by
chemical reaction R. W. Bunsen:
I2 + SO2 + 2 H2O → 2 HI + H2SO4

Karl Fischer discovered that this reaction could be used for water de-
terminations in a non-aqueous system containing an excess of sulfur
dioxide. Methanol proved to be suitable as a solvent. In order to achieve
an equilibrium shift to the right, it is necessary to neutralize the acids
that are formed during the process (HI and H2SO4). Karl Fischer used
pyridine for this purpose. Smith, Bryanz and Mitchell formulated a two-
step reaction:
1. I2 + SO2 + 3 Py + H2O → 2 Py-H+I – + Py•SO3

2. Py•SO3 + CH3OH → PyH-CH3SO4

According to these equations, methanol not only acts as a solvent but

also participates directly in the reaction itself. In an alcoholic solution,
the reaction between iodine and water takes place in the stoichiometric
ratio of 1:1. In an alcohol-free solution, the reaction between iodine and
water takes place in the stoichiometric ratio of 1:2:
1.I2 + SO2 + 3 Py + H2O → 2 Py-H+I – + Py•SO3

2.Py•SO3 + H2O → Py-H+HSO4 –

Further studies conducted by J. C. Verhoef and E. Barenrecht on the

subject of the Karl Fischer reaction have revealed that:
• Pyridine is not directly involved in the reaction, i.e., it only acts as a
­buffering agent and can therefore be replaced by other bases,
• The rate of the Karl Fischer reaction, described by the rate constant k,
depends on the pH value of the medium (see graphics below)

7
Karl Fischer chemical reaction

-d[I2]/dt = k • [I2] • [SO2] • [H2O]

One possible explanation for the influence of pH on the reaction rate is

that it is not the sulfur dioxide itself that is oxidized by iodine under the
influence of water, but rather the methyl sulfite ion. This is formed from
sulfur dioxide and methanol according to the equation:

-
2 CH3OH + SO2 → CH3OH2+ + CH3OSO2

The higher the pH of the solution, the more methyl sulfite is formed by
the capture of protons, and the faster the rate of the Karl Fischer reac-
tion.
In the pH range 5.5 to 8, all the sulfur dioxide is present as methyl
sulfite; the maximum reaction rate is reached here and cannot increase
further. At pH values above 8.5, the reaction rate increases due to side

8
 reactions between iodine and hydroxide or methylate ions; in a titration,
this results in a more sluggish endpoint and higher iodine consumption.
On the basis of this knowledge, E. Scholz developed a pyridine-free Karl
Fischer reagent with imidazole as base. This reagent not only replaced
the toxic, pungent pyridine, but also facilitated faster and more accurate
titrations because imidazole buffers in a more favorable pH range than
pyridine.
Studies by E. Scholz resulted in the following reaction scheme being
proposed for the Karl Fischer reaction:
1. ROH + SO2 + RN → (RNH) · SO3 R

2. (RNH) · SO3 R + 2 RN + I2 + H2O → (RNH) · SO4 R + 2 (RNH)I

This resulted in the general chemical equation:

ROH + SO2 + 3 RN + I2 + H2O → (RNH)•SO4 R + 2 (RNH)I

E. Scholz was also able to confirm the existence of basic methylsulfite

in methanol/SO2/I2 solutions during the titration. In 1988, A. Seubert
identified methylsulfite in KF solutions with the aid of IR spectroscopy
and isolated and identified methyl sulfate as the secondary product of
the KF reaction.
Experiments on the stoichiometry of the reaction showed that methanol
can in fact be replaced by other alcohols (e.g. ethanol, 2-propanol,
methoxyethanol, diethylene glycol monoethylether). This improves the
titer stability.

1.3 Consequences • Influence of pH on the Karl Fischer reaction

for practical Since the maximum rate of the Karl Fischer titration is in the pH range
­applications 5.5 to 8, pH values less than 4 and greater than 8 should be avoided
in practice. With acidic or basic samples, you should adjust the pH
value to the ideal range by adding buffering agents (for acids: imida-
zole, and for bases: salicylic acid).

• Influence of the solvent on the Karl Fischer reaction

The stoichiometry (molar ratio of H2O:I2) depends on the type of
­solvent:
Alcoholic solvent H2O:I2 = 1:1 (e.g. methanol)
Non-alcoholic solvent H2O:I2 = 2:1 (e.g. dimethylformamide)

9
Karl Fischer analyses

Studies by Eberius showed that iodine and water react in the ratio
of 1:1 if the percentage of methanol in the solvent is 20% or more.
­Methanol should therefore always be present in the minimum required
amount. If a methanol-free titrant has to be used (e.g. for determina-
tion in ­ketones or aldehydes), you can use other alkohols such as, for
instance, ­ethylene glycol monomethyl ether.

• Influence of the water content of the sample on the Karl Fischer

­reaction
The water content of the sample also influences the H2O:I2 molar
­ratio. J.C. Verhoef and E. Barendrecht observed an increase in the
titer with water contents greater than 1 mol/L. This, however, is not a
­serious ­limitation because the water concentration in the solvent is
usually much less.

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2. Volumetric and Coulometric
Karl Fischer Analyses
The determination of the water content according to Karl Fischer is
­nowadays performed by two different techniques:
–– Volumetric Karl Fischer Titration, where a solution containing iodine is
added using a motorized piston burette;
–– Coulometric Karl Fischer Analysis, where iodine is generated by elec-
trochemical oxidation in the cell
The selection of the appropriate technique is based on the estimated
­water content in the sample:

Volumetric Karl Fischer Titration

Iodine is added by a burette during titration.
Suitable for samples where water is present
as a major ­component: 100 ppm - 100%

Coulometric Karl Fischer Analysis

Iodine is generated electrochemically during
titration. Suitable for samples where water is
present in trace amounts: 1 ppm - 5%

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Volumetric KF analysis

2.1 Volumetric­ 2.1.1 One-component KF reagent

KF reagents The titrant contains iodine, sulfur dioxide and imidazole, dissolved in a
suitable alcohol. The solvent is methanol. You can also use a methanolic
solvent mixture specially adapted to the sample as the solvent.
The reagent can be stored for approximately two years. The drop in
titer, i.e., the decrease in concentration, is approximately 0.5 mg/mL
per year in a sealed bottle. The reagent is available in three different
­concentrations:
5 mg/mL for samples with a water content of 1000 ppm to 100%,
2 mg/mL for samples with a water content of less than 1000 ppm,
1 mg/mL for samples with a water content of less than 200 ppm.

2.1.2 Two-component KF reagent

The titrant contains iodine and methanol. The solvent contains sulfur
dioxide, imidazole and methanol.
A titration speed two or three times as high can be achieved with the
two-component reagent. Both the components are very stable in stor-
age. The titrant has a stable titer, provided that the bottle is tightly
sealed. It is available in two different concentrations:
• 5 mg/mL for samples with a water content of 1000 ppm to 100%,
• 2 mg/mL for samples with a water content of less than 1000 ppm.

Reagents + -
One-component Simple handling, fa- Titer less stable,
vorably priced. titration speed slower.

Two-component High titration speed, Solvent capacity

stable titer. restricted.

2.1.3 Pyridine-containing reagents

Despite the existence of pyridine-free reagents, which allow for fast and
accurate Karl Fischer titrations, reagents containing pyridine are still
used because they are cheaper and can be made in-­house.

One-component reagent:
The titrant contains iodine, sulfur dioxide and pyridine, dissolved in
methanol. The solvent is either methanol or consists of methanol mix-
tures. Some manufacturers have slightly increased the pyridine content

12
in the titrant to achieve a higher titration speed. This reagent is declared
as “rapid”. To improve stability, some manufacturers also sell the titrant
separated into solution A (sulfur dioxide, pyridine, methanol) and B
(­iodine, methanol). These solutions are mixed 1:1 just before use to
form the one-component titrant.

Two-component reagent:
The titrant contains iodine dissolved in an alcohol, e.g. methanol,
whereas the solvent consists of sulfur dioxide and a base, e.g. imida-
zole, dissolved in an alcohol (usually methanol), or an alcoholic mix-
ture. The separation into titrant and solvent improves stability of the KF
reagents, increases their lifetime, and results in higher titration speed.

2.1.4 Special reagents for aldehydes and ketones

Aldehydes (R-CHO) and ketones (R-CO-R) form acetals and ketals if
titrated with standard methanol-containing reagents. As a result, addi-
tional water is produced and titrated at the same time, leading to higher
water contents and a vanishing end point. Special methanol-free KF
one-component reagents such as e.g. HYDRANAL® (Composite 5K and
Working Medium K, from Sigma-Aldrich) and e.g. apura® (CombiTitrant
5 Keto with CombiSolvent Keto, from VWR/MERCK) are commercially
available to prevent this problem.

One-component reagent:
The titrant contains iodine, imidazole, sulfur dioxide and 2-methoxyetha-
nol, whereas the solvent contains 2-chloroethanol and trichlorometh-
ane. The titration takes slightly longer than with the standard KF reagent.
Note that it may be necessary to adapt the end point value in the titra-
tion method to these reagents. This special reagent is also suitable for
substanc­es that react with methanol, such as amines.

2.1.5 Karl Fischer reagents with ethanol

Since ethanol is less toxic than methanol, two-component, ethanol-
based reagents were launched in 1998. These reagents also allow for
titration of several ketones which form ketals considerably more slowly
in ethanol than in methanol.
The titrant contains iodine and ethanol, whereas the solvent contains
sulfur dioxide, imidazole, diethanolamine and ethanol.

13
Coulometric KF analysis

2.2 Coulometric 2.2.1 KF coulometry

KF analysis Coulometric KF determination of water is based on the standard reaction
equation for the KF reaction.
ROH + SO2 + 3 RN + I2 + H2O → (RNH)•SO4 R + 2 (RNH)I

In coulometry, however, the iodine is generated electrochemically by

anodic oxidation in the coulometric cell according to the following
­half-reaction:

2 I - → I2 + 2 e-

Iodine generation occurs at a generator electrode* (also called “genera-

tor cell”, or “inner burette”) incorporated into the glass titration cell. The
generator is close to the measuring electrode*, a double pin platinum
electrode, which is monitoring the potential of the sample solution by
voltametric technique during coulometric titration.
The classical coulometric cell consists of two parts, the anode com­
partment and the cathode compartment. Both parts are separated by a
diaphragm.

Measuring electrode
(double platinum pin)

Generator electrode

Catholyte
Anolyte
Diaphragm

* The terms “generator electrode” and “measuring electrode” mean an arrangement of two
electrodes (anode and cathode) to form an electrolytic cell.

14
 The anode compartment contains the anolyte, i.e. the KF electrolyte
which is required for the oxidation in order to generate iodine by apply-
ing a current at the generating electrode. This consists of sulfur dioxide,
­imidazole and iodide salts. Methanol or ethanol is used as solvent.
Depending on the application, other solvents such e.g. as chloroform,
octanol, hexanol or ethylene glycol may be added.
The cathode compartment contains the catholyte, i.e. the reagent that
enables completion of the whole electrochemical reaction: the oxidation
reaction in the anolyte compartment is completed by a reduction reac-
tion in the catholyte compartment. The catholyte is either a manufactur-
er-specific special reagent, or the same reagent as used in the anode
­compartment.

Anode reaction
At the anode, iodine is generated from
iodide by electrochemical oxidation. The
negative iodide ions release electrons at
the anode and form iodine, which subse-
quently reacts with water.

Cathode reaction
At the cathode, the positive hydrogen ions
are reduced to hydrogen. This is the main
product formed.
An ammonium salt is added to the catholyte in order to promote hydro-
gen production.
2 [RN]H + + 2e -→ H2 + 2 RN

Ammonium ions are then reduced with the formation of hydrogen and a
free amine.

Note that methyl sulfonic acid, CH3SO3H, present in the anolyte

c­ ompartment, can reach the cathode where it is reduced to a sulfide
compound (obnoxious smell!). This can cause the cathode to become
black.

To avoid this, the catholyte should be replaced once in two weeks.

15
Coulometric KF analysis

2.2.2 Stoichiometry of the coulometric KF rection

The amount of water titrated by coulometric technique is determined by
the amount of electrical current given in Coulomb, C, used to generate
iodine. To calculate it, it is necessary to first recall the definition of
1 ­Coulomb: one coulomb, C, is the quantity of charge that is transport-
ed by a current of 1 ampere in one second.
1C=1A·1s

On the other hand, it is known that to produce one mole of a chemical

substance that requires one electron, 96485 C of current are needed.
The value 96845 C/mol is known as the Faraday constant.
In Karl Fischer reaction two iodide ions, I-, are oxidized to one molecule
of iodine, I2, releasing two electrons at the anode. Subsequently, iodine
reacts with water.
2 I - → I2 → H2O

Therefore, 2 x 96485 C/mol is required for 1 mole of water. Since the

molar mass of water is 18.015 g/mol, this calculation can be expressed
as follows:
1 mg of water corresponds to a consumption of 10.712 C electrical ­current.

For an electrochemically optimized Karl Fischer cell, the current conver-

sion efficiency for iodine production is assumed to be 100%.
Since current and time can be accurately measured, no standardization
of the coulometric KF reagents is necessary. For this reason, coulometry
is designated as absolute method. As a consequence, it is used as a
reference method for the determination of water content.
Nevertheless, it is strongly recommended that the coulometer is regu-
larly checked by measuring a certified water standard.

2.2.3. Iodine generation

Iodine is generated by means of current pulses of 400, 300, 200 and
100 mA. The rate of iodine generation is adjusted by varying the pulse
duration and frequency and the pulse height (in mA). The maximum pulse
height depends on the following factors:
–– conductivity of the anolyte
–– voltage at the generator electrode
–– surface of the generator electrode

16
The voltage at the generator electrode and the surface of the electrode
depend on the type of coulometer. In addition, the conductivity of the
anolyte is influenced by the samples and additional solvents (chloro-
form, hexanol, etc.).
With standard conductivity values, the coulometer operates with
current pulses of 400 mA. This gives an iodine generation rate which
corresponds to a maximum of 2240 μg water/min.
At very low conductivities (i.e. less than 3–4 μS/cm), the maximum
­possible current applied by the instrument is 100 mA.

2.2.4 Generator electrode without diaphragm

with diaphragm without diaphragm

The first commercially available diaphragmless cell for KF coulometry

was introduced in 1989. The main advantages compared to cells with
diaphragms are:
–– no contamination or blockage of the diaphragm
–– easier cleaning
–– lower drift
Originally, the generator cell was first built with a porous ceramic frit
(diaphragm) separating the inner cathode compartment from the anode
compartment. The purpose is to prevent that iodine molecules gener-
ated at the anode are immediately reduced to iodide ions at the cathode
i­nstead of reacting with water.
To avoid this concurring reaction, a different geometry is used for the
generating cell without diaphragm. The cathode is smaller and made
in such a way that iodine cannot reach the cathode (i.e. the cathode is

17
More information

a small-size pin). In addition, a relatively high stirrer speed allows for

more rapid distribution of the generated iodine molecules. The latter can
thus react more efficiently with water.
Note that hydrogen gas generated at the cathode is forming gas bubbles
around its surface. Thus, this makes it almost impossible for iodine to
reach the cathode and be reduced to iodide. This effect is further en-
hanced by the small dimensions of the cathode.
Nevertheless, it is still possible for a very small amount of iodine to
reach the cathode. However, the relative error due to this effect can be
minimized by using a larger amount of sample.
In practice, for measurement of samples having a very low water con-
tent (e.g., lower than 50 μg water/sample), and for very accurate deter-
minations, the cell with diaphragm may yield more precise results.

2.2.5 Limitations for the use of the cell without diaphragm

The cell without diaphragm is not suitable for samples that are easily re-
duced. Nascent hydrogen is formed at the cathode. This is a very good
reducing agent, especially for nitro compounds such as nitrobenzene.
R-NO2 + 3 H2 → R-NH2 + 2 H2O

Besides this, other easily reducible substances such as unsaturated

fatty acids, etc. may also be reduced at the cathode by the formation of
water.

The diaphragmless cell is ideal for the determination of the water con-
tent of the following compounds:
–– hydrocarbons - chlorinated hydrocarbons
–– alcohols - phenols (most)
–– esters - ethers
–– ketones (with special reagent) - acetamides
–– etheric oils and essences - edible oils
–– petroleum oils

A cell with diaphragm is required for:

– samples with a very low water content (< 50 μg water/sample)
– very accurate determinations
– nitro compounds
– unsaturated hydrocarbons, especially when easily reducible.

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 3. Further Information

3.1 Literature HYDRANAL®-Manual, „Eugen Scholz Reagents for Karl FischerTitration“,

Sigma-Aldrich Laborchemikalien GmbH, D- 30918 Seelze / Germany,
2006.
SCHOLZ. E., „Karl Fischer Titration“, Springer Verlag Berlin, 1984.
WIELAND, G., „Wasserbestimmung durch Karl-Fischer-Titration –
Theorie und Praxis“, GIT Verlag GmbH, Darmstadt / Germany, 1985.

3.2 More guides METTLER TOLEDO has prepared a set of guides for the Karl Fischer titra-
tion. They explain basics, methods and techniques and provide tips and
hints for the daily practice.

1 Introduction to Karl Fischer Titration

2 Sample Preparation for Karl Fischer Titration
3 Taking Samples for Karl Fischer Titration
4 The Method at a Glance

3.3 Application Many more details regarding the Karl Fischer titration are published in the
brochure application brochure Good Titration Practice™ in Karl Fischer Titration
(ME 517252145).

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Good Measuring Practices
For Balances, Titrators and Pipettes

METTLER TOLEDO’s risk-based guidelines for ­titration, weighing and

pipetting empowers you to take the right decision when and where it
really matters. The five steps of Good Measuring Practices cover the
entire lifecycle of your instruments and provide you with practical guid-
ance to implement a sound quality management system.

1. Evaluation
5
Routine 1 Analyze your process flows
Operation Evaluation and its associated criteria to
consistently assure the high-
Good est quality of your application
Measuring and your data.
4
Calibration / Practices 2
Qualification Selection 2. Selection
Choose the ideal combination
3 of instrument and measuring
Installation / technology to best match your
Training
process needs.

3. Installation & Training

Enjoy every confidence in your new device and master it with full profes-
sional skills right from day one.

4. Calibration & Qualification

Trust the manufacturer-trained METTLER TOLEDO Service Team when it
comes to calibrating and qualifying your instruments.

5. Routine Operation
Benefit from tangible recommen­dations for optimal performance
­verification, calibration and ­maintenance.

www.mt.com/gtp

www.mt.com/karl-fischer
For more information

Mettler-Toledo AG, Analytical

CH-8603 Schwerzenbach, Switzerland
Tel. +41-44-806 77 11
Fax +41-44-806 73 50

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