Titration Application Note

Water Content Determination

Note

in 5% Karl Fischer Oven Standard

Titration Application

Improved time-to-results and boosting productivity are key drivers for automation. High degrees
of automation require robust analytical equipment, with appropriate instrument qualification,
calibration and maintenance procedures ensuring correct results. The equipment qualification of
the whole titration system guarantees customers the accuracy, precision, and uptime they need
in their daily workflow.

In addition to the hardware qualification, the verification of the InMotion Karl Fischer Oven
Autosampler is performed using certified reference materials containing a well-defined amount of
water. The HYDRANAL™ Water KF Oven Standard 150-160°C is one solid water standard used
to verify the suitability of the system.

In this application note, the water content of the KF Oven Standard 150-160°C is automatically
determined using an InMotion KF Oven Autosampler in combination with a C30S Compact KF
Coulometer. The InMotion KF Oven Autosampler produces consistent, repeatable results with high
precision that are comparable with the data obtained by direct KF titration using a V30S Compact
Volumetric Titrator.

METTLER TOLEDO Page 1 of 12

30785465 - M880 v1.0
 Introduction Sample preparation and procedures
Titration Application Note

Lactose is a sugar compound consisting of one

Gas phase extraction – InMotion KF Oven/C30SX
molecule of galactose and one molecule of glucose
bound together through an oxygen bridge, i.e., a 1. Place an empty 5 mL glass vial closed with a
disaccharide. It is present in milk, and its content screw cap into the drift position on the rack.
can vary between 2-8% by mass [1].
2. Water is adsorbed on the inner surface of the
Lactose monohydrate is a white, non-hygroscopic vials. Its amount i.e., the blank value, is
powder containing water and lactose in a fixed considered by determination with empty vials,
stoichiometric ratio of 1:1. The water content in e.g., three empty vials.
lactose monohydrate is approximately 5-5.1%:
3. Add 0.05 g lactose water standard into a 5mL
vial and close it with the screw cap.

4. Place all filled vials on the sample rack.

5. Start the method. The pretitration will be

performed, while the InMotion KF Oven is
heated up to the predefined temperature.

6. After reaching a drift value below the minimum

start limit, the analysis will begin:
©: Lactose monohydrate - 4-O-β-D-Galactopyranosyl-D-glucose, Milk sugar (sigmaaldrich.com)
Drift → 3 Blank values → Samples
Lactose monohydrate is a suitable solid water
standard than can be used to verify the performance Direct Titration – KF Volumetry
of a Karl Fischer (KF) titration system consisting of 1. Start the method. The pretitration will be
the InMotion KF Oven Autosampler connected to a performed, then the instrument will
C30S Compact Coulometric Titrator. subsequently switch to the standby mode.
In this application we describe the determination of 2. Place a weighing boat on the balance, tare it.
the water content in the lactose water standard by 3. Add 0.1 g of lactose water standard.
two selected sample preparation techniques, i.e.,
4. When the value is stable, press again "Tare".
by:
5. Press "Start Sample" on the titrator.
• gas phase extraction using an InMotion 6. Enter the sample in the titration vessel and
KF Oven Autosampler,
confirm it on the titrator display. The titration
and
automatically starts after the sample addition.
• direct KF volumetric titration.
7. Place the weighing boat back on the balance
and enter the sample weight on the terminal.

C30SX Compact Coulometric Titrator V30S Compact Volumetric KF Titrator

with InMotion KF Oven Autosampler

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30785465 - M880 v1.0
 Sample Instruments and accessories
Titration Application Note

HYDRANAL™ Water Standard KF Oven • C30SX Compact Coulometric KF Titrator

150-160°C, No. 34693, 0.05-0.1 g. (30252662)
or
T7/T9 Titration Excellence
Lot M004B
(30252675/30252676)
Certified content:
with Coulometric KF Kit Titration Excellence
(5.09 ± 0.05) % KF-Volumetry
(30267113)
(5.08 ± 0.05) % KF Oven 150°C
(5.07 ± 0.02) % KF Oven 160°C • InMotion KF Pro Autosampler (5 mL)
(30407503)
Lot I158A
Certified content: • Heated Transfer Tube
(5.11 ± 0.05) % KF-Volumetry (30407443)
• V30S Compact Volumetric KF Titrator
Analyte: (30252667)
Water, M = 18.02 g/mol, z = 1 or
T7/T9 Titration Excellence
(30252675/30252676)
Chemistry with Volumetric KF Kit for Titration Excellence
CH3OH + SO2 + 3 RNH + I2 + H2O ⇌ (51105605)
(RNH2)(SO4CH3) + 2 (RNH2)I
• DV1005 Glass Burette 5 mL
(51107500)
Reagents and Solutions • LabX Laboratory Software
• XPR205 SN Precision Balance
KF-Coulometry (C30SX + InMotion KF) (30355411)

KF Coulometer electrolyte: • Weighing boat

HYDRANAL™ Coulomat AG-Oven (00023952)

Standards: • Spatula
C30SX: • 10 mL syringe (00071482)
HYDRANAL™ Water Liquid Standard 1.0 mg/g with needle (00078483)
C30SX + InMotion KF:
HYDRANAL™ Water Standard KF Oven
220-230°C (5.55%), No 34748-10G,
0.05 g

KF-Volumetry (V30S)
Titrant:
HYDRANAL™ Composite 5,
5 mg H2O/mL
Solvent:
HYDRANAL™ Methanol Dry, CH3OH
80 mL
Standard:
HYDRANAL™ Water Liquid Standard 10.0 mg/g

METTLER TOLEDO Page 3 of 12

30785465 - M880 v1.0
 Results Coulometric KF (C30XS + InMotion KF):
Titration Application Note

Volumetric KF (V30S):
160°C DRIFT: Lot M004B
R1: Lot M004B 16.9 µg/min Certified value %
Water content (%) Certified value %
1 5.12 R1:
2 5.10 Blank (µg)
3 5.08 1 31.5
4 5.04 2 27.1
5 5.09 3 22.7
6 5.09 Mean 27.1
Mean 5.09 5.09 ± 0.05 s 4.4
s 0.03 Average ± srel 16.236 %
srel 0.523 % expanded uncertainty R2:
Water content (%)
• To achieve dissolution, the sample was
1 5.04
stirred during 180 s before titration.
2 5.09
• All samples have been titrated in the same 3 5.08
solvent batch i.e., 80 mL methanol. 4 5.08
• Several sample series have been 5 5.07
measured. For each sample series, a new 6 5.06
batch of fresh solvent has been used: Mean 5.07 5.07 ± 0.02
s 0.02 Average ±
Certificate of V30S
srel 0.353 % expanded uncertainty
Lot analysis (n=6)
Average ± Average ± • The samples have been heated at 150°C
exp. uncertainty std. deviation and 160°C during 1200 s with air as
M004B 5.08 ± 0.03 % carrier gas.

5.05 ± 0.09 % • Several series from both batches have

5.09 ± 0.05 % been measured:
5.11 ± 0.05 %
(KF Volumetry)
5.08 ± 0.04 % Certificate of V30S
Lot analysis (n=6)
5.09 ± 0.03 %
Average ± Average ±
exp. uncertainty std. deviation
I158A 5.10 ± 0.04 %
5.11 ± 0.05 % M004B T = 160°C 5.08 ± 0.05 %
5.06 ± 0.04 % 5.07 ± 0.02 %
(KF Volumetry) 5.07 ± 0.02 %
5.11 ± 0.08 %
T = 150°C 5.08 ± 0.03 %
5.08 ± 0.05 % 5.08 ± 0.03 %
• The solution in the titration cell becomes
turbid with increasing number of added I158A 160°C:
samples. 5.14 ± 0.04 %
5.11 ± 0.05 % 150°C:
• The samples cannot be completely
(KF Volumetry) 5.11 ± 0.04 %
dissolved; thus, water is accumulating
when adding more samples. This 150°C:
5.14 ± 0.04 %
explains the higher online drift before
starting the next titration.

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30785465 - M880 v1.0
 Measured values – Direct titration
Titration Application Note

Water amount (black) and online drift value (grey) of Water Standard KF Oven 150-160°C during volumetric KF titration (0.1 g).

Time V Meas. value Temperature Water Drift

[s] [mL] [mV] [°C] [mg] [ug/min]
0 0.00000 631.4 25.0 0.0000 0.0
0 0.00000 631.4 25.0 0.0000 0.0
1 0.00100 631.1 25.0 0.0052 0.0
… … … …
50 0.66700 126.7 25.0 3.4822 3585.5
51 0.66725 257.4 25.0 3.4836 3241.8
52 0.67925 220.5 25.0 3.5462 2936.9
53 0.69550 119.8 25.0 3.6310 3444.9
54 0.69550 192.2 25.0 3.6310 3149.1
55 0.69825 281.0 25.0 3.6454 2318.9
… … … …
178 0.96200 88.8 25.0 5.0224 101.7
179 0.96200 94.7 25.0 5.0224 93.8
180 0.96200 85.6 25.0 5.0224 93.8
181 0.96225 93.7 25.0 5.0237 86.0
182 0.96250 104.2 25.0 5.0250 93.8
183 0.96300 93.4 25.0 5.0276 101.6
184 0.96325 97.5 25.0 5.0289 85.9
185 0.96375 91.5 25.0 5.0315 93.8
186 0.96375 78.1 25.0 5.0315 78.2
187 0.96375 91.0 25.0 5.0315 62.5
189 0.96375 82.6 25.0 5.0315 54.7
190 0.96400 84.1 25.0 5.0328 62.5
191 0.96400 86.5 25.0 5.0328 54.9
192 0.96400 86.1 25.0 5.0328 46.9
193 0.96400 93.6 25.0 5.0328 31.4
194 0.96425 86.9 25.0 5.0341 31.3
195 0.96425 93.4 25.0 5.0341 15.6
EP 1.650249 91.1

METTLER TOLEDO Page 5 of 12

30785465 - M880 v1.0
 Measured values – KF Oven method
Titration Application Note

The measured potential (blue) and water amount (grey) of Water Standard KF Oven 150-160°C
using the gas-phase extraction method (0.05 g).

Time C Meas. value Water Drift

[s] [mC] [mV] [µg] [ug/min]
0 0.0 402.3 0.0 0.0
1 25.3 401.6 2.4 0.0
1 110.4 401.0 10.3 80.3
… … … …
113 17062.3 186.8 1592.8 1126.3
114 17190.6 194.5 1604.8 1104.9
115 17332.4 194.5 1618.0 1128.5
116 17472.6 181.8 1631.1 1158.8
117 17590.5 171.4 1642.1 1106.9
… … … …
1200 34289.5 103.9 3201.0 25.4
1200 34289.5 103.9 3201.0 25.4
EP 34289.528193 103.9

• The heating time is fixed to 1200 s, i.e., after 1200 s the titration is stopped.
• The fixed heating time is defined by setting both the minimum and the maximum time to the same value
t(min)=t(max)=1200 s.
• The use of a fixed heating time following a short stir time allows for immediate titration of the collected water
evaporated from the sample, thus avoiding loss of water due to the hot air flow stream into the titration cell.
• A fixed heating time improves accuracy (no evaporation of water from the titration cell) and reproducibility
(same conditions for all samples) of the results.

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30785465 - M880 v1.0
 Temperature scan
Titration Application Note

Temperature scan of Water Standard KF Oven 150-160°C (0.1 g).

The blue curve gives the measured potential together with the evaporated water amount (grey curve).

To better understand the water release process from the sample matrix, a temperature scan has been performed
between 50 and 220 °C at a heating rate of 2 °C/min and an air flow of 80 mL/min using the InMotion KF oven
Autosampler in combination with the C30SX Coulometer. As indicated in the figure above, water is completely
released in the region between 100 and 170 °C. Lactose starts to decompose for temperatures above 200 °C
leading to higher potential values, and the white powder in the glass vial becomes brown.

Melting point

Transmission curves of melting point ranges measured with a Melting Point System MP80 with a heating rate of 1°C/min. Both batches
with lot numbers I158A (solid lines) and M004B (dashed lines) have been measured using three capillaries (black, blue, and red lines).

The melting point ranges of the two batches I158A (solid lines) and M004B (dashed lines) slightly differs from
each other: batch I158B has a melting point range of 207.5-212 °C, which is slightly lower than the one of
batch M004B, i.e., 209-214 °C. This may be attributed to the slightly different nature of the materials used for
the preparation of the water standard.

METTLER TOLEDO Page 7 of 12

30785465 - M880 v1.0
 Thermogravimetric measurements
Titration Application Note

To get a complete picture of the behavior of the water standard under heating, further measurements have been
performed. Specifically, thermogravimetric measurements in combination with mass-spectrometry have been
carried out, as given in the figure below:

From top to bottom:

1) TGA (black solid line): Thermogravimetric analysis of lactose monohydrate. Water is completely evaporated at 150-160 °C.
2) DSC (black solid line): Differential scanning calorimetry analysis. The peak at 208-210 °C indicates the melting point.
3) DTG/MS: The mass-spectrometry (MS) violet curve identifies the peak at 140-160 °C as water.

The TGA/DSC/MS measurements confirm the previous results:

• The maximum of the water release of lactose monohydrate is at 150°C, and water is completely
evaporated above 160°C. Above 210°C, lactose is decomposing, as shown in the TGA-curve.
• The melting point of lactose monohydrate is located at 208-209°C (DSC curve).
• Decomposition is observable in the DTG curve for temperatures higher than 210°C, reaching a
maximum at 230°C.

METTLER TOLEDO Page 8 of 12

30785465 - M880 v1.0
 Remarks time as a termination criterium is used for
Titration Application Note

heating the sample and to stop the titration. This

Volumetric KF titration (V30S) is improving the accuracy and the precision of
the analysis.
• The concentration of the KF volumetric titrant 5
mg H2O/mL was determined prior to the sample • A fixed heating and titration time of 1200 s has
measurement using a 10.0 mg/g liquid water proven to be the most suitable:
standard.
o The fixed heating time is set when the
• The solubility of the water standard KF oven termination time is entered as
150-160°C is limited: after 3-4 samples of 0.1 t(min) = t(max) = 1200 s.
g each, the solvent in the titration cell becomes
o During this time, the sample is heated,
turbid. Therefore, it takes longer to achieve a
and the evaporated water collected in the
low and stable drift value e.g., 10 µg/min.
titration vessel is continuously titrated.
• Nevertheless, it was still possible to measure up
o This way, loss of water due evaporation of
to 6 samples in 80-100 mL KF solvent when
water collected in the titration vessel
waiting long enough to get a sufficiently low
caused by the hot air stream from the oven
online drift.
is avoided. Thus, the accuracy is
• Thus, it is recommended to always replace the increased.
KF solvent at the end of a sample series.
o Moreover, a long, fixed termination time
Recommended minimum volume for the KF
also improves the reproducibility since all
solvent: 80 mL.
samples are titrated under the same
• The results show that the deviation with respect conditions
to the certified values is small, i.e., smaller than
o A shorter heating time of e.g., 900 s or
0.04% (abs) for batch M004B and 0.05%
less gives too low water content results.
(abs) for batch I158A. This indicates that the
measured values are in very good agreement
with the values stated in the certificates.
Waste disposal and safety measures
Dispose the content of the beakers in the Karl
Coulometric KF titration
Fischer waste.
(C30XS-InMotion KF Oven)

• The performance of the C30SX KF Coulometer

was first tested with a 1.0 mg/g liquid water References and Literature
standard before connecting it to the InMotion KF [1] https://en.wikipedia.org/wiki/Lactose , 12.05.2022.
Oven Autosampler.
• Subsequently, the performance of the coupled
Further information
system C30XS KF Coulometer-InMotion KF
Titrator Compact V30S - Overview - METTLER TOLEDO (mt.com)
Oven Autosampler has been verified using a
Titrator Compact C30SX - Overview - METTLER TOLEDO (mt.com)
5.55 % KF Oven solid water standard.
InMotion KF Pro Oven Autosampler (5mL) - Overview - METTLER TOLEDO (mt.com)
• When adding the water standard into the glass
vials of the InMotion KF Oven Autosampler, care
must be taken to homogeneously distribute the
sample all over the whole surface of the vial's
bottom to improve the complete evaporation of
water.
• When using a KF Oven, a long, fixed titration

METTLER TOLEDO Page 9 of 12

30785465 - M880 v1.0
 Method – Direct Titration
Titration Application Note

001 Title
Control
Type KF Volumetric
Endpoint 100 mV
Compatible with V30S / T7 / T9
Control band 400 mV
ID M880A
Dosing rate (max) 5 mL/min
Title 5% H2O Std Lactose
Dosing rate (min) 80 µL/min
Start Normal
002 Sample Termination
Sample Type Drift stop relative
Sample Type KF Drift 15.0 µg/min
Number of IDs 1 At VMax 10.0 mL
ID1 Lactose monohydrate Min. time 0
Entry type Weight Max time Infinite
Lower limit 0.08 g
Upper limit 0.12 g 006 Calculation R1(Content)
Density 1 g/mL
Result type Predefined
Correction factor 1.0
Result Content
Temperature 25.0 °C
Result unit %
Autostart Yes
Formula R1 = (VEQ*CONC-
Entry After addition
TIME*DRIFT/1000)*C/m
Concentration
Constant C = 0.1
Titrant KF 1–comp 5
Decimal places 2
Nominal conc. 5 mg/mL
Result limits No
Standard Water Standard 10.0
Record statistics No
Entry Type Weight
Extra statistical functions No
Lower limit 0.8 g
Send to buffer No
Upper limit 1.2 g
Temperature 25.0 °C
Mix time 10 s 007 Record
Autostart Yes Summary No
Entry After addition Results Per sample
Conc. lower limit 4.5 mg/mL Raw results Per sample
Conc. upper limit 5.6 mg/mL Table of meas. values No
Result unit %
003 Titration stand Sample data No
Resource data No
Type KF stand
E-V No
Titration stand KF stand
E-t No
Source for drift Online
V-t No
Max. start drift 25 µg/min
H2O – t No
H2O- t & Drift – t Yes
004 Mix time V – t & Drift – t Yes
Duration 180 Method No
Series data No
005 Titration (KF Vol) [1]
Titrant 008 End of Sample
Titrant KF 1-comp 5
Nominal concentration 5 mg/mL
Reagent type 1-comp
Sensor
Type Polarized
Sensor DM143-SC
Unit mV
Indication Voltametric
Ipol 24.0 µA
Temperature Acquisition
Temperature Acquisition No
Stir
Speed 35%
Predispense
Mode None
Wait time 0s

METTLER TOLEDO Page 10 of 12

30785465 - M880 v1.0
 Method – KF Oven
Titration Application Note

001 Title
Decimal places 1
Type InMotion KF Coulometric
Result limits No
Compatible with C30S / T7 / T9
Record statistics No
ID M880B
Extra statistical functions No
Title 5% H2O Std Lactose

002 Drift 008 End of sample

Wait time 60 s
Duration 3 min
009 Blank
Name Blank IMKF Coul
003 Sample (Blank)
Value B = Mean[R1]
Sample Unit µg
Sample Type Blank Limits No
Number of IDs 1
ID1 Blank vial 5 mL
010 Sample
Entry type Fixed pieces
Pieces 1 pcs. Sample
Weight per piece 1.0 g Sample Type Sample
Correction factor 1.0 Number of IDs 1
Temperature 25.0 °C ID1 Lactose monohydrate
Analysis start Manual Entry type Weight
Lower limit 0.05 g
Upper limit 0.06 g
004 Titration stand (InMotion KF)
Density 1.0 g/mL
Type InMotion KF Correction factor 1.0
Titration stand InMotion KF/1 Temperature 25.0 °C
Temperature ramp No Entry Arbitrary
Oven temperature 150 °C (160°C)
Source for drift Determination
011 Titration stand (InMotion KF)
Max. start drift 25 µg/min
Carrier gas source InMotion KF Type InMotion KF
Flow rate 80 mL/min Titration stand InMotion KF/1
Transfer tube heating Yes Temperature ramp No
Oven temperature 150 °C
Source for drift Determination
005 Mix time
Max. start drift 25 µg/min
Duration 60 s Carrier gas source InMotion KF
Flow rate 80 mL/min
006 Titration (KF Coul) [1] Transfer tube heating Yes
Sensor
Type Polarized 012 Mix time
Sensor DM143-SC Duration 60 s
Unit mV
Indication Voltametric
013 Titration (KF Coul) [2]
Ipol 5.0 µA
Stir Sensor
Speed 45% Type Polarized
Control Sensor DM143-SC
Endpoint 100.0 mV Unit mV
Control band 300.0 mV Indication Voltametric
Rate Normal Ipol 5.0 µA
Generator current Automatic Stir
Termination Speed 45%
Type Delay time Control
Delay time 1s Endpoint 100.0 mV
Min. time 1200 s Control band 300.0 mV
Max. time 1200 s Rate Normal
Generator current Automatic
Termination
007 Calculation R1: InMotion blank value
Type Delay time
Result InMotion blank value Delay time 1s
Result unit µg Min. time 1200 s
Formula R1 = (ICEQ/10.712-TIME*DRIFT)/C Max. time 1200 s
Constant C=1

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30785465 - M880 v1.0
Titration Application Note
014 Calculation R2: Content blank comp. (B in µg)
Result Content blank comp.
(B in µg)
Result unit %
Formula R2 =
(ICEQ[2]/10.712-TIME[2]*DRIFT-B[Blank IMKF Coul])/(C*m)
Constant C = 10000
Decimal places 2
Result limits No
Record statistics No
Extra statistical functions No

015 Record
Summary No
Results Per sample
Raw results Per sample
Table of meas. values No
Sample data No
Resource data No
E-C No
E-t No
C-t No
H2O – t No
H2O- t & Drift – t Yes
C – t & Drift – t No
Method No
Series data No

016 End of Sample

METTLER TOLEDO Group

Analytical
Local contact: www.mt.com/contacts

Subject to technical changes

© 07/2022 METTLER TOLEDO. All rights reserved
30785465 / M880 v1.0

METTLER TOLEDO Page 12 of 12

30785465 - M880 v1.0