Competence Guide

Tips and Hints

Tips and Hints
For a Successful
For a Successful
Water Content
Water Content
Determination
Determination

Titration Excellence
KFTitration
Compact Titrators
Excellence
KF Compact Titrators

Water Content by Karl Fischer Titration

Essentials and Troubleshooting
 Editorial
The presence of water in petroleum products, pharmaceutical substances or foodstuffs is often undesirable.
A water content that is too high can reduce the shelf life of rice, make the processability of pharmaceutical
Editorial

products more difficult, or lead to quality problems with plastic products. The determination of the water content
in different materials is therefore crucial in production processes and quality control.

Chemical methods for the water content determination include techniques where a chemical reaction takes place
between water and a suitable reagent. The amount of water converted during the reaction is determined from the
amount of reagent consumed. The most well-known method based on this principle is the Karl Fischer titration.
The main advantages of this quantitative analytical technique are its

• high selectivity to water

• wide measuring range
• short analysis time
• accuracy and repeatability
• directness i.e. no additional reference method necessary

METTLER TOLEDO offers you a complete portfolio of solutions for successful water content determination by Karl
Fischer titration, including autosamplers and dedicated accessories for the unattended analysis of a high
number of samples. Detailed information can be found at Volumetric and Coulometric Karl Fischer Titrators – For
Accurate and Precise Water Determinations (mt.com).

This Competence Guide covers the most relevant points to be considered when performing a Karl Fischer titration
- without starting from basics. Beside a brief overview, this Guide will focus on three parts:

• The four key questions for a successful KF titration:

To achieve accurate and correct results by the KF titration it is necessary to first completely understand what
are the goal, the needs and the water content determination to be performed. Four specific questions will
support and guide you in the right direction.
• Sample preparation:
The water content can be successfully determined only if water is freely available to undergo the chemical
reaction according to Karl Fischer. If water is not freely available, dedicated sample preparation has first to be
applied to release water molecules from the sample matrix.
• Frequently asked questions:
A selection of the most relevant frequently asked questions

We are convinced that this document is a valuable support for your work on water content determination.

Daniel Buchmann Dr. Cosimo A. De Caro

Head Market Support Group Senior Applications Specialist
SBU Analytical Chemistry Market Support Group Analytical Chemistry

METTLER TOLEDO Competence Guide

 Table of Content
Overview 1
Karl Fischer (KF) Titration 1
Table of Content

Ready-to-use checklist for KF titrations 2

The four key questions in Karl Fischer titration 3
What is the expected water content of the sample? 3
How much sample has to be weighed? 4
How is water released from the sample matrix? 5
Which KF reagents should I use? 6
Sample preparation in Karl Fischer titration 8
Complete release of water from the sample matrix 8
Direct titration 9
Internal extraction 10
External extraction 11
External extraction in four steps 12
External extraction of lyophilized substances 14
External dissolution 15
Automated calculation for external extraction and dissolution 16
Titration workflow for external extraction/dissolution 17
Gas-phase extraction 18
Water content in gases 19
Troubleshooting: The most frequently asked questions 20
The drift does not decrease to a low and stable value even after 1-2 hours. Any hints? 20
What is the smallest amount of water that can be determined by Karl Fischer titration? 21
Is it mandatory to check the concentration of a new Karl Fischer titrant bottle? 22
Which water standards do you recommend for the concentration determination? 23
Are liquid water standards hygroscopic? 24
How often should the concentration of the KF titrant be determined? 25
How do you determine the titer for the coulometric KF reagent? 26
The verification with standards is outside the uncertainty in the certificate of analysis. Any suggestions? 27
What do you do if the result is not correct? 29
How long should the stand-by take until the online drift reaches a low level? 30
The online drift is still very high after titration end. How can this problem be solved? 31
Sometimes we get negative results when using the automated KF oven – Why? 32
Literature 33
Links 33

METTLER TOLEDO Competence Guide

 Overview
Karl Fischer (KF) Titration
Competence Guide

Titration is a straightforward, easy and affordable technique for the content determination of a specific analyte
in sample solutions. If the analyte is water, then the titration according to Karl Fischer is the method of choice.
Briefly, the water content is determined by addition of an iodine I2 solution of known concentration to the sample
dissolved in an alcoholic solution e.g. in methanol. Iodine reacts with water in a 1:1 ratio in a two-step reaction
involving methanol, an organic base and sulfur dioxide (SO2). This chemical reaction is designated according to
the German Chemist Karl Fischer who developed this analytical technique for the determination of the water
content in petroleum products i.e. the so-called Karl Fischer (KF) reaction.

During titrant addition, the potential difference is monitored with a polarized double pin platinum potentiometric
sensor (a redox electrode), such that the process can be visualized in a graph showing a titration curve. The
iodine titrant solution is added until a predefined potential endpoint is reached i.e. until a potential of 100 mV is
reached. The titrant consumption in mL up to this point allows for calculation of the water content in the sample
(Volumetric KF titration). Alternatively, the iodine needed for the KF reaction can be generated directly in the
titration vessel by electrochemical processes (Coulometric KF titration).

Figure 1: Titration curves showing the determination of water in cellulose by volumetric KF titration. 0.15 g cellulose was added directly
in the vessel of a volumetric KF titrator, stirred during 120 s to achieve complete dissolution, and subsequently titrated. The potential
(blue curve) and the titrant volume (red curve) are plotted versus time.

METTLER TOLEDO Competence Guide 1

 Ready-to-use checklist for KF titrations

The following points has to be verified to perform successful titrations:

Competence Guide

Environment − Temperature − Ambient temperature – i.e. 5-40 °C

− Humidity − non-condensing environment
− Pressure − Standard pressure approx. 1 atm
− Ventilation / air conditioning − Avoid exposition to air flows
− Sun exposure − Avoid exposition to direct sunlight
− Use − Inside / interior spaces
Equipment − Balance − Correct weighing range, connected
− Titration vessel / titration stand − Check tightness, grease the junctions if needed
− Molecular sieve / silica gel − Fresh / regenerated in the oven to eliminate absorbed water
− Burette, tubes − Eliminate gas bubbles before titration by rinsing once
− Sensor − Connected and defined in the titrator setup
− Automation and accessories − Connected and defined in the titrator setup
Chemicals and Samples − Titrant − Fresh, new titrant / tightly closed, not wet
− Solvent − Fresh, dry
− Coulometric electrolyte − Fresh, new electrolyte / tightly closed, not wet
− Water standards − Suitable concentration, use new standards
− Sample − Sufficient amount, homogeneous sample
− Auxiliary solvents and reagents − Very low water content
Users − Assistant − Administrator
− Operator − Check correct role
– Chemist − Verify access rights
– Lab manager
Documentation − Operating instructions − Available
− Qualification documents – Documented proof of correct working
− Standard operating procedures − Available and up-to-date
− Titration method validation − Documented evidence of method correctness
− Available and up-to-date

METTLER TOLEDO Competence Guide 2

 The four key questions in Karl Fischer titration
What is the expected water content of the sample?
Competence Guide

Two determination techniques are used in KF titration depending on the water content of the sample:

• Coulometric KF titration 1 ppm - 5%

• Volumetric KF titration 100 ppm - 100%

The techniques differ in the way they provide iodine for the Karl Fischer reaction with water. Based on the
expected water content, the most suitable instrument i.e. a coulometric or a volumetric KF titrator is selected:

Figure 2A: Coulometric (left, Coulometric KF Titration - Tutorial - YouTube) and volumetric (right, 13. Run titration - Karl Fischer Volumetric
Titrator - YouTube) KF titrators. Typical samples are indicated below each instrument.

When measuring several aliquots of the same sample in a sample series, the repeatability – expressed as
relative standard deviation Srel in % - is changing depending on the water content, as shown in figure 2 B. In
particular, decreasing water contents lead to poorer repeatability i.e. the relative standard deviation is increasing:

Figure 2B: Measurement range for coulometric and volumetric KF titration. The repeatability (rel. std. deviation Srel, %) is also given as a
function of the water content.

METTLER TOLEDO Competence Guide 3

 How much sample has to be weighed?

Select the suitable sample size for accurate and precise measurement i.e. small deviation from expected value
and low relative standard deviation:
Competence Guide

Coulometric KF titration

• The ideal coulometric titration sample contains 1 mg of water.

• The recommended practical working range is 0.1 to 5 mg.

Figure 3A: Optimal sample size for coulometric KF titration.

Volumetric KF titration

• The ideal volumetric titration uses ½ of the burette volume.

• The recommendations below use a standard burette size of 5 mL.

Figure 3B: Optimal sample size for volumetric KF titration.

METTLER TOLEDO Competence Guide 4

 How is water released from the sample matrix?

Obtaining accurate results requires extracting all water from the sample. Many liquid samples dissolve easily in
the classical KF solvent methanol, and thus the titration can be performed after direct injection of the liquid
Competence Guide

sample in the vessel. On the other hand, viscous oils, pastes or solid samples requires additional preparation
steps to release water from the sample matrix. These steps can include the use of auxiliary solvents, increased
dispersion e.g. longer (stirring times) or even higher temperatures to extract water completely:

Mixing
Many liquid or solid samples dissolve in methanol or in methanol
mixtures using a normal magnetic stirring bar:
• Methanol: liquid foods, cosmetics, organic solids
• Chloroform: petroleum products, fats, organic solids
• Long-chain alcohols (e.g. octanol): edible oils, fuels, creams
• Toluene/xylene: tars, waxes, crude oil
• Formamide: sugars, starches, solid foods, pharmaceuticals

Figure 4A: Dissolution by stirring with magnetic stirrer bar.

Dispersion
Thicker, viscous liquids, pastes or powders may require more intense
dispersion with a high-speed homogenizer to fully dissolve them.

Figure 4B: Dispersion using a high-speed homogenizer.

Warming
A thermostatable titration vessel allows warming up to 50°C with an
external silicon oil or water bath to improve dissolution and extraction
of water from the sample.

Figure 4C: Improving dissolution by warming up.

KF Oven
For solids or liquids causing side reactions or releasing water very
slowly, a drying oven heats samples up to 280°C to drive water out of
the sample and into the titration beaker.

Figure 4D: Water extraction in the gas phase by heating.

METTLER TOLEDO Competence Guide 5

 Which KF reagents should I use?

Easily soluble samples can be titrated by injection (addition) directly into the titration cell:
Competence Guide

Alcohols, ethers, aliphatic and aromatic hydrocarbons, etc.

One-component KF volumetric reagents Two-component KF volumetric reagents

Honeywell Titrant: HYDRANAL Composite 5 / 2 / 1 Titrant: HYDRANAL Titrant 5 / 2
Solvent: HYDRANAL Methanol Dry Solvent: HYDRANAL Solvent
Methanol Rapid HYDRANAL NEXTGEN Solvent FI
Merck Titrant: Aquastar CombiTitrant 5 / 2 / 1 Titrant: Aquastar Titrant 5 / 2
Solvent: Aquastar CombiMethanol Solvent: Aquastar Solvent
With ethanol (E) or methanol-free – KF kinetics with E-reagents is slower than with methanolic reagents
Honeywell Titrant: HYDRANAL Composite 5 / 2 / 1 Titrant: HYDRANAL Titrant 5 / 2 E
Solvent: HYDRANAL CompoSolver E Solvent: HYDRANAL Solvent E
HYDRANAL NEXTGEN Solvent E-FI
Merck Titrant: Aquastar CombiTitrant 5 / 2 / 1 --
Solvent: Aquastar CombiSolvent (methanol-free)
KF coulometric reagents
Honeywell Anolyte: HYDRANAL Coulomat A cell with diaphragm
Coulomat AG cell with and without diaphragm
Coulomat AD cell without diaphragm
NEXTGEN Coulomat AG-FI with and without diaphragm/free of imidazole
NEXTGEN Coulomat A-FA with diaphragm – Li-batteries, Ketones
Catholyte: HYDRANAL Coulomat CG cell with diaphragm
NEXTGEN Coulomat C-FA cell with diaphragm – Li-batteries, Ketones
Merck Anolyte: Aquastar CombiCouloma frit with diaphragm: anode and cathode compartment
Aquastar CombiCoulomat fritless cell with and without diaphragm
With ethanol (E) or methanol-free – KF kinetics with E-reagents is slower than with methanolic reagents
Honeywell Anolyte: HYDRANAL Coulomat E cell with and without diaphragm
Merck --

Ketones (e.g. acetone) and aldehydes react with methanol to form water. To suppress this undesired side
reaction, dedicated KF reagents are needed when dealing with these samples.

Ketones (e.g. acetone) and aldehydes

One-component KF volumetric reagents Two-component KF volumetric reagents

Honeywell Titrant: HYDRANAL Composite 5K --
Solvent: HYDRANAL KetoSolver (halogen-free)
Medium K (with chloroform)
Working Medium K
(CHCl3, CH2ClCH2OH)
Merck Titrant: Aquastar CombiTitrant 5 Keto --
Solvent: Aquastar CombiSolvent Keto
KF coulometric reagents
Honeywell Anolyte: HYDRANAL NEXTGEN Coulomat A-FA cell with diaphragm – Ketones, Li-batteries
Coulomat AK cell with diaphragm
Catholyte: HYDRANAL NEXTGEN Coulomat C-FA cell with diaphragm – Ketones, Li-batteries
Coulomat CG-K cell with diaphragm
Merck Anolyte: Aquastar Anolyte K cell with diaphragm
Catholyte: Aquastar Catholyte K cell with diaphragm

METTLER TOLEDO Competence Guide 6

 Many samples cannot be easily dissolved or are very difficult to be completely dissolved due to their non-polar
nature. Mixtures of organic solvents are used to improve the dissolution process. This is achieved either by
adding auxiliary solvents (e.g. decanol, formamide, chloroform) to methanol, or by using dedicated KF solvent
mixtures:
Competence Guide

Diesel oil, petroleum, edible oil, ethereal oils, ointments, butter, fats, etc.

Solvents for one-component KF volumetric reagents Solvents for two component KF volumetric reagents
Honeywell HYDRANAL Methanol Dry + max. 50% v/v decanol HYDRANAL Solvent Oil (with chloroform)
or hexanol
HYDRANAL LipoSolver CM (with chloroform) Solvent CM (with chloroform)
LipoSolver MH (with hexanol) Solver (Crude) Oil (xylene, chloroform)
Merck Aquastar CombiSolvent Fats (butyl acetate, Aquastar Solvent Oils & Fats (hexanol, 2-propanol)
decanol)
CombiSolvent Oils (with toluene)

Mineral oils are very difficult to be completely dissolved. Auxiliary solvents (chloroform, toluene, xylene and
mixtures thereof) added to methanol or dedicated KF solvents can cope with these challenging samples:

Mineral oil, transformer oil, silicon oil, grease, glue

Solvents for one-component KF volumetric reagents Solvents for two component KF volumetric reagents
Honeywell HYDRANAL Methanol Dry + up to 70% v/v HYDRANAL Solvent + up to 70% v/v chloroform
chloroform
Solver (Crude) Oil (xylene, chloroform) Solvent CM (with chloroform)
Solver (Crude) Oil (xylene, chloroform)
Merck Aquastar CombiMethanol + up to 70% v/v Aquastar Solvent + up to 70% v/v chloroform
chloroform
CombiSolvent Oils (with toluene) CombiSolvent Oils (with toluene)

Products containing sugar, starch or gelatin also need the use of formamide as auxiliary solvent to be
completely dissolved in the titration cell:

Sugars, sugar-and starch-based products

Solvents for one-component KF volumetric reagents Solvents for two component KF volumetric reagents
Honeywell HYDRANAL Methanol Dry + up to max. 50% v/v HYDRANAL Solvent + up to max. 50% v/v formamide
formamide
Merck Aquastar CombiMethanol + up to max. 50% v/v Aquastar Solvent + up to max. 50% v/v formamide
formamide

METTLER TOLEDO Competence Guide 7

 Sample preparation in Karl Fischer titration
Complete release of water from the sample matrix
Competence Guide

Water determination in liquid samples is generally straightforward and does not involve any challenges other
than improving the sample solubility to achieve a complete release of water into the KF solvent. The situation is
different when the water content has to be determined in solid samples.

In fact, water molecules present in solid samples are bound to the sample matrix in different ways i.e. water
molecules can be

• Entrapped in cavities of the sample structure, or

• Weakly bound in capillaries of biological samples, or
• Adsorbed on the sample surface, or
• Tightly bound in crystal structure.

Hence, water must be first completely released from solid samples to be available for the Karl Fischer titration:

SO2 + 3 RN + CH3OH + H2O + I2 ⇌ (RNH)SO4CH3 + 2 (RNH)I

The Karl Fischer reaction consists of two steps: first, sulfur dioxide SO2 reacts with a base RN and methanol
CH3OH to a methyl sulfite, (RNH)SO3CH3. Subsequently, methyl sulfite reacts with water H2O, iodine I2 and a
base giving methyl sulfate as a product.

The methods to efficiently release water from the sample matrix have been summarized in the following list:

• By dissolution in the Karl Fischer solvent,

• By mechanically crushing the sample in the titration vessel,
• By extraction from the sample matrix with a suitable solvent, or
• By evaporation in a dedicated KF oven connected to the titrator.

Based on these methods, different sample preparation techniques have been developed:

• Direct titration
• Internal extraction
• External extraction
• External dissolution
• Gas-phase extraction

The choice of the suitable technique depends on the sample to be titrated. The different tehcniques will be
presented in the next chapters.

METTLER TOLEDO Competence Guide 8

 Direct titration

The direct titration of a sample to determine its water content is possible if

• The sample is soluble,
Competence Guide

• It does not undergo side reactions with the KF reagents, and

• No additional sample preparation steps are needed.

Alcohols, esters, ethers, aliphatic and aromatic hydrocarbons:

These organic solvents such as do not cause any solubility issues when directly titrated.

Sugars, starch and starch containing products:

The dissolution is cumbersome or slow, hence an auxiliary solvent is used to improve it. In this case, formamide
is used in a volume ratio between 30% and max. 50% v/v with the KF solvent to achieve complete dissolution.

Hydrocarbons with long aliphatic chains:

They cannot be easily dissolved in the KF solvent – which is generally methanol. This is because they are mainly
non-polar substances - in contrast to methanol. Auxiliary solvents and warming up may become necessary.

Diesel fuel, petroleum edible oils and fats, ethereal oils, ointments:
These samples are difficult to dissolve in the KF solvent. The solubility is improved by mixing decanol or hexanol
up to 50% v/v with methanol. Both alcohols have long non-polar chains, thus improving the dissolution.

Mineral oils, transformer and silicon oils, greases and glues:

Chloroform is the auxiliary solvent used for complete dissolution of these samples. Note that the amount of
chloroform should not be higher than 70% v/v to avoid interferences during the Karl Fischer reaction.

pH value adjustment:
When the samples are acidic or basic, thee optimum pH for the Karl Fischer Titration i.e. between pH 5 and 7 is
achieved by adding buffers (imidazole, or benzoic acid) to the KF solvent before starting the titration.

Procedure for direct titration:

In the direct titration, the sample is introduced directly in the titration vessel using the back-weighing technique:
1. A sample aliquot is taken with the syringe/weighing boat.
2. The syringe/weighing boat is placed on the balance pan and the balance is tared.
3. Subsequently, the syringe/weighing boat is taken from the balance, and the sample is rapidly injected.
4. Back weighing of the empty syringe/weighing boat, the value entered in the instrument before starting titration.

Generator
electrode

Measuring electrode
(double platinium pin)

Diaphragm Anolyte
+

1 2 3 4
Figure 5: Back-weighing procedure

Summary :

Direct titration
Sample Soluble If required: Adjust pH value
No side reactions Improve solubility
- Auxiliary solvents
- Warming/heating
No specific sample preparation

METTLER TOLEDO Competence Guide 9

 Internal extraction

The internal extraction technique -more precisely: the direct titration with internal extraction- is applied when the
water is extracted directly in the titration vessel by means of e.g.
Competence Guide

• A long stir time (e.g. 5-10 min), and/or

• Speed (e.g. 80%), or
• High-speed homogenizer for a short but intensive dispersion action,
• Use of auxiliary solvents, or a combination of all previous steps.

The sample preparation by internal extraction is applied when

• the sample is not soluble,

• It does not undergo side reactions with the KF reagents, and
• The water release is fast.

Figure. 6A: The high-speed mixer is mounted on

the top of the titration stand of the instrument.

The use of a high-speed homogenizer allows for an efficient and quick dispersion of the sample. Consequently,
the water is released completely and fast from the sample matrix:

Figure. 6B/6C: Homogenizer before (left, B) and during action (right, C) in the titration vessel of the KF volumetric instrument.

Summary :

Internal extraction
Sample Insoluble e.g. almonds Extraction by Long stir time = 5-10 min
No side reactions e.g. instant coffee High–speed mixer
Fast water release e.g. tablets Improve solubility - Aux. solvents

METTLER TOLEDO Competence Guide 10

 External extraction

This sample preparation technique is needed when the water is extracted from the sample in an external vessel
containing a solvent of known water content during a defined time e.g. 1-2 hours. Subsequently, an aliquot of the
Competence Guide

extraction solvent is titrated by injection in the KF titration vessel. Briefly:

1 2 3 4
Blank Determination Weigh solvent and sample Extract water Settle and titration

Figure. 7: The sequence for the external extraction

To achieve accurate and precise results it is necessary that

• The amount of water present in the extraction solvent is very low.

• The extraction capacity of the solvent is large enough (typical values: chloroform can extract max. 350 ppm,
and toluene max. 600 ppm)
• The quantity of extraction solvent is large enough to allow for blank determination and for the extraction.

The sample preparation by external extraction is required when

• The samples are not soluble,

• The water release is slow,
• The water content of the samples is very low, thus a large sample size is needed
• The samples do not undergo side reactions with the KF reagents, and
• The samples cannot be titrated in the coulometric titration vessel since they are solid.

Summary :

External extraction
Sample Not soluble e.g. tobacco Extraction by Suitable solvent or water content << sample
solvent mixture water content
Slow water release e.g. cheese Long extraction time 1-2 h and more
Very low water content Large sample size Large sample size for the
e.g. surface water of sugar extraction to get a large
sample water amount with
respect to the solvent water
content
No side reactions e.g. wool
Solid sample – Direct coulometric NEVER enter solid sample Generator cell will be
titration not possible e.g. plastic pellets in a coulometric KF titration damaged!
vessel

METTLER TOLEDO Competence Guide 11

 External extraction in four steps

The workflow of the external extraction consists of four steps:

Competence Guide

1 • Place the extraction solvent in a flask

and close the flask with a septum
• Take an aliquot of the solvent with a
syringe
• Determine the water content (blank
value)

Solvent in bottle with Take an aliquat of the Determine water

septum solvent content of solvent =
Blank value
Figure. 8A: Blank determination
2 • Weigh the solvent
• Pour the solid sample in the flask
and close the flask with the septum
(dilution factor: 10-20, sample size:
5-20 g)
• Weigh the sample in the flask with
the septum

Weigh solvent Pour in solid sample Weigh sample

Figure. 8B: Weigh solvent and sample
3 Extract water from the sample
• overnight
• with shaking
• with ultrasonic and/or heating bath

Overnight With shaking With ultrasonic and/or

heating bath
Figure. 8C: Water extraction
4 • Wait for settling of the insoluble
sample
• Take an aliquot of the supernatant
liquid
• Titrate the aliquot to determine the
extracted water amount
Generator
electrode

Measuring electrode
(double platinium pin)

Diaphragm Anolyte
+

Settling Take a liquot of Titrate Sample

extraction sample

Figure. 8D: Water extraction

METTLER TOLEDO Competence Guide 12

 The calculation for the water content in ppm of the extracted sample is defined in the following formula:

-
Competence Guide

where
x Water content (ppm) of titrated sample/aliquot ppm
f1 Water content of extraction solvent (blank) ppm
f2 Weight of extraction solvent used grams
f3 Total weight of sample used for extraction grams

Similarly, the calculation for the water content in % of the extracted sample is defined in the following formula:

where
x Water content of titrated sample/aliquot %
f1 Water content of extraction solvent (blank) %
f2 Weight of extraction solvent used grams
f3 Total weight of sample used for extraction grams

METTLER TOLEDO Competence Guide 13

 External extraction of lyophilized substances

Lyophilized substances (or freeze-dried substances) have a very low water content in the order of e.g.
10-50 ppm. For this reason, they are very hygroscopic when exposed to air, i.e. they easily absorb moisture
Competence Guide

from the air as soon as the sealed vials are opened.

To avoid this, the extraction of water from the lyophilized sample is performed directly into the sealed vial by
adding dried coulometric anolyte during pretitration:

1. Fill the KF coulometer with fresh anolyte

2. Start the pretitration
3. The anolyte is dried and the instrument has reached stand-by status
4. While in stand-by, aspire 1-2 mL of dried anolyte with a 10 mL syringe from the cell
5. Shake the syringe to eliminate water present on the inner wall of the cylinder
6. Discard the aspired solution
7. Aspire 5-10 mL of dried anolyte with the same syringe
8. Inject the dried electrolyte in the sealed vial through the septum
9. Shake the vial to allow for water release from the lyophilized sample
10. Let the solution settle down
11. Aspirate an aliquot and titrate it.
12. Form the water amount determined with the KF coulometer and the sample mass given on the label of the
vial, the water content of the freeze-dried sample can be calculated.

↓
↓ ↓
Generator
electrode

Measuring electrode
(double platinium pin)

Diaphragm Anolyte
+

1. Pretitration is running Shake 1. Wait for settling

2. Inject dry anolyte into the septum bottle 2. Take an aliquot

Figure 9: External extraction of water in lyophilized (freeze-dried) substances

METTLER TOLEDO Competence Guide 14

 External dissolution

In the case when the water content is extremely low, a much too large sample mass (e.g. 30-40 g) has to be
used to achieve accurate and precise results. This will quickly fill up the titration vessel, and not more than
Competence Guide

1-2 samples could be titrated. Thus, the KF solvent must be often replaced. In the worst case, it has to be
replaced even after one single KF titration since the vessel is too full to accommodate other samples.

A large amount of the sample is dissolved in a flask containing a suitable solvent. Subsequently, an aliquot of
the extraction solvent is titrated by injection in the KF titration vessel. The whole procedure is similar to the
external extraction, however the calculations are slightly different:

)–(

where
x Water content of titrated sample/aliquot %, or ppm
f1 Water content of extraction solvent (blank) %, or ppm
f2 Weight of extraction solvent used grams
f3 Total weight of sample used for dissolution grams

The sample preparation by external dissolution is applied when

• The samples are soluble,

• The water distribution in the sample is inhomogeneous,
• The water content of the samples is very low, thus a large sample size is needed
• The samples do not undergo side reactions with the KF reagents, and
• The samples cannot be titrated in the coulometric titration vessel since they are solid.

As in the case for the external extraction, to achieve accurate and precise results it is necessary that

• The amount of water present in the dissolution solvent is very low.

• The dissolution capacity of the solvent is large enough
• The solvent amount used is large enough to allow for blank determination as well as for the dissolution.

Summary :

External dissolution
Sample Soluble e.g. phenol Dissolution Suitable solvent or water content << sample
solvent mixture water content
Non-homogeneous water distribution Long extraction time 1-2 h and more
e.g. sucrose
Very low water content Large sample size large water amount
e.g. petroleum products
No side reactions e.g. phenol
Solid sample e.g. sugar Direct coulometric titration not possible

METTLER TOLEDO Competence Guide 15

 Automated calculation for external extraction and dissolution

The specific calculations for both sample preparations are built-in in the high-end KF titrators to improve
easy-of-use and reduce errors when performing these techniques prior to the KF titration.
Competence Guide

In fact, these KF instruments have preprogrammed standard method templates as well as tested titration
methods to perform automatically the whole procedure including the calculations. Depending on the analysis to
be performed, the corresponding template is selected:

From the list of templates the method KFC002 is selected to develop a KF titration with external extraction (or
dissolution)

In the method function “Calculation” you can select the corresponding procedure sample preparation procedure,
as well as and the desired units i.e. “ppm” or “%”

METTLER TOLEDO Competence Guide 16

Competence Guide

Select the method function “Sample”

2.
1. Under the function “Sample”, select “Blank” to first
determine the water content (blank) of the 1.
extraction (dissolution) solvent.
2. Subsequently, the sample is determined
(select “sample”)

Briefly, the operator is guided through all steps of the analysis. In the case of an external extraction followed by
coulometric Karl Fischer Titration, the detailed workflow is given in the next page.

METTLER TOLEDO Competence Guide 17

 Titration workflow for external extraction/dissolution

1. Start blank determination

Competence Guide

1.1 Press “Start blank” when the corresponding button

is active.
1.2 After the blank has been determined, the value is
automatically stored in the instrument.

↓
2. Start sample titration

2.1 Press “Start sample” when the corresponding

button is active.

2.2 The solvent mass is requested to be entered in the

instrument.

2.3 The mass of the extraction sample is entered in the

instrument.

METTLER TOLEDO Competence Guide 18

 Gas-phase extraction

When the sample undergoes side reactions with the KF reagents, water has first to be released by heating the
sample in an oven with subsequent transfer into the titration vessel. The KF oven is connected to the KF titrator
Competence Guide

with a transfer tube with an inlet in the titration cell, and the evaporated moisture is transferred using a stream of
dry and inert gas such as nitrogen.

The gas-phase extraction is used when

• The samples are insoluble,

• The samples undergo side reactions with the KF reagents,
• The water release from the sample matrix is very slow, and
• The samples do not decompose when heated i.e. they are thermally stable.

Figure 10: A continuous flow of dry gas enter the sample vial in the oven and transport the evaporated water into the KF titration vessel.

Summary :

Gas-phase extraction
Sample Side reactions with the KF reagents e.g. Heating and No contact with reagents
motor oil collecting the
Insoluble e.g. plastic chips vapor
Slow water release e.g. polymers Heating time Recommendation:
• At least 10 min
• max. 30 min
Thermally stable – no decomposition Heating temperature • Select heating
e.g. polylactic acid (sutures) temperature 10-20°C
below melting point
• Avoid oxidation by using
nitrogen as inert transfer
gas

METTLER TOLEDO Competence Guide 19

 Water content in gases

To determine the water content of gases, the gas is purged through the titration vessel for a defined time period.
During this time, the KF solvent absorbs the gas humidity. First, the gas is passing through a flow meter to
Competence Guide

ensure a constant flow rate to achieve an accurate determination of the total gas volume. Subsequently, the gas
transfer is stopped, and the titration is started:.

Figure 11: The gas humidity is collected in the titration vessel during a specific time e.g. 5 min before titration.

The amount of water mwater determined by KF titration allows for the calculation of the water content:

1. The total gas volume V is determined by multiplying the gas flow rate (mL/min) with the time (min):
2. The water content is determined according to:

( )

1. The water content is determined according to:

Summary :

Determination of water in gases

Sample Gaseous samples e.g. nitrogen Purge the gas into Constant flow rate
the titration vessel
No side reaction with the KF reagents
e.g. liquefied petroleum gas

METTLER TOLEDO Competence Guide 20

 Troubleshooting: The most frequently asked questions
The drift does not decrease to a low and stable value even after 1-2 hours. Any hints?
Competence Guide

The air humidity is the big enemy of a Karl Fischer titration! Measures are implemented to reduce the influence of
air moisture on the KF reaction. More specifically, the penetration of air humidity in the titration vessel has to be
limited by checking the following points:

• Connections to openings of the KF titration vessel have to be greased i.e. tube inserts, electrode, adapters,
septum and generator cell (KF coulometer), as well as stoppers.
• Use dry and fresh molecular sieve and silica gel in the drying tubes to ensure an efficient absorption of
moisture from the air.
• KF coulometer: the ceramic frit of a generator cell with diaphragm has to be dry.

Figure 12: Grease and tighten all connections (green arrows)

Explanation:

During pretitration, the titrator add (generates) iodine to react with water present in the KF solvent. When the KF sol-
vent is dry, the titrator switches from pretitration in the stand-by mode. At this point, the online drift is still too high to
start a sample analysis. In general, it takes additionally 20-30 min to get a low and stable drift value of 10 μg/min.

To decrease the drift value it is also important to use fresh drying agents in the drying tubes. We recommend
exchanging molecular sieves every 6 weeks. The molecular sieves is regenerated by drying it in the oven
overnight at e.g. 250-300°C (check product data sheet)

Furthermore, if you use a cell with diaphragm, wash the diaphragm with dry methanol and store it for at least one
night in an oven at max. 80°C, or at a well-ventilated place to evaporate water trapped inside the ceramic diaphragm.

Solution:

How to quickly achieve low and stable online drift values

1. Work in suitable working spaces (low humidity, room temperature, no strong ventilation)
2. Check tightness of the system
3. Use fresh KF reagents, solvents and molecular sieve
4. Regularly renew and dry exhausted molecular sieve
5. Gently shake the titration vessel to collect traces of water from the glass walls
6. Coulometric generator cell: Dry it in the oven if the ceramic frit is too wet (max. 80°C)

What to do If after 20-30 min the online drift is still too high e.g. 20-30 μg/min
Check again points 1.-6. (1.-7.)

METTLER TOLEDO Competence Guide 21

 What is the smallest amount of water that can be determined by Karl Fischer titration?

In volumetric KF titration, this mainly depends on

Competence Guide

• The equipment used,

• The burette size,
• The KF titrant concentration, and
• The sample size.

A KF Compact Volumetric Titrator can determine the smallest amount (LOD) of 62.5 μg water in a 5 g sample,
using a 5 mL burette with a 5 mg/mL KF titrant. This corresponds to a water content of 12.5 ppm under optimal
conditions.

A KF Compact Coulometer can determine the smallest amount of 10 μg water. This corresponds to a water
content of 1 ppm for a 10 g sample – under optimal working conditions.

KF Volumetric Titrator KF Coulometer

Theoretical smallest increment Resolution: 20’000 burette steps 0.1 mC (100 mA x 1 ms)
5 mL burette: 0.25 μL Theoretical: 0.01 μg H2O
5 mg/mL: 1.25 μg H2O/step Resolution: < 0.1 μg H2O
Limit of detection (LOD) 50 x resolution 10 μg
5 mL burette: 12.5 μL Example:
5 mg/mL: 62.5 μg H2O 10 g sample → determ. of 1 ppm
Example: Assumptions:
5 g sample → determination of 12.5 ppm - Current is used for iodine generation
- Iodine is reacting with water only.

Explanation:

The highest dosing accuracy is reached when the titrant consumption is within 10-90% of the absolute burette volume.

Hence, the most accurate results will be obtained using half of the burette volume for one titration. For instance,
best results for a sample with an expected water content of 5% are achieved if you use a sample size of 0.25 g
in combination with a 5 mL burette and a 5 mg/mL KF one-component titrant.

If the water content is very low e.g. 0.1%, a much bigger sample size is required to obtain best results, or the
sample has to be titrated by a KF Coulometer.

Figure 13: Suitability of KF coulometric and volumetric titrators.

METTLER TOLEDO Competence Guide 22

 Is it mandatory to check the concentration of a new Karl Fischer titrant bottle?

Indeed, it is always necessary to determine the concentration of the Karl Fischer titrant since the value printed on
the label is a nominal value, and not the current concentration. Thus, if one calculate the water content using the
Competence Guide

nominal titrant concentration after a sample determination, the result will not be accurate. The determination
procedure is shown in a short film under Determine titrant concentration - Karl Fischer Titrator - YouTube.

Explanation:

The concentration of e.g. the one-component 5 KF titrant is always slightly higher than 5 mg/mL to allow for a
longer storage time of the bottle. Moreover, the titer is always changing with time even in a closed bottle due to
air moisture still penetrating into the bottle.

The concentration value indicated in the Certificate of Analysis is determined just after production of the reagent.
As soon as you open the bottle, air humidity reacts with it, thus decreasing its value. Hence, it is necessary to
perform a concentration determination value.

Note that the concentration determination using reference materials is a straightforward way to verify the correct
working of the whole titration instrument.

Solution:

Once the new KF titrant bottle has been connected to the titrator, a concentration determination is performed
using

• 1 mg H2O/g and 10 mg H2O/g liquid water standards

It is recommended to work with liquid water standards for the concentration determination of the KF titrants
due to their straightforward use.
• Sodium tartrate dihydrate (15.66 % water content)
Use a sample size of 50 mg for each titration, and exchange the KF solvent after performing three
determinations in the same solvent i.e. max. 150 mg.
• Pure water (100% water content)
It is not recommended to use pure water since the weighing uncertainty is too high due to the very small
amount of water needed (10-30 mg). Thus, the accuracy and repeatability are strongly affected by it.
The concentration value affects the speed of the Karl Fischer reaction: in fact, the titration time is increased with
decreasing titrant concentration.

To achieve optimum titration time, it is recommended to define tolerances for the concentration determination:

• 5 mg/mL: min. 4.5 mg/mL

• 2 mg/mL: min. 1.5 mg/mL

METTLER TOLEDO Competence Guide 23

 Which water standards do you recommend for the concentration determination?

The use of liquid water standards is strongly recommended since they allow for seamless determination of the
KF titrant concentration.
Competence Guide

Two liquid water standards for the volumetric KF titration are commercially available with a concentration
10 H2O mg/g and 1 mg H2O/g. Ready-to-use titration methods developed for these standards are stored in the
KF titrator, hence no additional steps are needed.

Explanation:

Beside liquid water standards, also certified solid standards are available for the concentration determination of
the KF titrants. The reference material of choice is sodium tartrate dihydrate, an organic compound with an exact
water content of 15.66%. Briefly, water is tightly bound in the crystal structure of sodium tartrate. The dissolution
in the KF solvent allows for release of water from the structure, and thus water can be subsequently titrated.

However, sodium tartrate dihydrate is only partially soluble in the KF solvent, hence the solvent must be replaced
as soon as it becomes turbid when titrating several samples:

• Max. 0.15 g sodium tartrate in 40 mL KF methanolic solvent i.e. max. three samples of each 0.05 g can be
titrated in the same solvent.
• Max. 0.05 g sodium tartrate in 40 mL KF methanolic i.e. after one sample of 0.05 g sodium tartrate the
solvent should be replaced.

Hence, the best results and most convenient use will be obtained with the liquid water standards. In fact, liquid
standards are soluble in most solvents without any problem when compared to sodium tartrate. When using
liquid standards, you can even perform six determinations in the same solvent. Moreover, you get a lower
standard deviation and the analysis time is much faster since no long stirring time is needed as for a solid water
standard, which requires a stir time of 120-180 s prior titration.

It is also possible to perform the concentration determination using pure water. However, the accuracy is strongly
affected due to very small sample size used i.e. 10-30 μL. Thus, the influence of sample handling will be highly
increased. As an example, if you lose e.g. 5 μL of your 10 μL pure water sample, then there will be a huge error
of 50%. On the other hand, the sample size with liquid standard is in the order of 1000 μL, and therefore a loss
of 5 μL is not so relevant as in the case of pure water.

Solution:

How to perform the titer determination of a KF titrant:

1. Flush the whole system i.e. the burette and tubes three times, and discard the dispensed reagent using the
manual operations of the titrator. This will remove any bubbles in the tubes and also guarantees that the titer
in the dosing unit and also in the tubes will be the same.
2. Use the 10 and/or 1 mg/g liquid water standard for the concentration determination.
3. If the ambient conditions are changing during the day, e.g. due to a humidity increase and/or a temperature
increase by more than 3-5°C, the concentration determination must be repeated.

METTLER TOLEDO Competence Guide 24

 Are liquid water standards hygroscopic?

Certified liquid water standards consist of an exact amount of water dissolved in a solvent mixture. The composi-
tion of the water standards has been optimized to reduce the absorption of air humidity. In any case, Good Titration
Competence Guide

Practice (GTP) strongly recommends not to leave the ampoules open with liquid standard inside. The liquid stan-
dard should be completely aspired and transferred from the ampoule in a gas-tight syringe (e.g. a glass syringe).

Explanation:

Every solvent that is exposed to air may absorb humidity until saturation is reached i.e. until chemical
equilibrium between the liquid and the gaseous phase is achieved. The amount of water absorbed from the air in
the liquid standard mainly depends on the

• Relative humidity of air,

• The polarity of the solvent, and
• The temperature

Solvents such as

• Ethanol, methanol and isopropanol i.e. alcohols (R-OH),

• Acetone CH3-CO-CH3 and methyl isobutyl ketone MIBK i.e. ketones (R1-CO-R2)
• Diethyl ether CH3CH2-CO-CH2CH3 and tetrahydrofuran (CH2)4O i.e. ethers (R1-O-R2)

easily absorb water from the air due to the presence of oxygen at-
oms in their chemical structure leading to electrical polarity. The
water molecule H2O is strongly polar, where the oxygen atom O
is negatively charged and the two hydrogen atoms H are positively
polarized. Thus, water is easily absorbed in a polar solvent because
polar substances tend to mix up.

On the other hand, solvent like toluene CH3C6H6 and chloroform

CHCl3 are weakly polar or even non-polar. Hence, the absorption
of water is extremely small.

Figure 15: The water molecule H2O contains two strong

By selecting suitable solvents for the production of the water standards, polar chemical bonds between hydrogen H and oxygen
the stability of these standards can be optimized. O atoms. The negative electrical charge shifts towards
the central red oxygen atom.

Solution:

How to use a liquid water standard

1. Open the glass vials containing 5-6 mL of water standard

2. Aspire 0.5-1 mL liquid standard into the syringe
3. Pull the piston to the maximum position
4. Shake the syringe to wash the inner wall
5. Discard the liquid standard
6. Aspire the whole content from the glass vial
7. Start the concentration determination.

Figure 16: Aspire the whole content in the syringe.

METTLER TOLEDO Competence Guide 25

 How often should the concentration of the KF titrant be determined?

The Karl Fischer titrant is very sensitive to air humidity; thus, its concentration must be determined every time
before determining the water content of a sample by volumetric Karl Fischer titration. The procedure of the
Competence Guide

concentration determination for KF titrants is shown in a short film under Determine titrant concentration -
Karl Fischer Titrator - YouTube.

Explanation:

The determination of the titrant concentration is a crucial step in titration workflow. Without knowing the exact
concentration it is not possible to achieve correct results. Thus, the determination must be performed at least
every time before starting a series of measurements. Moreover, KF titrants are sensitive to air humidity and their
concentration slowly decreases with time – even if the titrant bottle is tightly closed. It is estimated that the
decrease in concentration of the KF titrant is in the order of -0.1 mg/mL per week.

Therefore, depending on the ambient conditions, the concentration of the KF titration have to be determined more
frequently. For instance, if the temperature and humidity are changing a lot during the day, it is necessary to
repeat the determination even during the daily routine.

METTLER TOLEDO Competence Guide 26

 How do you determine the titer for the coulometric KF reagent?

It is not necessary to determine the concentration of the coulometric KF reagent since coulometry is an absolute
method because the amount of current used to generate iodine by electrochemical processes is directly
Competence Guide

measured by the electronics in the titrator. Thus, the generated current does not have to be compared to any
reference material.

Explanation:

The amount of electrical charges (Coulomb, C) needed for the iodine generation is used in the calculation of the
water content through the Faraday constant (96485 C/mol). It is required that the generator and electronics in
the instrument are correctly working to achieve accurate results.

Anode +

Generator
electrode

2H+ H2

Catholyte

Diaphragm Anolyte
I I I I I2 I2

Iodine I2 will be produced

just in time from iodide I
2I Anodic
I2 + 2 e
oxidation

Figure 17: Working principle of the coulometric determination of water by KF titration.

Calculation of the water content:

• 96485 C are required to produce 1mol of a chemical compound with 1 mol of electrons.
• Iodine I2 is generated from two iodide ions I- by oxidation at the anode of the generator cell.
• 1 mol I2 reacts with 1mol H2O, i.e. 1 mole H2O (18 g H2O) is equivalent to 2 x 96485 C.
This corresponds to 10.712 C/ mg H2O (Assumption: 100% electrochemical conversion).

It is recommended to periodically verify the performance of the KF coulometer. This is achieved by checking the
recovery (%) of certified liquid water standards 1 and 0.1 mg H2O/g.

METTLER TOLEDO Competence Guide 27

 The verification with standards is outside the uncertainty in the certificate of analysis. Any suggestions?

In general, if the results are outside the tolerances stated in the certificate of analysis (CoA) of the water
standards or they differ too much from the certified value, there is nothing left but to systematically check the
Competence Guide

whole titration system step-by-step to limit or suppress the influence of all possible sources of uncertainty.
Additionally, most errors come from handling problems, especially when working with the lowest water standard
with 0.1 mg/g (100 ppm).

Explanation:

The measurement uncertainty indicated on the CoA of the water standard e.g. 0.99% ± 0.02% only indicates the
uncertainty of the specific water standard measured just after production at the production site of the provider of
liquid standards. Thus, it indicates the calibration uncertainty of the water standard obtained at production plant
of the provider.

This means that the uncertainty of the CoA refers only to the conditions of content determination at the provider’s
site, i.e. to the analytical methods, instruments procedures, standard operating procedures SOPs, ambient
conditions etc. used at provider’s site. The uncertainty value is only valid under the same working conditions and
instruments at the provider’s site. However, this is never the case, and thus it is not related to the result
uncertainties of measurements. Therefore, it is not correct to state that a value is wrong if it outside the
tolerances of the CoA.

To assess if a measurement is acceptable or not, note that the providers of reference materials give
recommendations, as given below:

Figure 18: Certificates of analysis (CoAs) indicating both a) the calibration uncertainty of the water standard determined after production at
provider’s site, as well as b) recommendations for the assessment of tolerances for the determination of water by KF titration.

METTLER TOLEDO Competence Guide 28

 Solution:

How to reduce handling errors:

Competence Guide

Most errors come from handling problems, especially with the low water standards that have only 100 ppm. It
is recommended to use airtight glass syringes with long needles since glass is not absorbing air moisture as
plastic:

1. Shake the liquid standard glass ampoule before opening it.

2. Snap of the top tag of the ampoule.
3. Rinse the needle and the plunger by taking approx. 0.5 mL standard into the syringe. Avoid taking any air into
the syringe while doing this step.
4. Discard the solution and do not pull the plunger up and down.
5. Wipe the needle dry and draw the rest of the standard immediately into the syringe.
6. Remove any air bubble from the syringe and wipe the needle dry. Now your standard is well protected and
ready for use.
7. Add an aliquot to the titration vessel by back-weighing, and carry out the titration.
8. Before further use, remove few drops of standard from the tip of the syringe and wipe the needle dry again.

Suggestion:

How to define the tolerances for your measurements:

Use the limits recommended by the providers of the water standards as highlighted in yellow at the end of the
certificates in figure 18, and also of additional documents related to the standards.

Figure 19A: Definition of result limits in the method function “Calculation”.

Figure 19B: Warning message on the titrator display if the result is outside the limits.
METTLER TOLEDO Competence Guide 29
 What do you do if the result is not correct?

It is expected that the titrator has been adjusted and its performance verified with water standards. Thus, the
most important point to be checked is related with the applied sample preparation i.e. if the sample preparation
Competence Guide

has achieved a complete release of water from the sample matrix. In other words: Has all water been released
from the sample? Is it completely dissolved in the KF solvent?

Explanation:

For a successful water content determination by the Karl Fischer titration, the sample needs to be fully dissolved
or dispersed to completely release water. Moreover, no side reactions should take place, and the correct pH for
the KF reaction should be between 5.5 and 8.

For example, the determination of water content in lubricants requires the use of a KF oven to evaporate water,
since the additives present in these samples lead to side reactions that affect the result. Thus, a direct titration is
not the method of choice. In fact, the direct injection of oil samples containing additives such as thiols R-SH lead
to higher results because the thiol group R-SH reacts with iodine giving an erroneous higher consumption.

Note that there are also some oils and lubricants without additives, therefore they can be titrated by direct
injection in a mixture consisting of the KF solvent and auxiliary solvents such as e.g. chloroform facilitating the
dissolution of the oil samples.

Figure 20: Water is bound to the sample matrix and it has first to be released before starting the KF titration.

Solution:

Always select the suitable sample preparation first

In general, deviations from the expected results are due to incomplete release of water from the sample.
Depending on the sample nature, there are different preparation techniques for the complete release of water as
well as for the handling of the sample:

1. Direct titration
2. Internal extraction (inside titration cell)
3. External extraction
4. External dissolution
5. Gas-phase extraction
6. Water content in gases

METTLER TOLEDO Competence Guide 30

 How long should the stand-by take until the online drift reaches a low level?

In general, the stand-by (or conditioning) of the KF titration cell should take no longer than 20- 30 minutes to
reach low online drift values. This can be achieved if the KF coulometric electrolyte or the KF volumetric titrant
Competence Guide

and solvent used are fresh and the titrator is well tight, i.e. all apertures and connections of the cell are slightly
greased, correctly assembled and tighten.

To decrease the online drift value it is also important to use fresh drying agents in the drying tubes. We
recommend exchanging molecular sieves every 6 weeks. Furthermore, if you use a cell with diaphragm, wash
the porous ceramic diaphragm with dry methanol and store it at least for one night in an oven at max. 80°C, or
at a well-ventilated place to evaporate water trapped inside the ceramic diaphragm.

Explanation:

After assembling and filling the titration cell, the pretitration is started to eliminate the water present in the KF
solvent or the coulometric electrolyte. When the solvent is dried, the titrator switches from pretitration in the
stand-by mode. However, at this point the online drift is generally still too high to start a sample analysis.

During stand-by, the online-drift is slowly but continuously decreased since the titrator automatically reacts to
any potential change caused by the presence of humidity e.g. from air. This step may require 20-30 minutes in
general.

To speed up the process, it is beneficial to gently shake the titration vessel in order to efficiently collect traces of
water adsorbed on the glass wall of the titration cell.

Low drift values are necessary to achieve accurate and precise results. The following table gives indicative
values that can be reached with a KF titrator:

Suitable online drift for titration start

KF Volumetric Titration 1 - 10 μg/min
KF Coulometric Titration 1 - 5 μg/min

Solution:

How to achieve low drift values

1. Work in suitable working spaces (low humidity, room temperature, no strong ventilation)
2. Check tightness of the system
3. Use fresh KF reagents, solvents and molecular sieve
4. Regularly renew and/or dry exhausted molecular sieve
5. Gently shake the titration vessel to collect traces of water from the glass walls
6. Coulometric generator cell: Dry it again in the oven

What to do If after 20-30 min the online drift is still too high e.g. 20-30 μg/min
Check again points 1.-6. (1.-7.)

METTLER TOLEDO Competence Guide 31

 The online drift is still very high after titration end. How can this problem be solved?

A too high drift value e.g. 60 μg/min after sample titration can be attributed either to
Competence Guide

1. Side reactions with the KF reagents, or

2. The sample is not completely dissolved,
3. Water is not completely released.

Thus, either special KF reagents have to be used, or the sample must be heated with the KF oven, or the sample
preparation has to be improved to achieve the complete release of water.

Explanation:

Side reactions are caused by interfering chemical reactions between the sample and the KF reagents. Most of
these side reactions can be almost completely suppressed by using special KF reagents. For instance, in the
case of samples containing ketone groups e.g. R1-CO-R2, the use of so-called K-reagents allows to achieve
correct results by suppression of the side reactions.

Figure 21: KF titration of 5 mL acetone using two different solvent: blue line - classical KF solvent, green line - solvent for ketones and
aldehydes. The titration in methanol is not stopping even after 600 due to side reactions.

Another possibility to avoid side reactions is to heat up the sample in a KF oven and thus to evaporate water to
be titrated in the KF vessel.

In many cases, a longer stir time, a higher stir speed and the use of selected auxiliary solvents have proven to
be very beneficial to improve dissolution or extraction allow for complete release of water.

Solution:

Improve the sample preparation techniques

1. Select the correct KF reagent to avoid side reactions

2. Evaporate the water from the sample in a KF oven
3. Optimize the stir time, the stir speed and the use of auxiliary solvents to improve sample dissolution or water
extraction from sample matrix.

METTLER TOLEDO Competence Guide 32

 Sometimes we get negative results when using the automated KF oven – Why?

The KF instrument always determines the total amount of water present in the titration vessel, i.e. the water
present in the sample, the air humidity entering the vessel with time (drift) as well as adherent water on the
Competence Guide

glass surface of the sample vials (blank).

Drift and blank values are subtracted from the total amount of water. If the sum of both contributions are bigger
than the total amount of water, then the result becomes negative.

Explanation:

To calculate the water content of the sample, the contributions due to the air humidity (i.e. the drift) and – where
needed - of the adherent water on inner surfaces of the vials (i.e. blank value) have to be subtracted from the
determined water amount:

Volumetric KF titration:

VEQ: KF Titrant consumption mL

CONC: KF titrant concentration mg/mL
B[Blank]: Blank value μg
TIME: Titration time min
DRIFT: Drift value μg/min
m: sample mass g
1000: Conversion factor from μg to mg
0.1: Conversion factor from mg/g to %

Coulometric KF titration:

ICEQ: Current consumption mC

B[Blank]: Blank value μg
TIME: Titration time min
DRIFT: Drift value μg/min
m: sample mass g
10.712: Conversion factor from mC to μg
water ( 96485.33 C/mol · 2 (I2) ) / 18 g/mol / 1000 )
10000: Conversion factor from μg/g to %

Solution:

1. Drift:
a. Start the KF titration only when the online drift is very low e.g. below 5 μg/min. It may be necessary to wait
at least 15-20 min in stand-by to achieve low drift values.
b. Take into account or the drift by determining the blank using the same heating time as for the sample. In
this case, the drift value is set to 0 μg/min.
2. Blank value:
a. Use dry glass vials – previously dried in an oven,
b. Perform the blank determination for at least three vials, and calculate the average
c. Be sure that the inert gas used is correctly dried
3. Heating time:
Do not use too long heating time for the sample determination: a too long time increases the influence of the
drift value. Recommendation: 10-30 min.

METTLER TOLEDO Competence Guide 33

 Literature
“GTP Brochure – Good Titration Practice in Karl Fischer Titration”, METTLER TOLEDO Brochure No. 51725145,
October 2014.
Competence Guide

“METTLER TOLEDO Methods for Water Content Determination”, Titration Applications Brochure No. 38,
ME-51725075B, August 2012.

“Karl Fischer Titration with a Homogenizer – Practical Examples”, Titration Applications Brochure No. 27,
ME-51725053B, April 2013.

“Validation of Titration Methods - A Guideline for Customers”, Titration Applications Brochure No. 16,
ME-51724912C, March 2017.

“Selected Applications with Karl Fischer Oven Autosampler”, Titration Applications Brochure No. 49, 30452052,
November 2017.

Links
InMotion Karl Fischer Titration Oven Autosampler - METTLER TOLEDO – YouTube

Volumetric KF Titration - Tutorial - YouTube

Coulometric KF Titration - Tutorial - YouTube

Determine titrant concentration - Karl Fischer Titrator – YouTube

Automated Titration of the Water Content in Candies – YouTube

This application bulletin represents selected, possible application example. These have been tested with all
possible carein our lab with the analytical instrument mentioned in the bulletin. The experiments wre conducted
and the resulting data evaluated based on our current state of knowledge.

However, the application bulletin does not absolve you from personally testing its suitability for your intended
methods, instruments and purpose. As the use and transfer of an application example are beyond our control,
we can not accept responsibility therefore.

When chemicals and solvents are used, the general safety rules and the directions of the producer must be
observed.

METTLER TOLEDO Competence Guide 34

Titration Guide
Tips and hints for a successful water content determination by Karl Fischer Titration

This Titration Competence Guide is dedicated to the water content determination by

Karl Fischer (KF) titration. Focus has been put on the practical aspects rather than on
theoretical knowledge.

After a short introduction on Karl Fischer titration, the main points affecting result
accuracy such as low drift determination, tightness of the instrument, fresh KF reagents
and specific sample preparation will be presented in detail. In addition, frequently asked
troubleshooting questions will be presented and answered.

Several KF applications have been developed by the Market Support Group Analytical
Chemistry. They are available for download on the METTLER TOLEDO
website www.mt.com.

www.mt.com
For more information

Mettler-Toledo GmbH, Analytical

Heuwinkelstrasse 3
CH-8606 Nänikon, Switzerland
Tel +41-44 944 22 11
Fax +41-44 944 30 60

Subject to technical changes without notice.

©03/2023 Mettler-Toledo GmbH, 30736875
Market Support Group Analytical Chemistry