Titration

experiments
Karl Fischer

KF Guide 3
School

The Right Sample

Natural science laws experience “live” –

Taking Samples
learn
for Karl Fischer easily
Titration
2
Editorial

Dear Reader

Taking the right sample in the right way is a prerequisite to obtaining

accurate and reliable results. In the case of water and moisture content
determinations, avoiding water absorption from atmospheric moisture,
sampling tools or sample vials and vessels is a paramount sampling
objective.
Different sampling techniques have been developed to accommodate
the varying nature of samples since the first Karl Fischer titrations were
carried out in 1935.
This brochure explains some important rules and guidelines for draw-
ing samples to determine water content according to Karl Fischer.
Our tips and tricks incorporate the experience of METTLER TOLEDO’s
application chemists and are well proven through daily practice and
application.
Rely on good sampling techniques as the starting point of reliable re-
sults.

3
Content / Sampling

Content 4

1. Sampling 5
1.1 Taking the sample 5
1.2 Storing the sample 5
1.3 Amount of sample 6
2. Sample Addition 10
2.1 Liquid samples 10
2.2 Solid samples 12
3. The Influence of Atmospheric Humidity (Drift ­Determination) 15
3.1 Titration stand 16
3.2 The drift 16
4. More Information 18
4.1 Literature 18
4.2 More guides 18
4.3 Application brochures 18

4
 1. Sampling

1.1 Taking the When taking samples for water determination, you must be extremely
sample careful to exclude atmospheric moisture - the most common source of
error. If the water content of a sample changes during sampling due to
moisture being absorbed or desorbed, you will no longer be able to deter-
mine its true water content.
“An analysis cannot be better than the actual sample!”
When sampling, you should take into account the following points:
1. The sample must be representative, i.e. it must contain the same
­average amount of water as the material as a whole.
2. The sample should be taken quickly to exclude, or at least minimize,
the absorption or release of moisture.
3. Heterogeneous water distribution in samples:
In non-polar liquids, e.g. oils, the water is not uniformly dispersed. It
floats on the surface or sinks to the bottom. Liquids of this type must
be thoroughly mixed (by shaking) before a sample is taken.
In the case of non-polar solids, such as butter, which cannot be
mixed as thoroughly as liquids, the sample should be larger the more
­heterogeneous the distribution of the water.
4. Hygroscopic solids may exhibit higher water content on the surface
than inside if they have absorbed atmospheric moisture during
­storage.
5. Substances with very low water content:
Substances with a very low water content are frequently extremely
­hygroscopic. The sample must therefore be taken very quickly and with
a syringe or a spatula that is absolutely dry.

1.2 Storing the After you have taken the sample, you should determine its water content
sample as soon as possible. If you have to store the sample, keep it in a tightly
sealed bottle:

–– Glass bottles are preferable to plastic bottles because plastic is not

completely gas-tight, and thus air moisture can penetrate the plastic
and absorbed by the sample.
–– Use sample bottles with small openings to minimize the ingress of
moisture.
–– Use bottles with a septum stopper for liquids of very low water content.

5
Sampling

–– Use a bottle with an optimum volume for the amount of sample: the
smaller the gas space above the sample, the lower the amount of
­moisture.
– With liquid samples, rinse the bottles two or three times with the
­sample beforehand.

With liquid samples that do not dissolve water such as oils, water may
be separated from the sample if it is left to stand for a long time. This
can happen when a sample cools and the water solubility is decreased.
In such cases, the solubility of water in the sample can be increased us-
ing a solubility promotion agent such as 2-propanol.

1.3 Amount of The amount of sample used depends on

­sample • the expected water content, and the
• required accuracy and precision.

For coulometric analyses, the optimum amount of water is in the range

from 0.5 to 2 mg water per sample. Repeatable results can be obtained
even with 0.1 mg water per sample. Under optimum measurement
conditions, approx. 10-50 μg water can be detected if the demand for
repeatability is not too high.
In general, the accuracy is improved if larger sample amounts are used,
because the absorption of atmospheric moisture during sampling or
sample addition becomes less important.
For optimum accuracy with determinations in the range from 1 ppm to
1% water, it is recommended to use the minimum sample size given in
the following table:

Water content 1 10 50 100 500 1000 5000 10000

[ppm] = 1%
Min. sample 10 8 5 4 2 1 0.2 0.1
size [g]
Amount of 0.01 0.08 0.25 0.4 1.0 1.0 1.0 1.0
water [mg]

6
For volumetric titrations, the optimum amount of water is approximate-
ly 10 mg per sample. As a rule of thumb, the accuracy increases with
the amount of sample, because the absorption of air moisture during
sampling and sample addition becomes less important.

If a high level of accuracy is required, the amount of sample should re-

quire a titrant consumption between 30 and 70% of the nominal burette
volume. This corresponds to 7.5 to 17.5 mg water per sample for a
5 mL burette with a titrant concentration of 5 mg/mL.

Water content determinations below 1000 ppm are not required to be

quite as accurate: a relative standard deviation srel of 1 to 5% is gener-
ally considered as acceptable for such values. A titrant consumption of
0.1 to 0.05 mL is thus still acceptable when using a 5 mL burette.
This corresponds to 0.1 to 0.05 mg water per sample with a titrant
c­oncentration of 1 mg/mL.

For a more accurate determination of water traces (10 ppm to 1000 ppm)
by volumetric KF ttiration, a KF titrant with a lower concentration
(1 or 2 mg/mL) has to be used, and the following minimum sample
sizes should be used:

Water content [ppm] 10 50 100 200 500 1000

Min. sample size [g] 8 7 5 4 3 2

7
Sampling

Determination of the amount of sample for a water content in the

range from 1000 ppm to 100%

100% 1 µg

10% 10 µg

1% 0,01g 100 µg
0,1 g
1000 ppm 1g 1 mg
10 g
100 ppm 100 g 10 mg

10 ppm 100 mg

1 ppm 1000 mg

Principle:
The recommended sample size can be determined as a function of the
expected water con­tent for optimum KF analysis.
Procedure:
• Start from the optimum point (10 mg for volumetric KF, 1 mg for
­coulometric KF) or the recommended range
• The optimum point is con­nected by a straight line to the expected
­water content.
• The intersection point of this line with the “Amount of sample” scale
represents the recommended amount of sample to be used.

Note: Logarithmic scale!

Example:
Expected water content: 5000 ppm
Optimum amount of water: 10 mg/sample
Optimum sample amount: 2 g

8
 The Compact volumetric and colometric KF titrators support the user
with a sample size calculation routine, which is accessible in the online
­titrations screen:

Dependent on the expected water content, an optimum sample weight

range is recommended.

9
 2. Sample Addition
Sample addition

2.1 Liquid When adding liquid samples, you must take suitable precautions to pre-
samples vent atmospheric moisture from being absorbed, especially with samples
of low water content. The following procedures are suitable for the differ-
ent types of liquid samples:

Sample Examples Procedure

characteristics
High water content perfumes, Inject the sample into the titration vessel
aqueous either through a septum (KF coulometer)
Low viscosity emulsions, or through the needle hole in the three-
alcoholic hole adapter (volumetric KF titrator) using
beverages a 1 mL syringe with needle.
Low water content methanol, Store the sample in a bottle with a sep-
edible oils tum stopper to avoid moisture absorption
hexane, from the air.
Low water content toluene, Rinse a 10 mL syringe 2-3 times with
benzene the sample
Hygroscopic Inject the sample using a 1 mL or 10 mL
syringe through a septum cap.
Viscous glycerol, hy- Inject the sample using a 5 or 10 mL sy-
draulic oils, ringe with a thick needle into the titration
silicone oil, cell. Possibly warm the sample slightly to
mineral oils lower the viscosity.
massage oil In the case of a volumetric Karl ­Fischer titra-
tor, you can use a syringe without needle
since the sample can be added via the
larger hole of the three-hole adapter.
Very viscous ointments, Fill the 5 or 10 mL syringe with sample
creams, after removal of the piston. Inject it into the
yoghurt, cell using a wide-bore needle.
honey In the case of a volumetric Karl Fischer titra-
tor, you can use a syringe without needle
since the sample can be added via the
larger hole of the three-hole adapter.

a b c
Waxy candle wax, Liquefy the sample in an oven at approx.
paraffin, 50 °C and fill it into a syringe.
ski wax, The syringe is heated together with the wax.
supposito- This prevents the sample from solidifying in
ries the syringe during the weighing.

10
Taking samples from a bottle with septum stopper
After several samples have been taken, a vacuum devel-
ops in the bottle with septum stopper and it will no longer
be possible to take a sample aliquot. To avoid this, you
must aerate the bottle with dry air (equalize the pressure).
Fill a syringe without a plunger with molecular
sieves, seal it with cotton wool and insert it into
the septum bottle using a short needle. Air flows
through the molecular sieve into the b­ ottle when
you take a sample a­ liquot.

Sample addition of liquids with the back

weighing technique
– Fill the syringe to a quarter with sample. If
the sample is hygroscopic or has a low water
content (< 1000 ppm), use bottles with a
septum stopper and pressure equalization.
– Withdraw the plunger and rinse the syringe
with the sample by shaking it.
– Empty the syringe (into the waste bottle) and
repeat the rinsing two or three times.
– Fill the syringe with sample and wipe the
­needle with a paper tissue.
– Place the syringe (upside down) in a beaker on
the balance pan and tare the balance to “0”.
0.0000
– Start the titration method by pressing the 0

<Start> button.
– Inject the sample into the titration cell through
the septum stopper.
– Withdraw the plunger so that the drop at the
tip of the needle is sucked back into the nee-
dle. Otherwise when the syringe is removed,
the drop will adhere to the septum.
– Replace the syringe with the remaining sample
on the balance and back weigh it.
– Enter the sample size in the titrator or transfer
it automatically. -0.972
14
– Start the titration.

11
Sample addition

2.2 Solid It is not possible to directly titrate solid samples using KF coulometry
samples - when the titration cell is opened to add the sample, about 50-100 μg
water enter the anode compartment, depending on the ambient humidity.
With an optimum sample size of 1 mg water/sample, this would lead
to an error of 5% to 10%. For this reason, other methods have to be
used for the determination of low water content by KF coulometry in solid
samples:
• External extraction
• External dissolution
• Drying oven
On the other hand, in a volumetric KF titrator solids can be directly
transferred into the titration vessel. The sample should be quickly
weighed and added to minimize air exposure. If possible, it should be
added under the same conditions as it was transported and stored.
For instance, the storage of samples in a refrigerator may cause water
to condense; thus, it is necessary to warm up such samples to room
­temperature in a closed vessel before weighing.

Sample Procedure
characteristics
Brittle e.g. salts, crystalline samples:
Hard/soft Weighing boat
Pourable – Grind hard, coarse-grained samples
in a closed, cooled analytical mill;
pulverize less hard samples in a mortar.
– Add the sample with a weighing boat.

Finely powdered – Use a weighing boat with an attached flexible tubing to

Dusty add the sample: it prevents the sample from adhering
to the wall of the titration vessel or to the electrode.
Finely powdered e.g. salicylic acid, cellulose powder:
with very low water – Either weigh the sample in a dry box or extract it
content ­externally.
Soft e.g. jellied fruits, jelly bears, almond paste:
– Cut into small pieces with scissors or a knife and add the
sample with a spatula.

12
Sample Procedure
characteristics
Hard, fatty e.g. chocolate, solid fat:
– Grate the product and add the sample with a spatula.
Soft, fatty, inhomo- e.g. butter, margarine, edible fat:
geneous – Homogenize the sample well: the water is heterogene-
ously distributed. The water content is often lower atthe
surface than inside the sample.
– Add the sample with a spatula. Do not use a syringe,
because the water is released if it is pressurized
Waxy e.g. candles, paraffin, ski wax, suppositories:
– Liquefy the samples in a drying oven at approx. 50 °C
and fill them into a syringe.
– The syringe is heated up together with the wax. This
prevents the sample from hardening inside the syringe
during the weighing process
Creamy e.g. chocolate cream, honey, sugar products
Highly viscous – The METTLER TOLEDO Visco-Spoon™ simplifies the
handling with creamy samples since it can be directly
mounted into the titration stand.

13
Sample addition

Weight of solid samples with the b­ ack-weighing technique

–– Weigh the sample in the weighing boat.
–– Tare the balance to zero.
–– Add the sample into the titration vessel. Use a weighing boat with
­attached flexible tubing if necessary, to prevent the sample from
­adhering to the vessel wall or to the electrode.
–– Back-weigh the empty weighing boat.
–– Enter the weight in the titrator or transfer it automatically.
–– Start the titration.

1.3471 0.0000
5 0

14
3. The Influence of Atmospheric
­Humidity
Atmospheric humidity represents the most relevant source of error in Karl
Fischer titration. Moisture can enter the sample, the titrant and the titra-
tion stand. This problem is particularly relevant in tropical climates or in
coastal regions, where the relative humidity can achieve values of more
than 80%.
The assumption that air-conditioned rooms have a lower atmospheric
humidity is often not true: the majority of air-conditioning systems simply
cool the air. However, since cold air cannot absorb as much moisture,
the relative humidity increases. Briefly, the higher the atmospheric
­humidity in the laboratory, the greater is its influence on the results of
the Karl Fischer titration. The air-conditioning system should therefore be
equipped with a moisture condenser.
The Karl Fischer titrator should never be installed close to a ventilator of
the air conditioning system!

Water content in air Density of air

g/kg kg/m3

40.00 1.30

35.00
Water content in air
at 70% rH
30.00 1.25
Density of air
at 101.3 kPa at 50% rH
25.00

20.00 1.20

at 30% rH
15.00

10.00 1.15

5.00

0.00 1.10
10 ºC 15 ºC 20 ºC 25 ºC 30 ºC 35 ºC 40 ºC

Ambient air at 20 °C and 101.3 kPa contains water in the range of
5 - 10 g/m3 depending on the relative humidity (rH).

15
Drift determination

3.1 Titration stand The titration stand must be sealed as tightly as possible against atmos-
pheric moisture. The following rules should be observed:
• Close all openings in the titration stand.
• Condition the titration cell before use.
When you assemble the titration stand for the first time, there will be
moisture on the glass surface of the titration cell and the inserts. The
air within the titration vessel also contains moisture. After the anolyte
(coulometer) or the solvent (volumeter) has been added, it is titrated to
dryness during pretitration, i.e. until it is absolutely free of water.
The drift, however, remains high because the moisture present on the
glass walls and the air in the titration cell diffuse only slowly into the
anolyte (solvent). This can take 1 - 3 hours. The total moisture can
be titrated more quickly by gently moving the vessel from side to side
so that solvent swirls up the side of the vessel to pick up moisture ad-
sorbed on the glass walls. In this way, the residual moisture dissolves
more rapidly into the anolyte (solvent). Do not shake too vigorously to
avoid solvent gets on the cover of the vessel.
• Protect the titration cell with a desiccant (3 Å molecular sieve and
silica gel).
The desiccant absorbs the moisture and protects the titration cell against
the ingress of moisture. The drying capacity of the desiccant is limited
and it depends on the humidity and can be exhausted after 2 - 4 weeks
(indicated by a colour change of the indicator of the silica gel).
Silica gel can be regenerated over night at 150 °C, whereas molecular
sieves require temperatures up to 300 °C.

3.2 The drift No titration stand is completely water tight; traces of water will always
find a way into the titration cell. On the other hand, this water amount
is also titrated during analysis. Therefore it must be taken into account
when calculating the water content.
In this respect, it is necessary to quantify the amount of water entering
the titration cell during titration. Therefore, the drift indicates the quantity
of water that enters the titration stand over a defined period, t, and is
given in μg water/minute. This is achieved by titration of the dry solvent
for a defined time (drift determination).
In the standby titration, the KF titrator continuously titrates the water
that diffuses into the cell, and its value is displayed on the screen. At

16
the start of a sample titration, the last measured drift value is automati-
cally stored, if it is defined as the parameter “Source for drift: Online” in
method function “Titration stand (KF stand)”.

On the other hand, it is also possible to use the value of a previously

performed drift determination (“Determination”), or a fixed value can be
defined into the titration method (“Fix value”), or it can be entered by the
user (“Request”):

Source for Drift Explanation

Online Last measured value of standby
Determination The drift is determined and stored as raw result DRIFT
Fix value A fixed value is defined into method
Request The value can be entered immediately after starting
sample titration

The drift value is subsequently used in the calculation of the result

in order to compensate for the moisture that entered the titration cell
­according to the following formula:

Sample result = Total amount of water determined - (drift * titration

time).

For accurate results, the drift value should therefore be as low as

­possible and stable before the start of a titration!

17
 4. More Information
More Information

4 .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 G mbH, ­Darmstadt / Germany, 1985.

4 .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

4 .3 Application Many more details regarding the Karl Fischer titration are published in
brochure the application brochures
– Good Titration Practice™ in Karl Fischer Titration (ME 517252145).
– METTLER TOLEDO Methods for Water Content Determination,
­Application Brochure Nr. 38 (ME 51725075A).

18
19
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

Subject to technical changes

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