Foodpac e

Food Potentiometric Analysis Collection
Food PAC
6.6055.003
Methods for the Titrimetric/Potentiometric Analysis

of Foodstuffs
Dear User,
You have decided to purchase a Metrohm Titrator which, with its special collection of
methods for your own particular applications, is intended to meet all your requirements.
Metrohm always attempts to provide customers with as wide a range of application sup-
port as possible in order to make daily work easier.
In this Application File you will find descriptions of the analytical methods together with
the necessary comments and explanations and – specially for you – printouts of the instru-
ment parameters and examples of curves.
All these methods are loaded on the method memory card. All that you need to do is to
«feed» your titrator with the card, load the required method into the working memory and
off you go!!!
For Titrando users: a conversion program ensures that you can use Titrino parameters
in the Titrando without any problems. This conversion program is contained in the
6.6050.XXX PC Control program.
We wish you lots of pleasure and success in your work,
Your Metrohm
8.110.1911
1
Some additional information
– The methods described here have been drawn up taking the current state of knowledge into account.
– All the methods are formulated so that you can use them as SOPs (Standard Operating Procedures) in
your laboratory.
– Many of the methods described here can be automated even further; see the proposal given in the annex.
For details please consult your local Metrohm distributor, which can be found on the Internet under:
www.metrohm.com ⇒ Distributors
– The method memory card supplied can be used with the 798, 799, 785 and 751 Titrinos (from program
version 20). With the Metrodata VESUV Light 3.0 software (VESUV = Verification Support for Validation),
which is also supplied, you or your local Metrohm distributor can also transfer the sets of parameters to
716, 736, 794 or 751 Titrinos (<program version 20).
– Among other things, the supplied CDs contain:
• The VESUV backup file, which allows you to copy the 96 methods into 716, 736, 751, 785, 794, 798 and
799 Titrinos. For further information please consult the section «Restoring methods» in the Instructions
for Use supplied or contact your local Metrohm distributor.
If the VESUV software is used for data acquisition instead of the printer then the report «curve» must be
deleted at the Titrino, which is set instead to «mplist» (VESUV can only process measuring point lists).
• A conversion program for taking over Titrino parameters into the Titrando. This conversion program is
contained in the 6.6050.XXX PC Control program.
• Acrobat® Reader® for installation on your PC so that you can read PDF files.
• Application Bulletins nos. 25, 33, 53, 69, 84, 85, 86, 87, 94, 125, 129, 130, 139, 140, 141, 180, 188, 225,
235, 249 and 252.
– We recommend that you only load the methods that you require in your instrument.
– The method overview can be removed and kept near your instrument together with the Food PAC card.
Important symbols used in the methods

c(X) molar concentration of substance X in mol/L
M(X) molar mass of substance X in g/mol
w(X) mass fraction of substance X, e.g. w(NaCl) = 10%
σ(X) volume fraction of substance X, e.g. σ (EtOH) = 40% (formerly: % V/V, e.g.)
ρ ((X) mass concentration of substance X, e.g. ρ (NaOH) = 4.0 g/L
(β is also used – difference from density)
Literature
The procedures and methods have been drawn up based on the following publications:
– German Standard Methods for the Examination of Water, Waste Water and Sludge
– Official Methods of Analysis of the Association of Official Analytical Chemists (AOAC, USA)
– Swiss Foodstuffs Manual (Schweizerisches Lebensmittelbuch)
– U.S. Environmental Protection Agency (EPA)
Remarks
The determination of the water content of foodstuffs and semi-luxuries is not described in Food PAC. Please
consult the Metrohm monograph «Water determination by Karl Fischer titration», which can be obtained
free of charge from your local Metrohm distributor.
2
Contents
A pH value H Soft drinks, lemonades

A 1 Calibrating the pH glass electrode H 1 Citric acid, citrates
A 2 Measuring the pH value H 2 Phosphoric acid (cola drinks)
H 3 Potassium
B Preparing the most important titrants and H 4 Total phosphorus
determining their titer
J Fruit and vegetable juices, fruit nectars and
B 1 Alkaline titrants (NaOH, KOH)
B 2 Acidic titrants (HCl, H2SO4) jams
B 3 Iodine solution J 1 pH value and titratable total acidity
B 4 Thiosulfate J 2 Ascorbic acid (vitamin C)
B 5 Permanganate J 3 Sulfurous acid (sulfite)
B 6 Silver nitrate J 4 Chloride
B 7 Na2EDTA J 5 Total phosphorus
J 6 Sulfate
C Drinking water and mineral water J 7 Calcium and magnesium
C 1 pH value and acid capacity (carbonate hard- J 8 Potassium
ness) J 9 Ash alkalinity
C 2 Calcium and magnesium (Ca hardness, Mg J 10 Kjeldahl nitrogen, total protein
hardness and total hardness) J 11 Formol number
C 3 Chloride J 12 Reducing sugars in jams
C 4 Sulfate
C 5 Sulfides and hydrogen sulfide K Beer, vinegar, spirits and wine
C 6 Total and residual chlorine (free chlorine) K 1 Beer, pH value
C 7 Permanganate index (oxidizability) K 2 Beer, CO2 content
C 8 CO2 content of carbonated water K 3 Beer, ascorbic acid (vitamin C)
C 9 Oxygen content according to Winkler K 4 Beer, total sulfurous acid
K 5 Vinegar, pH value and total acidity
D Milk and dairy products K 6 Vinegar, volatile acidity
D 1 pH value K 7 Vinegar, ascorbic acid (vitamin C)
D 2 Titratable acidity, degree of acidity K 8 Vinegar, free sulfurous acid
D 3 Chloride K 9 Vinegar, total sulfurous acid
D 4 Calcium K 10 Vinegar, ash alkalinity
D 5 Vitamin C (ascorbic acid) K 11 Vinegar, chloride
D 6 Kjeldahl nitrogen K 12 Vinegar, sulfate
K 13 Spirits, pH value and total acidity
E Edible fats and oils K 14 Spirits, volatile acidity
K 15 Spirits, total esters
E 1 Acid number and free fatty acids (FFA)
E 2 Hydroxyl number L Coffee, cocoa and chocolate
E 3 Iodine number
E 4 Peroxide number L 1 pH value and degree of acidity (green and
E 5 Saponification number roasted coffee, coffee extracts)
L 2 Ash alkalinity (green and roasted coffee, cof-
F Cereals, flour milling products, dry pasta fee extracts)
products L 3 Chloride (green and roasted coffee)
L 4 Directly reducing sugars (roasted coffee,
F 1 pH value and degree of acidity coffee extracts)
F 2 Chloride and cooking salt content L 5 pH value (cocoa and chocolate powder)
F 3 Kjeldahl nitrogen, total protein L 6 Ash alkalinity (cocoa and chocolate pow-
F 4 Calcium and magnesium (in ash) der)
L 7 Kjeldahl nitrogen (cocoa and chocolate
G Honey, sugar and sweets powder)
G 1 pH value and free acids L 8 Free fatty acids in cocoa butter
G 2 Formol number L 9 Iodine number of cocoa butter
G 3 Reducing sugars L 10 Saponification number of cocoa butter
3
Contents
M Artificial sweeteners, gelling and thickening

agents
M 1 Methoxy and ethoxy groups in gelling and
thickening agents
M 2 Cyclamate in artificial sweeteners
M 3 Saccharin in artificial sweeteners
N Preserved fruits, vegetables and mushrooms

N 1 Oxalic acid in fruits and vegetables
N 2 Total sulfurous acid in dried fruits and veg-
etables
N 3 Cooking salt content of mushroom extracts
and concentrates
O Cooking salt, spices, pickling salt,

seasoning, herbal and flavored salts
O 1 Chloride content
O 2 Total iodine in cooking salt
O 3 Fluoride in cooking salt
O 4 Tricalcium phosphate in cooking salt
O 5 Nitrite in pickling salt
P Meat products, meat extracts, bouillon

preparations, aspic, seasonings, soups,
sauces
P 1 Chloride (NaCl) in meat products
P 2 Kjeldahl nitrogen and raw protein in meat
products
P 3 Sulfurous acid in meat products
P 4 Chloride (NaCl) in meat extracts, aspic and
bouillon
P 5 Kjeldahl nitrogen in meat extracts, aspic and
bouillon
P 6 Chloride (NaCl) in seasonings, soups, sauc-
es
P 7 Kjeldahl nitrogen in seasonings, soups,
sauces
Q Qualifying the titrator, validating a titration

method
Index
4
Method 1 –
A 1 Calibrating the pH glass electrode
Recommended • Comb. pH glass electrode with built-in Pt 1000 temperature sensor, e.g.
accessory 6.0258.000 Unitrode
Reagents • Buffer solutions pH = 4.00, 7.00 and 9.00, e.g. 6.2307.100, 6.2307.110 and
6.2307.120
General Free protons (H+ ions) occur in solutions just as little as free electrons. They
combine with water to form oxonium ions:
H+ + H2O → H3O+
The pH value is defined as the negative logarithm of the oxonium ion activity,
i.e. of the concentration of free, dissociated oxonium ions in mol/L: pH = –log
[H3O+]
Strictly speaking, the term pH only applies to purely aqueous solutions.
The pH scale ranges from 0 to 14 with the neutral point at pH = 7.0, where the
H3O+ and OH – ions are present in equilibrium. pH values below 7 result from
an H3O+ excess, pH values above 7 from an OH – ion excess. The more acidic a
A pH value
solution the higher its H3O+ ion concentration and the lower its pH value.
Weak acids, e.g. tartaric acid, do not dissociate completely. This means that
only a small fraction (approx. 2...3%) of their acid ions are released. This also
means that only on very rare occasions can the pH value be used as a measure
of the concentration of acids or bases.
As the pH scale is logarithmic this means that small differences in pH corre-
spond to large differences in the concentration of H3O+ ions. For example, at pH
= 3.0 there are ten times more H3O+ ions present than at pH = 4.0, and at pH =
3.1 there are twice as many H3O+ ions present than at pH = 3.4.
The measuring setup for potentiometric measurements always consists of two
electrodes – a measuring or indicator electrode and a reference electrode. For
practical reasons these two electrodes are usually contained in a single com-
bined electrode.
The indicator electrode (in this case the pH glass electrode) produces a poten-
tial that is dependent on the composition of the sample solution.
The reference electrode (usually Ag/AgCl) has the task of providing a potential
that is as independent as possible of the sample solution (reference potential).
The potential measurement itself takes place virtually current-free by using a
«voltmeter» (in this case a Titrino) with a high-impedance measuring input (this
is necessary to avoid unwanted potential drops). The measured potential U is
made up from the individual potentials produced by the indicator and reference
electrodes. The following illustration shows a schematic diagram with a sepa-
rate pH glass electrode (left) and a reference electrode (right):
U1: Galvani potential between measuring electrode and measuring solution

U2: Galvani potential between internal buffer and glass membrane
U3: Galvani potential between internal reference electrode and internal buffer
U4: Galvani potential of reference electrode
U5: Diffusion potential at the diaphragm
1
Method 1 –
The individual potentials U2, U3 and U4 are determined by the construction of

the electrodes and are therefore constant for a given electrode pair. The diffu-
sion potential U5 should be kept relatively constant and low by taking suitable
measures. These measures include an optimal and clean diaphragm, constant
stirrer speed during the measurements as well as a suitable reference electro-
lyte solution whose anions and cations have similar ionic mobilities – e.g. KCl.
In this way the potential U1 measured between the electrodes depends only on
the sample solution. In the pH measurement this potential is again dependent
on the activity ai of the measuring ion (H3O+ ion / OH – ion). This relationship is
described by the Nernst equation:
U = U0 + * log ai = U0 + UN * log a
where:
U measured difference in potential between indicator and reference electrode
U0 standard potential of the combined electrode (depends on its construction)
R gas constant (8.31441 J / (K mol)
T absolute temperature in K (273.15 + t / °C)
zi charge on the measuring ion i including its sign (+1 for H3O+ and –1 for OH–)
ai activity of measuring ion
UN Nernst slope (59.16 mV at 25 °C and z = 1)
2.303 conversion factor from natural to common logarithm
The Nernst slope UN describes the theoretical electrode slope and corresponds
to the change in potential produced by altering ai by a factor of ten. It depends
on the temperature and charge z of the measuring ion. Please note: The instru-
ment compensates the effect of temperature on UN but not on the pH value
of the solution!
The following table shows values of UN as a function of t / °C for z = 1:
Temperature t / °C U / mV An ideal pH glass electrode has a

slope of 1 (100% of the Nernst slope)
0 54.20 and an electrode zero point pHas of
10 56.18 7.0, the latter corresponding to Uas =
0 mV.
20 58.17
Things are different in practice. The
25 59.16 electrode zero point should have a
30 60.15 value for Uas of ±15 mV (corresponds
to pHas = 6.75...7.25) and the slope
38 61.74
should be >0.95 (>56.2 mV / pH at
40 62.14 25 °C).
50 64.12 In order to «inform» the instrument of
60 66.10 the true electrode data it is necessary
to calibrate the electrode. Buffer solu-
70 68.09 tions have a defined pH value which,
80 70.07 however, is temperature-dependent.
The relevant information about the
90 72.06 buffer solution is entered during the
calibration procedure. The following
table shows the pH values of Metrohm
buffer solutions as a function of the
temperature:
2
Method 1 –
Temperature t / °C pH = 4.00 ±0.02 pH = 7.00 ±0.02 pH = 9.00 ±0.02

10 3.99 7.06 9.13
20 3.99 7.02 9.04
25 4.00 7.00 9.00
30 4.00 6.99 8.96
38 4.02 6.98 8.91
40 4.02 6.98 8.90
50 4.04 6.97 8.84
60 4.07 6.97 8.79
70 4.11 6.98 8.74
80 4.15 7.00 8.71
90 4.20 7.01 8.68
Buffer solutions are not stable!! They can be decomposed by bacteria and/or
molds or – this applies to alkaline buffer solutions – alter their pH value by
absorbing CO2 from the atmosphere. This is why you should always use only
fresh buffer solutions and reject them after use, i.e. not pour them back into the
storage bottle.
Electrode calibration We recommend the following procedure for calibrating a pH glass electrode:
and – Remove the electrode from its storage vessel, attach a cable if necessary
electrode handling and connect it to the instrument.
– Open the electrolyte filling opening and, if necessary, top up the electrolyte
solution.
– Rinse the electrode thoroughly with dist. H2O and dab dry with a soft paper
tissue (do not rub).
– Fill pH = 7.0 buffer solution into a beaker and add a stirrer bar.
– Immerse the electrode into the buffer solution and stir for approx. 1 min.
Measure the temperature of the buffer solution and enter in the Titrino (not
necessary if temperature sensor is connected).
– On the Titrino enter the pH value of the buffer solution (at the corresponding
temperature) and start the calibration with buffer 1 under stirring.
– When the measured value has been accepted, remove the electrode from
the solution, rinse it thoroughly with dist. H2O and dab dry with a soft paper
tissue.
– Add pH = 4.0 or 9.0 buffer solution to a second beaker, add a stirrer bar,
immerse the electrode and stir for approx. 1 min (the second buffer solution
must have the same temperature as the first one).
– On the Titrino enter the pH value of the second buffer solution (at the corre-
sponding temperature) and continue the calibration under stirring.
– After the measured value has been accepted, end the calibration. Remove
the electrode from the solution, rinse the electrode thoroughly with dist. H2O
and dab dry with a soft paper tissue.
3
Method 1 –
What happens in the instrument (in this case Titrino) during calibration can be
seen in the following plot:
Handling
Laboratory electrodes should have a long lifetime. Their characteristics (slope,
response behavior, pHas / Uas) must lie within the given criteria. In order to en-
sure this a few basic rules must be observed:
• After use rinse the electrode thoroughly with dist. H2O and dab dry with a soft
paper tissue. Close off the electrolyte filling opening and store the electrode
by immersing it in electrolyte solution – usually c(KCl) = 3 mol/L – to an
adequate depth. Dry storage leads to delayed and poor response behavior.
The electrolyte solution may become concentrated and pHas / Uas could alter.
Storage in dist. H2O could result in the diaphragm being blocked by AgCl.
• If the electrode responds sluggishly and/or the slope is unsatisfactory then
the electrode membrane must be etched. This is done by immersing the
membrane in a 10% solution of ammonium difluoride (NH4HF2, use plastic
beaker) for 1 min, then swirling it for approx. 10 s in c(HCl) = 5 mol/L, rins-
ing it thoroughly with dist. H2O and then wiping off the silicate residue with
a moist tissue. In order to build up a new gel layer the electrode is placed in
c(KCl) = 3 mol/L for 24 h (or for 5 h in the same solution at 50 °C).
• If the diaphragm becomes blocked please refer to the electrode data sheet
that accompanies each electrode. Removing such a blockage is complicated
and time-consuming – it is better to send the electrode to your local Metrohm
distributor for this.
• Contamination by fats, oils, lacquers, paints, etc.: remove the contamination
with an organic solvent (acetone, petroleum benzine, toluene), rinse thor-
oughly with ethanol and dist. H2O, dab dry and place in electrolyte solution.
• Contamination by proteins: immerse the electrode in a solution of 5% pepsin
in c(HCl) = 0.1 mol/L for a few hours. Then rinse thoroughly with dist. H2O,
dab dry and place in electrolyte solution.
4
Method 1 –
Instrument parameters and calculation Result
tPA @CR
'044ITRINO '044ITRINO
DATE TIME $ATE 4IME
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PARAMETERS CALDATE
CALIBRATIONPARAMETERS P(5M6DP(
MEASINPUT BUFFER
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MEAN
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TEMPORARYVARIABLES

5
Method 2 –
A 2 Measuring the pH value
The pH value is extremely important for biological systems. It influences the

growth of microorganisms, the taste, color, redox potential and many other
properties.
The pH value of the sample also depends on the temperature. This temperature
dependency cannot be compensated by the instrument (e.g. Titrino), which
only adjusts the slope of the electrode. This means that information about the
pH value must always include the temperature at which it was measured.
Example of a red wine sample:
pH = 3.52 at 18.2 °C
The calibrated electrode is thoroughly rinsed with dist. H2O and dabbed dry
with a soft paper tissue. The electrode is immersed in the sample solution (or
pushed into the sample until the diaphragm is covered) and the pH is measured
(under stirring for liquid samples). When the drift criterion has been reached,
the pH value is displayed by the instrument or printed out (do not forget to mea-
sure the sample temperature). When the measurement has been completed the
electrode is thoroughly rinsed and dabbed dry with a soft paper tissue.
A pH value
1
Method 2 –
A 2 Measuring the pH value
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A pH value
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2
B Preparing the most important titrants and determining their titer
Preparing the most important titrants
and determining their titer
General Titrants are standard solutions, i.e. solutions that contain a defined content of
a reactant. This content is given as the molar concentration c in mol/L. The
«normality», which was frequently used previously, is no longer valid today and
should therefore not be used.
Examples:
• 0.1 N HCl ⇒ c(HCl) = 0.1 mol/L
• 0.1 N H2SO4 ⇒ c(H2SO4) = 0.05 mol/L
• 0.1 N iodine solution ⇒ c(I2) = 0.05 mol/L
• 0.1 N KMnO4 ⇒ c(KMnO4) = 0.02 mol/L [or c(1/5 KMnO4) = 0.1 mol/L]
Not all titrants have a stable titer, i.e. their concentration can vary with time.
Examples:
• Hydroxides absorb CO2 from the atmosphere to form carbonates
• Iodine solutions release iodine
• Thiosulfate solutions can decompose and deposit sulfur
• In the presence of organic substances, e.g. dust particles, permanganate
solutions deposit manganese dioxide (MnO2)
• The solvent may evaporate from non-aqueous titrants.
This all means that the titer of the titrant may alter as time passes. In order to
know the true titer concentration the titer must be determined at regular inter-
vals.
The so-called standard titrimetric substances are used for determining the titer.
Their content hardly changes, they are available with a defined degree of purity,
can be dried and can be traced back directly to standard reference materials
(e.g. National Institute of Standards and Technology – NIST, USA).
Such standard titrimetric substances /secondary standards are:
• For bases potassium hydrogen phthalate, M = 204.23 g/mol
• For acids tris(hydroxymethyl)-aminomethane, M = 121.14 g/mol
• For iodine arsenic trioxide, M = 197.841 g/mol
• For thiosulfate potassium iodate, M = 214.001 g/mol
• For permanganate disodium oxalate, M = 133.999 g/mol
• For AgNO3 sodium chloride, M = 58.443 g/mol
• For Na2EDTA calcium carbonate, M = 100.09 g/mol
Most standard solutions/titrants are commercially available as ready-to-use
solutions with a titer adjusted by the manufacturer at 20 °C to 1.000. We recom-
mend that you purchase such ready-to-use solutions and do not prepare them
yourselves.
In principle titer determinations should always be carried out at the same tem-
perature at which the analyses are later to be carried out. Please note that solu-
tions expand as their temperature increases.
1
B Preparing the most important titrants and determining their titer Preparing the most important titrants
and determining their titer
For aqueous solutions a temperature difference of 5 °C for a theoretical con-

sumption of 10.00 mL results in a difference in volume of 12.5 μL. This means
that a titer of 1.0000 at 20 °C becomes a titer of 0.9988 at 25 °C – and for non-
aqueous solutions this difference is even larger. In such a case a titer of 1.0000
at 20 °C becomes a titer of 0.9950 at 25 °C (possible error 0.5%)!
Normally the titer determination is carried out three times and the mean value is
used. The mean value of the titer** is best saved as a «Common Variable» (e.g.
C30) in the Titrino.
** In Europe a dimensionless factor, with at least 4 decimal places (e.g. 1.0015).

In the USA already multiplied by c, with at least 4 decimal places (e.g. 1.0015 mol/L).
2
Method 3 –
B 1 Alkaline titrants
Method 3 – Alkaline titrants do not have a stable titer. They may absorb CO2
from the atmosphere to form carbonates, e.g.:
2 NaOH + CO2 = Na2CO3 + H2O
This not only reduces the titer. The different strengths of the bases NaOH and
Na2CO3 may negatively influence the titration curves and therefore the results
– high-bias results are simulated, particularly for weak acids. In order to reduce
CO2 absorption as much as possible, soda lime (e.g. Merck no. 106839) is
placed in the drying/absorber tube of the Exchange Unit.
• c(NaOH) = 0.1 mol/L
If possible this titrant should be bought ready for use. Otherwise dissolve
4.0 g NaOH in CO2-free dist. H2O, make up to 1 liter and mix.
• c(KOH) = 0.1 mol/L in ethanol or isopropanol (IPA)
5.6 g KOH in approx. 25 mL CO2-free dist. H2O, allow to cool down and then
make up to 1 liter with ethanol or IPA and mix.
Recommended
• Comb. pH glass electrode for aqueous titrations and 6.0229.100 Solvotrode
accessories (electrolyte solution 6.2320.000) for non-aqueous titrations, with 6.2104.100
electrode cable
• 6.3026.220 Exchange Unit
Titer determination Potassium hydrogen phthalate is dried overnight in a drying oven at 105 °C and
allowed to cool down in a desiccator for at least 1 h.
Approx. 200 mg potassium hydrogen phthalate is weighed out into the titration
beaker with an accuracy of 0.1 mg and dissolved in approx. 50 mL dist. H2O. It
is then immediately titrated to after the first endpoint with c(base) = 0.1 mol/L.
1 mL c(base) = 0.1 mol/L corresponds to 20.423 mg KH-phthalate (C01)
Titer = C00 / C01 / EP1

EP1 = mL base up to endpoint
C00 = weight of KH-phthalate in mg
C01 = 20.423
1
B Preparing the most important titrants and determining their titer Method 3 –
B 1 Alkaline titrants
tPA tFR
DATE TIME $ATE 4IME
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2
Method 4 –
B 2 Acidic titrants
• c(HCl) = 0.1 mol/L in H2O or in ethanol

If possible this titrant should be bought ready for use. Otherwise place ap-
prox. 800 mL dist. H2O or ethanol in a 1000 mL volumetric flask, add 9.8 mL
w(HCl) = 32%, make up to the mark with dist. H2O or ethanol and mix.
• c(H2SO4) = 0.05 mol/L
prox. 800 mL dist. H2O in a 1000 mL volumetric flask and carefully add 2.7 mL
w(H2SO4) = 96% while swirling the flask. Make up to the mark with dist. H2O
and mix.
Recommended • Comb. pH glass electrode for aqueous titrations and 6.0229.100 Solvotrode
accessories (electrolyte solution 6.2320.000) for non-aqueous titrations, with 6.2104.020
electrode cable
• Exchange Unit 6.3026.220
Titer determination Tris(hydroxymethyl)-aminomethane (TRIS) is dried overnight in a drying oven

at 105 °C and allowed to cool down in a desiccator for at least 1 h. Approx. 100
mg TRIS is weighed out into the titration beaker with an accuracy of 0.1 mg and
dissolved in approx. 50 mL dist. H2O. It is then immediately titrated with HCl or
H2SO4 to after the first endpoint.
1 mL c(HCl) = 0.1 mol/L or 1 mL c(H 2SO 4) = 0.05 mol/L corresponds to
12.114 mg TRIS (C01)
Titer = C00 / C01 / EP1
EP1 = mL acid up to endpoint
C00 = weight of TRIS in mg
C01 = 12.114
1
B 2 Acidic titrants
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
TITRATIONPARAMETERS %0MLM6
MEASPTDENSITY 4ITER
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
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DOSELEMENTINTERNAL$
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STOPCONDITIONS
STOP6ABS
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REQIDENT/&&
REQSMPLSIZE/&&
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tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
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23TEXT4ITER
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
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MEAN
-.23
TEMPORARYVARIABLES

2
Method 5 –
B 3 Iodine solution
• c(I2) = 0.05 mol/L

If possible this titrant should be bought ready for use. Otherwise weigh out
25 g pure, iodate-free potassium iodide into a 1000 mL volumetric flask and
dissolve in 40 mL dist. H2O. Add 12.8 g iodine, seal the flask and shake it until
the iodine has dissolved completely. Make up to the mark with dist. H2O and
Recommended mix.
accessories
• 6.0431.100 Pt Titrode with 6.2104.020 electrode cable
Titer determination • 6.3026.220 Exchange Unit
Approx. 200...250 mg Na2S2O3 x 5 H2O is weighed out into the titration beaker
with an accuracy of 0.1 mg, treated with 5 mL c(CH3COOH) = 2 mol/L and di-
luted with dist. H2O to approx. 80 mL. The solution is then titrated with c(I2) =
0.05 mol/L to after the first endpoint.
1 mL c(I2) = 0.05 mol/L corresponds to 24.817 mg Na2S2O3 x 5 H2O (C01)
Titer = C00 / C01 / EP1
EP1 = mL iodine solution to endpoint
C00 = weight of Na2S2O3 x 5 H2O in mg
C01 = 24.817
1
B 3 Iodine solution
tPA tFR
DATE TIME $ATE 4IME
-%45- 5INIT M6-%45-
TITRATIONPARAMETERS %0ML M6
6STEPML 4ITER
DOSRATEMAXMLMIN STOP6REACHED
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6ABS
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tCF
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DATE TIME
-%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
-%45-
DEF
FORMULA
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23DECIMALPLACES
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TEMPORARYVARIABLES

2
B Herstellung und B Preparing the most important titrants and de-
Method 6 –
B 4 Thiosulfate
• c(Na2S2O3) = 0.1 mol/L

25 g Na 2S2O3 x 5 H2O into a 1000 mL volumetric flask, dissolve in CO2-free
dist. H2O, use this to make it up to the mark and mix.
Recommended • 6.0431.100 Pt Titrode with 6.2104.020 electrode cable

accessories • 6.3026.220 Exchange Unit
Titer determination Potassium iodate is dried overnight in a drying oven at 180 °C and allowed to
cool down in a desiccator for at least 2 h.
Approx. 50 mg KIO3 is weighed out into the titration beaker with an accuracy of
0.1 mg and dissolved in approx. 80 mL dist. H2O. Add approx. 1 g potassium
iodide and 10 mL w(H2SO4) = 25% and titrate immediately with c(Na2S2O3) =
1 mL c(Na2S2O3) = 0.1 mol/L corresponds to 3.567 mg KIO3 (C01)
Titer = C00 / C01 / EP1
EP1 = mL thiosulfate solution up to endpoint
C00 = weight of KIO3 in mg
C01 = 3.567
1
B 4 Thiosulfate
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY 4ITER
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
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REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
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tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
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23TEXT4ITER
23DECIMALPLACES
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-.23
TEMPORARYVARIABLES

2
Methode 7 –
B 5 Permanganate
• c(KMnO4) = 0.02 mol/L

If possible this titrant should be bought ready for use; this is particularly ad-
visable for this solution. Otherwise weigh out approx. 3.2 g KMnO4 into an
amber glass bottle and dissolve in 1 liter dist. H2O. The bottle is sealed and
allowed to stand for 1...2 weeks. The solution is then filtered through a glass
filter crucible into a carefully cleaned amber glass storage bottle (do not use
paper filters!).

Titer determination Disodium oxalate is dried overnight in a drying oven at 105 °C and allowed to
Approx. 70 mg Na2C2O4 is weighed out into the titration beaker with an accu-
racy of 0.1 mg and dissolved in approx. 50 mL dist. H2O. It is treated with 5 mL
w(H2SO4) = 25%, warmed to 50...60 °C and titrated with c(KMnO4) = 0.02 mol/L
to after the first endpoint. The titration is also possible at room temperature if
approx. 0.5 g MnSO4 is added to the solution as catalyst before the titration.
1 mL c(KMnO4) = 0.02 mol/L corresponds to 6.70 mg Na2C2O4 (C01)
Titer = C00 / C01 / EP1
EP1 = mL permanganate solution up to endpoint
C00 = weight of Na2C2O4 in mg
C01 = 6.7
1
B Preparing the most important titrants and determining their titer Methode 7 –
B 5 Permanganate
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY 4ITER
DOSRATEMAXMLMIN
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tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
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23TEXT4ITER
23DECIMALPLACES
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-.23
TEMPORARYVARIABLES

2
Methode 8 –
B 6 Silver nitrate
• c(AgNO3) = 0.1 mol/L

16.988 g AgNO3 into a 1000 mL volumetric flask and dissolve in approx. 600
mL dist. H2O. Add 0.1 mL c(HNO3) = 2 mol/L, make up to the mark with dist.
H2O and mix.
Recommended • 6.0430.100 Ag Titrode with Ag2S coating, 6.2104.020 electrode cable

Titer determination Sodium chloride is ground to a fine powder in a mortar, dried overnight in a
drying oven at 140 °C and allowed to cool down in a desiccator for at least 2 h.
Approx. 60 mg NaCl is weighed out into the titration beaker with an accuracy
of 0.1 mg and dissolved in approx. 50 mL dist. H2O. After the addition of 1 mL
c(HNO3) = 2 mol/L the solution is titrated with c(AgNO3) = 0.1 mol/L to after the
first endpoint.
1 mL c(AgNO3) = 0.1 mol/L corresponds to 5.844 mg NaCl (C01)
Titer = C00 / C01 / EP1
EP1 = mL AgNO3 up to endpoint
C00 = weight of NaCl in mg
C01 = 5.844
1
B Preparing the most important titrants and determining their titer Methode 8 –
B 6 Silver nitrate
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2
Method 9 –
B 7 Na2EDTA
• c(Na2EDTA) = 0.1 mol/L

37.224 g Na2EDTA x 2 H2O into a 1000 mL volumetric flask, dissolve in dist.
H2O, make up to the mark and mix.
Recommended • 6.0508.110 or 6.0504.100 Ca ISE with 6.2104.020 electrode cable and

accessories 6.0726.107 Ag/AgCl reference electrode with 6.2106.020 electrode cable
Titer determination Calcium carbonate is dried overnight in a drying oven at 140 °C and allowed to
Approx. 100 mg CaCO3 is weighed out into the titration beaker with an accuracy
of 0.1 mg and suspended in approx. 20 mL dist. H2O. Under stirring c(HCl) =
5 mol/L is added drop by drop until everything has dissolved completely. After
the addition of 30 mL dist. H2O and 5 mL buffer solution pH = 10** the solution
is titrated with c(Na2EDTA) = 0.1 mol/L to after the first endpoint.
** 54 g NH4Cl and 350 mL w(NH3) = 25% are dissolved in dist. H2O and made up to 1 liter with dist.
H2O.
1 mL c(Na2EDTA) = 0.1 mol/L corresponds to 10.009 mg CaCO3 (C01)

Titer = C00 / C01 / EP1
EP1 = mL Na2EDTA up to endpoint
C00 = weight of CaCO3 in mg
C01 = 10.009
1
B 7 Na2EDTA
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY 4ITER
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
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REQIDENT/&&
REQSMPLSIZE/&&
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tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
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4ITER##%0
23TEXT4ITER
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
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-.23
TEMPORARYVARIABLES

2
C Drinking water and mineral water
General Drinking water is the foodstuff par excellence, which is why the methods used
for examining it are placed at the start of this collection. Nine titrimetric analyses
– Food PAC covers only these – are described in subsections. Of course, these
methods can also be used for other types of water.

Other determinations that can be carried out in water with Metrohm instruments
are:
• Bromate, bromide, chloride, fluoride, nitrate, nitrite, phosphate, ammonium,
potassium, sodium, strontium, etc. by ion chromatography (IC)
• Aluminum, arsenic, lead, cadmium, chromium, iron, copper, manganese,
mercury, zinc, etc. by voltamperometry/polarography (VA)
• Electrical conductivity (and pH value)
Assessment of the analytical results has been deliberately ignored in Food
PAC. Please consult the relevant literature for assessment criteria.
1
Method 10 –
C 1 pH value and acid capacity
(carbonate hardness)
General The acid capacity of a water (K A) is the molar amount of oxonium ions that the
water can accept before reaching certain predefined pH values. It is given in
mmol/L.
For the acid capacity up to pH 8.2 (K A 8.2 , previously p value) HCl is used to

titrate to pH = 8.2; for the acid capacity up to pH 4.3 (K A 4.3 , previously m value)
the titration is to pH = 4.3.
Recommended • 6.0253.100 Aquatrode Plus with 6.2104.020 electrode cable

Reagents • Titrant: c(HCl) = 0.1 mol/L (Method 4). For very soft water possibly use
c(HCl) = 0.02 mol/L.
Analysis The Aquatrode has already been calibrated as per Method 1.

100 mL water sample is added to the titration beaker. The pH value is first mea-
sured under stirring. The sample is then titrated with c(HCl) = 0.1 mol/L using
pH = 3.5 as the stop criterion.
Calculations K A 8.2 = EP1* x C01 x C02 x C30

(Results given to two K A 4.3 = EP2** x C01 x C02 x C30
decimal places) EP1* = mL HCl until first endpoint is reached or a fixed EP at pH = 8.2
(e.g. C51)
EP2** = mL HCl until second endpoint is reached or a fixed EP at pH = 4.3
(e.g. C52)
C01 = concentration of HCl in mol/L (0.1 or 0.02)
C02 = 10 (conversion factor for mmol/L if 100 mL sample is used)
C30 = titer of HCl used (Method 4)
Remarks
• Many waters have an initial pH value below 8.2 – K A 8.2 is not found and there-
fore cannot be calculated (only EP1 at pH = 4.3).
• For calculating the carbonate hardness please refer to table under C 2.
• If the CO2 content is to be determined in carbonated waters (Method 17) then
K A 4.3 is saved in the Titrino as a Common Variable (e.g. C35).
1
Method 10 –
Instrument parameters and calculation
tPA tCF
DATE TIME DATE TIME
$%4P(-$%4 $%4P(-$%4
PARAMETERS # FMLA
TITRATIONPARAMETERS
MEASPTDENSITY
MININCR±L
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Titration curve Result
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2
Method 10 –
Instrument parameters and calculation – pH Result – pH
tPA tFR
DATE TIME $ATE 4IME
-%!3P(-P( -%!3P(-
PARAMETERS SMPLSIZEML
MEASURINGPARAMETERS #
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
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STATISTICS
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PRESELECTIONS Titration curve – pH
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
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tCF
'044ITRINO
DATE TIME
-%!3P(-P(
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-P(
DEF
FORMULA
SILOCALCULATIONS
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##

3
Method 10 –
Instrument parameters and calculation – TIP Result – TIP

tPA tFR
DATE TIME $ATE 4IME
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METHOD- #NOTVALID
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tCF
'044ITRINO
DATE TIME
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#

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DATE TIME
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TEMPORARYVARIABLES

4
Method 11 –
C 2 Calcium and magnesium
(Ca, Mg and total hardness)
General The total hardness is the sum of the hardness formers in a water (Ca, Mg, Ba
and Sr ions) in mmol/L. For information purposes, e.g. on washing powder
packages, expressions such as «soft», «hard», «°dH», «°fH», etc. still have a cer-
tain meaning; see the comparison table below.

As Al, Ba, Fe, Mn and Sr ions only occur in drinking water in small amounts (if
at all) in drinking and mineral water, the titrimetric determination is limited to
calcium and magnesium.
• 6.0508.110 or 6.0504.100 Ca ISE with 6.2104.020 electrode cable, and

Recommended • 6.0726.107 LL Ag/AgCl reference electrode with 6.2106.020 electrode cable
accessories • Exchange Unit 6.3026.220
Reagents • Titrant: c(Na2EDTA) = 0.1 mol/L (Method 9)

• Auxiliary complexing solution: c(acetylacetone) = 0.1 mol/L plus c(TRIS) =
0.2 mol/L. 20.4 g tris(hydroxymethyl)-aminomethane is weighed out into a
1000 mL volumetric flask and dissolved in approx. 500 mL dist. H2O. 12 mL
acetylacetone is added and the solution is made up to the mark with dist.
H2O and mixed.
Analysis 100 mL water sample is pipetted into the titration beaker. 15 mL auxiliary com-
plexing solution is added and titrated with c(Na2EDTA) = 0.1 mol/L to after the
second endpoint.
EP1 corresponds to the Ca content and the difference EP2 – EP1 to the Mg
content.
Calculations 1 mL c(Na2EDTA) = 0.1 mol/L corresponds to 4.008 mg Ca or 2.431 mg Mg

(The results are given to mg/L Ca = EP1 x C01 x C02 x C30 / C00
two decimal places.) mg/L Mg = (EP2 – EP1) x C03 x C02 x C30 / C00
Total hardness, mmol/L = EP2 x C04 x C02 x C30 / C00
EP1 = mL Na2EDTA up to first endpoint
EP2 = mL Na2EDTA up to second endpoint
C00 = sample volume in mL (100)
C01 = 4.008
C02 = 1000 (for 1 liter)
C03 = 2.431
C04 = 0.1 (concentration Na2EDTA in mol/L)
C30 = titer of Na2EDTA solution (Method 9)
Remarks The following table provides information about the different «degrees of hard-
ness»:
1
Method 11 –
mmol/L Total hardness

mg/L CaCO3 °fH °dH °USH «Hardness»
CaCO3 comparison table
0...0.7 0...70 0...7 0...3.92 0...3.77 very soft
0.7...1.5 70...150 >7...15 >3.92...8.4 >3.77...
soft
8.07
1.5...2.5 150...250 >15...25 >8.4... >8.07...
slightly hard
14.0 9.50
2.5...3.2 250...320 >25...32 >14.0... >9.50...
moderately hard
17.92 17.22
3.2...4.2 320...420 >32...42 >17.92... >17.22...
hard
23.52 22.60
>4.2 >420 >42 >23.52 >22.60 very hard
Conversion factors:
mmol/L x 100 mg/L CaCO3
mmol/L x 10 °fH (French degrees of hardness)
mmol/L x 5.6 °dH (German degrees of hardness)
mmol/L x 5.38 °USH (US degrees of hardness)
2
Method 11 –
tCF
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'044ITRINO DATE TIME
DATE TIME $%45-
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3
Method 12 –
C 3 Chloride


Reagents • Titrant: c(AgNO3) = 0.01 mol/L. Purchased ready-to-use or self-prepared
(Method 8): dilute c(AgNO3) = 0.1 mol/L 1:10 with dist. H2O.
• Nitric acid: c(HNO3) = approx. 2 mol/L; dilute w(HNO3) = 65% 1:5 with dist.
H2O.
Analysis 100 mL water sample is pipetted into the titration beaker. After adding 5 mL ni-
tric acid titrate with c(AgNO3) = 0.01 mol/L to after the first endpoint.
Calculation 1 mL c(AgNO3) = 0.01 mol/L corresponds to 0.3545 mg chloride

mg/L chloride = EP1 x C01 x C02 x C30 / C00
C00 = sample volume in mL
C01 = 0.3545
C02 = 1000 (for 1 liter)
C30 = titer of AgNO3 solution
Remarks • The Ag2S coating on the Ag Titrode provides short response times and in-
creases the long-term stability of this electrode.
• If a sample contains sulfides or hydrogen sulfide then these are comprised
in the first, very distinct endpoint. Chloride is then calculated from the differ-
ence EP2 – EP1.
1
Method 12 –
C 3 Chloride

tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY #HLORIDEMG,
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#HLORID%0
#
#
##
23TEXT#HLORID
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 13 –
C 4 Sulfate
General The method described here allows sulfate to be determined without interfer-
ence in the presence of Ca, Mg and chloride ions. An excess of Ba ions is added
to the sample to precipitate the sulfate. The barium fraction that has not been
precipitated out with the sulfate is then determined by back-titration with EGTA.

Two endpoints are obtained. EP1 corresponds to the Ca and the difference EP2
– EP1 to the Ba. Mg is not determined by EGTA and does not interfere.

6.0726.107 Ag/AgCl reference electrode with 6.2106.020 electrode cable
accessories
Reagents • Titrant: c(EGTA) = 0.05 mol/L

19.4 g ethylene glycol-bis-(2-aminoethyl)-tetraacetic acid (98%) is weighed
out into a beaker, a stirrer bar and approx. 200 mL dist. H2O are added and
the EGTA is suspended under stirring. Then c(NaOH) = 10 mol/L is added
until everything has dissolved completely. After cooling down, the solution is
transferred quantitatively to a 1000 mL volumetric flask with dist. H2O, made
up to the mark and mixed.
• Barium chloride solution: c(BaCl2) = 0.05 mol/L
12.34 g BaCl2 x 2 H2O (99%) is weighed out into a 1000 mL volumetric flask,
dissolved in c(HCl) = 0.01 mol/L, made up to the mark with it and mixed.
• Buffer solution pH = 10:
9 g NH4Cl is weighed out into a 1000 mL volumetric flask and dissolved in ap-
prox. 800 mL dist. H2O. After the addition of 60 mL w(NH3) = 25% the solution
is made up to the mark with dist. H2O and mixed.
• Hydrochloric acid: c(HCl) = approx. 2 mol/L Conc.
HCl is diluted 1:5 with dist. H2O.
• Calcium standard: c(Ca2+) = 0.1000 mol/L, e.g. Metrohm no. 6.2301.070
• Sulfate standard: 1 mL conc. H2SO4 / 1000 mL dist. water
Blank consumption Approx. 30 mL dist. H2O and 5.00 mL c(BaCl2) = 0.05 mol/L are placed in the
titration beaker. After the addition of 0.5 mL Ca standard and 5 mL buffer solu-
tion pH = 10 the solution is stirred for 3 min and then titrated with c(EGTA) =
0.05 mol/L to after the second endpoint. The titrant consumption (EP2 – EP1) is
saved in the titrator as a Common Variable (e.g. C31).
Analysis A water sample, which should not contain more than 20 mg sulfate, is pipetted
into the titration beaker and adjusted to pH <4 by adding HCl drop by drop.
0.75 mL sulfate standard is then added. After the addition of 5.00 mL c(BaCl2)
= 0.05 mol/L the reaction is allowed to take place under stirring for 3 min. After
the addition of 5 mL buffer solution pH = 10, the solution is again stirred for 3
min and then titrated with c(EGTA) = 0.05 mol/L to after the second endpoint.
EP1 corresponds to the Ca and the difference EP2 – EP1 to the excess Ba. The
sulfate addition must be subtracted.
1
Method 13 –
C 4 Sulfate
1 mL c(EGTA) = 0.05 mol/L corresponds to 4.803 mg sulfate and 2.004 mg Ca Calculations

RS1 = EP2 – EP1 (mL)
mg/L sulfate: RS2 = ((C31 – RS1) x C01 x C02 / C00)-C04
mg/L Ca: RS3 = EP1 x C03 x C02 / C00
EP1 = mL EGTA up to first endpoint
EP2 = mL EGTA up to second endpoint
C01 = 4.803
C02 = 1000 (for 1 liter)
C03 = 2.004
C04 = 260.781
C31 = mL EGTA for blank consumption (5 mL BaCl2)
• The water sample must be acidified so that no BaCO3 is precipitated. Remarks

• With this titration the Ca can be determined simultaneously, but not Mg or the
total hardness.
2
Method 13 –
C 4 Sulfate
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L #
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES tDE
PAUSES DATE TIME
MEASINPUT DEF
STOPCONDITIONS 3ULFAT# %0 %0
#
##
STOP6ABS 23TEXT3ULFAT
lLLINGRATEMAXMLMIN #A%0
#
##
STATISTICS 23TEXT#A
%0RECOGNITIONALL SILOCALCULATIONS
lX%0AT5/&&M6 MATCHID/&&
P+(.0/&& COMMONVARIABLES
PRESELECTIONS REPORT
REQIDENT/&& REPORT#/-FULLCURVE
REQSMPLSIZE/&& MEAN
LIMITSMPLSIZE/&& -.23
ACTIVATEPULSE/&& TEMPORARYVARIABLES

tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEML
%0ML M6
%0ML M6
3ULFATEMGL
#AMGL
STOP6REACHED

3
Method 14 –
C 5 Sulfides and hydrogen sulfide


Reagents • Titrant: c(AgNO3) = 0.001 mol/L
Prepare by diluting c(AgNO3) = 0.1 mol/L (Method 8) 1:100 with dist. H2O.
Add 0.1 mL w(HNO3) = 65% per liter.
• Sodium hydroxide: w(NaOH) = 30%
Analysis Place 1 mL NaOH in the titration beaker. Then add 100 mL water sample and
titrate immediately with c(AgNO3) = 0.001 to after the first endpoint.
Calculation 1 mL c(AgNO3) = 0.001 mol/L corresponds to 0.016 mg S2- or 0.017 mg H2S

mg/L sulfide = EP1 x C01 x C02 / C00
EP1 = mL AgNO3 up to first endpoint
C01 = 0.016 or 0.017
C02 = 1000 (for 1 liter)
Remarks – Sulfides and hydrogen sulfide are very volatile compounds. In order to avoid
losses we recommend that the NaOH is added at the time when the sample
is taken.
– The Ag2S coating on the Ag Titrode provides short response times and in-
creases the long-term stability of this electrode.
1
Method 14 –
C 5 Sulfides and hydrogen sulfide

tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L #
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES tDE
PAUSES DATE TIME
MEASINPUT DEF
STOPCONDITIONS (3%0
#
##
STOP6ABS 23TEXT(3
lLLINGRATEMAXMLMIN 3 %0
#
##
STATISTICS 23TEXT3
REQSMPLSIZE/&& MEAN

tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEML
%0ML M6
(3MGL
3 MGL
STOP6REACHED

2
Method 15 –
C 6 Total and residual chlorine (free chlorine)
General Free or residual chlorine is chlorine that is present in the form of hypochlorous
acid, hypochlorite ions or as dissolved elemental chlorine.
Bound chlorine is that fraction of the total chlorine that is present in the form of
chloramines or organic chloramines.

Total chlorine is that chlorine that is present in free or bound form, or in both
forms.
All the forms described above release a stoichiometric amount of elemental
iodine from the added potassium iodide; this iodine can be titrated with pheny-
larsine oxide. If the titration is carried out at pH = 4.0 the total chlorine is deter-
mined, at pH = 10 only the free chlorine (residual chlorine).
Recommended • 6.0451.100 comb. Pt-ring electrode with 6.2104.020 electrode cable

Reagents • Titrant: c(phenylarsine oxide) = 0.00282 mol/L

0.50 g C6H5AsO (97%) is weighed out into a 1000 mL volumetric flask, treated
with 1 mL c(NaOH) = 1 mol/L and 20 mL dist. H2O and dissolved as quickly
as possible by swirling. The solution is then made up to the mark with O2-free
dist. H2O and mixed.
• Buffer solution pH = 4.0:
243 g sodium acetate trihydrate is weighed out into a 1000 mL volumetric
flask and dissolved in 400 mL dist. H2O. After the addition of von 480 mL
glacial acetic acid the solution is made up to the mark with dist. H2O and
mixed.
• Sodium hydroxide: c(NaOH) = 1 mol/L (40 g/L NaOH)
• Sulfuric acid: w(H2SO4) = approx. 25%
800 mL dist. H2O is placed in a beaker and 120 mL w(H2SO4) = 96% is added
to it slowly and carefully under stirring. After cooling down, it is made up to 1
liter with dist. H2O and mixed.
• Potassium iodide: solid KI, iodate-free
• Iodate stock solution: c(1/6 KIO3) = 0.1 mol/L
3.5667 g KIO3 is weighed out into a 1000 mL volumetric flask, dissolved in
dist. H2O, made up to the mark and mixed.
• Iodate standard solution: c(1/6 KIO3) = 0.005 mol/L
5.00 mL iodate stock solution is placed in a 100 mL volumetric flask, made up
to the mark with dist. H2O and mixed. This solution must be freshly prepared
every day. It is used for determining the titer of the titrant.
Titer determination Approx. 1 g potassium iodide is dissolved in 100 mL dist. H2O in a beaker. 20
mL sulfuric acid and 3.00 mL iodate standard solution are added, the solution
mixed and kept in the dark for 5 min. After the addition of 100 mL dist. H2O it is
titrated with c(phenylarsine oxide) = 0.00282 mol/L to after the first endpoint.
The titer must be determined every day. The titer of the titrant is stored in the
titrator, e.g. as Common Variable C30.
Titer calculation Titer = C00 / EP1

EP1 = mL phenylarsine oxide solution up to endpoint
C00 = mL iodate standard solution (3.00 mL)
1
Method 15 –
200 mL water sample is treated in a beaker with approx. 1 g potassium iodide Total chlorine determi-
and 1 mL buffer solution pH = 4.0 and then titrated with c(phenylarsine oxide) nation
= 0.00282 mol/L to after the first endpoint.
200 mL water sample is treated in a beaker with approx. 1 g potassium iodide Free (residual) chlo-
and 1 mL c(NaOH) = 1 mol/L and then titrated with c(phenylarsine oxide) = rine determination
1 mL c(phenylarsine oxide) = 0.00282 mol/L corresponds to 0.2 mg chlorine Calculations

(Cl2)
mg/L chlorine = EP1 x C01 x C02 x C30 / C00
EP1 = mL phenylarsine oxide up to endpoint
C01 = 0.2
C02 = 1000 (for 1 liter)
C30 = titer of phenylarsine oxide solution
If the samples contain more than 5 mg/L chlorine (which should not be the case Remarks
with drinking water) then a smaller sample volume must be selected and made
up to 200 mL with dist. H2O.
2
Method 15 –

Total chlorine Total chlorine
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY 4OTAL#LMG,
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS Total chlorine
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#HLOR%0
#
#
##
23TEXT#HLOR
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 15 –

Residual chlorine Residual chlorine
tCF tFR
$%45- 5INIT M6$%45-
# FMLA SMPLSIZEML
# %0MLM6
# RESIDUAL#LMG,
STOP6REACHED

tDE
'044ITRINO
$ATUM :EIT
$%45-
DEF
&ORMEL
#HLOR%0
#
#
## Titration curve
234EXT#HLOR Residual chlorine
23.ACHKOMMASTELLEN
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#OMMON6ARIABLE
2EPORT
2EPORT#/-0ARAMVOLL-P,ISTE2ECHN
-ITTELWERT
-.23
4EMPORÛRE6ARIABLE

tPA
'044ITRINO
$ATUM :EIT
$%45-
PARAMETERS
4ITRATIONSPARAMETER
-ESSPKTDICHTE
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$OS'ESCHWMAXMLMIN
-ESSW$RIFTM6MIN
7ARTEZEITS
3TART6AUS
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P+(.0AUS
6ORWAHL
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%INMASSABFRAUS
'RENZW%INMASSAUS
!KTIVIERPULSAUS

4
Method 15 –

Titer Titer
tPA tFR
DATE TIME $ATE 4IME
$%45-4ITER 5INIT M6$%45-
MEASPTDENSITY 4ITER
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS Titer
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-4ITER
# FMLA

tDE
'044ITRINO
DATE TIME
$%45-4ITER
DEF
FORMULA
4ITER#%0
23TEXT4ITER
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

5
Method 16 –
C 7 Permanganate index (oxidizability)
General This method determines all the organic and inorganic components that can
be oxidized by permanganate under the selected conditions. However, some
organic compounds are only partly oxidized by permanganate or not at all. This
is why this method cannot be used as a measure for the theoretical oxygen

demand. It can be used for all waters whose chloride content is less than 500
mg/L.

Reagents • Permanganate stock solution: c(KMnO4) = 0.02 mol/L

Use the solution from Method 7.
Deionized water ob-
tained from organic ion • Titrant 1: c(KMnO4) = 0.002 mol/L
exchangers is not suit- Dilute 100 mL permanganate stock solution to 1 liter with dist. H2O. The solu-
able for preparing the tion has a shelf life of approx. 3 months.
reagents. Distilled water • Sodium oxalate stock solution: c(Na2C2O4) = 0.05 mol/L
or water from a reverse Sodium oxalate is dried overnight in a drying oven at 120 °C and allowed to
osmosis unit should be cool down in a desiccator for at least 2 h. 6.700 g Na2C2O4 is weighed out into
used instead. a 1000 mL volumetric flask, dissolved in dist. H2O, made up to the mark and
mixed.
• Titrant 2: c(Na2C2O4) = 0.005 mol/L
Dilute 100 mL sodium oxalate stock solution to 1 liter with dist. H2O. The solu-
tion has a shelf life of approx. 2 weeks.
• Sulfuric acid: c(H2SO4) = 3.75 mol/L
Approx. 500 mL dist. H2O is placed in a beaker and 55 mL w(H2SO4) = 98% is
added to it slowly and carefully under stirring. Permanganate stock solution
is added to it drop by drop until a permanent, slightly pink color is seen. After
cooling down, the solution is made up to 1 liter with dist. H2O and mixed.
Analysis 100 mL water sample is placed in a 300 mL wide-neck Erlenmeyer flask and
treated with 5 mL sulfuric acid and a few glass beads and quickly brought to
the boiling point. 10.00 mL c(KMnO4) = 0.002 mol/L (Titrant 1) is then added,
the solution brought back to the boiling point and kept under gentle boiling for
exactly 10 min from the start of boiling.
If a brown coloration or total discoloration occurs during the boiling process
then the determination must be repeated with a smaller amount of sample
(made up to 100 mL with dist. H2O).
After 10 min at the boiling point, 10.00 mL c(Na2C2O4) = 0.005 mol/L (Titrant 2)
is quickly added and mixed (if the solution does not turn colorless immediately,
it must again be heated briefly until total discoloration occurs). The warm solu-
tion is then titrated with c(KMnO4) = 0.002 mol/L (Titrant 1) to after the first end-
point. If the titrant volume exceeds 5 mL the determination must be repeated
with a correspondingly diluted water sample.
1
Method 16 –
1 mL c(KMnO4) = 0.002 mol/L corresponds to 0.316 mg KMnO4 Calculations

1 mg KMnO4 = 7.9 μmol O2
mg/L KMnO4 = EP1 x C01 x C02 x C30 / C00 (RS1)
Oxidizability; μmol/L O2 = EP1 x C01 x C02 x C03 x C30 / C00 or
RS1 x C03
EP1 = mL KMnO4 up to endpoint
C00 = sample volume in mL (normally 100)
C01 = 0.316
C02 = 1000 (for 1 liter)
C03 = 7.9
C30 = titer of permanganate solution (Method 7)
• Use only thoroughly cleaned glassware! Detergent residues falsify the result. Remarks
It is better to use the same Erlenmeyer flask each time.
• The boiling time (10 min) must be observed exactly.
• In order to accelerate the titration a small amount (approx. 0.1 g) of MnSO4
can be added to the solution as a catalyst. The titration could then be carried
out at room temperature.
2
Method 16 –
tPA tCF
DATE TIME DATE TIME
-%45- -%45-
PARAMETERS # FMLA
6STEPML #
DOSRATEMAXMLMIN #
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&& tDE
PAUSES '044ITRINO
DOSELEMENTINTERNAL$ DATE TIME
MEASINPUT -%45-
TEMPERATURE# DEF
STOPCONDITIONS FORMULA
STOP6ABS +-N/%0
#
#
##
STOP6ML 23TEXT+-N/
STOP5/&&M6 23DECIMALPLACES
STOP%0 23UNITMG,
lLLINGRATEMAXMLMIN 23LIMITCONTROL/&&
STATISTICS /23
#
STATUS/&& 23TEXT/
EVALUATION 23DECIMALPLACES
%0#M6 23UNIT±MOL,
%0RECOGNITIONGREATEST 23LIMITCONTROL/&&
lX%0AT5/&&M6 SILOCALCULATIONS
P+(.0/&& MATCHID/&&
PRESELECTIONS COMMONVARIABLES
REQIDENT/&& REPORT
REQSMPLSIZE/&& REPORT#/-FULLCURVE
LIMITSMPLSIZE/&& MEAN
ACTIVATEPULSE/&& -.23
TEMPORARYVARIABLES

tFR
'044ITRINO
$ATE 4IME
5INIT M6-%45-
SMPLSIZEML
%0ML M6
+-N/MG,
/±MOL,
STOPREACHED

3
Method 17 –
C 8 CO2 content of carbonated waters
General All the CO2 is bound as carbonate by the addition of NaOH and is released
again by back-titration with HCl. As the bicarbonates (acid capacity K A 4.3, Meth-
od 10) are also determined they must be taken into account when calculating
the CO2 content.

Recommended • 6.0253.100 Aquatrode Plus with 6.2104.020 electrode cable
Reagents • Titrant: c(HCl) = 1 mol/L

prox. 800 mL dist. H2O in a 1000 mL volumetric flask, add 100 mL w(HCl) =
32%, make up to the mark with dist. H2O and mix. Titer determination with
TRIS as per Method 4.
• Sodium hydroxide: c(NaOH) = 10 mol/L.
Weigh out 80 g NaOH into a beaker, add 110 mL dist. H2O and mix immedi-
ately until everything has dissolved. Caution, the mixture becomes very hot;
wear protective goggles! After cooling down, make up to 200 mL with dist.
H2O and mix.
Analysis The container is cooled down to 0 °C, opened with as little agitation as possible
and treated with 2 mL NaOH for each 100 mL it contains. It is closed immedi-
ately (seal cans with Scotch® tape) and mixed.
100 mL pretreated water sample is placed in the titration vessel and in a first
pre-titration (SET) titrated with c(HCl) = 1 mol/L to pH = 8.2. The titration is
continued with the same titrant in the DET mode to after the first endpoint (EP
at pH = approx. 4.3).
Calculation 1 mL c(HCl) = 1 mol/L corresponds to 1 mmol CO2 or 44 mg CO2

RS1 = EP1 x C01 x C02 x C30 / C00
mmol/L CO2: RS2 = RS1 – C35
mg/L CO2: RS3 = RS2 x C03
EP1 = mL HCl from pH = 8.2 to pH = approx. 4.3
C01 = 1 (1 mL HCl = 1 mmol CO2)
C02 = 1000 (for 1 liter)
C03 = 44 ( M(CO2) in g/mol)
C30 = titer of HCl solution (Method 4)
C35 = KA 4.3 (Method 10)
Remarks The HCl addition up to pH = 8.2 (SET) neutralizes the excess alkali and the
carbonates produced.
1
Method 17 –

DET DET
tPA tFR
DATE TIME $ATE 4IME
$%4P(-$%4 P(CINIT $%4P($%4
MEASPTDENSITY STOP6REACHED
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT Titration curve – DET
TEMPERATURE#
STOPCONDITIONS
STOP6ABS
STOP6ML
STOPP(/&&
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0ATP(/&&
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%4P(-$%4
# FMLA

tDE
'044ITRINO
DATE TIME
$%4P(-$%4
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
TEMPORARYVARIABLES
#%0

2
Method 17 –

SET SET
tPA tFR
DATE TIME $ATE 4IME
3%4P(-3%4 P(CINIT 3%4P(3%4
3%4 %0ML
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&& Titration vurve – SET
TITRATIONPARAMETERS
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-3%4
# FMLA

tDE
'044ITRINO
DATE TIME
3%4P(-3%4
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 17 –

TIP TIP
tPA tFR
DATE TIME $ATE 4IME
4)0-4)0 4)04)0
SEQUENCE #
METHOD3%4 23
METHOD$%4 23MMOLL
PAUSES 23MG,
STATISTICS 4)0TERMINATED
STATUS/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
MEASMODE/&&
TEMPERATURE#

tCF
'044ITRINO
DATE TIME
4)0-4)0
# FMLA
#
#
#

tDE
'044ITRINO
DATE TIME
4)0-4)0
DEF
SEQUENCE
METHOD3%4
METHOD$%4
PAUSES
FORMULA
23#
#
#
##
23TEXT23
23DECIMALPLACES
23UNIT
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2323 #
23TEXT23
23DECIMALPLACES
23UNITMMOLL
23LIMITCONTROL/&&
2323
#
23TEXT23
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
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COMMONVARIABLES
REPORT
REPORT#/-FULL
MEAN
-.23
TEMPORARYVARIABLES

4
Method 18 –
C 9 Oxygen content according to Winkler
General During the determination of the dissolved oxygen in water samples by the Win-
kler method the following chemical reactions occur:
• Fixation in the alkaline solution:
MnCl2 + 2 NaOH = Mn(OH)2+ 2 NaCl

2 Mn(OH)2 + O2 = 2 Mn(OH) 3 + 2 H+ (2 H+ + 2 OH – (excess) = 2 H2O)
• After acidification:
2 Mn(OH)3 + 6 HCl = 2 MnCl3 + 6 H2O
2 MnCl3 + 2 KI = 2 MnCl2 + 2 KCl + I2
• During the titration:
2 Na2S2O3 + I2 = Na2S4O6 + 2 NaI
From this it can be seen that one oxygen atom releases one iodine atom and
this then oxidizes one thiosulfate molecule.
Nitrite and Fe(III) ions could possibly interfere. Nitrite is made «harmless» with
sodium azide, Fe(III) with phosphoric acid.
As dilute solutions are titrated, potentiometric endpoint indication is less suit-
able. We therefore recommend bivoltametric endpoint indication with Ipol = 1 μA
at the double Pt electrode.
Recommended • 6.0308.110 double Pt-wire electrode with 6.2104.020 electrode cable

Reagents • Winkler I: 400 g MnCl2 x 4 H2O is weighed out into a 1000 mL volumetric flask,
dissolved in dist. H2O, made up to the mark and mixed.
• Winkler II: 500 g NaOH, 1 g NaN3 and 150 g potassium iodide are dissolved in
approx. 400 mL dist. H2O in a beaker while stirring with a glass rod. Caution
– mixture becomes very hot, wear protective goggles! After cooling down,
make up to 1 liter with dist. H2O and mix (use O2-free dist. H2O).
• Acid mixture: 350 mL each of w(H3PO4) = 85% and w(HCl) = 36% are mixed,
made up to 1 liter with dist. H2O and mixed again.
• Titrant: c(Na2S2O3) = 0.05 mol/L
12.5 g Na2S2O3 x 5 H2O into a 1000 mL volumetric flask, dissolve in CO2-free
dist. H2O, make up to the mark and mix. Titer determination with KIO3 (Meth-
od 6).
Sample preparation It is assumed that the water samples have been taken correctly and filled into
the special «Winkler bottles» (with oblique stoppers).
The sample bottle is opened, 2 mL each of Winkler I and Winkler II quickly
added and the bottle then immediately resealed without generating any air
bubbles. The contents are mixed and allowed to precipitate out for a few hours.
In the absence of O2 the precipitate is white; the more O2 the sample contains,
the browner the precipitate. The stopper is then removed and approx. 1/3 of
the supernatant clear solution aspirated off. 10 mL of the acid mixture is quickly
added and the bottle is sealed and mixed. This dissolves the precipitate and
forms a yellowish clear solution, the intensity of the yellow color again depend-
ing on the O2 content.
1
Method 18 –
If wide-neck bottles are used then the titration can be carried out in them di- Analysis
rectly (add stirrer bar).
Otherwise the solution is transferred quantitatively with dist. H2O into a beaker
and titrated with c(Na2S2O3) = 0.05 mol/L to a given endpoint that has previ-
ously been determined in a MET titration.
1 mL c(Na2S2O3) = 0.05 mol/L corresponds to 0.40 mg O2 Calculations

mg/L O2 = EP1 x C01 x C02 x C30 / (C00 – C03)
EP1 = mL thiosulfate up to endpoint
C00 = Winkler bottle content in mL
C01 = 0.4
C02 = 1000 (for 1 liter)
C03 = mL Winkler I + Winkler II (4)
C30 = titer of thiosulfate solution
2
Method 18 –
tPA tFR
DATE TIME $ATE 4IME
-%4)POL- 5INIT M6-%4)POL-
6STEPML /MG,
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
%0RECOGNITIONALL
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
/%0
#
#
## #
23TEXT/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Milk and dairy products
General This section describes methods that can be used routinely in the laboratory.
For special titrimetric methods, e.g. the determination of milk damage or the
acid number of milk fats please refer to specialist literature (e.g. Bosset, J.O.,
Forschungsanstalt für Milchwirtschaft (Swiss Federal Dairy Research Station),
Liebefeld-Bern).
In addition to titrators Metrohm manufactures a whole range of other analytical
and measuring instruments. For this section we would like to draw your particu-
lar attention to the 743 Rancimat, an instrument for determining the oxidation
stability, e.g. of baby foods, butter, powdered milk, etc. Nitrite, nitrate, chloride,
phosphate, sulfate or Ca, Mg, Na, K in milk, etc. can be determined by ion chro-
D Milk and dairy products

matography after dialysis. VA instruments are particularly suitable for determin-
ing traces of heavy or toxic metals.
1
Method 19 –
D 1 pH value
General For dairy products (containing protein) special electrodes are required that
a) have a special diaphragm suitable for protein-containing samples, and
b) contain a special electrolyte.
The shape of the electrode depends on the application. Needle electrodes are
used for cheese and/or butter.
Recommended • 6.0235.100 Porotrode or 6.0226.100 needle electrode, 6.2104.020 electrode
cable
accessories
• 6.1110.100 Pt 1000 temperature sensor with 6.2104.080 electrode cable

pH measurement It is assumed that the electrode has already been calibrated – see Method 1,
Section A.
Please note that only pH values that have been measured at the same tempera-
ture can be compared with one another. If no temperature sensor is connected
to the titrator then the temperature must be measured separately and entered
in the titrator.
Measurements in milk, cream and yogurt

These are carried out with the Porotrode. The electrode is immersed in the
sample so that its diaphragm is completely covered. The pH value will be shown
or printed out when the drift value set on the titrator has been achieved.
Measurements in young cheese, soft cheese, butter and curd cheese

The needle electrode is pushed into the sample while rotating it slightly.
After the measurement the electrode is wiped off with a soft paper tissue soaked
in alcohol and then rinsed thoroughly with dist. H2O.
1
Method 19 –
D 1 pH value

Pasteurized milk Pasteurized milk
tPA tFR
DATE TIME $ATE 4IME
-%!3P(- -%!3P(-
PARAMETERS #
MEASURINGPARAMETERS P(
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&&
PRESELECTIONS pH value
REQIDENT/&&
REQSMPLSIZE/&& Pasteurized milk
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-
DEF
FORMULA
P(#
23TEXTP(
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 19 –
D 1 pH value

Soft cheese Soft cheese
tPA tFR
-%!3P(- -%!3P(-
PARAMETERS #
-ESSPARAMETER P(
-ESSW$RIFTM6MIN
7ARTEZEITAUSS
-ESSEINGANG
4EMPERATUR#
:EITINTERVALLS
3TATISTIK
3TATUSAUS
6ORWAHL pH value
)DENTABFRAGENAUS
%INMASSABFRAUS Soft cheese
'RENZW%INMASSAUS
!KTIVIERPULSAUS

tCF
'044ITRINO
$ATUM :EIT
-%!3P(-
# FMLA

tDE
'044ITRINO
$ATUM :EIT
-%!3P(-
DEF
&ORMEL
P(#
234EXTP(
23.ACHKOMMASTELLEN
23%INHEIT
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6ERGLEICHS )DAUS
#OMMON6ARIABLE
2EPORT
2EPORT#/-VOLL0ARAM-P,ISTE2ECHN
-ITTELWERT
-.23
4EMPORÛRE6ARIABLE

3
Method 20 –
D 2 Titratable acidity, degree of acidity
General The sample is titrated with NaOH up to a pH of 8.3. This measures the natural
buffer substances present in the sample and the acidic substances produced
by microorganisms or added during the manufacturing process. Normally the
alkali consumption refers to a sample weight of 100 g. The titration must be
carried out slowly under very good stirring, as in these samples (suspensions/
emulsions) the acid-base reaction is inhibited.
Recommended • 6.0235.100 Porotrode with 6.2104.020 electrode cable


Reagents • Titrant 1: c(NaOH) = 0.1 mol/L. See Method 3, Section B
– Titrant 2: c(NaOH) = 0.25 mol/L
10.0 g NaOH in CO2-free dist. H2O, make up to 1 liter and mix. Titer determi-
nation with potassium hydrogen phthalate (Method 3).
Milk, yogurt, cream:

The Porotrode has already been or will be calibrated with a buffer solution ac-
cording to Method 1, Section A.
25 g sample (homogenized if necessary) is placed in a beaker. After the ad-
dition of approx. 25 mL dist. H2O the mixture is titrated with c(NaOH) = 0.25
mol/L to pH = 8.3 (SET mode).
Calculations Degree of acidity = mL c(NaOH) = 0.25 mol/L per 100 g sample

= EP1 x C01 x C30 / C00
EP1 = mL NaOH up to endpoint at pH = 8.3
C01 = 100 (for 100 g)
C30 = titer of NaOH (Method 3)
C00 = sample size in g (25)
Conserved milk products (powdered milk, condensed milk):

An amount of sample corresponding to 10 g fat-free dry weight is weighed out
into a beaker. After moistening if necessary it is stirred with 50 mL dist. H2O at
room temperature and allowed to stand for 20 min. The suspension should be
free from clumps. It is then titrated with c(NaOH) = 0.1 mol/L to pH = 8.3 (SET
mode).
For routine investigations the following sample amounts apply:
Whole milk powder 13 g
Skimmed milk powder 10 g
Condensed milk, sweetened 15 g
Condensed milk, unsweetened 43 g for 100 g/kg fat and 55 g for 80 g/kg fat
Calculations Titratable acidity = mL c(NaOH) = 0.1 mol/L per 10 g fat-free dry mass
= EP1 x C30
EP1 = mL NaOH up to endpoint at pH = 8.3
1
Method 20 –
Instrument parameters and calculation – Milk
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4
%0ATP(
DYNAMICS/&& tDE
MAXRATEMLMIN '044ITRINO
MINRATE±LMIN DATE TIME
STOPCRITDRIFT 3%4P(-
STOPDRIFT±LMIN DEF
3%4 FORMULA
%0ATP(/&& 4ITRB(%0
#
TITRATIONPARAMETERS 23TEXT4ITRB(
TITRDIRECTIONAUTO 23DECIMALPLACES
PAUSES 23UNIT
START6ABS 23LIMITCONTROL/&&
START6ML SILOCALCULATIONS
DOSRATEMAXMLMIN MATCHID/&&
PAUSES COMMONVARIABLES
EXTRTIMES REPORT
DOSELEMENTINTERNAL$ REPORT#/-FULLCURVE
MEASINPUT MEAN
TEMPERATURE# -.23
TIMEINTERVALS TEMPORARYVARIABLES
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – Milk Result – Milk
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEG
%0ML
DEGROFAC

2
Method 20 –
Instrument parameters and calculation – Condensed milk
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN tDE
STOPCRITDRIFT '044ITRINO
STOPDRIFT±LMIN DATE TIME
3%4 3%4P(-
%0ATP(/&& DEF
TITRATIONPARAMETERS FORMULA
TITRDIRECTIONAUTO 3AEUREGR%0
#
##
PAUSES 23TEXT3AEUREGR
START6/&& 23DECIMALPLACES
PAUSES 23UNIT
EXTRTIMES 23LIMITCONTROL/&&
DOSELEMENTINTERNAL$ SILOCALCULATIONS
MEASINPUT MATCHID/&&
TEMPERATURE# COMMONVARIABLES
TIMEINTERVALS REPORT
STOPCONDITIONS REPORT#/-FULLCURVE
STOP6ABS MEAN
STOP6ML -.23
lLLINGRATEMAXMLMIN TEMPORARYVARIABLES
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – Condensed milk Result – Condensed milk
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEG
%0ML
TITRABLEAC

3
Method 21 –
D 3 Chloride
General Proteins interfere with the determination, they are therefore removed from the
(dissolved/suspended) sample by precipitation with aluminum sulfate. We have
replaced the frequently proposed toxic titrant Hg(II) nitrate by silver nitrate,
which yields the same results.


Reagents • Titrant: c(AgNO3) = 0.1 mol/L. See Method 8, Section B
• Sodium hydroxide solution: c(NaOH) = 2 mol/L
• Nitric acid: c(HNO3) = 2 mol/L
• Aluminum sulfate: w(Al sulfate) = 20%
200 g Al2(SO4)3 x 18 H2O is dissolved in dist. H2O and made up to 1 liter.
Sample preparation Milk

20 mL milk and approx. 150 mL dist. H2O and 3...7 mL NaOH solution are placed
in a 200 mL volumetric flask. After thorough mixing 10 mL aluminum sulfate
solution is added, the mixture is made up to the mark with dist. H2O and then
shaken thoroughly. Allow to settle and then filter through a paper filter (if the
solution is turbid then too much NaOH has been added).
Cheese
1 to 2 g grated sample is treated with 7 mL NaOH and 50 mL warm dist. H2O
in a 200 mL volumetric flask and dissolved by shaking. After cooling down, 20
mL aluminum sulfate solution is added, the solution is made up to the mark with
dist. H2O and then shaken thoroughly. Then proceed as described above under
Milk.
Butter
1 to 2 g butter is weighed out into a 200 mL volumetric flask, treated with 7 mL
NaOH and 50 mL warm dist. H2O and thoroughly shaken. After cooling down,
20 mL aluminum sulfate solution is added, the solution is made up to the mark
with dist. H2O and then shaken thoroughly. Then proceed as described above
under Milk.
Analysis 100 mL of the filtered sample solution (1/2 of sample weight) is placed in the ti-
tration beaker, treated with 2.5 mL HNO3 and titrated with c(AgNO3) = 0.1 mol/L
to after the first endpoint.
Calculations 1 mL c(AgNO3) = 0.1 mol/L corresponds to 3.5453 mg chloride or 5.8443 mg

NaCl
Milk; mg/100 mL chloride = EP1 x C01 x C02 x C30 / C00
Butter/cheese; % NaCl = EP1 x C03 x C04 x C30 / C00
C00 = sample volume (mL) or sample weight (g) divided by 2
C01 = 3.5453
C02 = 10 (for 100 mL)
C03 = 5.8443
C04 = 0.1 (for %)
C30 = titer of AgNO3 solution (Method 8)
1
Method 21 –
D 3 Chloride
Instrument parameters and calculation – Milk
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES tDE
PAUSES DATE TIME
MEASINPUT DEF
STOPCONDITIONS -ILCH%0
#
#
##
STOP6ABS 23TEXT-ILCH
lLLINGRATEMAXMLMIN SILOCALCULATIONS
STATISTICS MATCHID/&&
STATUS/&& COMMONVARIABLES
EVALUATION REPORT
%0# REPORT#/-FULLCURVE
%0RECOGNITIONGREATEST MEAN
lX%0AT5/&&M6 -.23
P+(.0/&& TEMPORARYVARIABLES
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – Milk Result – Milk
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEML
%0MLM6
MILKMGL
STOP6REACHED

2
Method 21 –
D 3 Chloride
Instrument parameters and calculation – Cheese
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&& tDE
PAUSES '044ITRINO
MEASINPUT $%45-
TEMPERATURE# DEF
STOP6ABS .A#L%0
#
#
##
STOP6ML 23TEXT.A#L
STOP%0 23UNIT
STATISTICS SILOCALCULATIONS
STATUS/&& MATCHID/&&
EVALUATION COMMONVARIABLES
%0# REPORT
%0RECOGNITIONGREATEST REPORT#/-FULLCURVE
lX%0AT5/&&M6 MEAN
P+(.0/&& -.23
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – Cheese Result – Cheese
FR
'044ITRINO
$ATE 4IME
5INIT M6$%45-B
SMPLSIZEG
%0MLM6
.A#L
STOP6REACHED

3
Method 22 –
D 4 Calcium
General The method described here allows the calcium content of the sample to be de-
termined without ashing the sample. After a sample preparation that is easy to
carry out, the Ca is determined with EGTA using a Cu ISE.
Recommended • 6.0502.140 Cu ISE with 6.2104.020 electrode cable and 6.0726.107 Ag/AgCl
Reference electrode with 6.2106.020 electrode cable
accessories

38.04 g ethylene glycol-bis-(2-aminoethyl)-tetraacetic acid is dissolved in
250 mL c(NaOH) = 1 mol/L and, after cooling down, made up to 1 liter with
dist. H2O. Titer determination with CaCO3 (Method 9, Section B).
• Auxiliary solution: c(CuEGTA) = 0.05 mol/L
2.2 g NH4Cl and 3.751 g Cu(NO3)2 are dissolved in dist. H2O in a 200 mL volu-
metric flask, made up to the mark and mixed. 50.0 mL each of this solution
and c(EGTA) = 0.1 mol/L are mixed.
• Buffer solution pH = 10: 54 g NH4Cl is dissolved in approx. 400 mL dist. H2O,
treated with 300 mL w(NH3) = 25%, made up to 1 liter with dist. H2O and
mixed.
• Sulfuric acid: c(H2SO4) = 0.05 mol/L
• Sodium hydroxide: c(NaOH) = 0.1 mol/L
Sample preparation Milk, yogurt

Approx. 10 g sample is weighed out into the titration beaker and diluted with 90
mL dist. H2O.
Cheese
Approx. 1 g finely grated cheese is weighed out into the titration beaker and
treated with 10 mL sulfuric acid and approx. 50 mL dist. H2O. The mixture is
heated to 40 °C and then stirred at this temperature for 10 min. After cooling
down, the mixture is pre-neutralized with NaOH to pH = approx. 7.
Analysis 1 mL CuEGTA is added to the prepared sample solution and allowed to react for
30 s under stirring. After the addition of 5 mL buffer solution pH = 10, the titra-
tion is carried out with c(EGTA) = 0.1 mol/L to after the first endpoint.
Calculations 1 mL c(EGTA) = 0.1 mol/L corresponds to 4.008 mg Ca

% Ca = EP1 x C01 x C02 x C30 / C00
EP1 = mL EGTA up to endpoint
C00 = sample weight in g
C01 = 4.008
C02 = 0.1 (for %)
C30 = titer of EGTA solution (Method 9)
Remarks If magnesium is also to be determined then a second titration is carried out with
c(Na2EDTA) = 0.1 mol/L with the addition of CuEDTA. In this way the sum of
Ca + Mg is obtained. The Mg content can be determined from the difference
between the two titrant consumptions.
1 mL c(Na2EDTA) = 0.1 mol/L corresponds to 2.4305 mg Mg
1
Method 22 –
D 4 Calcium
Instrument parameters and calculation – Cheese
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&& tDE
PAUSES '044ITRINO
MEASINPUT $%45-
TEMPERATURE# DEF
STOP6ABS #A%0
#
#
##
STOP6ML 23TEXT#A
STOP%0 23UNIT
%0# REPORT
%0RECOGNITIONALL REPORT#/-FULLCURVE
lX%0AT5/&&M6 MEAN
P+(.0/&& -.23
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – Cheese Result – Cheese
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEG
%0ML M6
#A
STOP6REACHED

2
Method 23 –
D 5 Vitamin C (ascorbic acid)
General Vitamin C determination is carried out for milk, powdered milk and possibly
baby food. The titrant is 2,6-dichlorophenolindophenol and bivoltametry at the
double Pt-sheet electrode is used for endpoint indication. Metaphosphoric acid
is added to precipitate out interfering proteins. It also protects ascorbic acid
against autoxidation.
Recommended • 6.0309.100 double Pt-sheet electrode with 6.2104.020 electrode cable


Reagents • Titrant: c(DPIP) = 0.001 mol/L
330 mg 2,6-dichlorophenolindophenol-Na x 2 H2O and 100 mg NaHCO3 are
weighed out into a 1000 mL volumetric flask, completely dissolved in dist.
H2O, made up to the mark and mixed.
• Metaphosphoric acid: w [(HPO3)n] = 3%
15 g (HPO3)n is dissolved in dist. H2O, made up to 500 mL and mixed. The
solution is not stable. Even if kept in a refrigerator at 4 °C it has a shelf life of
only 1 to 2 days (hydrolyzes to H3PO4).
• Vitamin C standard: ρ (Vitamin C) = 1 mg/mL.
100.0 mg ascorbic acid is weighed out into a 100 mL volumetric flask, dis-
solved in metaphosphoric acid 3%, made up to the mark with it and mixed.
The solution is only stable for a few hours and must always be made up
freshly.
• Copper sulfate: ρ (CuSO4) = 100 g/L
15.6 g CuSO4 x 5 H2O is weighed out into a 100 mL volumetric flask, dissolved
in dist. H2O, made up to the mark and mixed. 1 mL = 100 mg CuSO4
Titer determination 15 mL metaphosphoric acid 3% and 5 mL dist. H2O are measured out into the
of titrant titration beaker. After the addition of 100 μL vitamin C standard (0.100 mg) the
titration is carried out immediately with c(DPIP) = 0.001 mol/L to after the first
endpoint. Titer determination must be carried out every day. The titer is stored
as a Common Variable (e.g. C30) in the Titrino.
Titer = C00 / C01 / EP1
EP1.= mL DPIP up to endpoint
C00.= «sample weight» vitamin C in mg (0.1)
C01.= 0.088 (vitamin C equivalent)
C31.= blank value for metaphosphoric acid in mL
Sample preparation 5.0 mL milk or prepared powder sample is pipetted into a centrifuge tube,
treated with 15 mL metaphosphoric acid and 1 mL dist. water and thoroughly
mixed/shaken. It is then centrifuged for 20 min at approx. 3000 min –1.
1
Method 23 –
In addition to vitamin C, milk contains other components that are oxidized by Determining the blank
DPIP and would simulate a too high vitamin C content. In order to determine this value of the sample
blank value, CuSO4 is added to the sample to block the vitamin C.
5.0 mL milk or prepared powder sample and 1 mL CuSO4 are pipetted into
a centrifuge tube, treated with 15 mL metaphosphoric acid and thoroughly
mixed/shaken. It is then centrifuged for 20 min at approx. 3000 min –1.15 mL of
the pretreated milk sample is then pipetted into the titration beaker and titrated
with c(DPIP) = 0.001 mol/L to after the first endpoint. This blank value (mL) is
also stored in the Titrino as a Common Variable (e.g. C31).
Blank value of sample = EP1 = C31
15 mL of the pretreated (centrifuged) sample solution and 5 mL dist. H2O are Analysis
pipetted into the titration beaker and titrated with c(DPIP) = 0.001 mol/L to after
the first endpoint.
1 mL c(DPIP) = 0.001 mol/L corresponds to 0.088 mg vitamin C Calculations

mg/L vitamin C = (EP1 – C31) x C01 x C02 x C03 x C30 / C00
EP1 = mL DPIP up to endpoint for sample solution
C01 = 0.088
C02 = 1.4 (dilution factor; only 15 mL out of 21 mL is titrated)
C03 = 1000 (for 1 liter)
C30 = titer of DPIP solution
C31 = blank value of sample
Titration curve – blank value Result – blank value
tFR
'044ITRINO
$ATE 4IME
5INIT M6-%4)POL-
SMPLSIZEML
%0MLM6
6IT#MG,
STOP6REACHED

2
Method 23 –
Instrument parameters and calculation Result – sample

sample
tPA tFR
DATE TIME $ATE 4IME
6STEPML 6IT#MG,
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
TEMPERATURE# Titration curve – sample
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
6IT#%0 #
#
#
#
##
23TEXT6IT#
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 23 –
Instrument parameters and calculation Result – titer

titer
tPA tFR
DATE TIME $ATE 4IME
-%4)POL-4ITER 5INIT M6-%4)POL-4ITER
PARAMETERS SMPLSIZEG
6STEPML 4ITER
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
TEMPERATURE# Titration curve – titer
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
%0RECOGNITIONLAST
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-4ITER
# FMLA
#

tDE
'044ITRINO
DATE TIME
-%4)POL-4ITER
DEF
FORMULA
4ITER##%0
23TEXT4ITER
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 24 –
D 6 Kjeldahl nitrogen (Kjeldahl N)
General The sample is digested with sulfuric acid in the presence of a catalyst. This
converts organically bound nitrogen to ammonium sulfate. In a distillation ap-
paratus the digestion solution is treated with excess NaOH and the released
ammonia (NH3) is distilled into a solution of boric acid. The ammonia absorbed
in the boric acid is titrated with HCl.
Sample preparation The amount of sample to be digested depends on the expected nitrogen con-
tent. The following table provides a guideline:
Kjeldahl N

g/kg N g/kg protein Weight in g
<10 <60 2...3
10 60 1.5
20 120 0.7
50 300 0.25
100 600 0.15
150 900 0.1
The thoroughly homogenized and, if necessary, pre-dried sample is weighed

out into the digestion tube, a catalyst tablet* and 10 mL w(H2SO4) = 98% are
added and heated in the digestion apparatus until the solution is clear. Heating
is continued for a further 10 min and the solution is then allowed to cool down.
* 5 g tablets of various compositions are commercially available. We recommend the use of those
without any Se and/or Hg additives.
Recommended • 6.0239.100 comb. pH glass electrode with 6.2104.020 electrode cable

Reagents • Titrant: c(HCl) = 0.1 mol/L. See Method 4, Section B

• Absorption solution: w(H3BO3) = 2%
20 g boric acid is dissolved in dist. H2O, made up to 1 liter and mixed.
Analysis The pH glass electrode is first calibrated with buffer solutions pH = 7.0 and pH
= 4.0 (Method 1, Section A).
Approx. 30 mL boric acid is placed in the titration vessel.
The digestion tube is connected to the distillation apparatus, the digestion solu-
tion is treated with NaOH and the distillation and Titrino are started at the same
time. The titration with c(HCl) = 0.1 mol/L takes place in the SET-pH mode with
the following parameter settings:
EP at pH 4.5
dynamics 6
max. rate 2 mL/min
min. rate 25 μL/min
titr. direction auto
1
Method 24 –
1 mL c(HCl) = 0.1 mol/L corresponds to 1.4007 mg N Calculation

g/kg N = EP1 x C01 x C30 / C00
EP1 = mL HCl up to endpoint
C01 = 1.4007
C30 = titer of HCl solution (Method 4)
2
Method 24 –

Milk powder Milk powder
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( .GKG
DYNAMICS
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&& Titration curve
TITRATIONPARAMETERS
TITRDIRECTIONAUTO Milk powder
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZEALL
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
.%0
#
##
23TEXT.
23DECIMALPLACES
23UNITGKG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Fat and oil numbers
General The parameters described are used to determine the quality of fats and oils.
The peroxide number is a good measure of the amount of damage suffered
by the sample (storage, oxidation by light or temperature effects). The iodine
number and the saponification number provide certain information about the
composition of the fat, whereas the hydroxyl number may indicate the presence
of certain (illicit) additives.
In this section we also propose methods for the determination of the hydroxyl,
iodine and peroxide numbers that require virtually no toxic reagents. These are
alternative methods that we have thoroughly tested.
In addition to titrators, Metrohm also manufactures other instruments for the
analysis of fats and oils. In particular, we would like to draw your attention to an
instrument for determining the oxidation stability, the 743 Rancimat. With the
E Edible fats and oils

797 VA Computrace you can determine traces of heavy metals (e.g. Ni, Pb, Cu,
etc.) and/or antioxidants (e.g. tocopherol, vitamin E) in your samples.
1
Method 25 –
Recommended • 6.0229.100 Solvotrode, 6.2320.000 electrolyte (tetraethylammonium bro-

accessories mide, c = 0.4 mol/L in ethylene glycol) with 6.2104.020 electrode cable
Reagents • Titrant: c(KOH) = 0.1 mol/L in IPA. See Method 3, Section B

• Solvent: ethanol/diethyl ether 1:1, neutralized with KOH against phenolphtha-
lein.
Analysis 5...10 g sample is weighed out into the titration beaker and dissolved in 50 mL
solvent. The solution is then titrated with c(KOH) = 0.1 mol/L to after the first
endpoint.

Calculations Acid number (AN) in mg KOH per g sample
FFA in mL c(KOH) = 0.1 mol/L per 10 g sample
AN = EP1 x C01 x C30 / C00
FFA = EP1 x C02 x C30 / C00
EP1 = mL KOH up to endpoint
C01 = 5.61 (equivalent weight of KOH)
C02 = 10 (for 10 g)
C30 = titer of KOH (Method 3)
1
Method 25 –
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN tDE
EQUILIBRTIMES '044ITRINO
START6/&& DATE TIME

PAUSES $%45-
DOSELEMENTINTERNAL$ DEF
MEASINPUT FORMULA
TEMPERATURE# 3:%0
#
##
STOPCONDITIONS 23TEXT3:
STOP6ABS 23DECIMALPLACES
STOP6ML 23UNIT
STOP5/&&M6 23LIMITCONTROL/&&
STOP%0 &&!%0
#
##
lLLINGRATEMAXMLMIN 23TEXT&&!
STATISTICS 23DECIMALPLACES
STATUS/&& 23UNIT
EVALUATION 23LIMITCONTROL/&&
%0# SILOCALCULATIONS
%0RECOGNITIONALL MATCHID/&&
lX%0AT5/&&M6 COMMONVARIABLES
P+(.0/&& REPORT
PRESELECTIONS REPORT#/-FULLCURVE
REQIDENT/&& MEAN
REQSMPLSIZE/&& -.23
LIMITSMPLSIZE/&& TEMPORARYVARIABLES
ACTIVATEPULSE/&&

Titration curve Result – Rapeseed oil
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEG
%0ML M6
!.
&&!
STOP6REACHED

2
Method 26 –
E 2 Hydroxyl number

Reagents • Titrant: c(TBAOH) = 0.1 mol/L in IPA/methanol.

You cannot prepare this titrant yourself, use e.g. Merck no. 109162. TBAOH
= tetrabutylammonium hydroxide. Titer determination with benzoic acid,
Section B. Approx. 120 mg benzoic acid is weighed out into the titration
beaker with an accuracy of 0.1 mg, dissolved in approx. 40 mL ethanol
and then titrated with c(TBAOH) = 0.1 mol/L to after the first endpoint.
1 mL c(TBAOH) = 0.1 mol/L corresponds to 12.212 mg C7H6O2
• TSI reagent:

Approx. 250 mL acetonitrile is treated with 20 mL toluene-4-sulfonyl-isocya-
nate (e.g. Fluka no. 89815) in a 500 mL volumetric flask. The solution is made
up to the mark with acetonitrile and mixed. The reaction solution has a shelf
life of approx. 1 month.
Analysis Depending on the expected hydroxyl number, 0.1...15 g sample* is weighed

out into the titration beaker and mixed with 3 mL toluene and 7 mL acetonitrile.
10 mL reaction solution is added, the beaker covered with a watch glass, and
the reaction is allowed to continue for 10 min under gentle stirring. After the ad-
dition of 0.5 mL dist. H2O the reaction is allowed to continue on the stirrer for
a further 5 min (this destroys the excess TSI reagent). After the addition of 30
mL acetonitrile the solution is titrated with c(TBAOH) = 0.1 mol/L to after the
second endpoint.
Rinsing is carried out after each titration, first with acetone and then with dist.
H2O. The electrode is then conditioned by immersing it in dist. H 2O for 1 min.
* The optimal sample weight is calculated as follows:
g sample = 40 / expected hydroxyl number (OHN)
Calculations Hydroxyl number (OHN) = mg KOH per g sample

OHN = (EP2 – EP1) x C01 x C02 x C30 / C00
EP1 = mL TBAOH up to first endpoint
EP2 = mL TBAOH up to second endpoint
C01 = 56.1 (M(KOH) in g/mol)
C02 = 0.1 (c(TBAOH) in mol/L)
C30 = titer of TBAOH
1
Method 26 –
E 2 Hydroxyl number
Instrument parameters and calculation Result – Safflower oil
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY %0ML M6
MININCR±L /(.
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
/(:%0 %0
#
#
##
23TEXT/(:
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 27 –
E 3 Iodine number

ccessories • 6.3026.220 Exchange Unit(s)
Reagents • Titrant: c(Na2S2O3) = 0.1 mol/L. See Method 6, Section B.

• Reaction solution: c(ICl) = 0.1 mol/L in glacial acetic acid
16.2 g iodine monochloride is dissolved in glacial acetic acid, made up to 1
liter with it and mixed.
• Glacial acetic: w(acetic acid) = 96...98%
• Catalyst solution: w(Mg acetate) = 3% in glacial acetic acid
45 g Mg(OOCCH3)2 x 4 H2O is dissolved in glacial acetic acid, made up to 1

• Potassium iodide solution: w(KI) = 10%
50 g KI is dissolved in dist. H2O, made up to 500 mL and mixed. It is stored in
a dark bottle and protected against light.
Blank value of 20 mL glacial acetic acid, 25.0 mL reaction solution and 10 mL catalyst solution
reaction solution are placed in an Erlenmeyer flask with an SGJ stopper. The flask is sealed and
placed in the dark for exactly 5 min. 15 mL KI is then added, the mixture is then
transferred quantitatively to the titration beaker with dist. H2O and immediately
titrated with c(Na2S2O3) = 0.1 mol/L to after the first endpoint.
The titrant consumption (EP1) is stored in the Titrino as a Common Variable
(e.g. C31).
BV = EP1 (mL) = C31
Analysis Depending on the expected iodine number, 0.1…1 g sample is weighed out into
an Erlenmeyer flask with an SGJ stopper and dissolved in 20 mL glacial acetic
acid. After the addition of 25.0 mL reaction solution and 10 mL catalyst solution
the flask is sealed and placed in the dark for exactly 5 min. 15 mL KI is then
added, the mixture is then transferred quantitatively to the titration beaker with
dist. H2O and immediately titrated with c(Na2S2O3) = 0.1 mol/L to after the first
endpoint.
Calculations Iodine number (IN) = g iodine per 100 g sample

IN = (C31 – EP1) x C01 x C30 / C00
EP1 = mL thiosulfate solution for sample
C01 = 1.269 (equivalent weight of iodine)
C30 = titer of thiosulfate solution (Method 6)
C31 = BV (mL thiosulfate solution)
Remarks • For vegetable fats and oils the iodine number lies between 75 and 150 g
iodine per 100 g. Exceptions are coconut fat with 6...11 and palm oil with ap-
prox. 50 g I2 / 100 g.
• Care must be taken that at least 25 mL thiosulfate are consumed by the
sample. If the consumption is lower than there is the risk that too low iodine
numbers will be obtained.
• For olive and sunflower oil we have found that the sample weights should be
less than 0.4 g.
1
Method 27 –
E 3 Iodine number
Instrument parameters and calculation Result – Safflower oil
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY ).
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
):# %0
#
##
23TEXT):
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 28 –
E 4 Peroxide number

Reagents • Titrant: c(Na2S2O3) = 0.01 or 0.001 mol/L

Freshly prepared every day from c(Na2S2O3) = 0.1 mol/L (Method 6, Section
B) by dilution with dist. H2O.
• Solvent: glacial acetic acid:
1-decanol 3 : 2, containing iodine 10...15 mg iodine is dissolved in 200 mL
1-decanol. 300 mL glacial acetic acid is added and the solution mixed.
• Potassium iodide solution: KI sat.
In an amber glass bottle approx. 100 mL dist. H2O is added to 130 g KI and

mixed. The solution must be stored protected against light.
Blank value of 10.0 mL solvent is pipetted into the titration beaker, treated with 200 μL potas-
sium iodide, thoroughly mixed and placed in the dark for 1 min. After the addi-
solvent tion of 50 mL dist. H2O the solution is titrated with c(Na2S2O3) = 0.01 mol/L or
0.001 mol/L to after the first endpoint. The titrant consumption (EP1) is stored in
the Titrino as a Common Variable (e.g. C31).
BV = EP1 (mL) = C31
Analysis Depending on the expected peroxide number, 1...2 g sample is weighed out
into the titration beaker, dissolved in 10.0 mL solvent, treated with 200 μL
potassium iodide, thoroughly mixed and placed in the dark for 1 min. After
the addition of 50 mL dist. H2O the solution is titrated with c(Na 2S2O3 ) = 0.01
mol/L or 0.001 mol/L to after the first endpoint.
Calculations Peroxide number (PON) in milliequivalents O2 per kg sample

PON = C01 x (EP1 – C31) x C30 / C00
EP1 = mL thiosulfate for sample
C01 = 10 for c(Na2S2O3) = 0.01 mol/L and 1 for c(Na2S2O3) = 0.001 mol/L
C31 = mL thiosulfate solution for BV
Remarks • For peroxide numbers below 1 c(Na2S2O3) = 0.001 mol/L is used.

• The blank value determination of the solvent normally causes great difficulty.
For this reason we have added a small amount of iodine to the solvent. A
precondition for this is that the same amount of solvent (10.0 mL) is always
used.
• The PON in refined products is normally in the range 0...1, in unrefined prod-
ucts in the range 5...20 meq. O2 / kg.
1
Method 28 –
E 4 Peroxide number
Instrument parameters and calculation Result – Sunflower oil
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY 0/8
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
0/:#
%0 #
##
23TEXT0/:
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 29 –
E 5 Saponification number

• 6.3026.220 Exchange Units
Reagents • Titrant: c(HCl) = 0.5 mol/L.

Ready to use or according to Method 4, Section B
• Reaction solution: c(KOH) = 0.5 mol/L in ethanol.
Ready to use or according to Method 3, Section B
• Solvent: ethanol, abs.

Blank value of 25.0 mL c(KOH) = 0.5 mol/L is pipetted into an Erlenmeyer flask with an SGJ
reaction solution and a stirrer bar is added. A reflux condenser is attached and the solution is
heated to gentle boiling for 30 min. After cooling down, it is diluted with 50 mL
ethanol and titrated with c(HCl) = 0.5 mol/L to after the first endpoint. The titrant
consumption (EP1) is stored in the Titrino as a Common Variable (e.g. C31).
BV = EP1 (mL) = C31
Analysis 0.5…3 g sample is weighed out with an accuracy of 0.5 mg into an Erlenmeyer
flask equipped with an SGJ. 25.0 mL c(KOH) = 0.5 mol/L and a stirrer bar are
added and the reflux condenser is attached. The solution is heated to gentle
boiling for 30 min. After cooling down, it is diluted with 50 mL ethanol and ti-
trated with c(HCl) = 0.5 mol/L to after the first endpoint.
Saponification number (SN) in mg KOH / g sample
Calculations SN = (C31 – EP1) x C01 x C30 / C00
EP1 = mL HCl for sample
C31 = mL HCl for BV
Remarks For most vegetable fats and oils the saponification number is around 180...195
mg KOH / g. Exceptions are coconut fat with 250...265 and palm oil with
190...210 mg KOH per g.
The back-titration produces two endpoints. EP1 corresponds to the excess
KOH and (EP2 – EP1) to the «potassium soap».
1
Method 29 –
E 5 Saponification number
Instrument parameters and calculation Result – Olive oil
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY %0MLM6
MININCR±L 3.
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
6:# %0
#
##
23TEXT6:
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Cereals, flour-milling products,
dry paste products
F Cereals, flour-milling products, dry paste products

Sample preparation, If necessary, the samples are finely ground in a laboratory cross-beater mill with
general a cooling jacket. The temperature must not exceed 40 °C!
1
F Cerealien, F Cereals, flour-milling products, dry paste pro-
Method 30 –
F 1 pH value and degree of acidity
Recommended • 6.0259.100 Unitrode with 6.2104.020 electrode cable

Reagents • Titrant: c(NaOH) = 0.1 mol/L. See Method 3, Section B

• Buffer solutions pH = 7.00 and pH = 4.00
Metrohm nos. 6.2307.110 and 6.2307.100
pH value It is assumed that the electrode has already been calibrated – see Method 1,
Section A.
Please note that the temperature at which the measurement was made must
always be mentioned together with the pH value. If no temperature sensor is
connected to the instrument then the temperature must be measured sepa-
rately and entered in the titrator. Recommended electrodes: 6.0258.000 Uni-
trode (with built-in Pt 1000) or 6.1110.100 temperature sensor with 6.2104.080
electrode cable.
90 mL dist. H2O is pipetted into a 250 mL beaker and 10 g sample is stirred in
or suspended. The electrode is immersed and the pH measurement is started
under stirring. As soon as the drift value set on the titrator has been reached,
the pH value is shown or printed out automatically.
Degree of acidity A further 100 mL dist. H2O is added to the sample suspension used for the pH
measurement. This suspension is then titrated with c(NaOH) = 0.1 mol/L in the
SET mode to pH 8.5 (delay time 60 s).
Calculation Degree of acidity = mL c(NaOH) = 0.1 mol/L per 10 g sample

Degree of acidity = EP1 x C30
EP1 = mL NaOH up to endpoint (pH = 8.5)
1
Method 30 –

pH value pH value
tPA tFR
DATE TIME $ATE 4IME
-%!3P(-P( -%!3P(-P(
PARAMETERS #
MEASURINGPARAMETERS
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&& pH value
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-P(
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-P(
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 30 –

Degree of acidity Degree of acidity – spaghetti
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
PARAMETERS %0ML
3%4 DEGROFACMLG
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&& Titration curve
TITRATIONPARAMETERS
TITRDIRECTIONAUTO Degree of acidity
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
3R%0
#
23TEXT3R
23DECIMALPLACES
23UNITMLG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 31 –
F 2 Chloride and cooking salt content


• Nitric acid: c(HNO3) = approx. 2 mol/L
w(HNO3) = 65% diluted 1:5 with dist. H2O.
• Ethanol: abs. 98%
Depending on the chloride content, 2...5 g finely ground sample is weighed out
Analysis into the titration beaker, treated with 50 mL ethanol, 50 mL dist. H2O and 5 mL
nitric acid and stirred on the magnetic stirrer for 10 min. The solution is then
titrated with c(AgNO3) = 0.1 mol/L to after the first endpoint.
1 mL c(AgNO3) = 0.1 mol/L corresponds to 3.5453 mg chloride or 5.8443 mg

Calculations NaCl
g/kg chloride = EP1 x C01 x C30 / C00
% NaCl = EP1 x C02 x C03 x C30 / C00
EP1 = mL AgNO3 solution up to endpoint
C01 = 3.5453
C02 = 5.8443
C03 = 0.1 (for %)
1
Method 31 –

Breadcrumbs Breadcrumbs
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY CHLORIDEGKG
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
TEMPERATURE# Breadcrumbs
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#HLORID%0
#
##
23TEXT#HLORID
23DECIMALPLACES
23UNITGKG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 31 –

Pasta Pasta
tPA tFR
DATE TIME $ATE 4IME
-%45- 5INIT M6-%45-
6STEPML .A#L
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
Titration curve
STOPCONDITIONS Pasta
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%45-
DEF
FORMULA
.A#L%0
#
#
##
23TEXT.A#L
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 32 –
F 3 Kjeldahl nitrogen, total protein

General With this type of sample the Kjeldahl method determines the total nitrogen. A
factor can be used to calculate the total protein content. This corresponds in
principle to the average content of «available» proteins.
The sample is digested with sulfuric acid in the presence of a catalyst. This
tent. The following table provides a guideline:
Kjeldahl-N
g / 100 g N g / 100 g protein Sample weight in g

<1 <6 2
2 12 1
5 30 0.5
The thoroughly homogenized and pulverized sample is weighed out into the
digestion tube, a catalyst tablet* and 10 mL w(H2SO4) = 98% are added and
heated in the digestion apparatus until the solution is clear. Heating is contin-
ued for a further 10 min.


EP at pH 4.5
dynamics 6
max. rate 2 mL/min
1
Method 32 –
1 mL c(HCl) = 0.1 mol/L corresponds to 1.4007 mg N Calculations

Total nitrogen (N) in g per 100 g dry weight
N = EP1 x C01 x C02 x C03 x C30 / C00 = RS1
Total protein (TP) in g per 100 g dry weight
TP = RS1 x C04**
** The factor C04 has the following values:
5.3 for oil seeds and shell fruits (e.g. linseed, sunflower seeds, etc.)
5.7 for cereal grains (e.g. wheat, rice, barley, etc.)
6.25 for heterogeneous foodstuffs (e.g. pasta products)
EP1 = mL HCl up to endpoint (pH = 4.5)

C01 = 1.4007
C02 = 0.1 (for 100 g)
C03 = correction factor for dry weight (100 / (100 – % H2O)
C04 = 5.3 or 5.7 or 6.25 (sample-dependent)
C30 = titer of HCl (Method 4)
2
Method 32 –
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS #
MAXRATEMLMIN #
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN tDE
3%4 '044ITRINO
%0ATP(/&& DATE TIME
TITRATIONPARAMETERS 3%4P(-
TITRDIRECTIONAUTO DEF
PAUSES FORMULA
START6/&& .%0
#
#
#
##
PAUSES 23TEXT.
EXTRTIMES 23DECIMALPLACES
DOSELEMENTINTERNAL$ 23UNITGG
MEASINPUT 23LIMITCONTROL/&&
TEMPERATURE# '023
#
TIMEINTERVALS 23TEXT'0
STOP6ABS 23UNITGG
CONDITIONING/&& REPORT#/-FULLCURVE
REQIDENT/&& MEAN
REQSMPLSIZEALL -.23
ACTIVATEPULSE/&&

Titration curve Result – Egg noodles
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEG
%0ML
.GG
TOTALPRGG

3
Method 33 –
F 4 Calcium and magnesium (in ash)

Reagents • Titrant: c(Na2EDTA) = 0.1 mol/L. See Method 9, Section B

• Auxiliary complexing solution:
c(acetylacetone) = 0.1 mol/L plus c(TRIS) = 0.2 mol/L. 20.4 g tris (hydroxy-
methyl)-aminomethane is weighed out into a 1000 mL volumetric flask and
dissolved in approx. 500 mL dist. H2O. 12 mL acetylacetone is added and the
solution is made up to the mark with dist. H2O and mixed.
• Hydrochloric acid: w(HCl) = 20%
Sample preparation 5…10 g sample is weighed out into a platinum or quartz crucible and heated
at 600 °C in a muffle furnace until only a pure white ash remains. After cooling
down, 2 mL HCl is added and the ash dissolved (possibly under warming). The
solution is transferred quantitatively to the titration beaker with dist. H2O.
Analysis The prepared sample solution is pre-neutralized with NaOH to pH 8 to 8.5. After
the addition of 15 mL auxiliary complexing solution it is titrated with c(Na2EDTA)
= 0.1 mol/L to after the second endpoint.
EP1 corresponds to Ca and the difference (EP2 – EP1) to Mg.

The results are given in g per 100 g dry weight.
g Ca / 100 g = EP1 x C01 x C03 x C04 x C30 / C00
g Mg / 100 g = (EP2 – EP1) x C02 x C03 x C04 x C30 / C00
C01 = 4.008
C02 = 2.431
C03 = 0.1 (for 100 g)
C04 = correction factor for dry weight (100 / (100 – % H2O)
Remarks This method can also be used for the determination of Ca and Mg in cream and
custard powders.
1
Method 33 –
F 4 Calcium and magnesium (in ash)
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L #
DOSRATEMAXMLMIN #
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&& tDE
PAUSES '044ITRINO
MEASINPUT $%45-
TEMPERATURE# DEF
STOP6ABS #A%0
#
#
#
##
STOP6ML 23TEXT#A
STOP%0 23UNITGG
STATISTICS -G%0 %0
#
#
#
##
STATUS/&& 23TEXT-G
EVALUATION 23DECIMALPLACES
%0# 23UNITGG
%0RECOGNITIONALL 23LIMITCONTROL/&&
lX%0AT5/&&M6 SILOCALCULATIONS
P+(.0/&& MATCHID/&&
PRESELECTIONS COMMONVARIABLES
REQIDENT/&& REPORT
REQSMPLSIZE/&& REPORT#/-FULLCURVE
LIMITSMPLSIZE/&& MEAN
ACTIVATEPULSE/&& -.23
TEMPORARYVARIABLES

Titration curve Result – Custard powder
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEG
%0ML M6
%0ML M6
#AGG
-GGG
STOP6REACHED

2
Honey, sugar and sweets
General This section also describes only those methods that can be carried out with a ti-
trator. We recommend the 712 Conductometer for measuring the conductivity.
Conductivity Honey
An amount of honey corresponding exactly to 20.0 g honey dry weight is dis-
solved in dist. H2O, made up to 100 mL and mixed. The conductivity (Κ) is
measured at 20 ±0.1 °C.
The result is given in mS/cm at 20 °C to two decimal places.
G Honey, sugar and sweets

Sugar
In this case the ash content can be calculated empirically from the conductiv-
ity.
5.0 g sugar is dissolved in dist. H2O, made up to 100 mL and mixed. The con-
ductivity (Κ) is measured at 20 ±0.1 °C → Κ5
The conductivity of the water used is also measured at the same temperature
→ ΚW
Κ5 = conductivity in μS/cm (of 5% sample solution)
% ash = Κ5 – (0.9 x ΚW ) x 0.0018
Or are you interested in different types of sugar? The Metrohm 817 Bioscan is a
well-proven analytical instrument for their determination.
1
Method 34 –
G 1 pH value and free acids


Metrohm nos. 6.2307.110 and 6.2307.100

It is assumed that the electrode has already been calibrated – see Method 1,
pH value Section A.
Please note that the temperature at which the measurement was made must al-
ways be mentioned together with the pH value. If no temperature sensor is con-
nected to the instrument then the temperature must be measured separately
and entered in the titrator (the 6.0258.000 Unitrode or the 6.1110.100 tempera-
ture sensor – with 6.2104.080 electrode cable – can be used).
10.0 g sample is weighed out into the titration beaker and dissolved in 75 mL
dist. H2O. The electrode is immersed and the pH measurement is started under
stirring. As soon as the drift value set on the titrator has been reached, the pH
value is shown or printed out automatically.
Free acid content The sample solution used for the pH measurement is titrated with c(NaOH) =
0.1 mol/L in the SET mode to pH 8.3.
Calculations Free acids in milliequivalents (meq) per kg sample

meq = EP1 x C01 x C30 / C00
C00 = sample weight in g (10)
C01 = 100 (conversion factor)
C30 = titer of NaOH
1
Method 34 –
Instrument parameters and calculation Result – pH

pH Forest honey
tPA tFR
DATE TIME $ATE 4IME
-%!3P(-P( -%!3P(
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&&
PRESELECTIONS
REQIDENT/&& pH value
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-P(
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-P(
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES
##

2
Method 34 –

SET SET
tPA tFR
DATE TIME $ATUM :EIT
3%4P(-3%4 P(CINIT 3%4P(-
PARAMETERS %INMASSG
3%4 %0ML
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRATIONPARAMETERS Titration curve – SET
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-3%4
# FMLA

tDE
'044ITRINO
DATE TIME
3%4P(-3%4
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES
#%0

3
Method 34 –
Instrument parameters and calculation Result – TIP

TIP Forest honey
tPA tFR
DATE TIME $ATE 4IME
4)0-4)0 4)0-4)0
SEQUENCE #
METHOD- #
METHOD- FREEACMEQKG
PAUSES P(
STATUS/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
MEASMODE/&&
TEMPERATURE#

tCF
'044ITRINO
DATE TIME
4)0-4)0
# FMLA
#

tDE
'044ITRINO
DATE TIME
4)0-4)0
DEF
SEQUENCE
METHOD-
METHOD-
PAUSES
FORMULA
FREIE(#
#
##
23TEXTFREIE(
23DECIMALPLACES
23UNITMEQKG
23LIMITCONTROL/&&
P(#
23TEXTP(
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULL
MEAN
-.23
TEMPORARYVARIABLES

4
Method 35 –
G 2 Formol number
General The formol number gives the number of amino groups that react with formalde-
hyde and split off a proton. This is a parameter that has no relationship to the
size of the molecule or the amount of amino acids.


• Formalin solution: w(HCHO) = approx. 35%
Adjusted to pH = 8.0 with NaOH.
Analysis 10 g sample (e.g. blossom honey) is weighed out into the titration beaker and
dissolved in approx. 30 mL dist. H2O. In a first SET titration the solution is titrat-
ed with c(NaOH) = 0.1 mol/L to pH = 8.0 (titrant consumption is not required).
15 mL formalin solution is then added, allowed to react under stirring for 1 min
and the solution again titrated in a second SET titration with c(NaOH) = 0.1
mol/L to pH = 8.0 (EP1).
Calculations Formol number (FN) in mL c(NaOH) = 0.1 mol/L per 100 g sample
FN = EP1 x C01 x C30 / C00
EP1 = mL NaOH for the second titration to the endpoint (pH = 8.0)
C01 = 100
1
Method 35 –
G 2 Formol number

SET 1 SET 1
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRATIONPARAMETERS Titration curve – SET 1
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 35 –
G 2 Formol number

SET 2 SET 2
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRATIONPARAMETERS Titration curve – SET 2
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES
#%0

3
Method 35 –
G 2 Formol number

TIP Blossom honey
tPA tFR
DATE TIME $ATE 4IME
4)0-4)0 4)0-4)0
SEQUENCE #
METHOD- FORMOL.B
METHOD-$/3 4)0TERMINATED
PAUSES
METHOD-
PAUSES
STATISTICS
STATUS/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
MEASMODE/&&
TEMPERATURE#

tCF
'044ITRINO
DATE TIME
4)0-4)0
# FMLA
#

tDE
'044ITRINO
DATE TIME
4)0-4)0
DEF
SEQUENCE
METHOD-
METHOD-$/3
PAUSES
METHOD-
PAUSES
FORMULA
&:#
#
##
23TEXT&:
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-PARAMFULLCALC
MEAN
-.23
TEMPORARYVARIABLES

4
Method 36 –
G 3 Reducing sugars
General Reducing sugars contain either aldehyde, ketone or hemiacetal groups.

In the method described here the sample is treated with an excess of Cu(II) ions
(Fehling’s solution). These are reduced to Cu(I) ions by the reducing sugars.
Potassium iodide is then added to the excess Cu(II) ions still present. This pro-
duces iodine and further Cu(I) ions. The released amount of iodine is titrated
with thiosulfate solution.
The oxidation of the sugars with Cu(II) – or the reduction of Cu(II) by sugars
– takes place neither stoichiometrically nor as a linear function of time. For this
reason it is not possible to carry out a titer determination. Rather, a so-called

calibration factor must be determined with glucose standards. In order to obtain
usable results for samples with different contents of reducing sugars it is neces-
sary to work with different sample weights. This is done so that under the same
conditions there is always approximately the same amount of reducing sugars
present in the titration beaker.

Reagents • Titrant: c(Na2S2O3) = 0.1 mol/L. See Method 6, Section B

• Sodium hydroxide: c(NaOH) = 2 mol/L
8 g NaOH is weighed out into a beaker and dissolved in approx. 60 mL dist.
H2O. After cooling down, it is made up to 100 mL with dist. H2O and mixed.
• Sulfuric acid: w(H2SO4) = 16%
90 mL w(H2SO4) = 98% is slowly and carefully added to approx. 800 mL dist.
H2O under stirring. After cooling down, it is made up to 1 liter with dist. H2O
and mixed.
150 g KI is weighed out into a 500 mL volumetric flask and dissolved in ap-
prox. 300 mL dist. H2O. 25 mL c(NaOH) = 2 mol/L is added, made up to the
mark with dist. H2O and mixed. The solution is stored in a dark bottle pro-
tected against light.
• Fehling’s solution I:
20.96 g CuSO4 x 5 H2O is weighed out into a 500 mL volumetric flask and
dissolved in approx. 250 mL dist. H2O. After the addition of 1 mL w(H2SO4) =
16% it is made up to the mark with dist. H2O and mixed.
• Fehling’s solution II:
173 g potassium sodium tartrate tetrahydrate is weighed out into a beaker
and dissolved in 200 mL dist. H2O. In a second beaker 40 g NaOH is dis-
solved in 200 mL dist. H2O and cooled down. Both solutions are rinsed into a
500 mL volumetric flask with dist. H 2O, made up to the mark and mixed.
• Glucose standards:
1.0 g and 2.0 g glucose are weighed out into two different 100 mL volumetric
flasks, dissolved in dist. H2O, made up to the mark and mixed. These stan-
dard solutions are not stable and must be prepared immediately before use.
They correspond to 10 g/L and 20 g/L glucose, respectively.
1
Method 36 –
G 3 Reducing sugars
Reducing sugars are given as invert sugar. The conversion factor glucose/invert Determining the
sugar is 1.8. calibration factor
Blank value of Cu solution
2.0 mL dist. H2O, 10.00 mL Fehling’s solution I and 5 mL Fehling’s solution II
are pipetted into a beaker. The mixture is heated to boiling and kept boiling for
exactly 30 s. It is then immediately cooled down under flowing water. 10 mL
each of dist. H2O, potassium iodide solution and sulfuric acid are then added
and the mixture titrated with c(Na2S2O3) = 0.1 mol/L to after the first endpoint.
The blank consumption (mL at EP1) is stored in the Titrino as a Common Vari-
able (e.g. C37).
Glucose standard 10 g/L

2.0 mL standard solution is pipetted into a beaker. The identical procedure to
that described under «blank value» is carried out.

that described under «blank value» is carried out.
Calculations Calculations
• g/L reducing sugars = (C37 – EP1) x C02
EP1 = mL thiosulfate for 2 mL glucose standard
C02 = 1.8 (conversion factor glucose/invert sugar)
C37 = mL thiosulfate for the blank value
• Calibration factor
This is obtained by taking the given values for glucose (10 and 20 g per liter)
and dividing them by the found or calculated value for invert sugar (g/L). The
mean value of the two glucose standards is stored in the Titrino as a Com-
mon Variable (e.g. C31). Example:
Glucose standard 10 g/L, 11.4 g/L invert sugar found 10 / 11.4 = 0.8772
Glucose standard 20 g/L, 21.8 g/L invert sugar found 20 / 21.8 = 0.9174
Calibration factor C31 = (0.8772 + 0.9174) / 2 = 0.8973
10 g sample is dissolved in dist. H2O, made up to 200 mL and mixed. This Sample preparation
means that the sample content is 50 g/L.
In order to always have approximately the same amount of reducing sugars in Analysis
the titration beaker it is necessary to work with different sample weights and to
use a dilution factor A (C03) for the final calculation.
Sample weight table
g/L invert sugar Sample volume Factor A

2 15 mL 0.1333
5 6 mL 0.3333
10...20 2 mL 1
25...30 1 mL 2
Samples containing more than 30 g/L invert sugar are diluted 1:1 with dist. H2O.
For a sample volume of 1 mL, A is then = 4.
2
Method 36 –
G 3 Reducing sugars
10.00 mL Fehling’s solution I, 5 mL Fehling’s solution II and 1...15 mL sample

solution (see table) are pipetted into a beaker. The mixture is heated to boiling
and kept boiling for exactly 30 s. It is then immediately cooled down under flow-
ing water. 10 mL each of dist. H2O, potassium iodide solution and sulfuric acid
are then added and the mixture titrated with c(Na2S2O3) = 0.1 mol/L to after the
first endpoint.
Calculations % reducing sugars = ((C37 – EP1) x C01 x C02 x C03 x C31) / C00
C00 = 50 (g/L sample)
C01 = 100 (for %)
C03 = dilution factor A
C31 = calibration factor
C37 = mL thiosulfate for blank value
Remarks • The boiling time of 30 s must be observed exactly, as otherwise incorrect re-
sults will be obtained. For the same reason the solution must be cooled down
immediately after the boiling time has elapsed.
• The method described here is an empirical method. It is never as exact as
other titration methods. Under the best conditions a relative error of ±2.2%
must be expected.
3
Method 36 –
G 3 Reducing sugars

Calibration Calibration – 10 g/L
tPA tFR
'044ITRINO '044ITRINO4IME:EIT
DATE TIME
$%45-+AL 5INIT M6$%45-+AL
PARAMETERS %0MLM6
TITRATIONPARAMETERS SUGARGL
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT Result
TEMPERATURE# Calibration – 20 g/L
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0 tFR
lLLINGRATEMAXMLMIN '044ITRINO
STATISTICS $ATE 4IME
STATUS/&& 5INIT M6$%45-+AL
EVALUATION %0MLM6
%0# SUGARGL
%0RECOGNITIONGREATEST STOP6REACHED
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-+AL
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-+AL
DEF
FORMULA
:UCKER# %0
#
23TEXT:UCKER
23DECIMALPLACES
23UNITGL
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 36 –
G 3 Reducing sugars
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L #
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&& tDE
PAUSES '044ITRINO
MEASINPUT $%45-
TEMPERATURE# DEF
STOP6ABS :UCKER# %0
#
#
#
# #
STOP6ML 23TEXT:UCKER
STOP%0 23UNIT
%0# REPORT
%0RECOGNITIONGREATEST REPORT#/-FULLCURVE
lX%0AT5/&&M6 MEAN
P+(.0/&& -.23
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – Candy Result – Candy
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEG,
%0MLM6
SUGAR
STOP6REACHED

5
Soft drinks, lemonades
General Soft drinks are more or less sweet-tasting, alcohol-free drinks. They are
made from fruit juices, the corresponding syrups or concentrates, natural
or artificial flavorings, milk components or plant extracts (e.g. cola nuts)
by dilution with drinking water or mineral water. They effervesce on pour-
ing because they usually contain added CO2.
In this section we have selected four titration methods that are frequently
used in practice. For the determination of the pH value, total acidity, and
calcium and magnesium please refer to Section J.
As this section also only describes methods that can be carried out with
a titrator, we would like to advise you to use Metrohm VA instruments for
H Softdrinks, lemonades
the determination of quinine. Application Bulletin no. 126 describes a de-
termination that is easy to carried out.
And not to be forgotten: our 817 Bioscan – with this instrument you can
rapidly and exactly determine different types of sugar and sugar alco-
hols.
1
Method 37 –
H 1 Citric acid, citrates
accessories reference electrode with 6.2106.020 electrode cable
Reagents • Titrant: c(CuSO4) = 0.05 mol/L

12.484 g CuSO4 x 5 H2O is weighed out into a 1000 mL volumetric flask, dis-
solved in dist. H2O, made up to the mark and mixed.
• Buffer solution pH = 8.0:
8.3 g boric acid and 6.4 g borax are weighed out into a 1000 mL volumetric
flask, dissolved in dist. H2O, made up to the mark and mixed.
• Methanol: CH3OH, reagent grade
Sample preparation 100 mL sample is degassed in an ultrasonic bath for 5 min (degassing can also
be carried out under vacuum).
50 mL of the treated sample is percolated through a strongly acidic cation ex-
changer (e.g. Dowex 50 WX8) into a beaker and the cation exchanger is rinsed
with 10 mL dist. H2O.
Analysis The prepared sample solution is treated with 50 mL each of methanol and buf-
fer solution pH = 8 and then titrated with c(CuSO4) = 0.05 mol/L to after the
first endpoint.
Calculations 1 mL c(CuSO4) = 0.05 mol/L corresponds to

9.606 mg citric acid
10.507 mg citric acid x H2O
16.221 mg tri-potassium citrate x H2O
14.705 mg tri-sodium citrate x H2O
g/L citric acid or citrate = EP1 x C01 / C00
EP1 = mL Cu sulfate up to endpoint
C01 = 9.606, 10.507, 16.221 or 14.705
Remarks The surface of the Cu ISE is polished with alumina powder after each titration.
1
Method 37 –
H 1 Citric acid, citrates
Instrument parameters and calculation Result – Lemonade
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY CITRICACGL
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#IT(%0
##
23TEXT#IT(
23DECIMALPLACES
23UNITGL
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 38 –
H 2 Phosphoric acid in cola drinks

Sample preparation 200 mL cola drink is degassed in an ultrasonic bath for 5 min (degassing can
also be carried out under vacuum).
Analysis 100 mL of the degassed sample solution is titrated with c(NaOH) = 0.1 mol/L to
after the first endpoint (first phosphoric acid step at pH = approx. 4.6).
Calculations 1 mL c(NaOH) = 0.1 mol/L corresponds to 9.7995 mg H3PO4

g/L H3PO4 = EP1 x C01 x C30 / C00
EP1 = mL NaOH up to endpoint
C01 = 9.7995
Remarks The determination only works in cola drinks that contain no citric acid, as the
citric acid is also determined. Such samples must be analyzed according to
Method 40, in which the sample is ashed (after previous neutralization to pH =
approx. 8 with NaOH).
The 761 SD Compact IC is an ion chromatography system that has been de-
veloped by Metrohm specially for the rapid and reliable determination of phos-
phoric acid (and citric acid) in these drinks. This instruments is recommended
worldwide by leading manufacturers of cola drinks.
1
Method 38 –
H 2 Phosphoric acid in cola drinks
tPA tFR
DATE TIME $ATE 4IME
$%4P(- P(CINIT $%4P(-
MEASPTDENSITY (0/GL
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOPP(/&&
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0ATP(/&&
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%4P(-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%4P(-
DEF
FORMULA
(0/%0
#
##
23TEXT(0/
23DECIMALPLACES
23UNITGL
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 39 –
H 3 Potassium
General The potassium content can provide information about the pure juice content of
soft drinks. A potassium content of 125...200 mg/L corresponds approximately
to a fruit juice content of 10%.
Recommended • 6.0504.110 K ISE with 6.2104.020 electrode cable and 6.0726.100 Ag/AgCl
reference electrode (inner electrolyte KCl, c = 3 mol/L and outer electrolyte
accessories NaCl, c = 1 mol/L) with 6.2106.020 electrode cable
Reagents • Titrant: c(STPB) = 0.05 mol/L
4.28 g sodium tetraphenylborate is weighed out into a 250 mL volumetric
flask and dissolved in approx. 200 mL dist. H2O. After the addition of 1 mL
c(NaOH) = 1 mol/L the solution is made up to the mark with dist. H2O and
mixed.
4 g NaOH is dissolved in dist. H2O and the solution made up to 100 mL after
cooling down.
55 mL w(HCl) = 36% is diluted to 100 mL with dist. H2O.
Sample preparation 25.0 mL sample is pipetted into a platinum or quartz crucible and evaporated
off at 140 °C in a drying oven. It is then heated in a muffle furnace at 700 °C until
only a white ash remains. After cooling down, 0.5 mL w(HCl) = 20% is added,
warmed to dissolve the ash and then rinsed into the titration beaker with dist.
H2O.
Analysis The treated sample solution is first neutralized to pH 6...8 with c(NaOH) = 1
mol/L and then titrated with c(STPB) = 0.05 mol/L to after the first endpoint.
Calculations 1 mL c(STPB) = 0.05 mol/L corresponds to 1.9549 mg K

mg/L K = EP1 x C01 x C02 / C00
EP1 = mL STPB up to endpoint
C01 = 1.9549
C02 = 1000 (for 1 liter)
Remarks No potassium was found in the «Orangina» (orange lemonade) sample. How-
ever, by means of a standard addition it could be proved that the determination
method functioned properly.
1
Method 39 –
H 3 Potassium
tPA FR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
TITRATIONPARAMETERS +MISSING%0
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6ABS
START6ML
DOSRATEMAXMLMIN
PAUSES
MEASINPUT
TEMPERATURE#
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONLAST
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA Titration curve with standard addition
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
+ALIUM%0
#
##
23TEXT+ALIUM
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLPARAMMPLISTCALC
MEAN
-.23
TEMPORARYVARIABLES

2
Methode 4 –
H 4 Total phosphorus
General The total phosphorus content can provide information about the pure juice con-
tent of soft drinks. A P2O5 content of 25...50 mg/L corresponds approximately to
a fruit juice content of 10%.


• Lanthanum nitrate: c(La nitrate) = 0.1 mol/L
21.7 g La(NO3)3 x 6 H2O is dissolved in approx. 450 mL dist. H2O in a beaker.
The pH value is adjusted to 4.2, the solution made up to 500 mL with dist.
H2O and mixed.
8 g NaOH is dissolved in dist. H2O and after cooling down, the solution is
made up to 100 mL with dist. H2O and mixed.
• Sulfuric acid: c(H2SO4) = 1 mol/L
5.4 mL w(H2SO4) = 96% is carefully added drop by drop to approx. 80 mL
dist. H2O in a beaker under stirring. After cooling down, the solution is made
up to 100 mL with dist. H2O and mixed.
• Digestion acids:
w(H2SO4) = 96% and w(HClO4) = 60%
Sample preparation 50.0 mL sample is pipetted into a platinum or quartz crucible and evaporated
off at 140 °C in a drying oven. It is then heated in a muffle furnace at 600...700
°C until only a white ash remains. After cooling down, the ash is treated with10
mL dist. H2O and 0.5 mL HClO4 and dissolved by warming. It is then rinsed into
a Kjeldahl flask with a little dist. H2O, 0.5 mL w(H2SO4) = 96% is added and the
solution boiled until acid fumes are liberated. After cooling down, the solution is
rinsed into the titration beaker with dist. H2O.
Analysis If necessary, the solution must be made up to approx. 50 mL with dist. H2O and
the pH value adjusted to 4.2 with c(NaOH) = 2 mol/L or c(H2SO4) = 2 mol/L
under stirring. 10 mL La(NO3)3 solution is added and the solution titrated in the
SET mode with c(NaOH) = 0.1 mol/L to pH = 4.2.
Calculations 1 mL c(NaOH) = 0.1 mol/L corresponds to 3.5486 mg P2O5

mg/L P2O5 = EP1 x C01 x C02 x C30 / C00
C01 = 3.5486
C02 = 1000 (for 1 liter)
Remarks The method is based on the following chemical reaction:

NaH2PO4 + La(NO3)3 = LaPO4 + 2 HNO3 + NaNO3
1
Method 40 –
H 4 Total phosphorus
Instrument parameters and calculation Result – «Orangina»
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( 0/MG,
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRATIONPARAMETERS
TITRDIRECTIONAUTO Titration curve
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
0/%0
#
#
##
23TEXT0/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Fruit and vegetable juices, fruit nectars
and jams
J Fruit and vegetable juices, fruit nectars and jams

General This section only describes methods that can be carried out with titrators.
However, Metrohm not only makes titrators but also a whole range of analytical
instruments for ion analysis.
For example, organic acids, chloride, nitrate, phosphate, sulfite and sulfate as
well as Na, K, Ca and Mg can be determined by ion chromatography and traces
of heavy metals (e.g. Cd, Cu, Fe, Mn, Pb, Zn) by VA methods.
With the 817 Bioscan various types of sugar and sugar alcohols can be deter-
mined quickly and accurately in a single run.
1
Method 41 –
J 1 pH value and titratable total acidity

Recommended • 6.0258.000 Unitrode with Pt 1000 temperature sensor
Reagents • Titrant I: c(NaOH) = 0.1 mol/L. See Method 3, Section B

• Titrant II: c(NaOH) = 1 mol/L. See Method 3, Section B
Metrohm nos. 6.2307.110 and 6.2307.100
Section A.
always be mentioned together with the pH value.
For jams the electrode is simply pushed into the sample; for juices the undiluted
juice is measured under stirring. When the drift value set on the titrator has been
reached, the pH value is shown or printed out automatically.
Titratable total acidity a) Jams

10.0 g sample, if necessary first comminuted in a food blender, is treated with
90 mL dist. H2O and heated until it starts to boil. After cooling down, the mixture
is titrated in the SET mode with c(NaOH) = 0.1 mol/L to pH = 8.5.
b) Fruit and vegetable juices

10.0 mL sample is treated with 40 mL dist. H2O and heated until it starts to boil.
After cooling down, the mixture is titrated in the SET mode with c(NaOH) = 0.1
mol/L to pH = 8.5.
Calculations For jams the results are given in milliequivalents (meq) per 100 g and for juices
in milliequivalents per liter.
a) Jams (meq / 100 g) = EP1 x C01 x C30 / C00
b) Juices (meq / L) = EP1 x C02 x C30 / C00
C00 = sample weight in g or sample volume in mL
C01 = 100 (conversion to 100 g)
C02 = 1000 (conversion to 1 liter)
C30 = titer of NaOH
For fruit juices the titratable acid content can also be given in g/L:
g/L acid = EP1 x C03 x C30
C03 = conversion factor for a particular acid:
0.75 for tartaric acid (grape juice)
0.67 for malic acid (apples, pears and stone fruits)
0.64 for citric acid (berries and citrus fruits)
1
Method 41 –
tPA
'044ITRINO tFR
DATE TIME '044ITRINO
-%!3P(-P( $ATE 4IME
PARAMETERS -%!3P(-P(
MEASURINGPARAMETERS SMPLSIZEML
SIGNALDRIFTM6MIN #
EQUILIBRTIME/&&S P(
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&& pH value
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-P(
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-P(
DEF
FORMULA
P(#
23TEXTP(
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 41 –
Instrument parameters and calculation – SET
tPA tCF
DATE TIME DATE TIME
3%4P(-3%4 3%4P(-3%4
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN tDE
3%4 '044ITRINO
%0ATP(/&& DATE TIME
TITRATIONPARAMETERS 3%4P(-3%4
PAUSES FORMULA
START6/&& 'ES(%0
#
##
PAUSES 23TEXT'ES(
DOSELEMENTINTERNAL$ 23UNITMEQ,
TEMPERATURE# 3AEURE%0
#
#
TIMEINTERVALS 23TEXT3AEURE
STOP6ABS 23UNITGL
REQIDENT/&& MEAN
REQSMPLSIZE/&& -.23
ACTIVATEPULSE/&&

Titration curve – SET Result SET – Orange juice
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-3%4
SMPLSIZEML
%0ML
TOTALACMEQ,
CITRICACGL

3
Method 42 –
J 2 Ascorbic acid (vitamin C)

Reagents • Titrant: c(I2) = 0.01 mol/L

Prepared from c(I2) = 0.05 mol/L by 1:5 dilution with dist. H2O
(Method 5, Section B).
• Glyoxal solution: w(glyoxal) = 40%
200 mL w(glyoxal) = 40% is adjusted with c(NaOH) = 1 mol/L to pH = 7.0.
The solution is stored away from light in a dark bottle in a refrigerator.
28 mL w(H2SO4) = 96% is carefully added to approx. 150 mL dist. H2O in a
beaker under stirring. After cooling down, the solution is made up to 200 mL
with dist. H2O and mixed.
Analysis 50 mL sample is pipetted into the titration beaker, treated with 2 mL glyoxal
solution, stirred briefly and allowed to stand for 5 min. After the addition of 5
mL sulfuric acid it is titrated with c(I2) = 0.01 mol/L up to the endpoint (EP1, to
be stored in the Titrino as Common Variable C31 as it is also required for the
determination of the sulfurous acid).
Calculations 1 mL c(I2) = 0.01 mol/L corresponds to 1.761 mg ascorbic acid

mg/L ascorbic acid = EP1 x C01 x C02 x C30 / C00
EP1 = mL iodine solution up to endpoint
C01 = 1.761
C02 = 1000 (for 1 liter)
C30 = titer of iodine solution (Method 5)
Remarks As this determination also includes the sulfite (SO2), this latter is eliminated by
the addition of an aldehyde (glyoxal). The aldehyde reacts with the sulfite to
form the addition compound aldehyde disulfite, which cannot be determined
titrimetrically:
R – CHO + H2SO3 → R – CH(OH) – O – SO3H
Please note that this method does not determine vitamin C alone. Under the
given titration conditions other «reductones» could be oxidized by the titrant.
This means that the method is not specific.
1
Method 42 –
J 2 Ascorbic acid (vitamin C)
Instrument parameters and calculation Result – Grape juice
tPA tFR
DATE TIME $ATE 4IME
6STEPML 6IT#MG,
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
6IT#%0
#
#
##
23TEXT6IT#
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 43 –
J 3 Sulfurous acid (sulfite)

Reagents • Titrant: c(I2) = 0.01 mol/L. See Method 42, J 2

• Sulfuric acid: w(H2SO4) = 25%. See Method 42, J 2
• Potassium iodide: KI solid, reagent grade
Analysis 50 mL juice (or 10 g homogenized jam plus 40 mL dist. H2O) is pipetted into the
titration beaker. Approx. 1 g potassium iodide and 5 mL sulfuric acid are added
and titrated with c(I2) = 0.01 mol/L to the endpoint.
Calculations 1 mL c(I2) = 0.01 mol/L corresponds to 0.641 mg SO2

mg/L SO2 = (EP1 – C31) x C01 x C02 x C30 / C00
C01 = 0.641
C02 = 1000 (for 1 liter)
C31 = mL iodine solution for ascorbic acid (Method 42, J 2)
Remarks A grape juice that did not contain any SO2 was used for the determination. Some
manufacturers do not use SO2 in their methods for the preparation of grape
juice. 1 mL c(Na2SO3)=0.1 mol/L was added to produce a detectable potential
jump.
See remarks concerning vitamin C in the previous method.
1
Method 43 –
tPA tFR
DATE TIME $ATE 4IME
TITRATIONPARAMETERS 3/MISSING%0
6STEPML MANUALSTOP
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
3/%0 #
#
#
##
23TEXT3/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 43 –

Standard addition Standard addition
tPA tFR
DATE TIME $ATE 4IME
6STEPML 3/MG,
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
STOPCONDITIONS
STOP6ABS Standard addition
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
3/%0 #
#
#
##
23TEXT3/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 44 –
J 4 Chloride


• Sulfuric acid: c(H2SO4) = 1 mol/L
5.4 mL w(H2SO4) = 96% is carefully added drop by drop to approx. 80 mL
dist. H2O in a beaker under stirring. After cooling down, the solution is made
up to 100 mL with dist. H2O and mixed.
Analysis 10.0 mL juice is pipetted into a beaker, 15 mL dist. H2O and 2 mL sulfuric acid
are added and the solution is titrated with c(AgNO3) = 0.1 mol/L to after the first
endpoint.
Calculations 1 mL c(AgNO3) = 0.1 mol/L corresponds to 3.545 mg chloride

mg/L chloride = EP1 x C01 x C02 x C30 / C00
C01 = 3.545
C02 = 1000 (for 1 liter)
1
Method 44 –
J 4 Chloride
Instrument parameters and calculation Result – Vegetable juice
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY CHLORIDEMGL
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#HLORID%0
#
#
##
23TEXT#HLORID
23DECIMALPLACES
23UNITMGL
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 45 –
J 5 Total phosphorus

Recommended – 6.0259.100 Unitrode with 6.2104.020 electrode cable
accessories – 6.3026.220 Exchange Unit
Reagents – Titrant: c(NaOH) = 0.1 mol/L. See Method 3, Section B

– Lanthanum nitrate: c(La nitrate) = 0.1 mol/L
21.7 g La(NO3)3 x 6 H2O is weighed out into a beaker and dissolved in approx.
450 mL dist. H2O. The pH is adjusted to 4.2, the solution made up to 500 mL
with dist. H2O and mixed.
– Sodium hydroxide: c(NaOH) = 2 mol/L
8 g NaOH is dissolved in dist. H2O, allowed to cool down, made up to 100 mL
and mixed.
– Sulfuric acid: c(H2SO4) = 1 mol/L. See Method 44, J 4
– Digestion acids:
w(H2SO4) = 96% and w(HClO4) = 60%
Sample preparation 10.0 mL or 10.0 g homogenized sample is evaporated in a platinum or quartz

crucible in a drying oven at 140 °C. It is then heated in a muffle furnace at
600...700 °C until only a white ash remains. After cooling down, the ash is
treated with10 mL dist. H2O and 0.5 mL HClO4 and dissolved by warming. It is
then rinsed into a Kjeldahl flask with a little dist. H2O, 0.5 mL w(H2SO4) = 96%
is added and the solution boiled until acid fumes are liberated. After cooling
down, the solution is rinsed into the titration beaker with dist. H2O.
Analysis If necessary, the solution must be made up to approx. 50 mL with dist. H2O and
the pH adjusted to 4.2 with c(NaOH) = 2 mol/L or c(H2SO4) = 2 mol/L under
stirring. 10 mL La(NO3)3 solution is added and titrated in the SET mode with
c(NaOH) = 0.1 mol/L to pH = 4.2.
Calculations The result is given in mg/L P2O5 for juices and in mg/100 g for jams.
1 mL c(NaOH) = 0.1 mol/L corresponds to 3.5486 mg P2O5
mg/L P2O5 = EP1 x C01 x C02 x C30 / C00
(mg/100 g P2O5 = EP1 x C01 x C03 x C30 / C00 – sometimes used for special
samples)
C00 = sample volume in mL or sample weight in g (10)
C01 = 3.5486
C02 = 1000 (for 1 liter)
C03 = 100 (for 100 g)
1
Method 45 –
J 5 Total phosphorus
Instrument parameters and calculation Result – Apple juice
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
PARAMETERS %0ML
3%4 0/MG,
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN Titration curve
3%4
%0ATP(/&&
TITRATIONPARAMETERS
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
0/%0
#
#
##
23TEXT0/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 46 –
J 6 Sulfate

accessories 6.0726.107Ag/AgCl-reference electrode with 6.2106.020 electrode cable
• 6.3026.220 Exchange Unit(s)

out into a beaker, approx. 200 mL dist. H2O and a stirrer bar are added and
20 g NaOH is dissolved in approx. 70 mL dist. H2O, allowed to cool down and
then made up to 100 mL with dist. H2O and mixed.
• Hydrochloric acid: w(HCl) = 20%. See Method 39, H 3
stored in the Titrino as a Common Variable (e.g. C31).
Sample preparation 20 mL sample is evaporated in a platinum or quartz crucible in a drying oven

at 115 °C. It is then heated in a muffle furnace at 600 °C until only a white ash
remains. After cooling down, 2 mL HCl is added and heated to dissolve the ash.
The solution is then transferred quantitatively to the titration beaker with dist.
H2O.
Analysis The prepared sample solution is treated with 5.00 mL c(BaCl2) = 0.05 mol/L. It
is allowed to react for 3 min under stirring and then treated with 10 mL buffer so-
lution pH = 10 and, if necessary, the pH is adjusted to pH = 10 with NaOH. The
solution is then titrated with c(EGTA) = 0.05 mol/L to after the second endpoint.
EP1 corresponds to Ca and the difference (EP2 – EP1) to the excess barium.
1
Method 46 –
J 6 Sulfate
The result is given in g/L K 2SO4. Calculations

1 mL c(BaCl2) = 0.05 mol/L corresponds to 8.713 mg K 2SO4
g/L K 2SO4: RS2 = (C31 – RS1) x C01 x C30 / C00
C01 = 8.713
C31 = mL EGTA for blank consumption
If only one endpoint is found then it must be assumed that the sulfate content of Remarks
the sample was too high (all the BaCl2 has been consumed). In this case the ti-
tration should be repeated with only half the previous sample volume (10 mL).
2
Method 46 –
J 6 Sulfate
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY
MININCR±L
DOSRATEMAXMLMIN
START6/&& DATE TIME
PAUSES $%45-
MEASINPUT FORMULA
TEMPERATURE# 23%0 %0
STOPCONDITIONS 23TEXT
STOP6ML 23UNITML
STOP%0 +3/# 23
#
##
lLLINGRATEMAXMLMIN 23TEXT+3/
STATUS/&& 23UNITGL
P+(.0/&& REPORT
REQIDENT/&& MEAN
REQSMPLSIZE/&& -.23
ACTIVATEPULSE/&&

Titration curve Result – Multivitamin juice
tFR
'044ITRINO
$ATE TIME
5INIT M6$%45-
SMPLSIZEML
%0ML M6
%0ML M6
23ML
+3/GL
STOP6REACHED

3
Method 47 –
J 7 Calcium and magnesium

Recommended • 6.0508.110 Ca ISE or 6.0504.100 with 6.2104020 electrode cable and
Reagents • Titrant: c(Na2EDTA) = 0.1 mol/L. See Method 9, Section B

• Auxiliary complexing solution:
c(acetylacetone) = 0.1 mol/L plus c(TRIS) = 0.2 mol/L
20.4 g tris(hydroxymethyl)-aminomethane is weighed out into a 1000 mL vol-
umetric flask and dissolved in approx. 500 mL dist. H2O. 12 mL acetylacetone
is added and the solution is made up to the mark with dist. H2O and mixed.
• Hydrochloric acid: w(HCl) = 20%. See Method 39, H 3

at 140 °C. It is then heated in a muffle furnace at 600 °C until only a white ash
remains. After cooling down, 2 mL HCl is added and heated to dissolve the ash.
The solution is then transferred quantitatively to the titration beaker with dist.
H2O.
Analysis The prepared sample solution is adjusted to pH = 8…8.5 with NaOH. After the
addition of 15 mL auxiliary complexing solution, it is titrated with c(Na2EDTA) =
0.1 mol/L to after the second endpoint. EP1 corresponds to Ca and the differ-
ence (EP2 – EP1) to the Mg.

mg/L Ca = EP1 x C01 x C02 x C30 / C00
mg/L Mg = (EP2 – EP1) x C03 x C02 x C30 / C00
C01 = 4.008
C02 = 1000 (for 1 liter)
C03 = 2.431
1
Method 47 –
J 7 Calcium and magnesium
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L #
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES tDE
PAUSES DATE TIME
MEASINPUT DEF
STOPCONDITIONS #A%0
#
#
##
STOP6ABS 23TEXT#A
lLLINGRATEMAXMLMIN -G%0 %0
#
#
##
STATISTICS 23TEXT-G
REQSMPLSIZE/&& MEAN

Titration curve Result – Grapefruit juice
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEML
%0ML M6
%0ML M6
#AMGL
-GMGL
STOP6REACHED

2
Method 48 –
J 8 Potassium

Recommended • 6.0504.110 K ISE with 6.2104.020 electrode cable and 6.0726.100 Ag/AgCl
accessories reference electrode (inner electrolyte KCl, c = 3 mol/L and outer electrolyte
NaCl, c = 1 mol/L) with 6.2106.020 electrode cable
Reagents • Titrant: c(STPB) = 0.05 mol/L

4.28 g sodium tetraphenylborate is weighed out into a 250 mL volumetric
flask and dissolved in approx. 200 mL dist. H2O. After the addition of 1 mL
c(NaOH) = 1 mol/L it is made up to the mark with dist. H2O and mixed.
4 g NaOH is dissolved in dist. H2O, allowed to cool down and then made up
to 100 mL with dist. H2O and mixed.
55 mL w(HCl) = 36% is diluted to 100 mL with dist. H2O.
Sample preparation 25.0 mL sample is evaporated in a platinum or quartz crucible in a drying oven
at 140 °C. It is then heated in a muffle furnace at 600 °C until only a white ash re-
mains. After cooling down, 2 mL w(HCl) = 20% is added and heated to dissolve
the ash. The solution is then transferred quantitatively to a 100 mL volumetric
flask with dist. H2O, made up to the mark and mixed.
Analysis 20 mL prepared sample solution (corresponding to 5 mL original sample) is

pipetted into the titration beaker and adjusted to pH 6…8 with c(NaOH) = 1
mol/L. It is then titrated with c(STPB) = 0.05 mol/L to after the first endpoint.
Calculations 1 mL c(STPB) = 0.05 mol/L corresponds to 1.9549 mg K

mg/L K = EP1 x C01 x C02 / C00
EP1 = mL STPB up to endpoint
C00 = sample volume in mL original sample (5)
C01 = 1.9549
C02 = 1000 (for 1 liter)
1
Method 48 –
J 8 Potassium
Instrument parameters and calculation Result – Orange juice
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY POTASSIUMMG,
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6ABS
START6ML
DOSRATEMAXMLMIN
PAUSES
MEASINPUT
TEMPERATURE#
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONLAST
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
+ALIUM%0
#
##
23TEXT+ALIUM
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 49 –
J 9 Ash alkalinity

accessories • 6.3026.220 Exchange Unit(s)
Reagents • Titrant I: c(HCl) = 0.1 mol/L. See Method 4, Section B

• Titrant II: c(NaOH) = 0.1 mol/L. See Method 3, Section B

at 120 °C. After the addition of a few drops of paraffin oil it is pre-ashed over a
naked flame. It is then heated in a muffle furnace at 540 °C. If any black carbon
particles are still present then, after cooling, the ash is pulverized, moistened
with dist. H2O and the ashing process repeated until only white ash remains. Af-
ter cooling down, 20.00 mL c(HCl) = 0.1 mol/L is added and the ash is dissolved
in it (gentle warming may be required) and the solution transferred quantitatively
to the titration beaker with dist. H2O. It is then heated on a boiling water bath for
15 min and allowed to cool down.
Analysis The treated sample solution is titrated in the SET mode with c(NaOH) = 0.1
mol/L to pH = 4.5.
Calculations The ash alkalinity is given in milliequivalents (meq) per liter.

meq/L = (C31 – EP1) x C01 x C30
C01 = 4 (conversion factor for 25 mL sample)
C31 = 20 (mL HCl added)
1
Method 49 –
J 9 Ash alkalinity
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT tDE
STOPDRIFT±LMIN '044ITRINO
3%4 DATE TIME
%0ATP(/&& 3%4P(-
TITRATIONPARAMETERS DEF
TITRDIRECTIONAUTO FORMULA
PAUSES !!LK# %0
#
#
START6ABS 23TEXT!!LK
START6ML 23DECIMALPLACES
DOSRATEMAXMLMIN 23UNITMEQ,
PAUSES 23LIMITCONTROL/&&
EXTRTIMES SILOCALCULATIONS
DOSELEMENTINTERNAL$ MATCHID/&&
MEASINPUT COMMONVARIABLES
TEMPERATURE# REPORT
TIMEINTERVALS REPORT#/-FULLCURVE
STOPCONDITIONS MEAN
STOP6ABS -.23
STOP6ML TEMPORARYVARIABLES
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve Result – Grape juice
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEML
%0ML
ASHALKMEQ,

2
Method 50 –
J 10 Kjeldahl nitrogen, total protein

General The Kjeldahl method determines the total nitrogen, from which the total protein
content can be calculated by multiplication with a factor. This corresponds in
principle to the average content of «available» proteins. This method is used for
vegetable juices and jams.
The sample is digested with sulfuric acid in the presence of a catalyst, whereby
organically bound nitrogen is converted to ammonium sulfate. In a distillation
apparatus the digestion solution is treated with excess NaOH and the released
10.0 g of the thoroughly homogenized sample is weighed out into a digestion
Sample preparation tube, a catalyst tablet* and 10 mL w(H2SO4) = 98% are added and heated in
the digestion apparatus until the solution is clear. Heating is then continued for
a further 10 min.


Analysis The pH glass electrode must first be calibrated with buffer solutions pH = 7.00
and pH = 4.00 (Method 1, Section A).
tion is treated with NaOH and the distillation and titration are started at the same
EP at pH 4.5
max. rate 2 mL/min
Calculations Total nitrogen (N) and total protein (TP) are given as «weight %».
1 mL c(HCl) = 0.1 mol/L corresponds to 1.4007 mg N
N = EP1 x C01 x C02 x C30 / C00 = RS1
TP = RS1 x C03
C01 = 1.4007
C02 = 0.1 (for %)
C03 = 6.25 (conversion factor to raw protein)
1
Method 50 –
J 10 Kjeldahl nitrogen, total protein
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS #
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN tDE
3%4 '044ITRINO
%0ATP(/&& DATE TIME
PAUSES FORMULA
START6/&& .%0
#
#
##
PAUSES 23TEXT.
DOSELEMENTINTERNAL$ 23UNIT
TEMPERATURE# '023
#
TIMEINTERVALS 23TEXT'0
STOP6ABS 23UNIT
REQIDENT/&& MEAN
REQSMPLSIZEALL -.23
ACTIVATEPULSE/&&

Titration curve Result – Vegetable juice
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPPLSIZEG
%0ML
.
TOTALPR

2
Method 51 –
J 11 Formol number

General The formol number gives the number of amino groups that react with formalde-
hyde and split off a proton. This is a parameter that has no relationship to the
size of the molecule or the amount of amino acids.
• 6.0259.100 Unitrode with 6.2104.020 electrode cable

Recommended • 6.3026.220 Exchange Unit(s)
accessories
Reagents • Titrant I: c(NaOH) = 1 mol/L. See Method 3, Section B
• Formalin solution: w(HCHO) = 35%
Adjusted to pH = 8.5 with NaOH.
Analysis 25 mL juice is pipetted into the titration beaker and diluted with approx. 20 mL
dist. H2O. In a first SET titration the mixture is titrated with c(NaOH) = 1 mol/L to
pH = 8.5. 15 mL formalin solution is then added, allowed to react under stirring
for 1 min and in a second SET titration the released acid is titrated with c(NaOH)
= 0.1 mol/L back to pH = 8.5 (EP1).
Calculations The formol number (FN) is given in mL c(NaOH) = 0.1 mol/L per 100 mL
sample.
FN = EP1 x C01 x C30 / C00
EP1 = mL c(NaOH) = 0.1 mol/L up to endpoint (pH = 8.5)
C01 = 100
1
Method 51 –
J 11 Formol number

SET 1 SET 1
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&& Titration curve – SET 1
TITRATIONPARAMETERS
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTEXTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 51 –
J 11 Formol number
Instrument parameters and calculation Result – SET 2

SET 2 Orange juice
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&& Titration curve – SET 2
TITRATIONPARAMETERS
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES
#%0

3
Method 51 –
J 11 Formol number

TIP Orange juice
tPA tFR
DATE TIME $ATE 4IME
4)0-4)0 4)0-4)0
SEQUENCE #
METHOD- FORMOL.B
METHOD-$/3 4)0TERMINATED
PAUSES
METHOD-
PAUSES
STATISTICS
STATUS/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
MEASMODE/&&
TEMPERATURE#

tCF
'044ITRINO
DATE TIME
4)0-4)0
# FMLA
#

tDE
'044ITRINO
DATE TIME
4)0-4)0
DEF
SEQUENCE
METHOD-
METHOD-$/3
PAUSES
METHOD-
PAUSES
FORMULA
&:#
#
##
23TEXT&:
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULL
MEAN
-.23
TEMPORARYVARIABLES

4
Method 52 –
J 12 Reducing sugars in jams

General Reducing sugars contain either aldehyde, ketone or hemiacetal groups. In
the method described here the sample is treated with an excess of Cu(II) ions
(Fehling’s solution). These are reduced to Cu(I) ions by the reducing sugars.
Potassium iodide is then added to the excess Cu(II) ions still present. This pro-
duces iodine and further Cu(I) ions. The released amount of iodine is titrated
with thiosulfate solution.
The oxidation of the sugars with Cu(II) – or the reduction of Cu(II) by sugars
– takes place neither stoichiometrically nor as a linear function of time. For this
reason it is not possible to carry out a titer determination. On the contrary, a so-
called calibration factor must be determined with glucose standards.
In order to obtain usable results for samples with different contents of reducing
sugars it is necessary to work with differing sample weights. This is done so that
under the same conditions there is always approximately the same amount of
reducing sugars present in the titration beaker.


8 g NaOH is weighed out into a beaker and dissolved in approx. 60 mL dist.
H2O. After cooling down, the solution is made up to 100 mL with dist. H2O and
mixed.
90 mL w(H2SO4) = 98% is slowly and carefully added to approx. 800 mL dist.
H2O under stirring. After cooling down, the solution is made up to 1 liter with
150 g KI is weighed out into a 500 mL volumetric flask and dissolved in ap-
prox. 300 mL dist. H2O. 25 mL c(NaOH) = 2 mol/L is added, made up to the
mark with dist. H2O and mixed.
• Fehling’s solution I:
20.96 g CuSO4 x 5 H2O is weighed out into a 500 mL volumetric flask and
dissolved in approx. 250 mL dist. H2O. After the addition of 1 mL w(H2SO4) =
16%, the solution is made up to the mark with dist. H2O and mixed.
• Fehling’s solution II:
173 g potassium sodium tartrate tetrahydrate is weighed out into a beaker
and dissolved in 200 mL dist. H2O. In a second beaker 40 g NaOH is dis-
solved in 200 mL dist. H2O and cooled down. Both solutions are rinsed into a
500 mL volumetric flask with dist. H 2O, made up to the mark and mixed.
• Glucose standards:
1.0 g and 2.0 g glucose are weighed out into two different 100 mL volumetric
flasks, dissolved in dist. H2O, made up to the mark and mixed. These stan-
dard solutions are not stable and must be prepared immediately before use.
They correspond to 10 g/L and 20 g/L glucose respectively.
1
Method 52 –
Reducing sugars are given as invert sugar. The conversion factor glucose/invert Determining the
sugar is 1.8. calibration factor
Blank value of Cu solution
2.0 mL dist. H2O, 10.00 mL Fehling’s solution I and 5 mL Fehling’s solution II are
pipetted into a beaker. The mixture is heated to boiling and kept boiling for ex-
actly 30 s. It is then immediately cooled down under flowing water. 10 mL each
of dist. H2O, potassium iodide solution and sulfuric acid are then added and
titrated with c(Na2S2O3) = 0.1 mol/L to after the first endpoint. The blank con-
sumption (mL at EP1) is stored in the Titrino as a Common Variable (e.g. C37).

that described under «blank value» is then carried out.

that described under «blank value» is then carried out.
– g/L reducing sugar = (C37 – EP1) x C02 Calculations

EP1 = mL thiosulfate for 2 mL glucose standard
– Calibration factor
This is obtained by taking the given values for glucose (10 and 20 g per liter)
and dividing them by the found or calculated value for invert sugar (g/L). The
mean value of the two glucose standards is stored in the Titrino as a Com-
mon Variable (e.g. C31). Example:
Glucose standard 10 g/L, 11.4 g/L invert sugar found 10/11.4 = 0.8772
Glucose standard 20 g/L, 21.8 g/L invert sugar found 20/21.8 = 0.9174
Calibration factor C31 = (0.8772 + 0.9174) / 2 = 0.8973
Jams – which are covered in this section – are thoroughly homogenized in a Sample preparation
blender. 10 g sample is weighed out into a 200 mL volumetric flask, approx. 100
mL dist. H2O is added, the flask is then sealed and thoroughly shaken. It is then
made up to the mark with dist. H2O and thoroughly mixed. The sample content
is therefore 50 g/L.
2
Method 52 –
Analysis In order to always have approximately the same amount of reducing sugars in
the titration beaker, it is necessary to work with different sample weights and to
use a dilution factor A for the final calculation.
Sample weight table
g/L invert sugar Sample volume Factor A

2 15 mL 0.1333
5 6 mL 0.3333
10...20 2 mL 1
25...30 1 mL 2
10.00 mL Fehling’s solution I, 5 mL Fehling’s solution II and 1...15 mL sample

solution (see table) are pipetted into a beaker. The mixture is heated to boiling
and kept boiling for exactly 30 s. It is then immediately cooled down under flow-
ing water. 10 mL each of dist. H2O, potassium iodide solution and sulfuric acid
are then added and the solution titrated with c(Na2S2O3) = 0.1 mol/L to after the
first endpoint.
Calculations g/L reducing sugars = (C37 – EP1) x C02 x C31 x A = RS1

% reducing sugars = RS1 x C01 / C00
C00 = 50 (g/L sample)
C01 = 100 (for %)
C31 = calibration factor
Remarks • The boiling time of 30 s must be observed exactly, as otherwise incorrect re-
sults will be obtained. For the same reason the solution must be cooled down
immediately after the boiling time has elapsed.
• The method described here is an empirical method. It is never as exact as
other titration methods. Under the best conditions a relative error of ±2.2%
must be expected.
3
Method 52 –
Instrument parameters and calculation Result – 10 g/L

Calibration Calibration
tPA tFR
DATE TIME $ATE 4IME
$%45-+AL 5INIT M6$%45-+AL
PARAMETERS %0MLM6
TITRATIONPARAMETERS SUGARGL
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT Result – 20 g/L
TEMPERATURE#
STOPCONDITIONS Calibration
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0 tFR
lLLINGRATEMAXMLMIN '044ITRINO
STATISTICS $ATE 4IME
STATUS/&& 5INIT M6$%45-+AL
EVALUATION %0MLM6
%0# SUGARGL
%0RECOGNITIONGREATEST STOP6REACHED
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-+AL
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-+AL
DEF
FORMULA
:UCKER# %0
#
23TEXT:UCKER
23DECIMALPLACES
23UNITGL
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 52 –
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
PARAMETERS SMPLSIZEG,
MEASPTDENSITY SUGAR
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
:UCKER# %0
#
#
#
# #
23TEXT:UCKER
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

5
Beer, vinegar, spirits and wine
General In this section only those methods are described that can be carried out with
titrators. However, Metrohm not only manufactures titrators but also a whole
range of analytical instruments for ion analysis.
For example, organic acids, chloride, nitrate, phosphate, sulfite and sulfate as
well as Na, K, Ca and Mg can be determined by ion chromatography, and traces
of heavy metals (e.g. Cd, Cu, Fe, Mn, Pb, Zn) by VA methods.
K Beer, vinegar, Spirits und Wine

With the 817 Bioscan various types of sugar and sugar alcohols can be deter-
mined quickly and accurately in a single run.
For the analysis of wine we have drawn up 6.6043.00X Wine PAC (Wine Po-
tentiometric Analysis Collection). Wine PAC contains all the usual methods (Au,
CH, EU, Israel, RSA, USA) for pH value, total and volatile acids, free and total
sulfurous acid, ascorbic acid, reducing sugars, CO2, ash alkalinity, Ca, Mg,
chloride, total P, sulfate, ammonium, K, Na and fluoride.
Wine PAC comprises a file with the printed-out methods and a memory card
from which you can load the methods in to your titrator. For this reason we have
not treated wine analysis separately in this section.
1
Method 53 –
K 1 pH value in beer

accessories
Reagents • Buffer solutions pH = 7.00 and pH = 4.00

Metrohm nos. 6.2307.110 and 6.2307.100
K Beer, vinegar, Spirits and Wine

Measurement It is assumed that the electrode has already been calibrated – see Method 1,
Section A.
CO2 is removed from the sample (ultrasonic bath or vacuum); the pH is normally
measured at 20 °C under stirring. When the drift value set on the titrator has
been reached the pH value is automatically shown on the display or printed
out.
1
Method 53 –
Calibration protocol – dark beer Result
tPA tFR
DATE TIME $ATE 4IME
-%!3P(- -%!3P(-
PARAMETERS #
MEASURINGPARAMETERS
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&& pH value
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

Result
Result
2
Method 53 –
Calibration protocol – light beer Result
tPA tFR
DATE TIME $ATE 4IME
-%!3P(- -%!3P(-
PARAMETERS #
MEASURINGPARAMETERS
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&& pH value
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 54 –
K 2 CO2 content in beer
General All the CO2 is bound as carbonate by the addition of NaOH and is converted
to bicarbonate by back-titration with HCl (if titration were to be continued the
organic acids would also be determined in addition to the CO2).

Reagents • Titrant: c(HCl) = 1 mol/L

prox. 800 mL dist. H2O in a 1000 mL volumetric flask, add 100 mL w(HCl) =
32%, make up to the mark with dist. H2O and mix. Titer determination with
TRIS as per Method 4.
• Sodium hydroxide: c(NaOH) = 10 mol/L.
Weigh out 80 g NaOH into a beaker, add 110 mL dist. H2O and mix imme-
diately until everything has dissolved. Caution, mixture becomes very hot;
wear protective goggles! After cooling down, make up to 200 mL with dist.
H2O and mix.
Analysis The container is cooled down to 0 °C, opened with as little agitation as possible
and treated with 1 mL NaOH for each 100 mL it contains. It is closed immedi-
ately (seal cans with Scotch tape) and mixed thoroughly.
100 mL pretreated sample is placed in the titration vessel and titrated with
c(HCl) = 1 mol/L to past the second endpoint.
Calculations 1 mL c(HCl) = 1 mol/L corresponds to 44 mg CO2

mg/L CO2 = (EP2 – EP1) x C01 x C02 x C30 / C00
EP1 = mL HCl up to first endpoint (NaOH)
EP2 = mL HCl up to second endpoint (Na2CO3)
C01 = 44
C02 = 1000 (for 1 liter)
1
Method 54 –
K 2 CO2 content in beer
tPA tFR
DATE TIME $ATE 4IME

$%45- 5INIT M6$%45-
MININCR±L #/MG,
DOSRATEMAXMLMIN 3TOP6REACHED
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#/%0 %0
#
#
##
23TEXT#/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 55 –
K 3 Ascorbic acid (vitamin C) in beer


Prepared from c(I2) = 0.05 mol/L by diluting 1:5 with dist. H2O (Method 5,

Section B)
beaker under stirring. After cooling down, it is made up to 200 mL with dist.
H2O and mixed.
Analysis 50 mL degassed sample (ultrasonic bath or vacuum) is pipetted into the titra-
tion beaker, treated with 2 mL glyoxal solution, stirred briefly and allowed to
stand for 5 min. After the addition of 5 mL sulfuric acid it is titrated with c(I2) =
0.01 mol/L up to the endpoint (EP1, to be stored in the Titrino as Common Vari-
able C31 as it is also required for the determination of the sulfurous acid).

C01 = 1.761
C02 = 1000 (for 1 liter)
Remarks As this determination also measures the sulfite (SO2) this latter is eliminated
by the addition of an aldehyde (glyoxal). The aldehyde reacts with the sulfite to
form the addition compound aldehyde-disulfite, which cannot be determined
titrimetrically:
given titration conditions other «reductones» can be oxidized by the titrant. This
means that the method is not specific.
1
Method 55 –
K 3 Ascorbic acid (vitamin C) in beer
tPA tFR
DATE TIME $ATE 4IME

PARAMETERS SMPLSIZE ML
6STEPML 6IT#MG,
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
6IT#%0
#
#
##
23TEXT6IT#
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 56 –
K 4 Total sulfurous acid (sulfite) in beer

Reagents • Titrant: c(I2) = 0.01 mol/L. See Method 55, K 3

• Sulfuric acid: w(H2SO4) = 25%. See Method 55, K 3

20 g NaOH is weighed out into a beaker and dissolved in approx. 300 mL
dist. H2O. After cooling down, the solution is made up to 500 mL with dist.
H2O and mixed.
Analysis 50 mL degassed sample (ultrasonic bath or vacuum) is pipetted into the titra-
tion beaker, 25 mL c(NaOH) = 1 mol/L is added, briefly stirred and allowed to
react for 10 min. 10 mL w(H2SO4) = 25% and approx. 1 g potassium iodide are
then added and titrated with c(I2) = 0.01 mol/L up to the endpoint.

mg/L total SO2 = (EP1 – C31) x C01 x C02 x C30 / C00
C01 = 0.641
C02 = 1000 (for 1 liter)
C31 = mL iodine solution for ascorbic acid (Method 55, K 3)
1
Method 56 –
K 4 Total sulfurous acid (sulfite) in beer
tPA tFR
DATE TIME $ATE 4IME

6STEPML TOTAL3/MG,
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
'ES3/%0 #
#
#
##
23TEXT'ES3/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 57 –
K 5 pH value and total acidity in vinegar

Reagents • Titrant: c(NaOH) = 1 mol/L

40.0 g NaOH is dissolved in CO2-free dist. H2O, allowed to cool down and

then made up to 1000 mL and mixed. Titer determination see Method 3, Sec-
tion B.
Metrohm nos. 6.2307.110 and 6.2307.100
Section A.
The pH is measured in the undiluted sample under stirring (normally at 20 °C).
When the drift value set on the titrator has been reached, the pH value is auto-
matically shown or printed out.
Titratable total acidity 10.0 mL vinegar is placed in the titration beaker, diluted with approx. 50 mL CO2-
free dist. H2O and titrated with c(NaOH) = 1 mol/L to after the first endpoint.
Calculations The acetic acid content is given in g per 100 mL.

1 mL c(NaOH) = 1 mol/L corresponds to 0.060 g acetic acid
g acetic acid / 100 mL = EP1 x C01 x C02 x C30 / C00
C01 = 0.06
C02 = 100 (for 100 mL)
1
Method 57 –

pH pH
tPA tFR
DATE TIME $ATE 4IME
-%!3P(-P( -%!3P(-P(
PARAMETERS #
MEASURINGPARAMETERS P(
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&&
PRESELECTIONS
REQIDENT/&& pH value
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-P(
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-P(
DEF
FORMULA
P(#
23TEXTP(
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 57 –

DET DET
tPA tFR
DATE TIME $ATE 4IME
$%45-$%4 5INIT M6$%45-$%4
MEASPTDENSITY ACETICACG,
MININCR±L 3TOP6REACHED
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE# Titration curve – DET
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-$%4
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-$%4
DEF
FORMULA
%SSIGS%0
#
#
##
23TEXT%SSIGS
23DECIMALPLACES
23UNITG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 58 –
K 6 Volatile acids in vinegar
Recommended See Method 57, K 5

accessories
Reagents • Titrant: c(NaOH) = 1 mol/L. See Method 57, K 5

Sample preparation 10.0 mL vinegar sample is treated with 10 mL dist. H2O in a round-bottom flask
and analysis and subjected to steam distillation until approx. 400 mL liquid has distilled over.
During the distillation the volume of liquid in the distillation flask should be kept
at approx. 20 mL.
The distillate is heated until it starts to boil, cooled down and then titrated in the
SET mode with c(NaOH) = 1 mol/L to pH = 8.5.
Calculations The volatile acids are given in g acetic acid per 100 mL sample.
1 mL c(NaOH) = 1 mol/L corresponds to 0.060 g acetic acid
Volatile acids; g acetic acid / 100 mL = EP1 x C01 x C02 x C30 / C00
C01 = 0.06
C02 = 100 (for 100 mL)
1
Method 58 –
K 6 Volatile acids in vinegar
tPA tFR
DATE TIME $ATE 4IME

3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( #(#//(G,
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRATIONPARAMETERS Titration curve
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
#(#//(%0
#
#
##
23TEXT#(#//(
23DECIMALPLACES
23UNITG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 59 –
K 7 Ascorbic acid (vitamin C) in vinegar


Prepared from c(I2) = 0.05 mol/L by diluting 1:5 with dist. H2O (Method 5,

Section B).
beaker under stirring. After cooling down, it is made up to 200 mL with dist.
H2O and mixed.
Analysis 50 mL vinegar sample is pipetted into the titration beaker, treated with 2 mL
glyoxal solution, stirred briefly and allowed to stand for 5 min. After the addition
of 5 mL sulfuric acid it is titrated with c(I2) = 0.01 mol/L up to the endpoint (EP1,
to be stored in the Titrino as Common Variable C31 as it is also required for the
determination of the sulfurous acid).

C01 = 1.761
C02 = 1000 (for 1 liter)
Remarks As this determination also measures the sulfite (SO2), this latter is eliminated
by the addition of an aldehyde (glyoxal). The aldehyde reacts with the sulfite to
form the addition compound aldehyde-disulfite, which cannot be determined
titrimetrically:
given titration conditions other «reductones» could be oxidized by the titrant.
This means that the method is not specific
1
Method 59 –
K 7 Ascorbic acid (vitamin C) in vinegar
tPA tFR
DATE TIME $ATE 4IME

6STEPML 6IT#MG,
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
6IT#%0
#
#
##
23TEXT6IT#
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 60 –
K 8 Free sulfurous acid (sulfite) in vinegar
General In contrast to wine, the addition of sulfurous acid to vinegar is only used to pro-
tect the color and aroma from oxidation. Without SO2 addition the vinegar as-
sumes a brownish color in the course of several months and spiced and herbal
vinegars lose their taste.


– Potassium iodide: KI solid, reagent grade
Analysis 50 mL vinegar is pipetted into the titration beaker, approx. 1 g potassium iodide
and 5 mL H2SO4 are added and the solution then titrated with c(I2) = 0.01 mol/L
up to the endpoint.

mg/L free SO2 = (EP1 – C31) x C01 x C02 x C30 / C00
C01 = 0.641
C02 = 1000 (for 1 liter)
Remarks See remarks about vitamin C in the previous method.
1
Method 60 –
K 8 Free sulfurous acid (sulfite) in vinegar
Instrument parameters and calculation Titration curve
tPA
'044ITRINO
DATE TIME
-%4)POL-
PARAMETERS
TITRATIONPARAMETERS
6STEPML
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
TEMPERATURE#
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
3/%0 #
#
#
##
23TEXT3/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 61 –
K 9 Total sulfurous acid (sulfite) in vinegar
Recommended See Method 60, K 8

accessories


20.0 g NaOH is weighed out into a beaker, dissolved in approx. 300 mL dist.
H2O, allowed to cool down and then made up to 500 mL and mixed.
Analysis 50 mL vinegar is pipetted into the titration beaker, 25 mL c(NaOH) = 1 mol/L is

added, briefly stirred and allowed to stand for 10 min. 10 mL w(H2SO4) = 25%
and approx. 1 g potassium iodide are added and the solution titrated with c(I2)
= 0.01 mol/L up to the endpoint.

mg/L total SO2 = (EP1 – C31) x C01 x C02 x C30 / C00
C01 = 0.641
C02 = 1000 (for 1 liter)
Remarks See remarks about vitamin C in Method 59.
1
Method 61 –
K 9 Total sulfurous acid (sulfite) in vinegar
Instrument parameters and calculation Titration curve
tPA
'044ITRINO
DATE TIME
-%4)POL-
PARAMETERS
TITRATIONPARAMETERS
6STEPML
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
)POL ±!
ELECTRODETEST/&&
TEMPERATURE#
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%4)POL-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%4)POL-
DEF
FORMULA
'ES3/%0 #
#
#
##
23TEXT'ES3/
23DECIMALPLACES
23UNITMG,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 62 –
K 10 Ash alkalinity of vinegar



Sample preparation 25 mL sample is evaporated in a platinum or quartz crucible in a drying oven at
120 °C. After the addition of one drop of paraffin oil it is pre-ashed over a naked
flame. It is then heated in a muffle furnace at 600 °C. If any black carbon par-
ticles are still present then, after cooling, the ash is pulverized, moistened with
dist. H2O and the ashing process repeated until only white ash remains. After
cooling down, 20.00 mL c(HCl) = 0.1 mol/L is added and the ash is dissolved
in it (gentle warming may be required) and then transferred quantitatively to the
titration beaker with dist. H2O. The solution is then heated on a boiling water
bath for 15 min and allowed to cool down.
Analysis The prepared sample solution is titrated in the SET mode with c(NaOH) = 0.1
mol/L to pH = 4.5.
Calculations The ash alkalinity is given in milliequivalents (meq) per liter sample.
meq/L = (C31 – EP1) x C01 x C30
C01 = 4 (conversion factor for 25 mL sample)
C30 = titer of NaOH (Method 3, Section B)
1
Method 62 –
K 10 Ash alkalinity of vinegar
tPA tCF
DATE TIME DATE TIME

3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT tDE
STOPDRIFT±LMIN '044ITRINO
3%4 DATE TIME
%0ATP(/&& 3%4P(-
TITRATIONPARAMETERS DEF
TITRDIRECTIONAUTO FORMULA
PAUSES !!LK# %0
#
#
START6ABS 23TEXT!!LK
START6ML 23DECIMALPLACES
DOSRATEMAXMLMIN 23UNITMEQ,
PAUSES 23LIMITCONTROL/&&
EXTRTIMES SILOCALCULATIONS
DOSELEMENTINTERNAL$ MATCHID/&&
MEASINPUT COMMONVARIABLES
TEMPERATURE# REPORT
TIMEINTERVALS REPORT#/-FULLCURVE
STOPCONDITIONS MEAN
STOP6ABS -.23
STOP6ML TEMPORARYVARIABLES
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SAMPLSIZEML
%0ML
ASHALKMEQ,

2
Method 63 –
K 11 Chloride in vinegar


• Protective colloid: polyvinyl alcohol solution

40 mg polyvinyl alcohol is dissolved in 100 mL hot dist. H2O and cooled
down.
Analysis 1.0 mL vinegar sample is pipetted into the titration beaker and treated with ap-
prox. 30 mL dist. H2O and 5 mL polyvinyl alcohol solution. It is then titrated with
c(AgNO3) = 0.1 mol/L to after the first endpoint.
Calculations The chloride content is given in g/L NaCl.

1 mL c(AgNO3) = 0.1 mol/L corresponds to 5.84 mg NaCl
g/L NaCl = EP1 x C01 x C30
C01 = 5.84
1
Method 63 –
K 11 Chloride in vinegar
Instrument parameters and calculation Result – spiced vinegar
tPA tFR
DATE TIME $ATE 4IME

$%45- 5INIT M6$%45-
PARAMETERS SMPLSIZE ML
MEASPTDENSITY .A#LGL
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
.A#L%0
#
#
23TEXT.A#L
23DECIMALPLACES
23UNITGL
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Methode 64 –
K 12 Sulfate in vinegar
General Any sulfurous acid that may be present in the sample is driven off by boiling un-
der nitrogen. Sulfates are precipitated with excess BaCl2 and the excess BaCl2
is back-titrated with EGTA.
Recommended acces- • 6.0508.110 or 6.0504.100 Ca ISE with 6.2104.020 electrode cable and

sories 6.0726.107 Ag/AgCl reference electrode with 6.2106.020 electrode cable

out into a beaker, a stirrer bar and approx. 200 mL dist. H2O are added and
• Hydrochloric acid conc.: w(HCl) = 36%
Sample preparation This work must be carried out under a fume hood!
50 mL dist. H2O and 1 mL HCl are placed in a 300 mL Erlenmeyer flask and the
solution is heated to boiling while nitrogen is passed through it. 100 mL vinegar
sample is added to the boiling solution and boiling is continued until the vol-
ume has been reduced to approx. 100 mL. After cooling down, the solution is
transferred to a 200 mL volumetric flask with dist. H2O, made up to the mark and
mixed.
stored in the Titrino as a Common Variable (e.g. C31).
Analysis 40.0 mL of the prepared sample solution (corresponding to 20 mL original

sample) is pipetted into the titration beaker. 5.00 mL c(BaCl2) = 0.05 mol/L is
added and allowed to react under stirring for 3 min. 10 mL buffer solution pH
= 10 is added and, if necessary, the pH is adjusted to pH = 10 with NaOH. It is
then titrated with c(EGTA) = 0.05 mol/L to after the second endpoint. EP1 cor-
responds to Ca and the difference (EP2 – EP1) to the excess barium.
1
Methode 64 –
The results are given in g/L K 2SO4. Calculations

1 mL c(BaCl2) = 0.05 mol/L corresponds to 8.713 mg K 2SO4
g/L K 2SO4: RS2 = (C31 – RS1) x C01 x C30 / C00
C00 = sample volume in mL original sample (20)
C01 = 8.713
C31 = mL EGTA for blank consumption
If only one EP is found then it must be assumed that the sulfate content of the Remarks
sample was too high (all the BaCl2 has been consumed). In this case the titra-
tion should be repeated with half the previous sample volume (10 mL original
sample or 20 mL prepared sample solution).
2
Methode 64 –
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L
DOSRATEMAXMLMIN
START6/&& DATE TIME
PAUSES $%45-
MEASINPUT FORMULA
TEMPERATURE# 23%0 %0
STOPCONDITIONS 23TEXT
STOP6ML 23UNITML
STOP%0 +3/# 23
#
#
##
lLLINGRATEMAXMLMIN 23TEXT+3/
STATUS/&& 23UNITGL
P+(.0/&& REPORT
REQIDENT/&& MEAN
REQSMPLSIZE/&& -.23
ACTIVATEPULSE/&&

tFR
'044ITRINO
$AT% 4IME
5INIT M6$%45-
SMPLSIZEML
%0ML M6
%0ML M6
23ML
+3/GL
3TOP6REACHED

3
Method 65 –
K 13 pH value and total acidity in spirits



Metrohm nos. 6.2307.110 and 6.2307.100
Section A.
The pH is measured in the undiluted sample. When the drift value set on the
titrator has been reached the pH value is automatically shown on the display or
printed out.
Titratable total acidity 50 mL sample is pipetted into an Erlenmeyer flask and, if necessary, diluted
with dist. H2O to give an alcohol volume fraction of σ (EtOH) = 40%. A reflux
condenser equipped with a CO2 absorber (e.g. soda lime) is attached and the
solution is heated to gentle boiling and kept boiling for 10 min to drive off the
CO2 from the sample. After cooling down, the solution is titrated with c(NaOH)
= 0.1 mol/L in the SET mode to pH = 8.2.
Calculations The titratable total acidity is given in mg acetic acid per liter alcohol.
1 mL c(NaOH) = 0.1 mol/L corresponds to 6.005 mg acetic acid
mg/L acetic acid = EP1 x C01 x C02 x C30 / (C00 x C03)
C01 = 6.005
C02 = 1000 (for 1 liter)
C03 = X (X = e.g. 0.4 for σ (EtOH) = 40%)
1
Method 65 –
Instrument parameters and calculation Result – pH
tPA tFR
DATE TIME $ATE 4IME
-%!3P(-P( -%!3P(-P(
PARAMETERS #
MEASURINGPARAMETERS
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&& pH value
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-P(
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-P(
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 65 –
tPA tCF
DATE TIME DATE TIME
3%4P(-3%4 3%4P(-3%4
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS/&& #
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4 tDE
%0ATP(/&& '044ITRINO
TITRATIONPARAMETERS DATE TIME
TITRDIRECTIONAUTO 3%4P(-3%4
PAUSES DEF
START6/&& FORMULA
PAUSES #(#//(%0
#
#
##
#
EXTRTIMES 23TEXT#(#//(
DOSELEMENTINTERNAL$ 23DECIMALPLACES
MEASINPUT 23UNITMG,
TEMPERATURE# 23LIMITCONTROL/&&
TIMEINTERVALS SILOCALCULATIONS
STOPCONDITIONS MATCHID/&&
STOP6ABS COMMONVARIABLES
STOP6ML REPORT
lLLINGRATEMAXMLMIN REPORT#/-FULLCURVE
STATISTICS MEAN
STATUS/&& -.23
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – SET Result SET – cognac
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEML
%0ML
#(#//(MG,

3
Method 66 –
K 14 Volatile acids in spirits
Recommended See Method 65 K 13

accessories
Reagents • Titrant: c(NaOH) = 0.1 mol/L. See Method 65, K 13

Sample preparation 50.0 mL sample is placed in a round-bottom flask and subjected to steam distil-
lation until approx. 200 mL liquid has distilled over. The distillation is carried out
and analysis so that the sample is first concentrated to approx. 15 mL and then this volume is
maintained. The distillate is heated until it starts to boil, cooled down and then
titrated in the SET mode with c(NaOH) = 0.1 mol/L to pH = 8.2.
Calculations The volatile acids are given in mg acetic acid per liter alcohol.
1 mL c(NaOH) = 0.1 mol/L corresponds to 6.005 mg acetic acid
mg/L acetic acid = EP1 x C01 x C02 x C30 / (C00 x C03)
C01 = 6.005
C02 = 1000 (for 1 liter)
C03 = X (X = e.g. 0.4 for σ (EtOH) = 40%)
1
Method 66 –
K 14 Volatile acids in spirits
tPA tCF
DATE TIME DATE TIME

3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS/&& #
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4 tDE
TITRDIRECTIONAUTO 3%4P(-
PAUSES DEF
START6/&& FORMULA
PAUSES #(#//(%0
#
#
##
#
EXTRTIMES 23TEXT#(#//(
MEASINPUT 23UNITMG,
STOP6ML REPORT
STATISTICS MEAN
STATUS/&& -.23
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve Result – Kirsch
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEML
%0ML
#(#//(MG,

2
Method 67 –
K 15 Total esters in spirits
General Spirits contain various esters, the majority of which are acetic acid ethyl es-
ters.
The esters are saponified by boiling with excess alkali and the unconsumed
alkali back-titrated with HCl.

Recommended See Method 65, K 13 (two Exchange Units)
accessories
Reagents • Titrant I: c(NaOH) = 0.1 mol/L. See Method 3, Section B

• Titrant II: c(HCl) = 0.1 mol/L. See Method 4, Section B
Sample preparation 50 mL sample is pipetted into an Erlenmeyer flask and titrated in the SET mode
with c(NaOH) = 0.1 mol/L to pH = 8.2 (see Method 65, K 13).
A few boiling stones are added and the solution is treated with an excess** of
c(NaOH) = 0.1 mol/L. A reflux condenser is attached and the solution is heated
to gentle boiling and kept boiling for 30 min. It is then allowed to cool down.
** The following guidelines apply for c(NaOH) = 0.1 mol/L:
• Rum and brandy 10 mL
• Pomaceous spirits 15 mL
• Kirsch, plum brandy, Williams, Gentian schnapps 25 mL
Analysis The prepared sample is titrated in the SET mode with c(HCl) = 0.1 mol/L to pH
= 7.9.
Calculations The total esters are given in mg acetic acid ethyl ester per liter alcohol.
1 mL c(NaOH) = 0.1 mol/L corresponds to 8.81 mg acetic acid ethyl ester
mg/L acetic acid ethyl ester = (C31 – EP1) x C01 x C02 / (C00 x C03)
C01 = 8.81
C02 = 1000 (for 1 liter)
C03 = X (X = e.g. 0.4 for σ (EtOH) = 40%)
C31 = mL c(NaOH) = 0.1 mol/L added
1
Method 67 –
Instrument parameters and calculation – pretitration
tPA tCF
DATE TIME DATE TIME
3%4P(-3%4 3%4P(-3%4
PARAMETERS # FMLA
3%4
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN tDE
3%4 3%4P(-3%4
%0ATP(/&& DEF
TITRDIRECTIONAUTO SILOCALCULATIONS
PAUSES MATCHID/&&
START6/&& COMMONVARIABLES
PAUSES REPORT
EXTRTIMES REPORT#/-FULLCURVE
DOSELEMENTEXTERNAL$ MEAN
MEASINPUT -.23
TEMPERATURE# TEMPORARYVARIABLES
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

2
Method 67 –
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS/&& #
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4 tDE
TITRDIRECTIONAUTO 3%4P(-
PAUSES DEF
START6/&& FORMULA
PAUSES %STER# %0
#
##
#
EXTRTIMES 23TEXT%STER
MEASINPUT 23UNITMG,
STOP6ML REPORT
STATISTICS MEAN
STATUS/&& -.23
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve Result – Williams
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEML
%0ML
%STERMG,

3
Coffee, cocoa and chocolate
General In addition to titrators Metrohm manufactures a whole range of other analyti-

cal instruments. In particular, we would like to mention the 743 Rancimat, an
instrument for determining the oxidation stability of oils and fats (or samples
containing oils and fats such as nuts or chocolate). Metrohm also manufactures
IC instruments for the determination of sodium and potassium as well as the
«Bioscan» for various types of sugars.
L Coffee, cocoa and chocolate
1
Method 68 –
L 1 pH value and degree of acidity
(green and roasted coffee, coffee extracts)
Recommended – 6.0258.000 Unitrode with Pt 1000 temperature sensor

Reagents – Titrant: c(NaOH) = 0.1 mol/L. See Method 3, Section B

– Buffer solutions pH = 7.00 and pH = 4.00
Metrohm nos. 6.2307.110 and 6.2307.100

Sample preparation Green coffee beans are dried in a drying oven for 2 h at 103 °C before grind-
ing.
Green and roasted coffee are ground up so that the fine powder completely
passes through a 0.63 mm mesh sieve, and is immediately collected in a seal-
able airtight container.
Green and roasted coffee

10.0 g sample is weighed out into an Erlenmeyer flask and 200 mL dist. H2O is
poured over it. The mixture is stirred with a glass rod while heating to boiling in
less than 5 min and then allowed to boil for 5 min with occasional stirring. It is
allowed to cool down and filtered through a paper filter into a 250 mL volumetric
flask. It is made up to the mark with dist. H2O and mixed.
Coffee extract
1.0 g dry coffee extract is dissolved in 50 mL CO2-free dist. H2O.
Section A.
50 mL of the freshly prepared extract is pipetted into the titration beaker. The
electrode is immersed and the pH measured under stirring. When the drift value
set on the titrator has been reached the pH value is automatically shown on the
display or printed out.
Degree of acidity The extract already used in measuring the pH is used again (50 mL, corre-
sponding to 2 g green or roasted coffee or 1 g coffee extract).
The extract is titrated with c(NaOH) = 0.1 mol/L in the SET mode to pH = 8.0.
Calculations The degree of acidity is given in millival per 100 g sample.

Degree of acidity = EP1 x C01 x C30 / C00
C00 = sample weight in g (2 for green and roasted coffee, 1 for coffee extract)
C01 = 10 (conversion factor; 0.1 x 100)
1
Method 68 –
tPA tFR
DATE TIME $ATE 4IME
-%!3P(-P( -%!3P(-P(
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&&
PRESELECTIONS pH value
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-P(
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-P(
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES
##

2
Method 68 –
tPA tCF
DATE TIME DATE TIME
3%4P(-3%4 3%4P(-3%4
PARAMETERS # FMLA
3%4
%0ATP(
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN tDE
3%4 3%4P(-3%4
%0ATP(/&& DEF
TITRDIRECTIONAUTO SILOCALCULATIONS
PAUSES MATCHID/&&
START6/&& COMMONVARIABLES
PAUSES REPORT
EXTRTIMES REPORT#/-FULLCURVE
DOSELEMENTINTERNAL$ MEAN
MEASINPUT -.23
TEMPERATURE# TEMPORARYVARIABLES
TIMEINTERVALS #%0
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – SET Result SET – Instant coffee
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-3%4
SMPLSIZEG
%0ML

3
Method 68 –
Instrument parameters and calculation – TIP Result – TIP
tPA tFR
DATE TIME $ATE 4IME
4)0-4)0 4)0-4)0
SEQUENCE #
METHOD- #
METHOD- DEGROFAC
PAUSES P(
STATUS/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
MEASMODE/&&
TEMPERATURE#

tCF
'044ITRINO
DATE TIME
4)0-4)0
# FMLA
#

tDE
'044ITRINO
DATE TIME
4)0-4)0
DEF
SEQUENCE
METHOD-
METHOD-
PAUSES
FORMULA
3R#
#
##
23TEXT3R
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
P(#
23TEXTP(
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULL
MEAN
-.23
TEMPORARYVARIABLES

4
Method 69 –
L 2 Ash alkalinity



Sample preparation 6.0 g green or roasted coffee or 2.0 g dry coffee extract is weighed out into
a platinum or quartz crucible and heated in a muffle furnace at 540 °C. If any
black carbon particles are still present then, after cooling, the ash is pulverized,
moistened with dist. H2O and the ashing process repeated until only white ash
remains.
After cooling down, 20.00 mL c(HCl) = 0.1 mol/L is added and the ash is dis-
solved in it (gentle warming may be required) and then transferred quantita-
tively to the titration beaker with dist. H2O. It is then heated on a boiling water
bath for 15 min and allowed to cool down.
mol/L to pH = 4.5.
Calculations The total ash alkalinity is given in milliequivalents (meq) per 100 g dry sub-
stance.
meq / 100 g = (C31 – EP1) x C01 x C30 / C00
5
Method 69 –
L 2 Ash alkalinity
Instrument parameters and calculation Result – Roasted coffee
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
PARAMETERS SMOLSIZEG
3%4 %0ML
%0ATP( ASHALKMEQ,
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
!!LK# %0
#
#
23TEXT!!LK
23DECIMALPLACES
23UNITMEQ,
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

6
Method 70 –
L 3 Chloride (green and roasted coffee)
General The chloride content is approx. 300...400 mg/kg. Higher contents may be
caused by penetration of seawater into ships’ holds or treating the green coffee
with fumigants containing halogens (e.g. methyl bromide).
The pulverized sample is combusted with oxygen according to the Schoeniger
method and the chloride is titrated with AgNO3. This method determines the
total chloride content (chloride plus organically bound chlorine).

Reagents • Titrant: c(AgNO3) = 0.001 mol/L in glacial acetic acid

10.0 mL c(AgNO3) = 0.1 mol/L (see Method 8, Section B) is pipetted into a
1000 mL volumetric flask, made up to the mark with glacial acetic acid and
mixed.
• Glacial acetic acid: w(acetic acid) = 96%, reagent grade
• Sodium hydroxide: c(NaOH) = 0.1 mol/L. See Method 3, Section B
Sample preparation Approx. 40 mg finely ground sample is weighed out exactly onto a chloride-free
(Schoeniger combus- filter paper, the filter paper is folded and attached to the ignition device of the
tion) combustion apparatus.
10 mL c(NaOH) = 0.1 mol/L is placed in the 500 mL combustion flask, which
is then filled with pure oxygen. The flask is sealed with the ignition device and
combustion is started by ignition. The sealed flask is then shaken until all the
fumes produced have been completely absorbed by the liquid.
The flask contents are rinsed quantitatively into the titration beaker with two por-
tions of 20 mL glacial acetic acid.
Analysis The prepared sample solution is titrated with c(AgNO3) = 0.001 mol/L to after
the first endpoint.
Calculations The chloride content is given in mg chloride per kg sample (ppm).

1 mL c(AgNO3) = 0.001 mol/L corresponds to 35.45 μg chloride
ppm chloride = EP1 x C01 x C02 / C00
C00 = sample weight in mg
C01 = 35.45
C02 = 1000 (for 1 kg)
3
Method 70 –
L 3 Chloride (green and roasted coffee)
Instrument parameters and calculation Result – Green coffee
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY CHLORIDEPPM
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#HLORID%0
#
##
23TEXT#HLORID
23DECIMALPLACES
23UNITPPM
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 71 –
L 4 Directly reducing sugars
(roasted coffee, coffee extracts)
Reagents • Titrant: c(Na2EDTA) = 0.05 mol/L

Prepared by 1:1 dilution of c(Na2EDTA) = 0.1 mol/L (Method 9, Section B)

with dist. H2O.
• Alkaline CuEDTA solution:
286 g Na2CO3 x 10 H2O and 38 g Na2EDTA are weighed out into a 1000
mL volumetric flask and dissolved in approx. 500 mL dist. H 2O (lukewarm).
The solution is treated with a mixture of 25 g CuSO4 x 5 H2O in 100 mL dist.
H2O, allowed to cool down and then made up to the mark with dist. H2O and
mixed. The solution is allowed to stand for several days and, if necessary, is
filtered before use.
35 mL w(HNO3) = 65% is placed in a 500 mL volumetric flask, made up to the
• Ammonia solution: c(NH3) = approx. 1 mol/L
38 mL w(NH3) = 25% is placed in a 500 mL volumetric flask, made up to the
• Carrez solution I:
75 g K4Fe(CN) 6 x 3 H2O is weighed out into a 500 mL volumetric flask, dis-
solved in dist. H2O, made up to the mark and mixed.
• Carrez solution II:
150 g ZnSO4 x 7 H2O is weighed out into a 500 mL volumetric flask, dissolved
in dist. H2O, made up to the mark and mixed.
4.0 g NaOH is dissolved in dist. H2O, allowed to cool down and then made up
to 100 mL and mixed.
• Bromothymol blue: 0.1% in σ (ethanol) = 20%
• Extraction solution: σ (ethanol) = 80%, reagent grade
• Auxiliary reagents: sea sand, cleaned, and CaCO3 reagent grade
Sample preparation Extraction

5.00 g finely ground roasted coffee or 2.00 g dry coffee extract is homoge-
neously mixed in a mortar with 1 g CaCO3. A coffee spoonful of sea sand is
added, mixed and transferred quantitatively to the extraction thimble of the
extraction apparatus (e.g. Soxhlet). The mixture is extracted for 2 h with σ (etha-
nol) = 80%. The brown turbid extract is then concentrated in a rotary evapora-
tor to a volume of 10...20 mL.
Carrez clarification
The solution obtained is transferred quantitatively to a 50 mL volumetric flask by
rinsing several times with dist. H2O, treated with 1 mL Carrez solution I and thor-
oughly mixed. 1 mL Carrez solution II is then added, mixed again and «neutral-
ized» by the drop-by-drop addition of c(NaOH) = 1 mol/L using bromothymol
blue as the indicator. It is then made up to the mark with dist. H2O, thoroughly
mixed and then filtered through a dry folded filter paper.
The filtrate must not contain any zinc ions. If some crystals of Na 2HPO4 are
added there should be no precipitate formed. Any zinc phosphate that forms is
centrifuged off.
3
Method 71 –
Reduction of Cu(II) by the sugars

10.00 mL of the clear filtrate (corresponding to 1/5 of the sample weight) is
treated with 10 mL alkaline CuEDTA solution in a round-bottom flask and sev-
eral boiling stones are added. A reflux condenser is attached and the mixture is
brought to boiling point in less than 2 min. From this point in time onwards the
mixture is gently boiled for exactly 10 min. Boiling is then stopped by the addi-
tion of 25 mL cold dist. H2O (through the condenser) and the flask contents are
cooled down under flowing water.
The contents are now filtered through a glass filter crucible and rinsed thor-
oughly with dist. H2O. (The filtrate must still be slightly blue, otherwise the re-
duction must be repeated with less sample solution.) The filter residue (Cu2O)
is dissolved with a few drops of w(HNO3) = 65% and rinsed into the titration
beaker with approx. 5 mL c(HNO3) = 1 mol/L and then with dist. H2O.
The prepared acidic sample solution is neutralized with c(NH3) = 1 mol/L un- Analysis
til a weakly blue-colored turbidity is seen. A further 2.5 mL c(NH3) = 1 mol/L
solution is then added, the solution is diluted to approx. 100 mL with dist. H2O
and titrated in the MET mode with c(Na2EDTA) = 0.05 mol/L to after the first
endpoint.
The results are given as glucose mass fraction, w(glucose). Calculations

1 mL c(Na2EDTA) = 0.05 mol/L corresponds to 1.95 mg glucose
w(glucose) in % = EP1 x C01 x C02 / C00
EP1 = mL Na2EDTA up to endpoint
C00 = 1/5 sample weight in g (1 g roasted coffee, 0.4 g coffee extract)
C01 = 1.95
C02 = 0.1 (for %)
4
Method 71 –
Instrument parameters and calculation Result – Instant coffee
tPA FR
DATE TIME $ATE 4IME
-%45- 5INIT M6-%45-
6STEPML 'LUCOSE
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
STOPCONDITIONS Titration curve
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%45-
DEF
FORMULA
'LUCOSE%0
#
##
23TEXT'LUCOSE
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

5
Method 72 –
L 5 pH value of cocoa and chocolate powder

accessories
Reagents • Buffer solutions pH = 7.00 and pH = 4.00

Metrohm nos. 6.2307.110 and 6.2307.100
• Dist. H2O and acetone

Section A.
10 g powdered sample is stirred with boiling dist. H2O in a beaker to form a sus-
pension. The mixture is quickly cooled down to 20 °C while stirring is continued
and the pH value is measured (also under stirring). When the drift value set on
the titrator has been reached the pH value is automatically shown on the display
or printed out.
After each measurement the electrode is first rinsed with acetone (to remove
adhering fat residues) and then thoroughly rinsed with dist. H2O.
5
Method 72 –
L 5 pH value of cocoa and chocolate powder
Instrument parameters and calculation Result – Chocolate powder
tPA tFR
DATE TIME $ATE 4IME
-%!3P(- -%!3P(-
PARAMETERS #
MEASURINGPARAMETERS
SIGNALDRIFTM6MIN
EQUILIBRTIME/&&S
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STATISTICS
STATUS/&& pH value
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%!3P(-
# FMLA

tDE
'044ITRINO
DATE TIME
-%!3P(-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

6
Method 73 –
L 6 Ash alkalinity of cocoa and chocolate
powder



Sample preparation 2.0 g cocoa mass or cocoa powder or 5.0 g chocolate powder is weighed out
into a platinum or quartz crucible, initially ashed over a naked flame and then
heated in a muffle furnace at 600 °C. After cooling down, 20.00 mL c(HCl) = 0.1
mol/L is added and the ash is dissolved in it (gentle warming may be required)
and then transferred quantitatively to the titration beaker with dist. H2O. The
solution is then heated on a boiling water bath for 15 min and allowed to cool
down.
mol/L to pH = 4.5.
Calculations The total ash alkalinity is given in milliequivalents (meq) per 100 g sample.
meq / 100 g = (C31 – EP1) x C01 x C30 / C00
C00 = sample weight in g (2 or 5)
3
Method 73 –
L 6 Ash alkalinity of cocoa and chocolate
powder
Instrument parameters and calculation Result – Cocoa powder
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( MEQG
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
MEQ# %0
#
##
23TEXTMEQ
23DECIMALPLACES
23UNITG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 74 –
L 7 Kjeldahl nitrogen in cocoa and chocolate
powder
General With these samples the Kjeldahl method determines the total nitrogen.

tent:
<1 g N / 100 g 2...3 g

1 g N / 100 g approx. 1.5 g
2 g N / 100 g approx. 0.7 g
The powdered sample is weighed out into a digestion tube, a catalyst tablet*
and 10 mL w(H2SO4) = 98% are added and heated in the digestion apparatus
until the solution is clear. Heating is then continued for a further 10 min.


30 mL boric acid is placed in the titration beaker. The digestion tube is con-
nected to the distillation apparatus, the digestion solution is treated with NaOH
and the distillation and titration are started at the same time. The titration with
c(HCl) = 0.1 mol/L takes place in the SET-pH mode with the following param-
eter settings:
EP at pH 4.5
dynamics 6
max. rate 2 mL/min
Calculations The total nitrogen is given in g N per 100 g sample.

g N / 100 g = EP1 x C01 x C02 x C30 / C00
C01 = 1.4007
C02 = 0.1 (for 100 g)
3
Method 74 –
L 7 Kjeldahl nitrogen in cocoa and chocolate
powder
Instrument parameters and calculation Result – Cocoa powder
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( .GG
DYNAMICS
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZEALL
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
.%0
#
#
##
23TEXT.
23DECIMALPLACES
23UNITGG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 75 –
L 8 Free fatty acids (FFA) in cocoa butter

Reagents • Titrant: c(KOH) = 0.1 mol/L in IPA. See Method 3, Section B

• Solvent: ethanol/diethyl ether 1:1

Neutralized with KOH against phenolphthalein.
Analysis 5...10 g sample is weighed out into the titration beaker and dissolved in 50 mL
solvent mixture. It is then titrated with c(KOH) = 0.1 mol/L to after the first end-
point.
Calculations The free fatty acids (FFA) are given in mL c(KOH) = 0.1 mol/L per 10 g sam-
ple.
FFA = EP1 x C01 x C30 / C00
EP1 = mL KOH up to endpoint
C01 = 10 (for 10 g)
3
Method 75 –
L 8 Free fatty acids (FFA) in cocoa butter
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY &&!
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
&&!%0
#
##
23TEXT&&!
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 76 –
L 9 Iodine number in cocoa butter


• Reaction solution: c(ICl) = 0.1 mol/L in glacial acetic acid
16.2 g iodine monochloride is dissolved in glacial acetic acid, made up to 1

• Glacial acetic acid: w(acetic acid) = 96...98%
• Catalyst solution: w(Mg acetate) = 3%
45 g Mg(OOCCH3)2 x 4 H2O is dissolved in glacial acetic acid, made up to 1
50 g KI is dissolved in dist. H2O, made up to 500 mL and mixed. It is stored in
a dark bottle and protected against light.
Blank value of 20 mL glacial acetic acid, 25.0 mL reaction solution and 10 mL catalyst solu-
reaction solution tion are placed in an Erlenmeyer flask with an SGJ stopper. The flask is sealed
and placed in the dark for exactly 5 min. 15 mL KI is then added, the mixture
transferred quantitatively to the titration beaker with dist. H2O and immediately
titrated with c(Na2S2O3) = 0.1 mol/L to after the first endpoint.
The titrant consumption (EP1) is stored in the Titrino as a Common Variable
(e.g. C31).
BV = EP1 (mL) = C31
Analysis Depending on the expected iodine number, 0.1…1 g sample is weighed out into
an Erlenmeyer flask with an SGJ stopper and dissolved in 20 mL glacial acetic
acid. After the addition of 25.0 mL reaction solution and 10 mL catalyst solution
the flask is sealed and placed in the dark for exactly 5 min. 15 mL KI is then
added, the mixture is then transferred quantitatively to the titration beaker with
dist. H2O and immediately titrated with c(Na2S2O3) = 0.1 mol/L to after the first
endpoint.
Calculations The iodine number (IN) is given in g iodine per 100 g sample.
IN = (C31 – EP1) x C01 x C30 / C00
EP1 = mL thiosulfate solution for sample
C01 = 1.269 (equivalent weight of iodine)
C31 = blank value (mL thiosulfate solution)
1
Method 76 –
L 9 Iodine number in cocoa butter
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY ).
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
):# %0
#
##
23TEXT):
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 77 –
L 10 Saponification number of cocoa butter

Reagents • Titrant: c(HCl) = 0.5 mol/L

Purchased ready for use or prepared as per Method 4, Section B

• Reaction solution: c(KOH) = 0.5 mol/L in ethanol
Purchased ready for use or prepared as per Method 3, Section B
• Solvent: ethanol, abs.
Blank value of 25.0 mL c(KOH) = 0.5 mol/L is pipetted into an Erlenmeyer flask with an SGJ
and a stirrer bar is added. A reflux condenser is attached and the solution is
reaction solution heated to gentle boiling for 30 min. After cooling down, it is diluted with 50 mL
ethanol and titrated with c(HCl) = 0.5 mol/L to after the first endpoint. The titrant
consumption (EP1) is stored in the Titrino as a Common Variable (e.g. C31).
The titrant consumption (EP1) is stored in the Titrino as a Common Variable.
BV = EP1 (mL) = C31
Analysis 0.5…3 g sample is weighed out into an Erlenmeyer flask with an SGJ with an
accuracy of 0.5 mg. 25.0 mL c(KOH) = 0.5 mol/L and a stirrer bar are added
and the reflux condenser is attached. The solution is heated to gentle boiling
for 30 min. After cooling down, it is diluted with 50 mL ethanol and titrated with
c(HCl) = 0.5 mol/L to after the first endpoint.
Calculations The saponification number (SN) is given in mg KOH per g sample.

SN = (C31 – EP1) x C01 x C30 / C00
C31 = mL HCl for blank value
3
Method 77 –
Result – blank value Titration curve – blank value
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-"7
SMPLSIZEG
%0ML M6
%0MLM6
"6
3TOPP6REACHED

Result
4
Method 77 –
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MININCR±L SAP.B
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
6:# %0
#
##
23TEXT6:
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

5
Artificial sweeteners, gelling and thickening
agents
M Artifical sweeteners, gelling and thickening agents

General In addition to titrators, Metrohm also manufactures a whole range of other ana-
lytical instruments. VA instruments are particularly suitable for the determina-
tion of traces of heavy metals or toxic metals and/or saccharin.
5
Method 78 –

thickening agents
Principle The methoxy and ethoxy groups are split off from the ether molecule by boil-
ing hydriodic acid and converted to the corresponding alkyl iodide. Nitrogen
is used as the carrier gas to transport the alkyl iodides into a bromine solution
where they are oxidized to iodic acid. After elimination of the excess bromine
with formic acid, potassium iodide is added and the released iodine titrated with
thiosulfate.
Gelling agent-OCH3 + Hl → Gelling agent-OH + CH3I
CH3I + Br2 → CH3Br + BrI
BrI + Br2 + 3 H2O → HIO3 + 5 HBr
HIO3 + 5 KI + 5 HCOOH → 3 I2 + 5 HCOOK + 3 H2O
3 I2 + 6 Na2S2O3 → 3 Na2S4O6 + 6 NaI

• Zeisel reaction/distillation apparatus, consisting of a conical-bottom reaction
flask with gas inlet tube, condenser, three gas washers and delivery tube with
distillation receiver.
Reagents • Titrant: c(Na2S2O3) = 0.1 mol/L. See Method 6, Section B.

• Washing solution:
6.15 g cadmium sulfate hydrate (3 CdSO4 x 8 H2O) is dissolved in dist. H2O
in a beaker and made up to 100 mL. In a second beaker 7.85 g Na 2S2O3 x 5
H2O is dissolved in dist. H2O and made up to 100 mL. The solutions are then
combined and mixed. A yellow precipitate may be formed; this does not in-
terfere.
• Potassium acetate solution:
10 g potassium acetate is dissolved in a mixture of 180 mL w(acetic acid) =
99% and 20 mL acetic acid anhydride.
• Bromine solution:
5 mL bromine is dissolved in 145 mL potassium acetate solution. This solu-
tion has a shelf life of only 1 day!
• Sodium acetate solution:
22 g sodium acetate is dissolved in dist. H2O, made up to 100 mL and
mixed.
• Sulfuric acid: w(H2SO4) = approx. 10%
5.7 mL w(H2SO4) = 96% is carefully added to 90 mL dist. H2O under stirring
and allowed to cool down.
• Methyl red indicator:
0.1 g methyl red is dissolved in 30 mL ethanol, treated with 20 mL dist. H2O
and mixed.
• Formic acid:
w(HCOOH) = 98% and w(HCOOH) = 49% (1:1 dilution)
• Hydriodic acid: w(HI) = 57%
• Potassium iodide: KI, reagent grade
• Phosphorus: red, amorphous
• Nitrogen: ultrapure, from cylinder
1
Method 78 –
thickening agents
The samples are dried in a drying oven at 105 °C until their weight is constant Sample preparation
and the Zeisel apparatus is placed under a fume hood.
The washing solution (absorption of I2 and HI) is placed in the first washing
bottle, 10 mL bromine solution is placed in both the second and third washing
bottles and 10 mL each of sodium acetate solution and w(HCOOH) = 98% is
placed in the receiver (for binding the bromine vapors).
Depending on the degree of substitution, 0.05 g...0.5 g sample is placed in the
reaction flask, 0.2 g phosphorus and two to three boiling stones are added and
mixed. After the addition of 5 mL hydriodic acid nitrogen is passed through (gas
flow approx. 2 bubbles/second) and the flask is heated in a heating mantle for
1 h at 160 °C.
When ether cleavage is complete the contents of the second and third wash-
ing bottles are transferred quantitatively with dist. H2O into a beaker into which
10 mL sodium acetate solution has already been pipetted, and then diluted to
approx. 100 mL with dist. H2O. The solution is stirred while w(HCOOH) = 49%
is added drop by drop until all the free bromine has been bound (the added
methyl red indicator must not become discolored).
The prepared sample solution is slightly acidified by adding sulfuric acid drop Analysis
by drop until the methyl red indicator has clearly changed color to red. Approx.
1 g potassium iodide is then added and immediately titrated with c(Na2S2O3) =
1 mL c(Na2S2O3) = 0.1 mol/L corresponds to 0.000517 g CH3O – Calculations

0.000751 g CH3CH2O –
g / 100 g CH3O – = EP1 x C01 x C03 x C30 / C00
g / 100 g CH3CH2O- = EP1 x C02 x C03 x C30 / C00
C01 = 0.000517
C02 = 0.000751
C03 = 100 (for 100 g)
C30 = titer of thiosulfate solution (Method 6)
2
Method 78 –
thickening agents
tPA tCF
DATE TIME DATE TIME
$%45- $%45-
PARAMETERS # FMLA
MEASPTDENSITY #
MININCR±L #
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES tDE
PAUSES DATE TIME
MEASINPUT DEF
STOPCONDITIONS -ETHOXY%0
#
#
##
STOP6ABS 23TEXT-ETHOXY
STOP5/&&M6 23UNITGG
lLLINGRATEMAXMLMIN %THOXY%0
#
#
##
STATISTICS 23TEXT%THOXY
EVALUATION 23UNITGG
REQSMPLSIZE/&& MEAN

Titration curve Result – Carboxymethyl cellulose
tFR
'044ITRINO
$ATE 4IME
5INIT M6$%45-
SMPLSIZEG
%0MLM6
-ETHOXYGG
%THOXYGG
3TOP6REACHED

3
Method 79 –

Principle The titration is based on the fact that the sulfamine group of the cyclamate is
oxidized to sulfate by sodium nitrite under very acidic conditions:
R–NHSO3 – + NO2– + 3 H3O+ → R–SO42– + N2 + 4 H2O
Recommended • 6.0451.100 comb. Pt-ring electrode with 6.2104.020 electrode cable

Reagents • Titrant: c(NaNO2) = 0.2 mol/L

13.8 g NaNO2 is weighed out into a 1000 mL volumetric flask, dissolved in
dist. H2O, made up to the mark and mixed. The titer is determined against
sodium cyclamate that has been dried at 105 °C.
• Hydrochloric acid: c(HCl) = approx. 2 mol/L
165 mL w(HCl) = 36% is added to approx. 800 mL dist. H2O in a 1000 mL
volumetric flask, made up to the mark and mixed.
Analysis A sample amount containing approx. 200...400 mg cyclamate is weighed out

into the titration beaker and dissolved in approx. 50 mL dist. H2O. After the addi-
tion of 50 mL c(HCl) = 2 mol/L the mixture is titrated with c(NaNO2) = 0.2 mol/L
to after the first endpoint.
Calculations The cyclamate content is given in %.

1 mL c(NaNO2) = 0.2 mol/L corresponds to 40.42 mg sodium cyclamate
39.66 mg calcium cyclamate
% cyclamate = EP1 x C01 x C02 x C30 / C00
EP1 = mL NaNO2 solution up to endpoint
C01 = 40.42 or 39.66
C02 = 0.1 (for %)
C30 = titer of NaNO2 solution
1
Method 79 –
tPA tFR
DATE TIME $ATE 4IME
-%45- 5INIT M6-%45-
6STEPML #YCLAMATE
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
STOPCONDITIONS
STOP6ABS Titration curve
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%45-
DEF
FORMULA
#YCLAMAT%0
#
#
##
23TEXT#YCLAMAT
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 80 –

Principle The saccharin sample is dissolved in dist. H2O, acidified and extracted with eth-
yl acetate. After evaporation it is dissolved in ethanol and titrated with NaOH.


• Hydrochloric acid: c(HCl) = approx. 2 mol/L. See Method 79, cyclamate
• Ethyl acetate: acetic acid ethyl ester, reagent grade
• Ethanol: absolute
Sample preparation The sample is pulverized in a mortar. An amount of sample that contains ap-
prox. 400 mg sodium saccharinate is weighed out into a separating funnel and
dissolved in approx. 20 mL dist. H2O. 5 mL c(HCl) = 2 mol/L is added and ex-
tracted initially with 30 mL, followed by 5 portions each of 20 mL ethyl acetate.
The combined extracts are evaporated to dryness in a rotary evaporator.
Analysis The dry extract is transferred quantitatively from the evaporation flask to the
titration beaker with ethanol (approx. 50 mL). After the addition of approx. 40 mL
dist. H2O it is titrated with c(NaOH) = 0.1 mol/L to after the first endpoint.
Calculations The saccharin content is given in %.

1 mL c(NaOH) = 0.1 mol/L corresponds to 20.52 mg Na saccharin
20.22 mg Ca saccharin
% saccharin = EP1 x C01 x C02 x C30 / C00
C01 = 20.52 or 20.22
C02 = 0.1 (for %)
C30 = titer of NaOH
Remarks The weight of sodium saccharinate must be such that a precipitate is formed
after treatment with HCl (approx. 400 mg).
1
Method 80 –
tPA tFR
$%45- 5INIT M6$%45-
4ITRATIONSPARAMETER %0MLM6
-ESSPKTDICHTE %0ML M6
-IN)NKREMENT±L .A 3ACC
$OS'ESCHWMAXMLMIN STOP6REACHED
-ESSW$RIFTM6MIN
7ARTEZEITS
3TART6AUS
0AUSES
$OSELEMENTINTERN$
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4EMPERATUR#
!BBRUCHBEDINGUNGEN Titration curve
3TOPP6ABS
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%INMASSABFRAUS
'RENZW%INMASSAUS
!KTIVIERPULSAUS

tCF
'044ITRINO
$ATUM :EIT
$%45-
# FMLA
#
#

tDE
'044ITRINO
$ATUM :EIT
$%45-
DEF
&ORMEL
.A 3ACC%0
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234EXT.A 3ACC
23.ACHKOMMASTELLEN
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-ITTELWERT
-.23
4EMPORÛRE6ARIABLE

2
Preserved fruits, vegetables and mushrooms

General In addition to titrators, Metrohm also manufactures a whole range of other
analytical instruments. VA instruments are particularly suitable for the determi-
nation of traces of heavy metals or toxic metals. Or are you interested in deter-
mining different types of sugars? With the Metrohm 817 Bioscan a well-proven
analytical instrument is available. In addition, nitrite, nitrate, phosphate, sulfate,
etc. can be determined simultaneously by ion chromatography.
1
Method 81 –
N1 Oxalic acid in fruits and vegetables

Principle The sample is pulverized and the oxalic acid is extracted with water. An aliquot
is acidified and titrated with permanganate.

Reagents • Titrant I: c(KMnO4) = 0.02 mol/L. See Method 7, Section B

• Manganese sulfate hydrate: MnSO4 x H2O, reagent grade
• Sea sand, reagent grade
Sample preparation 10 g sample is mixed with approx. 5 g sand and thoroughly pulverized in a
mortar. The mixture is rinsed into a 250 mL volumetric flask with dist. water and
made up to the mark. After vigorous shaking the solids are allowed to settle.
Analysis 25 mL of the prepared, slightly green sample solution is pipetted into a beaker
and treated with 2 mL H2SO4. After the addition of 0.5 g MnSO4 x H2O it is titrated
in the MET mode with c(KMnO4) = 0.02 mol/L.
Calculations The oxalic acid content is given in %.

1 mL c(KMnO4) = 0.02 mol/L corresponds to 4.5018 mg oxalic acid
% oxalic acid = EP1 x C01 x C02 / C00
C00 = 1 (sample weight in g /10)
C01 = 4.5018
C02 = 0.1 (for %)
1
Method 81 –
N1 Oxalic acid in fruits and vegetables
Instrument parameters and calculation Result – Spinach leaves
tPA tFR
DATE TIME $ATE 4IME
-%45- 5INIT M6-%45-
PARAMETERS %0ML M6
TITRATIONPARAMETERS OXALICAC
6STEPML STOP6REACHED
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6ABS
START6ML
DOSRATEMAXMLMIN
PAUSES
MEASINPUT
TEMPERATURE#
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#M6
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
-%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
-%45-
DEF
FORMULA
/XALS%0
#
##
23TEXT/XALS
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 82 –
N 2 Total sulfurous acid in dried fruits and
vegetables

Principle The sample is treated with methanol and phosphoric acid and the SO2 is dis-
tilled in a stream of nitrogen into a receiver containing H2O2. This oxidizes the
SO2 to H2SO4 which can then be titrated with NaOH. This method is specific.
SO2 + H2O2 = H2SO4

• Cylinder containing ultrapure nitrogen
• Special apparatus for SO2 distillation (see figure below)
Reagents • Titrant: c(NaOH) = 0.01 mol/L

Prepared from c(NaOH) = 0.1 mol/L by dilution with CO2-free dist. H2O (1:10,
Method 3, Section B)
• Phosphoric acid: w(H3PO4) = 85%
• Methanol: approx. 99%, reagent grade
• Absorption solution: w(H2O2) = 0.3%
1 mL w(H2O2) = 30% is made up to 100 mL with dist. H2O and the pH is ad-
justed to pH = 8.1 with c(NaOH) = 0.01 mol/L.
Sample preparation In these samples the SO2 is usually very unevenly distributed and in apricots,
for example, varies from one dried fruit to the other. For this reason an aver-
age value must be obtained. This is done by pouring liquid nitrogen over 20
dried fruit and pulverizing them when deep-frozen (liquid nitrogen must also be
added during the grinding process).
10.0 g of the still cold, ground sample is weighed out into a round-bottom flask
and allowed to thaw slightly. 50 mL each of methanol and H3PO4 are added, ni-
trogen is bubbled through the mixture (1 mL/min), which is then heated to boil-
ing. For dried fruit this process is ended after 30 min and for dried vegetables
after 1 h.
Analysis The absorption solution is titrated in the SET mode with c(NaOH) = 0.01 mol/L
to pH = 8.1.
Calculations The total sulfurous acid is given in mg SO2 per 100

g sample.
1 mL c(NaOH) = 0.01 mol/L corresponds to
0.32 mg SO2
mg SO2 / 100 g = EP1 x C01 x C02 x C30 / C00
C01 = 0.32
C02 = 100 (for 100 g)
Apparatus for determining SO2
1
Method 82 –
N 2 Total sulfurous acid in dried fruits and
vegetables
Instrument parameters and calculation Result – Dried apricots
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( 3/MGG
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
3/%0
#
#
##
23TEXT3/
23DECIMALPLACES
23UNITMG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 83 –
and concentrates


w(HNO3) = 65% diluted 1:5 with dist. H2O 1.
10.0 g sample is weighed out into a beaker and suspended with approx. 50
Sample preparation mL dist. H2O. The mixture is heated to boiling, allowed to cool down and then
transferred to a 100 mL volumetric flask with dist. H2O, made up to the mark and
mixed. It is then filtered through a chloride-free filter paper.
Analysis 10.0 mL filtrate (1/10 of sample weight) is pipetted into the titration beaker. It is
diluted to approx. 50 mL with dist. H2O, 2 mL c(HNO3) = 2 mol/L is added and
the solution titrated with c(AgNO3) = 0.1 mol/L to after the first endpoint.
Calculations The cooking salt content is given in %.

% NaCl = EP1 x C01 x C02 x C30 / C00
C00 = 1/10 sample weight in g (1)
C01 = 5.8443
C02 = 0.1 (for %)
1
Method 83 –
and concentrates
Instrument parameters and calculation Result – Powdered mushrooms
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
PARAMETERS %0MLM6
TITRATIONPARAMETERS .A#L
MININCR±L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
TEMPERATURE#
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
#HLORID%0
#
##
23TEXT#HLORID
23DECIMALPLACES
23UNITGKG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Cooking salt, spices, pickling salt, seasoning,
herbal and flavored salts
tion of traces of heavy metals or toxic metals (e.g. Cd, Co, Cu, Fe, Mo, Ni, Pb,
Zn, etc.). For cooking/common salt and sea salt the samples do not need to be
prepared – just dissolve in dist. H2O and analyze!
1
Method 84 –


40 mg polyvinyl alcohol is dissolved in 100 mL hot dist. H2O and allowed to
cool down.
Sample preparation 1.00 g sample is weighed out into a beaker, approx. 50 mL dist. H2O is poured
over it and brought to the boil. After cooling down, the mixture is transferred to
a 100 mL volumetric flask with dist. H 2O, made up to the mark and mixed.
Analysis 10.0 mL of the prepared sample solution (1/10 of the sample weight) is pipet-
ted into the titration beaker. It is made up to approx. 50 mL with dist. H2O, 2
mL c(HNO3) = 2 mol/L and 5 mL protective colloid are added and the mixture
Calculations The chloride content is given in % NaCl.

% NaCl = EP1 x C01 x C02 x C30 / C00
C00 = 1/10 sample weight in g (0.1)
C01 = 5.8443
C02 = 0.1 (for %)
1
Method 84 –
Instrument parameters and calculation Result – Seasoning mixture
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY .A#L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
.A#L%0
#
#
##
23TEXT.A#L
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 85 –
Principle Iodide is oxidized to iodate by bromine water. After elimination of the excess
bromine by formaldehyde the solution is treated with potassium iodide and the
released iodine titrated with thiosulfate.
I – + 3 Br2 + 3 H2O → IO3 – + 6 HBr
IO3 – + 5 I – + 6 H3O+ → 3 I2 + 9 H2O

Reagents • Titrant: c(Na2S2O3) = 0.01 mol/L

Freshly prepared by 1:10 dilution of c(Na2S2O3) = 0.1 mol/L (Method 6, Sec-
tion B) with CO2-free dist. H2O.
• Bromine solution:
Approx. 3.6 g bromine is dissolved in 100 mL dist. H 2O.
then made up to 100 mL and mixed.
• Formaldehyde solution: w(HCHO) = 36%
• Acetic acid: w(CH3COOH) = 96%
• Potassium iodide: KI, reagent grade
Sample preparation 50.0 g sample is weighed out into a beaker and dissolved in approx. 180 mL
dist. H2O. 2 mL bromine solution and 10 mL NaOH are added and allowed to
react for 2 min under stirring. The solution is then treated in succession with
10 mL each formaldehyde solution and acetic acid and allowed to react for a
further 5 min under stirring.
Analysis Approx. 1 g potassium iodide is added to the prepared sample solution, which
is then titrated with c(Na2S2O3) = 0.01 mol/L to after the first endpoint.
Calculations The total iodine content is given in mg/kg (ppm).

1 mL c(Na2S2O3) = 0.01 mol/L corresponds to 0.2115 mg iodine
mg/kg total iodine = EP1 x C01 x C02 x C30 / C00
C01 = 0.2115
C02 = 1000 (for 1 kg)
1
Method 85 –
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY TOTAL)MGKG
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
'ES)%0
#
#
##
23TEXT'ES)
23DECIMALPLACES
23UNITMGKG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 86 –
General The fluoride content is too low for titrimetric determination. This is why direct
potentiometry using an F ISE is used. However, the following points must be
observed:
• Selectivity
The F ISE responds not only to fluoride ions. In too strongly acidic solutions,
in which HF or even H2F2 is present, no measurements can be made. On the
other hand the F ISE has a cross-sensitivity to OH – ions. This is why it is best
to measure at pH = 5...6.5.
• Ionic activity
The concentration cannot be measured directly with ion-selective electrodes.
It is always the ionic activity that is measured. The activity coefficient (f) de-
pends on the total ionic activity and the temperature of the solution. In order
to avoid measuring errors the measurements are always made at a constant
ionic strength and temperature.
• Electrode slope
This obeys (as in pH measurement) the Nernst equation. This means that for
a negatively charged fluoride ion at 25 °C a change in potential of –59.2 mV
occurs when the activity (concentration) is increased by the factor 10. The
table below shows the electrode slope as a function of temperature:
Theoretical electrode slope of 1.00 (100%) as a function of the temperature
15.0 °C 57.2 mV
16.0 °C 57.4 mV
17.0 °C 57.6 mV
18.0 °C 57.8 mV
19.0 °C 58.0 mV
20.0 °C 58.2 mV
21.0 °C 58.4 mV
22.0 °C 58.6 mV
23.0 °C 58.8 mV
24.0 °C 59.0 mV
25.0 °C 59.2 mV
• Interfering ions
In addition to OH – ions, metal ions that form sparingly soluble fluoride salts
also interfere. These are primarily Al, Ca and Fe and a complexing agent is
added (in our case citrate) to release the fluoride ions again.
• TISAB solution
TISAB = Total Ionic Strength Adjustment Buffer. By the addition of this solu-
tion the sample solution is given a practically constant ionic strength and is
buffered to an optimal pH value. As the TISAB solution also contains citrate,
the Ca and Fe contained in the sample are masked.
Recommended • 6.0502.150 F ISE with 6.2104.020 electrode cable and 6.0726.107 Ag/AgCl
1
Method 86 –
• TISAB solution: Reagents

129 g trisodium citrate and 20 g potassium nitrate are weighed out into a
beaker and dissolved in approx. 600 mL dist. H2O. The pH is adjusted to pH
= 5.7 by adding c(HCl) = 2 mol/L. The solution is then made up to 1 liter with
• Fluoride standard I: ρ (fluoride) = 1000 mg/L
2.210 g NaF is weighed out into a 1000 mL volumetric flask and dissolved in
100 mL TISAB solution. The solution is then made up to the mark with dist.
H2O and mixed. It must be stored in a plastic bottle.
• Fluoride standard II: ρ (fluoride) = 10 mg/L
1.00 mL fluoride standard I is pipetted into a 100 mL plastic volumetric flask,
made up to the mark with dist. H2O and mixed. This solution has a limited
shelf life and must be made up freshly each week.
25 mL fluoride standard II (10 mg/L F) and 25 mL TISAB solution are placed in Determining the
a plastic beaker. The electrode is immersed in the solution, which is stirred. The
mV value is read off and noted at a constant potential. The electrode is then
electrode slope
rinsed with dist. H2O, dabbed dry with a soft paper tissue and the above proce-
dure is repeated with fluoride standard I (1000 mg/L F).
The mV difference is obtained from the two mV measured values and divided by
2. This gives the absolute electrode slope in mV per concentration factor of 10.
For the relative electrode slope the following applies:
relative electrode slope = absolute electrode slope

theoretical electrode slope
10.0 g sample is dissolved in dist. H2O, made up to 100 mL and mixed (corre- Analysis
sponds to 100 g sample/L).
25 mL each of TISAB solution and sample solution and a stirrer bar are placed
in a plastic beaker. The electrode is immersed in the solution, which is stirred.
The mV value is read off and noted at a constant potential.
Small portions of fluoride standard I (1000 mg/L F) are now added until an
optimal mV difference of approx. 15 mV is obtained (note the exact volume of
standard added). At constant potential, the mV value is read off and noted. The
difference between the two measured values is calculated (in mV).
The fluoride content is calculated according to the following equation: Calculations

CX = VSt × CSt The fluoride content is
dU/S given in mg/kg.
10 × (V0 + VSt) –V0
where:
CX = required fluoride concentration in mg/L
Vst = volume of added standard solution in mL
Cst = concentration of added standard solution in mg/L (1000)
V0 = volume of sample solution in mL (25)
dU = mV difference, sample before and after spiking with standard
S = absolute electrode slope at measuring temperature (mV per concentration factor of 10)
As the fluoride content in mg/L refers to the dilute sample (100 g sample/L) the
result must be multiplied by 10 to obtain mg/kg.
2
Method 86 –
tPA MEAS
'044ITRINO
DATE TIME tFR
-%!35- '044ITRINO
PARAMETERS $ATE 4IME
MEASURINGPARAMETERS -%!35-
SIGNALDRIFTM6MIN #M6
EQUILIBRTIMES
MEASINPUT
TEMPERATURE#
TIMEINTERVALS MEAS
STATISTICS
STATUS/&& tFR
PRESELECTIONS '044ITRINO
REQIDENT/&& $ATE 4IME
REQSMPLSIZE/&& -%!35-
LIMITSMPLSIZE/&& #M6
ACTIVATEPULSE/&&

tCF DOSVOLM,
'044ITRINO
DATE TIME
-%!35- #ALCULATION
# FMLA
#X
>

tDE MG,
MGKG
'044ITRINO #XMGKG
DATE TIME
-%!35-
DEF
FORMULA
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 87 –
Reagents • Titrant I: c(Na2EDTA) = 0.1 mol/L. See Method 9, Section B

• Titrant II: c(CuSO4) = 0.1 mol/L
25.2 g CuSO4 x 5 H2O is weighed out into a 1000 mL volumetric flask and dis-
solved in approx. 800 mL dist. H2O. One drop of w(H2SO4) = 96% is added
and the solution made up to the mark with dist. H2O and mixed.
• Ammonia solution: w(NH3) = 25%
Analysis 5.0 g salt is weighed out into the titration beaker and dissolved in approx.
100 mL dist. H2O. After the addition of 2 mL hydrochloric acid the solution is
heated to approx. 50 °C and then allowed to cool down to room temperature.
The solution is treated with 20.00 mL c(Na2EDTA) = 0.1 mol/L and 8 mL ammo-
nia solution under stirring and then titrated with c(CuSO4) = 0.1 mol/L to after
the first endpoint.
Calculations The tricalcium phosphate content is given in g per 100 g.

1 mL c(Na2EDTA) = 0.1 mol/L corresponds to 10.047 mg tricalcium phosphate
g / 100 g = (C31 – EP1) x C01 x C02 x C30 / C00
EP1 = mL CuSO4 solution up to endpoint
C01 = 10.047
C02 = 0.1 (conversion factor for g/100 g)
C31 = 20 (mL Na2EDTA solution added)
1
Method 87 –
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY 23GG
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
%0RECOGNITIONALL
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
23# %0
#
#
##
23TEXT
23DECIMALPLACES
23UNITGG
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 88 –
Principle Nitrite is oxidized to nitrate by permanganate. The excess permanganate is

back-titrated with iron(II) solution.
2 MnO4 – + 5 NO2– + 6 H3O+ → 5 NO3 – + 2 Mn2+ + 9 H2O
MnO4 – + 5 Fe2+ + 8 H3O+ → Mn2+ + 5 Fe3+ + 12 H2O

Reagents • Titrant I: c(1/5 KMnO4) = 0.1 mol/L. See Method 7, Section B

• Titrant II: Fe(II) solution, c = 0.1 mol/L
39.2 g (NH4)2Fe(SO4)2 x 6 H2O is weighed out into a 1000 mL volumetric flask,
treated with 25 mL w(H2SO4) = 25%, dissolved in dist. H2O, made up to the
mark and mixed.
71 mL w(H2SO4) = 96% is carefully added under stirring to approx. 400 mL
dist. H2O in a beaker. After cooling down, the solution is made up to 500 mL
with dist. H2O.
Determining the 25.00 mL c(1/5 KMnO4) = 0.1 mol/L is pipetted into the titration beaker, 100 mL
blank value dist. H2O and 20 mL w(H2SO4) = 25% are added and the solution titrated with
Fe(II) solution c = 0.1 mol/L to after the first endpoint.
The titrant consumption (EP1) is stored in the Titrino as Common Variable
C31.
BV = EP1 = C31
Sample preparation 50.0 g pickling salt is dissolved in dist. H2O, made up to 500 mL and mixed. If
any insoluble residues are present then these are filtered off through a paper
filter.
Analysis 100 mL filtrate (corresponding to 10 g sample) is pipetted into the titration bea-
ker and, under stirring, first treated with 25.00 mL c(1/5 KMnO4) = 0.1 mol/L and
then with 20 mL w(H2SO4) = 25%. After stirring for 3 min the solution is titrated
with Fe(II) solution c = 0.1 mol/L to after the first endpoint.
Calculations The NaNO2 content is given in %.

1 mL c(1/5 KMnO4) = 0.1 mol/L corresponds to 3.488 mg NaNO2
% NaNO2 = (C31 – EP1) x C01 x C02 x C30 / C00
EP1 = mL Fe(II) solution up to endpoint
C00 = sample weight in g / 100 mL (10)
C01 = 3.488
C02 = 0.1 (for %)
C30 = titer of KMnO4 solution (Method 7)
C31 = blank value (mL Fe(II) solution)
Remarks If the solution already looses its color during the nitrite oxidation then a smaller
amount of sample must be used.
3
Method 88 –
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY .A./
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
.A./# %0
#
#
##
23TEXT.A./
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
P Meat products, meat extracts, bouillon preparations, aspic, seasonings, soups, sauces
Meat products, meat extracts, bouillon
preparations, aspic, seasonings, soups, sauces
tion of traces of toxic metals (e.g. As, Cd, Hg, Pb, Tl) and/or vitamin C, sulfite
and nitrite. Ion chromatography (IC) is particularly suitable for the simultaneous
determination of anions (e.g. chloride, nitrate, nitrite, sulfate, sulfite) and with
the 817 Bioscan different types of sugar can be determined (e.g. in spices and
soups).
1
Method 89 –

• Mixer or high-frequency homogenizer (e.g. Polytron or Ultra-Turrax)

w(HNO3) = 65% is diluted 1:5 with dist. H2O.
The sample is first comminuted (e.g. with a knife). 10.0 g sample is placed in
Sample preparation an Erlenmeyer flask, 190 mL dist. H2O is added and homogenized (e.g. with a
high-frequency homogenizer for 1...2 min).
50 g homogenized sample mixture (corresponding to ¼ of the sample weight)

Analysis is weighed out into the titration beaker, 50 mL dist. H2O and 2 mL nitric acid are
added and titrated with c(AgNO3) = 0.1 mol/L to after the first endpoint.
Calculations The chloride content is given in % NaCl (g/100 g).

% NaCl = EP1 x C01 x C02 x C30 / C00
C00 = ¼ sample weight in g (2.5)
C01 = 5.8443
C02 = 0.1 (for %)
Remarks For a NaCl content of more than 7% we recommend that less of the homog-
enized sample mixture is weighed out and dist. H2O is added to make up to 100
mL.
1
Method 89 –

Smoked salmon Smoked salmon
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY .A#L
MININCR±L STOPP6REACHED
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE# Titration curve – Smoked salmon
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
.A#L%0
#
#
##
23TEXT.A#L
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

2
Method 89 –

Salami Salami
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY .A#L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE# Titration curve – Salami
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
.A#L%0
#
#
##
23TEXT.A#L
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

3
Method 90 –
products
General With this type of sample the Kjeldahl method determines the total nitrogen. A
factor can be used to calculate the raw protein content. The conversion factor is
obtained from the average nitrogen content in meat protein of 16%.
Sample preparation The samples often have a very heterogeneous composition and must therefore
be well homogenized. This is best carried out in a blender.
The amount of sample to be digested depends on the expected nitrogen or raw
protein content. The following table provides a guideline:
g / 100 g N g / 100 g protein Sample weight, g

<1 <6 2
2 12 1
5 30 0.5
The homogenized sample is weighed out into the digestion tube, a catalyst
tablet* and 10 mL w(H2SO4) = 98% are added and heated in the digestion ap-
paratus until the solution is clear. Heating is continued for a further 10 min.


EP at pH 4.5
dynamics 6
max. rate 2 mL/min
1
Method 90 –
products
1 mL c(HCl) = 0.1 mol/L corresponds to 1.4007 mg N Calculations

Total nitrogen (N) in % (g / 100 g)
N = EP1 x C01 x C02 x C30 / C00 = RS1
Raw protein (RP) in % (g / 100 g)
RP = RS1 x C03
C01 = 1.4007
C02 = 0.1 (for %)
C03 = 6.25 (conversion factor for raw protein)
2
Method 90 –
products
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS #
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN tDE
3%4 '044ITRINO
%0ATP(/&& DATE TIME
PAUSES FORMULA
START6/&& .%0
#
#
##
PAUSES 23TEXT.
DOSELEMENTINTERNAL$ 23UNIT
TEMPERATURE# 2023
#
TIMEINTERVALS 23TEXT20
STOP6ABS 23UNIT
REQIDENT/&& MEAN
REQSMPLSIZEALL -.23
ACTIVATEPULSE/&&

Titration curve Result – Frying sausage
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEG
%0ML
.
RAWPR

3
Method 91 –
Principle The sample is treated with methanol and phosphoric acid and the SO2 distilled
in a stream of nitrogen into a receiver containing H 2O2. This oxidizes the SO2 to
H2SO4, which can then be titrated with NaOH. This method is specific, but can
only be used in the absence of volatile acids (e.g. acetic acid).
SO2 + H2O2 = H2SO4

• Gas cylinder containing ultrapure nitrogen
• Special apparatus for SO2 distillation (see Method 82, N 2)
Reagents • Titrant: c(NaOH) = 0.01 mol/L

Prepared from c(NaOH) = 0.1 mol/L by dilution with CO2-free dist. H2O (1:10,
Method 3, Section B)
• Phosphoric acid: w(H3PO4) = 85%
• Methanol: approx. 99%, reagent grade
• Absorption solution: w(H2O2) = 0.3%
1 mL w(H2O2) = 30% is made up to 100 mL with dist. H2O and the pH ad-
justed with c(NaOH) = 0.01 mol/L to pH = 8.1.
Sample preparation 10 mL absorption solution is diluted with 10 mL dist. H2O in the absorption ves-
sel. Before the start the distillation apparatus is purged with nitrogen for 5...10
min.
10.0 g homogenized sample and 50 mL each of methanol and H3PO4 are placed
in the round-bottom flask. Nitrogen is bubbled through the solution (1 mL/min)
which is heated to boiling and kept boiling for 15 min.
Analysis The absorption solution is titrated in the SET mode with c(NaOH) = 0.01 mol/L
to pH = 8.1 (first calibrate the electrode with buffer solutions pH = 7.00 and pH
= 4.00).
Calculations The sulfurous acid is given in mg SO2 per kg sample.

1 mL c(NaOH) = 0.01 mol/L corresponds to 0.32 mg SO2
mg/kg SO2 = EP1 x C01 x C02 x C30 / C00
C01 = 0.32
C02 = 1000 (for 1 kg)
Remarks The determination of the blank value and the sample produced the same result.
This means that it can be assumed that the sample no longer contains any
SO2.
1
Method 91 –
Instrument parameters and calculation – blank value
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN tDE
3%4 '044ITRINO
%0ATP(/&& DATE TIME
PAUSES FORMULA
START6/&& 3/%0
#
#
##
PAUSES 23TEXT3/
DOSELEMENTINTERNAL$ 23UNITMG
TEMPERATURE# SILOCALCULATIONS
TIMEINTERVALS MATCHID/&&
STOPCONDITIONS COMMONVARIABLES
STOP6ABS REPORT
STOP6ML REPORT#/-FULLCURVE
lLLINGRATEMAXMLMIN MEAN
STATISTICS -.23
STATUS/&& TEMPORARYVARIABLES
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve – blank value Result – blank value
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEG
%0ML
3/MGG

2
Method 91 –
tPA tCF
DATE TIME DATE TIME
3%4P(- 3%4P(-
PARAMETERS # FMLA
3%4 #
%0ATP( #
DYNAMICS/&&
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN tDE
3%4 '044ITRINO
%0ATP(/&& DATE TIME
PAUSES FORMULA
START6/&& 3/%0
#
#
##
PAUSES 23TEXT3/
DOSELEMENTINTERNAL$ 23UNITMG
TEMPERATURE# SILOCALCULATIONS
TIMEINTERVALS MATCHID/&&
STOPCONDITIONS COMMONVARIABLES
STOP6ABS REPORT
STOP6ML REPORT#/-FULLCURVE
lLLINGRATEMAXMLMIN MEAN
STATISTICS -.23
STATUS/&& TEMPORARYVARIABLES
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

Titration curve Result Air-dried beef
tFR
'044ITRINO
$ATE 4IME
P(CINIT 3%4P(-
SMPLSIZEG
%0ML
3/MGG

3
Method 92 –
P 4 Chloride (NaCl) in meat extracts, aspic
and bouillon


cool down.
Sample preparation 10.0 g sample is dissolved in approx. 500 mL hot dist. H2O, allowed to cool
down, made up to 1 liter with dist. H2O and mixed.
Analysis 25.0 mL (1/40 of sample weight) of the prepared sample solution is pipetted
into the titration beaker and made up with dist. H2O to approx. 50 mL. After the
addition of 2 mL c(HNO3) = 2 mol/L and 5 mL protective colloid it is titrated with
c(AgNO3) = 0.1 mol/L to after the first endpoint.

% NaCl = EP1 x C01 x C02 x C30 / C00
C00 = 1/40 sample weight in g (0.25)
C01 = 5.8443
C02 = 0.1 (for %)
5
P Meat products, meat extracts, bouillon preparations, aspic, seasonings, soups, sauces Method 92 –
P 4 Chloride (NaCl) in meat extracts, aspic
and bouillon
Instrument parameters and calculation Result – Vegetable bouillon
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY .A#L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
.A#L%0
#
#
##
23TEXT.A#L
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

6
Method 93 –
P 5 Kjeldahl nitrogen in meat extracts, aspic
and bouillon
General With this type of sample the Kjeldahl method determines the total nitrogen.
Sample preparation The weight of the sample to be digested depends on the fat content. With a
high fat content 1.5...2 g sample is weighed out, with a low fat content 2...2.5 g
sample.
Recommended – 6.0239.100 comb. pH glass electrode with 6.2104.020 electrode cable

Reagents – Titrant: c(HCl) = 0.1 mol/L. See Method 4, Section B

– Absorption solution: w(H3BO3) = 2%
EP at pH 4.5
dynamics 6
max. rate 2 mL/min
Calculations The total nitrogen is given in % N.

% N = EP1 x C01 x C02 x C30 / C00
C01 = 1.4007
C02 = 0.1 (for %)
3
P 5 Kjeldahl nitrogen in meat extracts, aspic
and bouillon
Instrument parameters and calculation Result – Aspic powder
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( .
DYNAMICS
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZEALL
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
.%0
#
#
##
23TEXT.
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 94 –
P 6 Chloride (NaCl) in seasonings, soups,
sauces


w(HNO3) = 65% is diluted 1:5 with dist. H2O.
cool down.
Sample preparation Powdered, granulated and pasty samples: 10.0 g is dissolved in approx. 500
mL hot dist. H2O, allowed to cool down, made up to 1 liter and mixed. Liquid
seasonings: 25.0 g is diluted to1 liter with dist. H2O.
Analysis 25.0 mL (1/40 of sample weight) of the prepared sample solution is pipetted
into the titration beaker and diluted with dist. H2O to approx. 50 mL. After the
addition of 2 mL c(HNO3) = 2 mol/L and 5 mL protective colloid the mixture is

% NaCl = EP1 x C01 x C02 x C30 / C00
C00 = 1/40 sample weight in g (0.25 or 0.625)
C01 = 5.8443
C02 = 0.1 (for %)
3
P 6 Chloride (NaCl) in seasonings, soups,
sauces
Instrument parameters and calculation Result – Packet soup
tPA tFR
DATE TIME $ATE 4IME
$%45- 5INIT M6$%45-
MEASPTDENSITY .A#L
DOSRATEMAXMLMIN
SIGNALDRIFTM6MIN
EQUILIBRTIMES
START6/&&
PAUSES
DOSELEMENTINTERNAL$
MEASINPUT
STOPCONDITIONS
STOP6ABS
STOP6ML
STOP5/&&M6
STOP%0
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
EVALUATION
%0#
lX%0AT5/&&M6
P+(.0/&&
PRESELECTIONS
REQIDENT/&&
REQSMPLSIZE/&&
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
$%45-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
$%45-
DEF
FORMULA
.A#L%0
#
#
##
23TEXT.A#L
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Method 95 –
sauces
General With this type of sample the Kjeldahl method determines the total nitrogen.
Sample preparation The weight of the sample to be digested depends on the fat content. With a
high fat content 1.5...2 g sample is weighed out, with a low fat content 2...2.5 g
sample.
Recommended – 6.0239.100 comb. pH glass electrode with 6.2104.020 electrode cable

Reagents – Titrant: c(HCl) = 0.1 mol/L. See Method 4, Section B

– Absorption solution: w(H3BO3) = 2%
EP at pH 4.5
dynamics 6
max. rate 2 mL/min
Calculations The total nitrogen is given in % N (g/100 g).

% N = EP1 x C01 x C02 x C30 / C00
C01 = 1.4007
C02 = 0.1 (for %)
3
sauces
Instrument parameters and calculation Result – Gravy
tPA tFR
DATE TIME $ATE 4IME
3%4P(- P(CINIT 3%4P(-
3%4 %0ML
%0ATP( .
DYNAMICS
MAXRATEMLMIN
MINRATE±LMIN
STOPCRITDRIFT
STOPDRIFT±LMIN
3%4
%0ATP(/&&
TITRDIRECTIONAUTO
PAUSES
START6/&&
PAUSES
EXTRTIMES
DOSELEMENTINTERNAL$
MEASINPUT
TEMPERATURE#
TIMEINTERVALS
STOPCONDITIONS
STOP6ABS
STOP6ML
lLLINGRATEMAXMLMIN
STATISTICS
STATUS/&&
PRESELECTIONS
CONDITIONING/&&
REQIDENT/&&
REQSMPLSIZEALL
LIMITSMPLSIZE/&&
ACTIVATEPULSE/&&

tCF
'044ITRINO
DATE TIME
3%4P(-
# FMLA
#
#

tDE
'044ITRINO
DATE TIME
3%4P(-
DEF
FORMULA
.%0
#
#
##
23TEXT.
23DECIMALPLACES
23UNIT
23LIMITCONTROL/&&
SILOCALCULATIONS
MATCHID/&&
COMMONVARIABLES
REPORT
REPORT#/-FULLCURVE
MEAN
-.23
TEMPORARYVARIABLES

4
Q Qualifying the titrator,
validating a titration method
General Titration is, in contrast to HPLC, for example, an absolute method and requires
no reference method. The reactions (stoichiometry) are known and normally
take place very quickly. If a titrant with a known titer is used then the content of
the sample can be determined directly.
There is no point in validating the components of a titration system individually.
It is always the complete system (titrator, burets, electrode, evaluation, possibly
a sample changer) that is validated. The qualification of the titrator and the
validation of the titration method are carried out simultaneously!
1
Method 96

Reagents • Titrant: c(NaOH) = 0.1 mol/L (CO2-free, Method 3)

• «Sample»: L(+)-tartaric acid, pulverized, ultrapure
• Solvent: dist. or deionized water
Remarks • Sodium hydroxide solution is very sensitive to CO2-absorption from the atmo-
sphere. This is why the absorber tube of the Exchange Unit must be filled with
a CO2 absorber (e.g. soda lime).
• The balance must previously have been validated.
• The measuring series should be carried out without any interruption and with
as constant a room temperature as possible.
General procedure The sample – in our example tartaric acid – is weighed out into the titration bea-
ker with an accuracy of 0.1 mg and dissolved in approx. 50 mL dist. H2O. It is
then titrated with c(NaOH) = 0.1 mol/L to after the first endpoint.
For example, 5 different sample weights are selected whose titrant consump-
tions lie between 2.5 mL and approx. 15 mL, and a 20 mL buret is used – refilling
the buret cylinder during the titration must be avoided at all cost!
Approx. 40 mg, 70 mg, 90 mg, 100 mg and 120 mg tartaric acid are weighed
out.
Calculations 1 mL c(NaOH) = 0.1 mol/L corresponds to 7.5045 mg C4H6O6

% tartaric acid = (EP1 x C30) x C01 x C02 / C00
C00 = weight of tartaric acid in mg
C01 = 7.5045
C02 = 100 (for %)
C30 = titer of NaOH
Special remarks The evaluation criteria given in the following sections can only be achieved un-
der optimal conditions (e.g. titer determination with a secondary standard). Dif-
ferent criteria apply to non-certified sample materials. In such cases it is up to
the user to decide the limits within which a method can be recognized as being
robust and accurate enough and can be carried out within a reasonable time.
a) Linear regression: titrant consumption as a function of tartaric acid

sample weight
mL / sample weight = EP1 x C01 x C30
C01 = equivalent weight of titrated substance (7.5045 for tartaric acid)
C30 = titer of titrant (NaOH, Method 3)
For uncovering systematic errors, e.g. of method-specific interference, a lin-
ear regression of titrant consumption (in mL) x titer against mg per weight of
«sample» can be calculated. This should be done by using a powerful pocket
calculator or a statistics package or spread-sheet software on a PC.
The quantity «mg per sample weight» is plotted as the x coordinate (indepen-
dent variable) and the titrant consumption times the titer as the y coordinate
1
Method 96
(dependent variable).
The linear regression draws a straight line through the measuring points so
that the sum of the squares of the individual deviations is at its minimum. The
regression curve is described by the equation: y = bx + a, where a is the y-axis
intercept and b the slope of the curve (see diagram below).
Systematic errors in the titration method can be recognized by a significant
deviation of the zero point coordinate of the y axis (intercept), i.e. the regression
line calculated from the pairs of values titrant consumption and sample weight
does not intercept the y axis exactly at the origin of the coordinate system.
asys as a measure for the systematic error is calculated from the mean value
of the x values, the mean value of the y values and the regression coefficient
(slope).
The calculation formulae for this are:
asys = y axis intercept = y –bx
If asys lies outside a range of ±0.010 mL (or ±10 μL) then it must be assumed that Evaluation
there is a systematic error. It is then essential to check the titration method and
other system-specific interferences. If it is not possible to further optimize the
method then the individual mL values (titrant consumption) must be corrected
by the value of asys (consumption minus asys in mL), so that the systematic er-
ror of the method is not included in the evaluation of the titrator. As a result the
relevant characteristic data for the precision and correctness of the results must
be recalculated using the corrected consumption values.
2
Method 96
b) Linear regression: percentage content against titrant consumption

Mass fraction w in % = EP1 x C01 x C02 x C30 / C00
C00 = sample weight in mg (e.g. tartaric acid)
C01 = equivalent weight of titrated substance (7.5045 for tartaric acid)
C02 = 100 (for %)
C30 = titer of titrant (e.g. NaOH, Method 3)
A further possible method for uncovering systematic errors is the presentation
of the regression curves (scatter diagram) of the pairs of values percentage
content against titrant consumption. As this diagram makes the scattering of the
results readily visible its use is to be recommended.
Significant positive and negative slopes of the regression lines indicate an
apparent dependency of the percentage content on the amount of the titrant
consumption or the sample weight. This can also be an indication of method-
specific, systematic interferences.
The slope b%/Vol (regression coefficient, see above for calculation formula) of the
linear equation y = bx + a should ideally have a value of 0.000, i.e. the curve
should be a horizontal straight line passing through y = 1.000.
If b%/Vol lies outside a range of ±0.005 then a systematic, method-specific error

Evaluation must also be assumed. A correction of the mL values by asys (consumption in
mL minus asys in mL) and a subsequent recalculation of the percentage content
will show a dramatic improvement in the renewed presentation of the regression
curve «percentage content against mL consumption».
c) Precision and accuracy

For the validation the precision (degree of scatter) and the accuracy of the
measured results are relevant. The evaluation of these quantities is carried out
as follows:
From the (5) determinations the values obtained (w %) are used for the calcula-
tion of the mean value x and the absolute standard deviation Sabs. These calcu-
lations can be carried out directly in the instrument with the included statistics
function or by using suitable software (PC, pocket calculator).
3
Method 96
The precision (repeatability) of the measurement is expressed by the relative

standard deviation Srel:
Srel = Sabs 100 / x
The relative standard deviation should be <0.3%. It should normally be easily
possible to observe this practical limit. Under optimal conditions relative stan-
dard deviations of 0.1% can be achieved.
The accuracy of the results obtained is based on the content guaranteed by the
manufacturer. Strictly speaking, comparisons should be made with primary or
secondary standards (standard titrimetric substances). The maximum variation
should not exceed ±1%.
• Sample Possible sources

Unsuitable, contaminated, moist, inhomogeneous.
of error
• Balance / weighing
Balance too inaccurate, drafts, temperature influences, balance contami-
nated, temperature difference between titration beaker and balance, care-
less weighing, sample too small. (If the sample weight is too low then the
measuring points of the linear regression – percentage content against titrant
consumption – will lie outside the regression line.)
• Titrant
Contains carbonate, contaminated.
• Dosing device
Leaking valve, leaking tubing connections, leaking antidiffusion valve, leak-
ing piston (film of liquid or crystals beneath the piston), dirty cylinder (visible
etched rims), filling rate too fast, air bubbles in system.
• Measurement
Poor stirring/mixing, electrode response too slow, electrode or its diaphragm
contaminated or blocked, loose connection (plug, cable).
• Titrator
Unsuitable titration mode, incorrect measuring parameters, titration speed
too high or too low.
• Temperature
Temperature variations during the titration series.
Application Bulletin no. 252 Additional literature

«Validation of Metrohm titrators (potentiometric) according to GLP/ISO 9000»
4
Method 96
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5
Method 96
Validation Record Titrators Company : Metrohm AG

Division : Appl.Lab.
Temperature in °C : 22.90 Instrument : 799 GPT Titrino
Titrant : c(NaOH) Electrode : 6.0259.100
Conc. in mol/L : 0.1 mol/L Slope: 0.976
Lot/ date of manufact : 30440.00 pHas : 7.02
L(+)-
Primary standard tartaric acid Exchange Unit : 6.3026.220
Molar mass in g/mol 150.09 Buret volume : 20 mL
Titration parameters: Mode : DET U
Meas. Pt density 4
Min. increment 10.0
Stop V 20 mL
Meas. Value drift 50 mV/min
Smpl. size : Volume : Content: No. Remark :

110.4 mg 14.851 ml 100.04% 1
61.7 mg 8.291 ml 99.93% 2
50.0 mg 6.699 ml 99.64% 3
120.6 mg 16.181 ml 99.78% 4
66.7 mg 8.995 ml 100.29% 5
42.2 mg 5.676 ml 100.03% 6
100.0 mg 13.420 ml 99.80% 7 Mean = 99.9040 sabs = 0.1929
85.4 mg 11.480 ml 99.97% 8 Content theor = 99.9994 srel = 0.19 %
55.3 mg 7.427 ml 99.88% 9 Delta = -0.10 % asys = 0.0074 ml
81.0 mg 10.857 ml 99.68% 10 bT/Vol = -0.0052
Result :
Dat : 15.04.2004

: Visum : bh
6
Method 96
7
Index
Keywords Section Method

Acid capacity, water C 10
Acid number, AN E 25
Ascorbic acid, beer K 55
Ascorbic acid, fruit juice J 42
Ascorbic acid, milk D 23
Ascorbic acid, vinegar K 59
Ash alkalinity, chocolate L 73
Ash alkalinity, cocoa L 73
Ash alkalinity, coffee L 69
Ash alkalinity, fruit juice J 49
Ash alkalinity, vegetable juice J 49
Ash alkalinity, vinegar K 62
Beer K
Bouillon P 92
Bread F
Ca, custard F 33
Ca, dairy products D 22
Ca, dry pasta F 33
Ca, fruit juice J 47
Ca, vegetable juice J 47
Ca, water C 11
Carbon dioxide, water C 17
Carbonate hardness, water C 10
Cereals F
Cheese D
Chloride, coffee L 70
Chloride, dairy products D 21
Chloride, dry pasta F 31
Chloride, fruit juice J 44
Chloride, vegetable juice J 44
Chloride, vinegar K 63
Chloride, water C 12
Chocolate L
Citric acid, soft drinks H 37
CO2 content, beer K 54
Cocoa L
Coffee L
Condiments O
Cooking salt O
Cooking salt, dry pasta F 31
1
Index

Custard, cream powder F
Cyclamate, sweeteners M 88
Degree of acidity, coffee L 68
Degree of acidity, dairy products D 20
Degree of acidity, dry pasta F 30
Drinking water C
Dry pasta F
Edible fats, edible oils E
Ethoxy groups, gelling or thickening agents M 78
Flour mill products F
Fluoride, cooking salt O 86
Formol number, fruit juice J 51
Formol number, honey G 35
Formol number, vegetable juice J 51
Free acids, honey G 34
Free fatty acids, cocoa butter L 75
Free fatty acids, FFA E 25
Fruit N
Fruit juice J
Gelling or thickening agents M
Honey G
Hydrogen sulfide, water C 14
Hydroxyl number, OHN E 26
Iodine number, cocoa butter L 76
Iodine number, IN E 27
Iodine, cooking salt O 85
Jams J
K, fruit juice J 48
K, vegetable juice J 48
Kjeldahl N, aspic P 93
Kjeldahl N, bouillon P 93
Kjeldahl N, cocoa L 74
Kjeldahl N, dairy products D 24
Kjeldahl N, dry pasta F 32
Kjeldahl N, fruit juice J 50
Kjeldahl N, jams J 50
Kjeldahl N, meat extracts P 93
Kjeldahl N, meat products P 90
Kjeldahl N, sauces P 95
Kjeldahl N, soups P 95
2
Index

Kjeldahl N, spices P 95
Kjeldahl N, vegetable juice J 50
Lemonade H
Meat extracts P
Meat products P
Methoxy groups, gelling or thickening agents M 78
Mg, custard F 33
Mg, dry pasta F 33
Mg, fruit juice J 47
Mg, vegetable juice J 47
Mg, water C 11
Milk powder D
Milk, dairy products D
Mineral water C
Mushrooms N
NaCl, aspic P 92
NaCl, bouillon P 92
NaCl, cooking salt O 84
NaCl, dairy products D
NaCl, meat extracts P 92
NaCl, meat products P 89
NaCl, preserved mushrooms O 84
NaCl, sauces P 94
NaCl, seasonings P 94
NaCl, soups P 94
NaCl, spices O 84
NaCl, vinegar K 63
Nitrite, pickling salt O 88
Nitrogen: see Kjeldahl
Oxalic acid, fruit and vegetables N 82
Oxidizability, water C 16
Oxygen, water C 18
Pasta (dry) F
Peroxide number, PON E 28
pH, beer K 53
pH, calibration A 1
pH, chocolate L 72
pH, cocoa L 72
pH, coffee L 68
pH, confectionery G 34
3
Index

pH, dairy products D 19
pH, dry pasta F 30
pH, fruit juice J 41
pH, general A
pH, honey G 34
pH, jams J 41
pH, measurement A 2
pH, spirits K 65
pH, sweets G 34
pH, vegetable juice J 41
pH, vinegar K 57
pH, water C 10
Phosphoric acid, cola drinks H 38
Pickling salt O
Potassium, soft drinks H 39
Qualification, titrators Q 96
Raw protein, meat products P 90
Reducing sugars, coffee L 71
Reducing sugars, confectionery G 36
Reducing sugars, honey G 36
Reducing sugars, jams J 52
Residual chlorine, water C 15
Saccharin, sweeteners M 80
Saponification number, cocoa butter L 77
Saponification number, SN E 29
Sauces P
Sea salt O
Seasonings P
Sodium nitrite, pickling salt O 88
Soft drinks H
Soups P
Spices O
Spirits K
Starch F
Sugar, confectionery G
Sulfate, fruit juice J 46
Sulfate, vegetable juice J 46
Sulfate, vinegar K 64
Sulfate, water C 13
Sulfide, water C 14
4
Index

Sulfite, fruit juice J 43
Sulfite, jams J 43
Sulfurous acid, beer K 56
Sulfurous acid, dried fruit N 82
Sulfurous acid, dried vegetables N 82
Sulfurous acid, fruit juice J 43
Sulfurous acid, jams J 43
Sulfurous acid, meat products P 91
Sulfurous acid, vinegar K 60
Sulfurous acid, vinegar K 61
Sweeteners M
Table salt O
Titer determination, general B
Titer H2SO4 B 4
Titer HCl (in ethanol) B 4
Titer iodine solution B 5
Titer KOH in ethanol B 3
Titer Na2EDTA B 9
Titer NaOH B 3
Titer permanganate B 7
Titer phenylarsine oxide C 15
Titer silver nitrate B 8
Titer thiosulfate B 6
Titrant B
Titratable acids, dairy products D 20
Total acidity, fruit juice J 41
Total acidity, jams J 41
Total acidity, spirits K 65
Total acidity, vegetable juice J 41
Total acidity, vinegar K 57
Total chlorine, water C 15
Total esters, spirits K 67
Total iodine, cooking salt O 85
Total phosphorus, fruit juice J 45
Total phosphorus, jams J 45
Total phosphorus, soft drinks H 40
Total phosphorus, vegetable juice J 45
Total protein, dry pasta F 32
Total protein, flour F 32
Total protein, fruit juice J 50
5
Index

Total protein, jams J 50
Total protein, vegetable juice J 50
Tricalcium phosphate, cooking salt O 87
Validation, titration methods Q 96
Vegetable juice J
Vegetables N
Vinegar K
Vitamin C, fruit juice J 42
Vitamin C, milk D 23
Volatile acids, spirits K 66
Volatile acids, vinegar K 58
Water hardness C 11
Wine K
Winkler method C 18
Yogurt D
6
Beispiel eines vollautomatisierten Analysensystems
für Trinkwasser (MATI 1)
Analysierte Parameter
• pH-Wert
• Elektrische Leitfähigkeit
• Säurekapazitäten/Carbonathärte
• Ca-, Mg- und Gesamthärte
1
Method overview 6.6055.003 Food PAC
Card DIR 1 Method 1 A1 pH value – calibrating the pH glass electrode

Method 2 A2 pH value – measuring the pH value
Method 3 B1 Preparing the titrants and determining their titer – alkaline titrants (NaOH, KOH)
Method 4 B2 Preparing the titrants and determining their titer – acidic titrants (HCl, H2SO4)
Method 5 B3 Preparing the titrants and determining their titer – iodine solution
Method 6 B4 Preparing the titrants and determining their titer – thiosulfate
Method 7 B5 Preparing the titrants and determining their titer – permanganate
Method 8 B6 Preparing the titrants and determining their titer – silver nitrate
Method 9 B7 Preparing the titrants and determining their titer – Na2EDTA
DIR 2 Method 10 C1 Drinking water and mineral water – pH value and acid capacity (carbonate hardness)
Method 11 C2 Drinking water and mineral water – calcium and magnesium (Ca hardness, Mg hardness
and total hardness)
Method 12 C3 Drinking water and mineral water – chloride
Method 13 C4 Drinking water and mineral water – sulfate
Method 14 C5 Drinking water and mineral water – sulfides and hydrogen sulfide
Method 15 C6 Drinking water and mineral water – total and residual chlorine (free chlorine)
Method 16 C7 Drinking water and mineral water – permanganate index (oxidizability)
Method 17 C8 Drinking water and mineral water – CO2 content of carbonated water
Method 18 C9 Drinking water and mineral water – oxygen content according to Winkler
Method 19 D1 Milk and dairy products – pH value
Method 20 D2 Milk and dairy products – titratable acidity, degree of acidity
Method 21 D3 Milk and dairy products – chloride
Method 22 D4 Milk and dairy products – calcium
Method 23 D5 Milk and dairy products – vitamin C (ascorbic acid)
Method 24 D6 Milk and dairy products – Kjeldahl nitrogen
DIR 3 Method 25 E1 Edible fats and oils – acid number and free fatty acids (FFA)
Method 26 E2 Edible fats and oils – hydroxyl number
Method 27 E3 Edible fats and oils – iodine number
Method 28 E4 Edible fats and oils – peroxide number
Method 29 E5 Edible fats and oils – saponification number
Method 30 F1 Cereals, flour milling products, dry pasta products – pH value and degree of acidity
Method 31 F2 Cereals, flour milling products, dry pasta products – chloride and cooking salt content
Method 32 F3 Cereals, flour milling products, dry pasta products – Kjeldahl nitrogen, total protein
Method 33 F4 Cereals, flour milling products, dry pasta products – calcium and magnesium (in ash)
Method 34 G1 Honey, sugar and sweets – pH value and free acids
Method 35 G2 Honey, sugar and sweets – formol number
Method 36 G3 Honey, sugar and sweets – reducing sugars
Method 36 G3 Honey, sugar and sweets – reducing sugars
DIR 4 Method 37 H1 Soft drinks, lemonades – citric acid, citrates
Method 38 H2 Soft drinks, lemonades – phosphoric acid (cola drinks)
Method 39 H3 Soft drinks, lemonades – potassium
Method 40 H4 Soft drinks, lemonades – total phosphorus
Method 41 J1 Fruit and vegetable juices, fruit nectars and jams – pH value and titratable total acidity
Method 42 J2 Fruit and vegetable juices, fruit nectars and jams – ascorbic acid (vitamin C)
Method 43 J3 Fruit and vegetable juices, fruit nectars and jams – sulfurous acid (sulfite)
Method 44 J4 Fruit and vegetable juices, fruit nectars and jams – chloride
Method 45 J5 Fruit and vegetable juices, fruit nectars and jams – total phosphorus
Method 46 J6 Fruit and vegetable juices, fruit nectars and jams – sulfate
Method 47 J7 Fruit and vegetable juices, fruit nectars and jams – calcium and magnesium
Method 48 J8 Fruit and vegetable juices, fruit nectars and jams – potassium
Method 49 J9 Fruit and vegetable juices, fruit nectars and jams – ash alkalinity
Method 50 J 10 Fruit and vegetable juices, fruit nectars and jams – Kjeldahl nitrogen, total protein
Method 51 J11 Fruit and vegetable juices, fruit nectars and jams – formol number
Method 52 J12 Fruit and vegetable juices, fruit nectars and jams – reducing sugars in jams
Method overview 6.6055.003 Food PAC Page 2
Card DIR 5 Method 53 K1 Beer, vinegar, spirits and wine – beer, pH value
Method 54 K2 Beer, vinegar, spirits and wine – beer, CO2 content
Method 55 K3 Beer, vinegar, spirits and wine – beer, ascorbic acid (vitamin C)
Method 56 K4 Beer, vinegar, spirits and wine – beer, total sulfurous acid
Method 57 K5 Beer, vinegar, spirits and wine – vinegar, pH value and total acidity
Method 58 K6 Beer, vinegar, spirits and wine – vinegar, volatile acidity
Method 59 K7 Beer, vinegar, spirits and wine – vinegar, ascorbic acid (vitamin C)
Method 60 K8 Beer, vinegar, spirits and wine – vinegar, free sulfurous acid
Method 61 K9 Beer, vinegar, spirits and wine – vinegar, total sulfurous acid
Method 62 K10 Beer, vinegar, spirits and wine – vinegar, ash alkalinity
Method 63 K11 Beer, vinegar, spirits and wine – vinegar, chloride
Method 64 K12 Beer, vinegar, spirits and wine – vinegar, sulfate
Method 65 K13 Beer, vinegar, spirits and wine – spirits, pH value and total acidity
Method 66 K14 Beer, vinegar, spirits and wine – spirits, volatile acidity
Method 67 K15 Beer, vinegar, spirits and wine – spirits, total esters
DIR 6 Method 68 L1 Coffee, cocoa and chocolate – pH value and degree of acidity (green and roasted coffee,
coffee extracts)
Method 69 L2 Coffee, cocoa and chocolate – ash alkalinity (green and roasted coffee, coffee extracts)
Method 70 L3 Coffee, cocoa and chocolate – chloride (green and roasted coffee)
Method 71 L4 Coffee, cocoa and chocolate – directly reducing sugars (roasted coffee, coffee extracts)
Method 72 L5 Coffee, cocoa and chocolate – pH value (cocoa and chocolate powder)
Method 73 L6 Coffee, cocoa and chocolate – ash alkalinity (cocoa and chocolate powder)
Method 74 L7 Coffee, cocoa and chocolate – Kjeldahl nitrogen (cocoa and chocolate powder)
Method 75 L8 Coffee, cocoa and chocolate – free fatty acids in cocoa butter
Method 76 L9 Coffee, cocoa and chocolate – iodine number of cocoa butter
Method 77 L10 Coffee, cocoa and chocolate – saponification number of cocoa butter
DIR 7 Method 78 M1 Artificial sweeteners, gelling and thickening agents – methoxy and ethoxy groups in
gelling and thickening agents
Method 79 M2 Artificial sweeteners, gelling and thickening agents – cyclamate in artificial sweeteners
Method 80 M3 Artificial sweeteners, gelling and thickening agents – saccharin in artificial sweeteners
Method 81 N1 Preserved fruits, vegetables and mushrooms – oxalic acid in fruits and vegetables
Method 82 N2 Preserved fruits, vegetables and mushrooms – total sulfurous acid in dried fruits and
vegetables
Method 83 N3 Preserved fruits, vegetables and mushrooms – cooking salt content of mushroom
extracts and concentrates
Method 84 O1 Cooking salt, spices, pickling salt, seasoning, herbal and flavored salts – chloride content
Method 85 O2 Cooking salt, spices, pickling salt, seasoning, herbal and flavored salts – total iodine
in cooking salt
Method 86 O3 Cooking salt, spices, pickling salt, seasoning, herbal and flavored salts – fluoride in
cooking salt
Method 87 O4 Cooking salt, spices, pickling salt, seasoning, herbal and flavored salts – tricalcium
phosphate in cooking salt
Method 88 O5 Cooking salt, spices, pickling salt, seasoning, herbal and flavored salts – nitrite in
pickling salt
DIR 8 Method 89 P1 Meat products, meat extracts, bouillon preparations, aspic, seasonings, soups, sauces
– chloride (NaCl) in meat products
Method 90 P2 Meat products, meat extracts, bouillon preparations, aspic, seasonings, soups, sauces
– Kjeldahl nitrogen and raw protein in meat products
– sulfurous acid in meat products
– chloride (NaCl) in meat extracts, aspic and bouillon
– Kjeldahl nitrogen in meat extracts, aspic and bouillon
– chloride (NaCl) in seasonings, soups, sauces
– Kjeldahl nitrogen in seasonings, soups, sauces
DIR 9 Method 96 Q Qualifying the titrator, validating a titration method

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Food Potentiometric Analysis Collection

Methods for the Titrimetric/Potentiometric Analysis

Important symbols used in the methods

A pH value H Soft drinks, lemonades

M Artificial sweeteners, gelling and thickening

N Preserved fruits, vegetables and mushrooms

O Cooking salt, spices, pickling salt,

P Meat products, meat extracts, bouillon

Q Qualifying the titrator, validating a titration

U1: Galvani potential between measuring electrode and measuring solution

The individual potentials U2, U3 and U4 are determined by the construction of

Temperature t / °C U / mV An ideal pH glass electrode has a

Temperature t / °C pH = 4.00 ±0.02 pH = 7.00 ±0.02 pH = 9.00 ±0.02

Instrument parameters and calculation Result

The pH value is extremely important for biological systems. It influences the

Instrument parameters and calculation Result

For aqueous solutions a temperature difference of 5 °C for a theoretical con-

** In Europe a dimensionless factor, with at least 4 decimal places (e.g. 1.0015).

Titer = C00 / C01 / EP1

Instrument parameters and calculation Result

• c(HCl) = 0.1 mol/L in H2O or in ethanol

Titer determination Tris(hydroxymethyl)-aminomethane (TRIS) is dried overnight in a drying oven

Instrument parameters and calculation Result

• c(I2) = 0.05 mol/L

Instrument parameters and calculation Result

• c(Na2S2O3) = 0.1 mol/L

Recommended • 6.0431.100 Pt Titrode with 6.2104.020 electrode cable

Instrument parameters and calculation Result

• c(KMnO4) = 0.02 mol/L

Recommended • 6.0431.100 Pt Titrode with 6.2104.020 electrode cable

Instrument parameters and calculation Result

• c(AgNO3) = 0.1 mol/L

Recommended • 6.0430.100 Ag Titrode with Ag2S coating, 6.2104.020 electrode cable

Instrument parameters and calculation Result

• c(Na2EDTA) = 0.1 mol/L

Recommended • 6.0508.110 or 6.0504.100 Ca ISE with 6.2104.020 electrode cable and

1 mL c(Na2EDTA) = 0.1 mol/L corresponds to 10.009 mg CaCO3 (C01)

Instrument parameters and calculation Result

C Drinking water and mineral water

C Drinking water and mineral water

Recommended • 6.0253.100 Aquatrode Plus with 6.2104.020 electrode cable

Analysis The Aquatrode has already been calibrated as per Method 1.

Calculations K A 8.2 = EP1* x C01 x C02 x C30

Instrument parameters and calculation

Titration curve Result

Instrument parameters and calculation – pH Result – pH

Instrument parameters and calculation – TIP Result – TIP

C Drinking water and mineral water

• 6.0508.110 or 6.0504.100 Ca ISE with 6.2104.020 electrode cable, and

Reagents • Titrant: c(Na2EDTA) = 0.1 mol/L (Method 9)

Calculations 1 mL c(Na2EDTA) = 0.1 mol/L corresponds to 4.008 mg Ca or 2.431 mg Mg

mmol/L Total hardness

Instrument parameters and calculation

Titration curve Result

Recommended • 6.0430.100 Ag Titrode with Ag2S coating, 6.2104.020 electrode cable

C Drinking water and mineral water

Calculation 1 mL c(AgNO3) = 0.01 mol/L corresponds to 0.3545 mg chloride

Instrument parameters and calculation Result

C Drinking water and mineral water