Basics of Titration

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Outline

 Overview
 Terminology and Definitions
 Basic Concept
 Titration Types and Calculation
 Reaction Types
 Automated Titration
 Tips and Hints
 Literature

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Outline

 Overview
 Terminology and Definitions
 Basic Concept
 Titration Types and Calculation
 Reaction Types
 Automated Titration
 Tips and Hints
 Literature

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Overview

 Titration is an analytical method which is widely used in many different

segments and for many different samples.

 Chemical industry: Water content in solvents, acid

value in resins and polymers, hydroxyl value in polyols,
surfactant content in raw materials, …
 Food and beverage: Acid and vitamin C content in fruit
juices, chloride content in spicy sauces, nitrite content in
food products, …
 Electroplating: Nickel content in electroless bath, gold
and silver content in alloys, chromium content in
electroplating baths, …

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Overview

 Pharmaceutical: Water contents in raw products, chondroitin sulfate sodium

in tablets, purity and content of different active pharmaceutical ingredients like
benzylnicotinate, clotrimazol and diclofenac sodium, …
 Petrochemical: Acid and base number in oils, mercaptane sulfur content in
kerosene, chloride content in motor oil, …
 Environmental: Total hardness of water, copper content in aqueous solution,
residual chlorine in bleach, fluoride content in water, …

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Outline

 Overview
 Terminology and Definitions
 Basic Concept
 Titration Types and Calculation
 Reaction Types
 Automated Titration
 Tips and Hints
 Literature

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Terminology and Definitions

 Qualitative chemical analysis: Determine the chemical composition in a

sample using chemical reactions and interactions.
Example: Test for halides with Beilstein test (green flame if halides are
present).
 Quantitative chemical analysis: Determine the content in a sample of one or
more chemical substances using chemical reactions and interactions.
Example: Vitamin C content in orange juice by titration.
 Sample: A small representative portion of the entity that should be analyzed.
Example: Orange juice
 Analyte: The chemical substance to be determined with the analysis.
Example: Vitamin C
 Matrix: Everything in the sample besides the analyte(s).
Example: Water, pulp, sugar, ... in vitamin C analysis of orange juice
 Titrant: Solution of known concentration which is added to the sample
(titration) and reacts with the analyte. The analyte content is calculated from
the consumption of the titrant.

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Outline

 Overview
 Terminology and Definitions
 Basic Concept
- Reaction
- Titrant
- Indication
- Titration curve
- Titration modes

 Titration Types and Calculation

 Reaction Types
 Automated Titration
 Tips and Hints
 Literature

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Basic Concept

 Titration is the determination of the quantity of a specific substance

(analyte) contained in a sample by controlled addition of a reagent
(titrant) of known concentration based on a complete chemical reaction
between the substance and the reagent.

 Example:
Determination of acetic acid (CH3COOH) by titration with sodium hydroxide
(NaOH)

CH3COOH + NaOH CH3COONa + H2O

(analyte) (titrant) (reaction products)

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Basic Concept

 Practical Procedure:
- The sample (e.g. vinegar) is diluted  sample solution with low pH value (= acidic).
- A NaOH solution of known concentration (titrant) is carefully added to the acidic
solution (titration).
- The analyte (CH3COOH) reacts with the titrant (NaOH)  pH rises slowly (from
acidic to basic)
- As soon as all analyte has reacted, the pH rises immediately (excess of NaOH in the
solution) equivalence point

 Content calculation:
-Sample size is known.
-Concentration of titrant is known.
-Titrant consumption (volume) until the equivalence point is measured during titration.
 The acetic acid content in the sample can be calculated from these three variables.

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Basic Concept

 Apparatus: The standard equipment for a manual titration

includes the following :
- Burette: for accurate titrant addition Burette
- Titration beaker
- Stirrer

 Several prerequisites have to be fulfilled in order to perform a

titration:
- Suitable reaction needed (reaction between the titrant and the
analyte)
- Titrant concentration and used volume must be known very
accurately
- Equivalence point must be detectable
Titration beaker

Stirrer

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Basic Concept – Reaction

 Selective:
The chemical reaction between the analyte and the titrant must be selective
(only the analyte should react with the titrant).

 Fast:
The reaction should be fast in order to guarantee that the added titrant reacts
immediately with the analyte.

 Complete:
The equilibrium of the reaction should lie strongly on the product side of the
reaction to guarantee a complete reaction.

 Unambiguous:
The stoichiometry of the reaction must be known and unambiguous.

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Basic Concept – Reaction

 For calculation of the result the equivalent number z has to be known.

 The equivalent number of an analyte is determined by the stoichiometry. In
general the equivalent number is the number of moles "monovalent" titrant
(e.g. NaOH, HCl, AgNO3, Na2S2O3) which are needed for the reaction with one
mole of analyte.
 For acid/base titrations the equivalent number is the number of protons (H+)
an acid releases by complete dissociation or the number of protons a base
gains by reaction with an acid.
 For redox titrations the equivalent number is given by the number of
electrons which are released or gained by a reductant or oxidant.
 Example 1: Sulphuric acid is a diprotic acid
H2SO4 + 2 NaOH Na2SO4 + 2 H2O 𝑧=2
 Example 2: For the reduction of Cr6+ to Cr3+ three electrons are needed
Cr6+ + 3 e- Cr3+ 𝑧=3

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Basic Concept – Reaction

 If a titrant is "polyvalent" the equivalent concentration (normality) has to be

used for calculation.
 The equivalent concentration of titrant A is calculated by the following
formula, where z is the equivalent number of the titrant:
𝑐(1/z A) = 𝑐(A) ∙ 𝑧
 Example 1: Sulphuric acid (c(H2SO4) = 0.05 mol/L) as titrant:
H2SO4 SO42- + 2 H+
mol mol
𝑐(1/2 H2 SO4 ) = 𝑐(H2 SO4 ) ∙ 𝑧 = 0.05 ∙ 2 = 0.1
L L
 Example 2: Potassium permanganate (c(KMnO4) = 0.02 mol/L) as titrant:

KMnO4 + 8 H+ + 5 e- Mn2+ + 4 H2O + K+

mol mol
𝑐(1/5 KMnO4 ) = 𝑐(KMnO4 ) ∙ 𝑧 = 0.02 ∙ 5 = 0.1
L L

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Basic Concept – Titrant

 To calculate the content of the analyte, the titrant concentration must be

known very accurately.
 To determine the exact titrant concentration, a titer determination is carried
out prior to the sample analysis.
 For the titer determination, a substance of high purity (titrimetric standard) is
titrated.
 Example: The titer of sodium hydroxide is determined by titrating potassium
hydrogen phthalate.

KOOC-C6H4-COOH + NaOH KOOC-C6H4-COONa + H2O

 The actual titrant concentration (cact) is the nominal concentration (cnom) times
the titer (t)
𝑐act = 𝑐nom ∙ 𝑡

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Basic Concept – Titrant

 The key point of a good titration is the accurate determination of the

volume of titrant used until the equivalence point.

 For this reason the following two requirements have to be fulfilled:

- A titrant addition in small quantities must be possible.
- An accurate reading for the volume used is needed.

 A manual burette fulfills these requirements

- Stopcock: add small volumes
- Grading: accurate volume reading

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Basic Concept – Indication

 The equivalence point (when all analyte has reacted with the titrant) of a
titration should be easily visible or measurable.
 The two basic indication principles are:
- Indicator
- Electrode

 Indicator:
- An indicator is a substance which changes its color at the equivalence point.
- Small amounts of an indicator are added to the solution prior to the analysis.

- Example:
Phenolphthalein is an indicator which is pink under basic conditions and colorless
under acidic conditions. At the equivalence point of an acetic acid titration by sodium
hydroxide, the color will immediately change from colorless to pink.

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Basic Concept – Indication

 Electrode:
- An electrode is an instrument that measures a specific property of a solution
electrochemically.
- A suitable electrode has to be chosen for each type of reaction. The electrode must
measure a property which is related to the titration reaction.
- The whole titration procedure can be followed with an electrode  titration curve

- Example:
For the titration of acetic acid by sodium hydroxide a pH electrode is used. The pH
electrode measures the H+ concentration related to acetic acid that hasn't reacted.
The equivalence point is reached when the pH value changes suddenly from acidic
to basic.

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Basic Concept – Titration curve

 If an electrode is used for a titration the signal vs. titrant volume can be
plotted.
 Such a plot is called a titration curve.
 The equivalence point can be determined directly from a titration curve and is
the inflection point of the curve.

 Example: Titration curve (pH vs. volume) of acetic acid titration with sodium
hydroxide. The equivalence point is clearly visible.

pH

Equivalence point

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Basic Concept – Titration modes

 Two different titration modes are used:

- Equivalence point titration (EQP)
- Endpoint titration (EP)

Signal
 EQP:
- The EQP is reached as soon as all analyte has
reacted with the titrant. EQP
- In a titration curve the EQP is the inflection point
of the curve.
- The titration is carried out over the EQP and
evaluated afterwards.
Volume
Signal
 EP:
- The EP is reached as soon as the signal reaches
a predefined value.
- The titration is usually stopped at the EP
EP

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Outline

 Overview
 Terminology and Definitions
 Basic Concept
 Titration Types and Calculation
- Direct titration
- Titer determination
- Blank compensated titration
- Back titration
 Reaction Types
 Automated Titration
 Tips and Hints
 Literature

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Titration Types and Calculation – Direct

 The most common and basic titration type is: direct titration.
 In a direct titration the analyte reacts directly with the titrant.
 Example: In the titration of acetic acid with sodium hydroxide the acetic acid
reacts directly with sodium hydroxide by exchanging a proton. The content of
acetic acid can be calculated directly from the used sodium hydroxide (titrant)
volume (consumption).

CH3COOH + NaOH CH3COONa + H2O

(analyte) (titrant) (reaction products)

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Titration Types and Calculation – Direct

 The analyte content in a direct titration is calculated from the titrant

consumption at the equivalence or end point using the following formula:

𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅=
𝑚

R: Result
VEQ: Titrant consumption at the equivalence or end point (in mL)
c: Titrant concentration (in mol/L)
t: Titrant titer (no unit)
C: Constant for unit conversion (unit part, unit is dependent on the result
needed)
m: Sample size (in g or mL)
 The constant C is dependent on the result unit wanted and on the unit of the
sample size.

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Titration Types and Calculation – Direct

 Examples of constants:
Sample size
g mL
entry unit
𝑀
% 𝐶=
10 ∙ 𝑧
𝑀
mg/g g/L 𝐶=
Result unit

𝑧
𝑀 ∙ 1000
ppm mg/L 𝐶=
𝑧
1000
mmol/kg mmol/L 𝐶=
𝑧
1
mol/kg mol/L 𝐶=
𝑧

M: Molar mass of the analyte (in g/mol)

z: Equivalent number (no unit)

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Titration Types and Calculation – Direct

 Example: For the titration of the acetic acid (M = 60.04 g/mol) in a 1 g sample
of vinegar, 5 mL sodium hydroxide (c = 0.1 mol/L, titer: 0.9900) were
consumed until the equivalence point. To calculate the acetic acid content in %
you have to use the following formula:

𝑀 60.04 g/mol g∙%

𝐶= = mg = 6.004
10 ∙ 𝑧 10 g ∙ % ∙ 1 mmol

mol g∙%
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 5 mL ∙ 0.1 ∙ 0.9900 ∙ 6.004
𝑅= = L mmol = 2.97 %
𝑚 1g

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Titration Types and Calculation – Titer

 The titer is usually determined by a direct titration, but the calculation is a little
different.
 The titer (t) is defined as the actual concentration (cact) divided by the nominal
concentration (cnom) of a titrant:
𝑐act
𝑡=
𝑐nom

 The following formula can be used to calculate the titer of a titrant, if this titrant
was used to titrate a titer standard by a direct titration:
𝑚
𝑡=
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝐶
t: Titrant titer (no unit)
m: Titer standard sample size (in g or mL)
VEQ: Titrant consumption at the equivalence or end point (in mL)
c: (Nominal) titrant concentration (in mol/L)
C: Constant for unit conversion (unit part, unit is dependent on the titer
substance form)
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Titration Types and Calculation – Titer

 If the titer standard substance is a solid, use the following formula for the
constant C:
𝑀
𝐶=
10 ∙ 𝑝 ∙ 𝑧

 For liquid titer standards the following constant is used:

1
𝐶=
𝑐𝑠𝑡 ∙ 𝑧

M: Molar mass of the analyte (in g/mol)

p: Purity of the solid titer standard (in %)
z: Equivalent number of the titer standard (no unit)
cst: Concentration of the liquid titer standard (in mol/L)

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Titration Types and Calculation – Titer

 Example: Potassium hydrogen phthalate (KHP, M= 204.23 g/mol) was used

as a solid titer standard to determine the titer of sodium hydroxide (c = 0.1
mol/L). 0.0931 g of KHP (purity: 99 %) was weighed into a titration beaker
and diluted with deionized water. This solution was titrated with sodium
hydroxide. The equivalence point was detected at a titrant consumption of
4.5238 mL. For this measurement the titer can be calculated as follows:

g
𝑀 204.23 g
𝐶= = mol = 0.206
10 ∙ 𝑝 ∙ 𝑧 10 mg ∙ 99 % ∙ 1 mmol
g∙%
𝑚 0.0931 g
𝑅= = = 0.9990
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝐶 4.5238 mL ∙ 0.1 mol ∙ 0.206 g
L mmol

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Titration Types and Calculation – Blank

 In some titrations it may happen that the solvent itself also reacts with the
titrant. The amount of titrant used for the solvent is called the blank value.
 The blank value has to be compensated to get the correct result.
 To determine the blank value a titration of the solvent without any sample
has to be performed. The blank value is the volume of the titrant used until the
equivalence or endpoint is reached.
 For a titration where a blank value is used, take care to always use the same
solvent volume!

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Titration Types and Calculation – Blank

 Example: The acid number of motor oil is determined by a non-aqueous

acid/base titration with potassium hydroxide (in 2-propanol) as titrant. As
solvent a mixture of toluene, 2-propanol and water is used. Before titrating
the sample, the solvent mixture without any sample is titrated. The titrant
consumption of this blank measurement is compensated in the calculation of
the sample measurement.

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Titration Types and Calculation – Blank

 The calculation of a blank value compensated titration is similar to the direct

titration:
(𝑉𝐸𝑄 − 𝐵) ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅=
𝑚

R: Result
VEQ: Used titrant volume until the equivalence or end point (in mL)
B: Blank value, titrant volume used for the titration of the solvent (in mL)
c: Titrant concentration (in mol/L)
t: Titrant titer (no unit)
C: Constant for unit conversion (unit part, unit is depending on the result
needed)
m: Sample size (in g or mL)
 The constant C is the same as for the direct titration

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Titration Types and Calculation – Back

 In a back titration a known excess amount of reagent (titrant A with known

concentration) is added to the sample. This titrant reacts with the analyte.
Afterwards the amount of non-reacted titrant A will be titrated with a second
titrant (titrant B).
 The added volume of titrant A has to be known very precisely.
 For accurate measurements a so called back value is determined by titrating
the added volume of titrant A without any sample. The result is based on the
used volume of titrant B without any sample.
 Such titrations are often used when the reaction between the analyte and the
first titrant is slow.

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Titration Types and Calculation – Back

 Example: For the titration of nitrite in soya sauce a known volume of

potassium permanganate (titrant A) is added to the sample. Potassium
permanganate will react with the nitrite and the excess of the permanganate is
titrated with ammonium ferrous sulfate(II) (titrant B).

5 NO2- + 2 MnO4- + 6 H+ 2 Mn2+ + 5 NO3- + 3 H2O

(analyte) (titrant A)
MnO4- + 5 Fe2+ + 8 H+ Mn2+ + 5 Fe3+ + 4 H2O
(titrant A) (titrant B)

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Titration Types and Calculation – Back

 The analyte content in a back titration is calculated using the following formula:
(𝐵𝑘 − 𝑉𝐸𝑄) ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅=
𝑝∙𝑚
R: Result
Bk: Back value, volume of titrant B used for the titration of titrant A without
sample (in mL)
VEQ: Volume of titrant B used to back titrate the excess of titrant A after
sample addition (in mL)
c: Concentration of titrant B (in mol/L)
t: Titer of titrant B (no unit)
C: Constant for unit conversion (unit part, unit is depending on the result
needed)
p: Stoichiometric proportion between titrant A and titrant B of the second
reaction, stoichiometric factor of titrant B divided by factor of titrant A (no
unit)
m: Sample size (in g or mL)
 The constant C is the same as for the direct titration
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Outline

 Overview
 Terminology and Definitions
 Basic Concept
 Titration Types and Calculation
 Reaction Types
- Acid / Base
- Precipitation
- Redox
- Karl Fischer
- Complexometric
 Automated Titration
 Tips and Hints
 Literature
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Reaction Types – Acid / Base

 Reaction:
- This type of titration is based on the reaction between an acid (AH) and a base (B),
where a proton (H+) is exchanged.
AH + B A- + BH+
- If the analyte is an acid the titrant will be a base and vice versa.
 Analytes:
- Weak or strong acid / base
- Examples: Hydrochloric acid, sulphuric acid, acetic acid, sodium hydroxide,
ammonia, …
 Titrants:
- Strong base / acid solution
- Examples: Sodium hydroxide 0.1 mol/L, hydrochloric acid 0.1 mol/L
 Indication:
- pH indicators: pH dependent dyes (for manual titration), examples: phenolphthalein,
methyl orange, …
- pH sensor (for automated titration)

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Reaction Types – Acid / Base

 The parameter which changes during a acid / base titration is the pH value.
 The pH value is defined as the negative decadic logarithm of the H+ (or H3O+)
concentration in a solution:

𝑝𝐻 = − log 𝑐 H + = − log 𝑐 H3 O+

 Even in pure deionized or distilled water a small amount of ions is present due
to the amphoteric properties of water (it can react as a acid or a base). Water
molecules react with each other to build H3O+ and OH-:

H2O + H2O H3O+ + OH-

 In pure deionized water the concentration of H3O+ and OH- ions are both 10-7
mol/L. The product of these two concentrations is called the ionic product of
water (Kw) and this value is a constant:
mol mol 𝑚𝑜𝑙 2
𝐾w = 𝑐 H3 O+ ∙𝑐 OH − = 10−7 ∙ 10−7
= 10−14
L L 𝐿2
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Reaction Types – Acid / Base

 If we calculate the pH value for pure water (c(H3O+) = 10-7 mol/L) it is 7.

A pH value of 7 is neutral.
 If an acid (AH) is added to an aqueous solution, it will react (dissociation) with
water to form more H3O+ ions (and less OH- ions due to Kw):

AH + H2O A- + H3O+

The H3O+ concentration will increase and the pH value will decrease.
A pH value below 7 is acidic.
 If a base (B) is added to an aqueous solution, it will react with water to form
more OH- ions (and less H3O+ ions due to Kw):

B + H2O BH+ + OH-

The H3O+ concentration will decrease and the pH value will increase.
A pH value above 7 is alkaline.

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Reaction Types – Acid / Base

Orange juice
Egg white
Coca Cola
Cheese Water Antacida ( Mg(OH)2 )
Lemon juice Milk
Beer Borax

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
acidic alkaline
Sulfuric acid Acetic acid Ammonia solution Caustic
4.9 % (1 M) 0.6% (0.1 M) 0.017% (0.01 M) soda 4%

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Reaction Types – Acid / Base

 For a manual titration various pH indicators exist. The selection must be based
on the pH range where the equivalence point occurs: the indicator has to be
chosen such that its color change occurs at the equivalence point.
 The following graph shows common pH indicators and their color change
range:
Thymol blue

Congo red

Methyl orange

Bromcresol green

Methyl red

Bromthymol blue

Phenol red

Phenolphthalein

Thymolphthalein

0 2 4 6 8 10 12
pH
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 Reaction Types – Acid / Base

 For automated titration the pH value is measured by a pH sensor (usually

combined sensor), which consists of a reference system (usually Ag/AgCl) and
a glass membrane which is sensitive to H+ (or H3O+).
 Since the measured signal is a potential (mV) and not pH, the sensor has to
be calibrated (correlate the mV signal to a pH value).
 The calibration is done by measuring the potential of various calibration buffers
with defined pH value. From these measurements (calibration points) a
calibration line is calculated.
 Theoretically (at 25°C) the slope of the calibration line is -59.16 mV/pH and
the zero point is pH 7.00.
E [mV]
zero point
hydrated layer membrane glass

0 pH
1 14
slope

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Reaction Types – Acid / Base

Example 1: Titration of acetic acid in vinegar with sodium hydroxide

 The acetic acid concentration of a vinegar sample should be determined.
Since acetic acid is an acid this will be done by titrating with sodium hydroxide.
The reaction for this titration is
CH3COOH + NaOH CH3COONa + H2O

 For this sample an acetic acid content of 5 % is expected. From this value one
can calculate the sample size by re-arranging the calculation formula so that
the sample size can be calculated
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅= → 𝑚=
𝑚 𝑅
 If we use a 20 mL burette, a titrant consumption of about 10 mL (half of burette
volume) would be perfect. We use a sodium hydroxide solution with c = 0.1
mol/L and assume a titer of 1. Form these values the approximate sample size
can be calculated.

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Reaction Types – Acid / Base

𝑀 60.04 g/mol g∙%

𝐶= = mg = 6.004
10 ∙ 𝑧 10 g ∙ % ∙ 1 mmol

mol g∙%
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 10 mL ∙ 0.1 ∙ 1 ∙ 6.004
𝑚= = L mmol = 1.2 g
𝑅 5%

 Based on this calculation 1.3134 g of vinegar was weighed on an analytical

balance into a titration beaker and diluted in 50 mL deionized water. This
solution was titrated with sodium hydroxide (c = 0.1 mol/L and t = 0.9877) on
an automated titrator equipped with a pH sensor until the equivalence point
was recognized.

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Reaction Types – Acid / Base
Titration Curve
12

10

8
pH

2
0 2 4 6 8 10
V [mL]
VEQ

 From the titrant consumption (8.3784 mL) the content of acetic acid in the
sample was calculated:
mol g∙%
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 8.3784 mL ∙ 0.1 ∙ 0.9877 ∙ 6.004
𝑅= = L mmol = 3.78 %
𝑚 1.3134 g

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Reaction Types – Acid / Base

Example 2: p-value of tap water

 The p-value (or total acidity) of water indicates how much mmol of sodium
hydroxide is needed to neutralize 1 L water until the color from
phenolphthalein (color change range: pH 8.0 – 9.6) indicator changes.
 To perform this titration with an automated titrator an endpoint (EP) titration to
pH 8.2 is used. Before performing the endpoint titration the pH sensor is
calibrated using three pH buffer solutions: pH 4.01, 7.00 and 9.21.
 100 mL of tap water must be accurately pipetted into a titration beaker and
titrated with sodium hydroxide (c = 0.01 mol/L and t = 0.9887) to pH 8.2. The
consumed titrant until the endpoint is e.g. 2.3554 mL.
 Using this value the following p-value is calculated:
mL
1000 1000 L mL
𝐶= = = 1000
𝑧 1 L
mol mL
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 2.3554 mL ∙ 0.01 ∙ 0.9887 ∙ 1000
𝑅= = L L = 0.233 mmol
𝑚 100 mL L
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Reaction Types – Acid / Base

Example 3: Simultaneous titration of sodium hydroxide and carbonate

 If a sodium hydroxide solution is not closed and not protected against carbon
dioxide intake, carbon dioxide will be dissolved and will react to carbonate. If
one performs a titration, this carbonate will also be titrated.
 During the titration three different bases are present: sodium hydroxide
(NaOH), sodium carbonate (Na2CO3) and sodium hydrogen carbonate
(NaHCO3). Since NaOH is the strongest base this species will react first with
the titrant (hydrochloric acid, HCl). Afterwards Na2CO3 will react to NaHCO3
and this base will react at the end to CO2.

1. NaOH + HCl NaCl + H2O

2. Na2CO3 + HCl NaHCO3 + NaCl
3. NaHCO3 + HCl CO2 + NaCl + H2O

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Reaction Types – Acid / Base

 Theoretically, we should observe three equivalence points but the first point
will be very flat and only two points will be clearly visible:
- 1. EQP: Reaction of NaOH and Na2CO3
- 2. EQP: Reaction of NaHCO3

 From these two EQP's the NaOH and Na2CO3 concentrations can be
determined.

 To perform this titration 7 mL of sodium hydroxide sample is added with a

accurate pipette into a titration beaker and the solution is diluted with 50 mL of
deionized water. This solution is titrated with hydrochloric acid (c = 0.1 mol/L
and t = 0.9932) until two equivalence points are recognized (automatic titrator
with pH sensor).

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Reaction Types – Acid / Base

Titration Curve

12

10

8
pH

2
0 2 4 6 8 10 12 14 16
V [mL] VEQ1 VEQ2

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Reaction Types – Acid / Base

 In the titration curve the two equivalence points at VEQ1 = 10.6847 mL and
VEQ2 = 14.0042 mL are clearly visible.
 The titrant consumption from the first to the second EQP value corresponds to
the amount of NaHCO3 that reacted to CO2. From the consumption between
these two EQP's the Na2CO3 concentration can be calculated directly:

𝑉𝐸𝑄 = 𝑉𝐸𝑄2 − 𝑉𝐸𝑄1 = 14.0042 mL − 10.6847 mL = 3.3195 mL

1 1
𝐶= = =1
𝑧 1

mol
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 3.3195 mL ∙ 0.1 L ∙ 0.9932 ∙ 1 mol
𝑅= = = 0.047
𝑚 7 mL L

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Reaction Types – Acid / Base

 The NaOH concentration can be calculated from the first EQP, but the Na2CO3
reaction to NaHCO3 is also included. To get the correct titrant consumption,
the titrant volume for the Na2CO3 reaction has to be subtracted. This volume is
exactly the same (difference between the first and the second EQP) as
calculated before.

𝑉𝐸𝑄 = 𝑉𝐸𝑄1 − 𝑉𝐸𝑄2 − 𝑉𝐸𝑄1 = 10.6847 mL − (14.0042 mL − 10.6847 mL)

= 7.3652 mL
1 1
𝐶= = =1
𝑧 1
mol
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 7.3652 mL ∙ 0.1 L ∙ 0.9932 ∙ 1 mol
𝑅= = = 0.105
𝑚 7 mL L

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Reaction Types – Precipitation

 Reaction:
- The analyte (A) and the titrant (T) react and form a compound which precipitates.

A + T AT
- Usually silver nitrate (AgNO3) is used as a titrant, which forms hardly soluble silver
salts (e.g. AgCl)
 Analytes:
- Ions which form hardly soluble silver salts
- Examples: chloride, cyanide, thiocyanate
 Titrants:
- Silver nitrate
 Indication:
- Indicators: Potassium dichromate (for manual titration)
- Silver ring electrode (for automated titration)

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Reaction Types – Precipitation

Example: Titer determination of silver nitrate

 The titer of silver nitrate (AgNO3) is determined by sodium chloride (NaCl) as
standard substance.

NaCl + AgNO3 AgCl + NaNO3

 To determine the titer, 43.2 mg of NaCl standard (purity: 99.5 %) is weighed

into a titration beaker and dissolved in 50 mL nitric acid (c = 0.02 mol/L). The
nitric acid is needed to keep a pH value of 4.5 in the sample solution, since the
precipitation reaction takes place under slightly acidic conditions.
 The sample solution is titrated with AgNO3 (c = 0.1 mol/L) while the signal is
measured with a silver ring sensor (measures Ag+ concentration). The titration
is stopped as soon as the equivalence point is recognized.
 During the titration the precipitation of AgCl is clearly visible.

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Reaction Types – Precipitation

Titration Curve
200

150

100
E [mV]

50

-50

-100
0 1 2 3 4 5 6 7 8 9 10
V [mL] VEQ

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Reaction Types – Precipitation

 The equivalence point is clearly visible and the titrant consumption at this point
is 7.3796 mL. From this value the titer of the silver nitrate titrant solution can
be calculated as follows:
g
𝑀 58.44 g
𝐶= = mol = 0.059
10 ∙ 𝑝 ∙ 𝑧 10 mg ∙ 99.5 % ∙ 1 mmol
g∙%

𝑚 0.0432 g
𝑅= = = 0.9922
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝐶 7.3796 mL ∙ 0.1 mol ∙ 0.059 g
L mmol

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Reaction Types – Redox

 Reaction:
- The analyte and the titrant exchange electrons during this reaction and the oxidation
state of both changes.
- If the analyte is an oxidizing agent (O) the titrant is a reducing agent (R) and vice
versa. Oxidation: R R+ + e-
Reduction: O + e- O-

Redox reaction: R + O O- + R+
 Analytes:
- Oxidizing and reducing agents
- Examples: Sulphurdioxide, Nitrite, Iron, …
 Titrants:
- Oxidizing and reducing agents
- Examples: Iodine, Sodium thiosulfate, potassium permanganate, …
 Indication:
- Color change due to colored titrant, starch solution (for Iodine)
- Platinum sensor (for automated titration)
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Reaction Types – Redox

Example 1: Titer determination of potassium permanganate

 The titer of potassium permanganate (KMnO4) is determined by using Di-
sodium oxalate (Na2C2O4) as the standard. During this titration Na2C2O4 is
oxidized to CO2 and KMnO4 is reduced to Mn2+:

Oxidation: Na2C2O4 2 CO2 + 2 Na+ + 2 e-

Reduction: KMnO4 + 8 H+ + 5 e- Mn2+ + 4 H2O + K+

Redox reaction:
5 Na2C2O4 + 2 KMnO4 + 16 H+ 10 CO2 + 10 Na+ + 2 Mn2+ + 8 H2O + 2 K+

 Since two electrons are released by the oxidation of Na2C2O4 the equivalent
number of this substance is z = 2.
 Since 5 electrons are needed for the reduction of the titrant KMnO4 the
concentration of the titrant will be given as c(1/5 KMnO4) which is five times
higher than the concentration c(KMnO4).
𝑐 = 𝑐 1/5 KMnO4 = 𝑐 KMnO4 ∙ 5
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Reaction Types – Redox

 To perform this titration 32.4 mg of Na2C2O4 (purity: 99 %) were weighed into a

titration beaker using an analytical balance. The standard was dissolved by
adding 50 mL of 5 % sulphuric acid solution. The sulphuric acid is needed
since this reaction needs strong acidic conditions (see H+ in the reaction).
 The sample solution was titrated with KMnO4 solution (c(1/5 KMnO4) = 0.1
mol/L) until the equivalence point was detected. A combined platinum ring
electrode was used to measure the signal during the titration.
 The titrant consumption (4.8196 mL) from the titration curve was used to
calculate the titer of the KMnO4 titrant solution:
g
𝑀 134.01 g
𝐶= = mol = 0.068
10 ∙ 𝑝 ∙ 𝑧 10 mg ∙ 99 % ∙ 2 mmol
g∙%
𝑚 0.0324 g
𝑅= = = 0.9886
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝐶 4.8196 mL ∙ 0.1 mol ∙ 0.068 g
L mmol

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Reaction Types – Redox

Example 2: Iodometric determination of Cr6+

 Cr6+ should be determined in an electroplating bath. This is done in a two step
reaction. First Cr6+ is reduced to Cr3+ using iodide (I-) which is oxidized to
iodine (I2). The iodine produced is then titrated with sodium thiosulfate
(Na2S2O3):
1. 2 Cr6+ + 6 I- 2 Cr3+ + 2 I2
2. 2 Na2S2O3 + I2 Na2S4O6 + 2 I-

 Since three electrons are needed to reduce Cr6+ to Cr3+ the equivalent number
is z = 3.
 5 mL of a 1:100 diluted bath sample (m = 0.05 mL) is added into a titration
beaker. The sample is diluted in
- 5 mL of 50 % H2SO4 solution to acidify the solution
- 40 mL deionised water.
 To have an excess of iodide in the solution 5 mL of 10 % potassium iodide (KI)
solution was added. This iodide reacts with the Cr6+ and a corresponding
amount of iodine is produced.
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Reaction Types – Redox

 After mixing the sample solution it is finally titrated with Na2S2O3 (c = 0.1
mol/L, t = 0.9954) until the equivalence point is reached using an automated
titrator equipped with a platinum ring electrode.

 The Cr6+ content in the bath is calculated from the titrant consumption at the
equivalence point (VEQ = 4.2612 mL):

g
𝑀 52.00
𝐶= = mol = 17.33 g
𝑧 3 mol

mol g
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 4.2612 mL ∙ 0.1 ∙ 0.9954 ∙ 17.33
𝑅= = L mol = 147.01 g
𝑚 0.05 mL L

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Reaction Types – Karl Fischer

 Reaction:
- Karl Fischer titration is a special type of redox titration for water determination.

ROH + SO2 + 3 R'N + I2 + H2O (R'NH)·SO4R + 2 (R'NH)I

- The Karl Fischer reaction needs an alcohol (ROH, usually methanol), sulfur dioxide,
a base (imidazole or pyridine), iodine and the analyte water.
 Analyte: Water
 Titrants and Solvents:
- One-component reagent: contains iodine, sulfur dioxide and a base dissolved in
methanol. The solvent is methanol.
- Two-component reagent: Contains iodine dissolved in methanol. The solvent
contains sulfur dioxide, a base and methanol.
 Indication:
- Polarized double pin platinum electrode (voltammetry)

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Reaction Types – Karl Fischer

 The Karl Fischer reaction is a two step reaction:

1. ROH + SO2 + R'N (R'NH)·SO3R
2. (R'NH)·SO3R + I2 + H2O + 2 R'N (R'NH)·SO4R + 2 (R'NH)I
 From this reaction it can be seen that water reacts with iodine attaining 1:1
stoichiometry.
 The solvent ROH (usually methanol) is also involved in the reaction, this
means that at least 50 % of the solvent should always be methanol.
 A suitable base (R'N, usually imidazole, in the past: pyridine) keeps the pH
value between 5 and 7. This is the optimal pH range so that the reaction is fast
and yet no side reactions occur.

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 Reaction Types – Karl Fischer

 Karl Fischer titration needs a special setup: The titration vessel has to be
protected from humidity as it has an influence on the result. For this reason
Karl Fischer titrations are almost always done using an automatic titrator.
 A typical setup for a Karl Fischer titration is shown below:

Dosing tube for titrant Sensor

Molecular sieve

Glass vessel
Burette filled with titrant

Solvent
Magnetic stirrer

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Reaction Types – Karl Fischer

 Karl Fischer titration is an endpoint titration.

 The endpoint is detected with a polarized double platinum pin sensor. A fixed
polarization current (Ipol) is applied to this sensor and the voltage (U in mV),
which is used to maintain this current is measured.

 During the titration:

The iodine from the titrant reacts directly with
the water
 No free iodine in the solution
 High voltage is needed to maintain the
polarization current

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Reaction Types – Karl Fischer

 At the end of the titration:

No water is available for the reaction
with iodine
 Free iodine in the solution causes
ionic conduction
 Low voltage is needed, signal drop

E
 As soon as the voltage signaldrops below a
defined endpoint the titration stops

t
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Reaction Types – Karl Fischer

 A small amount of water will always enter the titration vessel, even in a tightly
closed titration vessel which is protected against humidity. This amount is
given by the drift value (usually in µg/min).
 The drift value is used in the calculation of the sample water content to
compensate the water which has entered the titration during the sample
measurement:
𝑡𝑖𝑚𝑒 ∙ 𝑑𝑟𝑖𝑓𝑡
𝑉𝐸𝑄 ∙ 𝑐 − ∙𝐶
1000
𝑅=
𝑚
R: Result, water content
VEQ: Titrant consumption (in mL)
c: Titrant concentration (in mg/mL)
time: Titration time (in min)
drift: Drift value (in µg/min)
C: Constant (dependent on the unit)
m: Sample size (in g)

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Reaction Types – Karl Fischer

 Since the concentration of the titrant is given in µg/mL, the C constant contains
no molar mass and is only used to change the unit of the water content:

% 𝐶 = 0.1
Result unit

ppm 𝐶 = 1000

mg/g 𝐶=1

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Reaction Types – Karl Fischer

 The titrant concentration is given in µg water that reacts with one mL of titrant.
The concentration has to be determined by titrating a water standard:

𝑐𝑠𝑡 ∙ 𝑚
𝑐=
𝑑𝑟𝑖𝑓𝑡 ∙ 𝑡𝑖𝑚𝑒
𝑉𝐸𝑄 −
𝑐(𝑜𝑙𝑑) ∙ 1000

c: Titrant concentration (in mg/mL)

cst: Water content of the water standard (in mg/g)
m: Sample size (in g)
VEQ: Titrant consumption (in mL)
drift: Drift value (in µg/min)
time: Titration time (in min)
c(old): Old titrant concentration that was used to determine the drift value (in
mg/mL)

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Reaction Types – Karl Fischer

 Due to the closed titration vessel the measurement procedure of a Karl Fischer
titration is a little different. Multiple samples can be measured in the same
titration vessel, using the same solvent:
 Pretitration: Pretitration
- The water which is present in the solvent is titrated.
 Standby: Standby
- The water which enters the titration vessel is constantly
titrated. The titrator is ready for the analysis.
 Inject sample Inject sample

 Sample titration:
- Water from the sample is titrated, the content is determined. Sample titration
Once the determination is finished, the titrator goes back into
standby mode and is ready for the next sample.
 Stop analysis: Stop analysis
- Occurs if all samples have been run, the titration vessel is full,
the solvent is contaminated or after a predefined number of samples.
- The solvent is then changed and the procedure starts again (pretitration).
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Reaction Types – Karl Fischer

Example: Water content determination in ethanol

 The titration is started. Once the pretitration is done the titrator goes into
standby mode. A stable and low drift value (typically < 25 µg/min) is awaited.
 The drift value read just before the sample determination starts is noted.
 Since ethanol is completely soluble in methanol, the sample can be directly
inserted into the titration vessel. For this purpose about 5 mL of ethanol is
drawn up into a syringe, placed on a balance and tared.
 The sample determination is started (drift value: d = 11.3 µg/min, titrant
concentration: c = 4.8976 mg/mL). About 1 mL of ethanol is added with the
syringe into the titration beaker. The syringe is once again placed on the
balance. The sample size is the difference between the weight of the syringe
before and after the sample addition (back weighing): 1.0348 g

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Reaction Types – Karl Fischer

 The titration ends (endpoint was set to 100 mV) after 2.1865 mL and a titration
time of 2.293 min.

Titration Curve

600

400
E [mV]

EP
200

0
0 20 40 60 80 100 120 140
t [s]

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Reaction Types – Karl Fischer

 The water content in ethanol is determined using the following calculation:

𝑡𝑖𝑚𝑒 ∙ 𝑑𝑟𝑖𝑓𝑡
𝑉𝐸𝑄 ∙ 𝑐 − 1000 ∙𝐶
𝑅=
𝑚
μg
mg 2.293 min ∙ 11.3
2.1865 mL ∙ 4.8976 mL − min ∙1
μg
1000 mg
=
1.0348 g
mg
= 10.323
g

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Reaction Types – Complexometric

 Reaction:
- Analyte (A) and titrant (T) react together to build a complex ([AT]).

A + T [AT]

 Analytes:
- Metal ions with a charge > 1
- Examples: Zinc ion, magnesium ion, calcium ion, …
 Titrants:
- Ligands which build complexes with the analyte
- Example: Ethylenediaminetetraacetic acid (EDTA)
 Indication:
- Indicators (also ligands) that change color if they are not bound to the analyte,
examples: Murexide, Eriochrome Black T
- Indicators and a photometric sensor (for automated titration)

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Reaction Types – Complexometric

 In a manual titration the color change of the indicator is observed visually with
the naked eye.
 In an automated titration an indicator is also added, but the color change is
observed by a photometric sensor.

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Reaction Types – Complexometric

Example: Total hardness of water

 Total hardness of water is expressed as the CaCO3 (M = 100.09 g/mol, z = 1)
content in water. It is a typical analysis done as a complexometric titration of
calcium and magnesium at pH 10.
 50 mL of tap water, 10 mL of 5 % ammonia solution (for pH 10), and 1 mL of 1
% Eriochrome Black T indicator solution were added into a titration beaker.
This solution was titrated by EDTA (c = 0.1 mol/L, t = 0.9763) and the color
change, from violet to blue, was observed with a photometric sensor at pH 10.
The equivalence point was found at a titrant consumption of 0.7392 mL.
 The hardness (as CaCO3 content) can be calculated as follows:
g mg
𝑀 ∙ 1000 100.09 ∙ 1000 g mg
mol
𝐶= = = 100090
𝑧 1 mol
mol mg
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 0.7392 mL ∙ 0.1 ∙ 0.9763 ∙ 100090
𝑅= = L mol = 144.47 ppm
𝑚 50 mL

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Outline

 Overview
 Terminology and Definitions
 Basic Concept
 Titration Types and Calculation
 Reaction Types
 Automated Titration
 Tips and Hints
 Literature

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Automated Titration

 An automatic titrator adds the titrant automatically while measuring the signal
with the sensor. No operator interaction is needed for the titration itself.
 Comparison between manual and automated titration:

Manual Automated
Titration Titration
Burette Graduated glass Glass cylinder
burette
Indication Visual (Indicators) Sensor
Titration addition Manual Automatically
controlled
Data recorded Only used titrant Titration curve
volume

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Automated Titration

 Accurate and precise results - due to the automatic titrant addition and the
possibility of adding very small amounts of titrant (lower than 0.001 mL).

 Fast but still precise titrations - due to the controlled titrant addition determined
by an algorithm (small increments near the equivalence point).

 Highly reproducible results - since no operator interaction is needed the

results are highly reproducible even if another operator performs the analysis.

 Full titration curve is recorded, which allows a titration to the equivalence

point and post-processing.

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Automated Titration

 The results can be calculated and printed automatically by the titrator.

 Traceability - the titration curve and the results can be stored (e.g. on a
computer if the titrator is connected) or printed directly.

 Operator has minimal contact with chemicals since the titrant and the
reagents can be added directly from the bottle.

 Depending on the titrator, the analysis can be fully automated thus saving
time.

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Outline

 Overview
 Terminology and Definitions
 Basic Concept
 Titration Types and Calculation
 Reaction Types
 Automated Titration
 Tips and Hints
 Literature

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Tips and Hints

 Titration beakers
- Use absolutely clean titration beakers.
- Clean titration beakers in a dishwasher and rinse at least 2 times
with deionised water.
- The material of the beakers (plastic or glass) should be resistant
to all chemicals used for the titration.
- If the analytes are sensitive to light use special beakers which
protect the sample from UV radiation.

 Burette
- Choose the appropriate burette size. The titrant consumption
should be within 20 % to 90 % of the burette volume for an
accurate measurement
- Estimate the burette size by calculating the titrant consumption
from the expected value or perform a fast test titration.
- Avoid bubbles in the burette or remove them before starting the
titration (rinsing) as they will produce results that are too high. De-
gas the titrant or reduce the titration speed if bubbles start coming
from the titrant.
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Tips and Hints

 Tubes (automated titration)

- Remove all bubbles from the tubes (or the results will be too high) by tapping
gently during rinsing.
- Use a siphon to avoid diffusion of titrant into the titration solution.

 Indicator (manual titration)

- Choose an indicator with a color change range within the range of the
equivalence jump.
- Don't add too much indicator; the indicator may influence the analyte/titrant
reaction. Just a few drops of indicator solution usually suffice.

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Tips and Hints

 Sensor (automated titration)

- Store and maintain the sensor as described in the operating
instructions.
- Ensure that the electrolyte level is high enough, namely
higher than internal elements and titration solution
- Poor sensor response results in titrant consumption and
results that are too high.
- Perform a sensor test periodically to check slope, offset and
drift value of sensors which need calibrating (pH sensor).
- Perform a sensor calibration, especially before endpoint
titrations and after every maintenance action.
- Some sensors need conditioning (see operating
instructions) before they can be used or after they have not
been used for a long time.
- Reference electrodes (which are also part of a combined
electrode) need to be in contact with the sample solution by a
junction (diaphragm). Avoid junction blockage for correct and
stable results. When required clean the junction using an
appropriate solution (see operating instructions).

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Tips and Hints

 Sample solution
- The overall sample solution (sample, reagents and solvent) should be enough to
cover the sensor, typically 50 mL. Add solvent (water for aqueous titrations) if the
volume is too small.
- The amount of sample should be chosen such that the titrant consumption is within
20 % to 90 % of the burette volume.
- Use accurate measuring tools (analytical balances, pipettes, …) to measure the
sample size.
- Perform a blank value determination if the solvent isn't pure.
- Depending on the titration reaction (e.g. redox and precipitation titration) make sure
that the pH value of the sample solution is within the requested range.

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Tips and Hints

 Titrant
- Perform a titer determination to know the titrant concentration accurately.
- Use high purity, stable, non-hygroscopic substances with a high molecular weight
(for a high sample size) as the titer standard substance.
- A titer determination interval (usable life) and a lifespan should be defined
depending on the stability of the titrant.
- Protect titrant from CO2, UV radiation, humidity or other factors which may have an
influence on the concentration.

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 METTLER TOLEDO

Basics of Titration

Definition

■ Quantitative chemical determination

of a compound in a sample….

….by the controlled addition of a

reagent (titrant) of known
concentration

2
Conditions for Reaction (1)

■ Simple and instantaneous

■ The endpoint (equivalence point) must be clear
„ stoichiometric

Conditions for Reaction (2)

■ Known titrant concentration - titer value

■ Observable end point

4
Titration Techniques (1)

■ Manual

Titration Techniques (2)

■ Automatic

Indicator
Burette (sensor)

Titrant

Sample

6
Major Measurement Techniques

Electrochemical
Potentiometric
Voltametric
Amperometric

Photometric
Turbidimetric
Colorimetric

Conductometric

Measurement Techniques (1)

■ Electrochemical Measurement
„ Electrode behaviour in Galvanic cell
„ Nernst Equation U = Potential difference

E = Eo + 2.3 RT/nF x ln aH+ glass

electrode

reference
electrode

8
Measurement Techniques (2)

■ Electrochemical Measurement Techniques

„ Potentiometric
■ Nernst equation
■ Electrodes in chemical and electrical equilibrium
■ Acid/base, precipitation, redox
„ Voltametric
■ Potential difference with constant current
■ E - V curve
■ KF
„ Amperometric
■ Current difference with constant voltage (potential)
■ I - V curve
■ Vit C

Measurement Techniques (3)

■ Electrochemical sensors
„ combined glass electrodes (DGxxx)
„ aqueous and non-aqueous acid/base
„ half-cell reference electrode (DX200)
„ metal electrodes (DMxxx)
„ platinum and silver ring
„ redox and precipitation titrations
„ ion selective electrodes (DXxxx)
„ surfactant sensitive electrode (DS500)

10
Measurement Techniques (4)
■ Photometric Measuring Principle
„ Turbidimetric
„ decrease in light intensity
„ Colorimetric
Detector
„ change in light transmission
„ Bouguer- Beer- Lambert Law
Photodiode
„ Applications
„ metal determinations
Light guide
„ surfactants

Concave
mirror
11

Measurement Techniques (5)

■ Photometric sensors – DP550/660

Phototrode DP550/660

12
Measurement Techniques (6)

■ Principle of Measurement
„ migration of ions
- Current +
„ Faraday’s Law
- +
- +
- - +
+
+
- +
χ = G.Z -
G = conductance, µS or mS Ionic Mobility in a Solution
Z = cell constant, cm-1

13

Measurement Techniques (7)

■ Conductometric Sensor
„ 4 ring platinum rings

Induced current by outer rings

is measured by inner rings
Removable
plastic sheath
Main sensor body

Voltage applied to outer

Platinum rings rings generating a current

14
Major Titration Reactions

Titrimetric Analyses

Acid-Base Oxidation-Reduction
Aqueous (Redox)
Non-aqueous Diazotizations
Manganometry
Complexometric Precipitation Iodometry
Turbidimetry Dichromate
Cerimetry

15

Types of Titrimetric Analyses (1)

■ Acid/Base Titration - Aqueous

„ DG111,101,114,115
„ Titration of an acid HA with a strong base
(eg NaOH)

HA + H2O H3O+ + A-
H3O+ + OH- 2 H2O

„ fast reaction, rapid equilibrium

„ weak acids difficult to titrate
„ for not too dulute solutions, titration curve depends on Ka (Kb)
„ titrate separate protons : at least 2 pK units difference
„ Applications : Acid content in wine, ketchup, milk
16
Types of Titrimetric Analyses (2)

■ Acid/Base Titration - Non- Aqueous

„ DG113 – movable sleeve diaphragm
„ analyte is only sparingly soluble in water
„ analyte or titrant enter into an undesired reaction with water
„ pKa values of analytes are too close together
„ analyte is too weak an acid or base in water
„ alcoholic titrants : check titer frequently (eg KOH in ethanol)
„ Applications : TAN/TBN

Solvent HS acts both as an acid and a base

Acid : HS H+ + S-
Base : HS + H+ SH2+
Sum : 2 HS SH2+ + S-
17

Types of Titrimetric Analyses (3)

■ Redox Titration (DM140)

„ gain or loss of electrons
„ conjugate redox couple
„ reduction → entity gains electrons
„ oxidation → entity gives up electrons
„ eg Titration of copper with sodium thiosulphate in excess KI

2Cu2+ + 4I- <===> 2CuI↓ + I2

I2 + I- <===> I3- (reactive species)
2S2O32- + I3- <===> S4O62- + 3I-
2Cu2+ ≡ I2 ≡ 2S2O32-
Applications : Copper, chromium and nickel content in electroplating baths
18
Types of Titrimetric Analyses (4)

■ Precipitation Titration (DM141)

„ formation of a sparingly soluble precipitate between
titrant and analyte
„ classic example : determination of halogens with silver
nitrate

Ag+ + X- AgX (solid) X = Cl-, Br-, I-

„ reaction may be quite slow under certain conditions

„ solution becomes supersaturated before precipitation takes place
„ coprecipitation in mixed halides
„ too concentrated solutions → inclusion of sample and titrant in
precipitation → errors, counter with rapid stirring
„ Applications : NaCl in chips, silver content of coins
19

Types of Titrimetric Analyses (5)

■ Turbidimetric Titration
„ similar to precipitation
„ colloidal formations
„ phototrode
„ DP550/660 : for precipitation during titration
„ Applications : anionic content of metal coolants and
lubricants used in metal cutting processes

20
Types of Titrimetric Analyses (6)

■ Complexometric Titration (DP550/660)

„ colorimetric
„ metal-indicator complex → metal-titrant complex MY
„ most important - EDTA (Na2H2Y.2H20)
„ others : DCTA, EGTA, etc
CO
„ metal to ligand ratio - 1 : 1
O CH2
CO CH2
O N
M2+ + H2Y2- MY2- + 2H+ CH2

M3+ + H2Y2- MY- + 2H+

M
CH2
N
O
CO CH2
CH2
„ Applications : Total hardness in water O CO

Calcium content in cheese

21

Types of Titrimetric Analyses (7)

■ 2-phase Titration
„ colorimetric, DP550, special beaker with lateral port
„ mixed anionic- cationic dye in chloroform

dye
H2O H2O
H2O
CHCl3 CHCl3 CHCl3
Surfactant-dye Titrant-dye
Surfactant-Titrant
Before EQP At EQP After EQP
Pink titrant-anionic dye Blue titrant-cationic dye
complex in chloroform complex in chloroform

Applications : Liquid detergents and shower foam

22
Types of Titrimetric Analyses (8)

■ 2-phase titration setup with DP550

„ Application brochure 22

23

Types of Titrimetric Analyses (9)

■ Conductometeric
„ Water analyses - conductivity, pH, alkalinity
„ α-acids in hobs
„ Acid mixtures (segmented curve)

24
Who Titrates?

„ Pharmaceuticals
„ Food
„ Plastics
„ Cosmetics
„ Beverages
„ Toiletries
„ Petrochemical
„ Chemicals
„ Etc

25
Potentiometric Titration
Excellence T5/T7/T9
Theory and Basic Operation
Agenda 2

1 Basic theory

2 Introduction of the Titrator Excellence

3 Method creation

4 Running sample

5 Q&A

For internal use - Confidential

Definition of Titration 3

Determination of the quantity of a chemical substance in a sample...

Examples
▪ Salt in chips / cheese / eggs
▪ Vitamin C in orange juice
▪ Acid content in milk, vines
▪ Free fatty acid in edible oils
▪ Water hardness
▪ Nickel, copper in electroplating bath
Definition of Titration 4

By controlled addition of a reagent (titrant)

of known concentration ...

with…
▪ Glass burette with stopcock
▪ Dosing instrument / Dispenser
▪ Automatic titrator
Definition of Titration 5

based on a complete chemical reaction between

substance (S) and reagent (R) ...

A
▪ simple (S + R → SR)
▪ fast
▪ stoichiometric

chemical reaction…
Definition of Titration 6

…to an endpoint, which is appropriately detectable.

Visually with chemical indicator OR Electrochemically with a Sensor

Definition of Titration: Summary 7

▪ Determination of the quantity of a chemical substance in a sample ...

▪ by controlled addition
of a reagent (titrant) of known concentration ...

▪ based on a complete chemical reaction between

substance and reagent ...

▪ to an endpoint, which is appropriately detectable.

Manual Titration 8

Burette

Titrant

Sample
& Indicator

Color change
= endpoint Result
calculation
Manual Titration – Automatic Titration 9

Burette

Titrant

Indicator
(sensor)

Sample
Chemical reactions for titrations 10

Requirements for the chemical reactions

▪ High reaction rate at ambient conditions

(short titration time)
▪ Easy experimental conditions
(temperature, pressure, solvent, pH, etc.)
▪ Specific reaction
(no side reactions with other components)
▪ Strictly stoichiometric course of reaction
▪ Course of reaction visible
Types of chemical reactions for titration 11

Acid / Base titration

HCl + NaOH → NaCl + H2O

Examples
▪ Acid content in wine, milk
▪ Acid content in oranges juice
▪ Acid content in ketchup
▪ Free fatty acid in edible oil and fats
▪ Concentration of HCl
▪ Concentration of Ammonia
▪ Acid number of oils

METTLER TOLEDO Electrodes: pH sensitive membrane

Types of chemical reactions for titration 12

Precipitation titration

NaCl + AgNO3 → AgCl (s) + NaNO3

Examples
▪ Sodium chloride content in food:
ketchup, chips, cheese, eggs
▪ Chloride content in water and waste water
▪ Silver content in coins and jewelry

METTLER TOLEDO Electrode: Silver ring electrode

Types of chemical reactions for titration 13

Complexometric titration

Mg2+ + EDTA4- → [Mg-EDTA] 2-

Examples
▪ Total hardness in water (Ca & Mg)
▪ Calcium content in cheese and milk
▪ Cement Analysis (Al, Fe, Mg, Ca)
▪ Electroplating bath (Cu, Zn, Cd)

METTLER TOLEDO Sensor: Phototrode

Types of chemical reactions for titration 14

Redox titration

SO2 + I2 + 2 H2O → H2SO4 + 2 HI

Examples
▪ Sulfuric dioxide in Wine
▪ Vitamin C content in orange juice
▪ Gold content in alloys
▪ Electroplating bath (Ag, Cu, Ni, Cr)
▪ Determination of peroxides

METTLER TOLEDO Electrodes:

Potentiometric: Pt ring electrode
Voltametric and Amperometric: Pt pin electrode
DM140 DM143
Types of chemical reactions for titration 15

Karl Fisher titration for water determination

I2 + SO2 + 2 H2O → 2 HI + H2SO4

A special type of titration
▪ Specific for water, which is in many samples
▪ Fast (1 – 2 minutes)
▪ No decomposition
▪ Wide measuring range (0,001% – 100 %)
METTLER TOLEDO Electrode: Pt pin electrode
For Karl Fisher titration a separate generation of instruments exist. For more
information take a look to the Karl Fisher presentation or to our specific brochures.

Volumetric KF- Coulometric KF-

titration titration
 Types of chemical reactions for titration 16

Precipitation for surfactant determination

[surfactant] – + [surfactant] + → Complex

anionic surfactant cationic surfactant colloidal complex

Clear solution → turbid solution

Examples
▪ Washing powder
▪ Softener
▪ Toothpaste Colloidal complex
▪ Cosmetics (cross section)

▪ Paper

▪ METTLER TOLEDO Sensors: Phototrode

Major titration reactions 17

Titration reactions

Acid / Base
Oxidation / Reduction
Aqueous
(Redox)
Non-aqueous
Diazotizations
Manganometry
Complexometric Precipitation Iodometry
Turbidimetry Cerimetry
Karl Fischer
Agenda 27

1 Basic theory

2 Introduction of the Titrator Excellence

3 Preparation for titration

4 Running sample

5 Q&A

For internal use - Confidential

 Titrator Excellence 28

Stirrer
Titrant-in line Titrant-out line

Electrode

Burette

Sample beaker

Status light
▪ Burette locker
Power button

Touch screen terminal

For internal use - Confidential

Titrator Excellence 29

User interface on touch screen terminal

Simple Touchscreen operation

▪ Shortcut buttons for easy start of methods and
manual operations
▪ For edit and start methods
▪ For sample series
▪ For view and edit results
▪ For view and define SETUP
▪ For define and start manual operation

For internal use - Confidential

Agenda 30

1 Basic theory

2 Introduction of the Titrator Excellence

3 Preparation for titration

4 Running sample

5 Q&A

For internal use - Confidential

Preparation for titration 31

Before start titrating, we need…

▪ Method:
- Creation, checking
▪ Resources
- Titrant
- Sensor
- Stirrer
- Burette
▪ Sample
- Weighing
- Create sample series
- Prepare to titrate

For internal use - Confidential

Preparation for titration 32

Method creation (for EQP titration)

Method/ New/ Standard method templates: choose EQP

For internal use - Confidential

Preparation for titration 33

Method creation (for EQP titration)

1/ Title: give a name

For internal use - Confidential

Preparation for titration 34

Method creation (for EQP titration)

2/ Sample

Type ID of sample

Usually choose Weight

For internal use - Confidential

Preparation for titration 35

Method creation (for EQP titration)

3/ Titration stand: default if no more titration stand

For internal use - Confidential

Preparation for titration 36

Method creation (for EQP titration)

4/ Stir: mixing before titration

For internal use - Confidential

Preparation for titration 37

Method creation (for EQP titration)

5/ Titration (EQP)

Choose titrant

Choose sensor

Set speed for titration

▪ Dispense a volume of
titrant before start
titration

For internal use - Confidential

Preparation for titration 38

Method creation (for EQP titration)

5/ Titration (EQP)

Titration control: normal, fast,

cautious or user (fully manual)

Setting for Equivalent point evaluation

Preparation for titration 39

Method creation (for EQP titration)

6/ Calculation: click to Result proposals to choose result type

For internal use - Confidential

Preparation for titration 40

Method creation (for EQP titration)

7/ Record

For internal use - Confidential

Preparation for titration 41

Method creation (for EQP titration)

After finished, click save/ start, click to AddToHome to create a shortcut

For internal use - Confidential

Preparation for titration 42

Method creation (for EQP titration)

Name of shortcut
(maximum 10
characters)

Position at home
screen

For internal use - Confidential

Preparation for titration 43

Resources preparation
▪ Titrant:
- Determine titer (correction factor)
- Prepare burette for titrant drawing

▪ Sensor:
- Uncover the cap on sensor's body
- Verify and calibrate sensor before use.
- Make sure it is in good condition.

▪ Burette:
- Choose right burette with right titrant (each burette has a chip placed in)
- Ensure no air bubble in it.
- Check burette locker.

For internal use - Confidential

Agenda 44

1 Basic theory

2 Introduction of the Titrator Excellence

3 Preparation for titration

4 Running sample

5 Q&A

For internal use - Confidential

 Running sample 45

▪ Go to Method, choose method to run, click Start or click on Shortcut created

Number of samples want to run

Type ID of
sample

Input weight value

Click Start when

finishing input

Note: need to type number of samples first. If it is more than 1, then we need to type sample ID
and sample size in Sample window (mark as red)
For internal use - Confidential
Running sample 46

▪ Prepare sample and attach it on the titration arm.

▪ Check the availability of sensor, stirrer, burette-out line and add sample as instruction
showed on screen

For internal use - Confidential

 Running sample 47

At titration window:
Status of titration

Realtime values

Titration curve

Axes for changing Suspend titration

Sample series: user can process
displayed units, axis
check or adjust sample
series in here
For internal use - Confidential
 Running sample 48

After finishing titration process, results will be calculated and printed out automatically

Note: due to compliant and regulation, only one result can be stored. When running a new
sample, the previous result will be replaced.
For internal use - Confidential
Running sample 49

Cleaning
▪ Remove the titration beaker from titration arm
▪ Rinse sensor, stirrer by sample's solvent, make sure the limit
▪ Dry it by tissue, carefully absorb solvent remaining on sensor membrane.
▪ The system is ready for new analysis.

Note:
- If solvent used for titration is non polar, the sensor should be reactivated in pH 4.01 buffer
before starting new titration.
- After completing titration, cleaning the sensor, cover the cap on sensor body and preserve
it in solution attached in sensor box.

For internal use - Confidential

Agenda 50

1 Basic theory

2 Introduction of the Titrator Excellence

3 Preparation for titration

4 Running sample

5 Q&A

For internal use - Confidential

Question? 51

For internal use - Confidential

 Good Titration Practices (GTP)

Agenda 2

1 Basic of Titration

2 Reaction types in Titration

3 Manual VS Automatic Titration

4 Titrator Instrument

5 Aplication in F&B

6 Tips and Hints

Basic of Titration 3

Terminology and Definition

 Qualitative chemical analysis: Determine the chemical composition in a sample

using chemical reactions and interactions.
Example: Test for halides with Beilstein test (green flame if halides are present).
 Quantitative chemical analysis: Determine the content in a sample of one or
more chemical substances using chemical reactions and interactions.
Example: Vitamin C content in orange juice by titration.
 Sample: A small representative portion of the entity that should be analyzed.
Example: Feed
 Analyte: The chemical substance to be determined with the analysis.
Example: Protein
 Matrix: Everything in the sample besides the analyte(s).
Example: Fat, Carbohydrate, Additive, ... in Protein analysis of feed
 Titrant: Solution of known concentration which is added to the sample (titration)
and reacts with the analyte. The analyte content is calculated from the consumption
of the titrant.
Basic of Titration 4

What is titration?

 Titration is the determination of the quantity of a specific substance (analyte)

contained in a sample by controlled addition of a reagent (titrant) of known
concentration based on a complete chemical reaction between the substance and
the reagent.

 Example:
Determination of acetic acid (CH3COOH) by titration with sodium hydroxide (NaOH)

CH3COOH + NaOH CH3COONa + H2O

(analyte) (titrant) (reaction products)

Basic of Titration 5

 Practical Procedure:
- The sample (e.g. vinegar) is diluted  sample solution with low pH value (= acidic).
- A NaOH solution of known concentration (titrant) is carefully added to the acidic solution
(titration).
- The analyte (CH3COOH) reacts with the titrant (NaOH)  pH rises slowly (from acidic to
basic)
- As soon as all analyte has reacted, the pH rises immediately (excess of NaOH in the
solution) equivalence point

 Content calculation:
- Sample size is known.
- Concentration of titrant is known.
- Titrant consumption (volume) until the equivalence point is measured during titration.
 The acetic acid content in the sample can be calculated from these three variables.

Basic of Titration 6

 Apparatus : The standard equipment for a manual titration includes the

following :
- Burette: for accurate titrant addition Burette
- Titration beaker
- Stirrer

 Several prerequisites have to be fulfilled in order to perform a titration:

- Suitable reaction needed (reaction between the titrant and the analyte)
- Titrant concentration and used volume must be known very
accurately
- Equivalence point must be detectable

Titration beaker

Stirrer
Basic of Titration 7

Reaction must be …

 Selective:
The chemical reaction between the analyte and the titrant must be selective (only the
analyte should react with the titrant).

 Fast:
The reaction should be fast in order to guarantee that the added titrant reacts immediately
with the analyte.

 Complete:
The equilibrium of the reaction should lie strongly on the product side of the reaction to
guarantee a complete reaction.

 Unambiguous:
The stoichiometry of the reaction must be known and unambiguous.

Basic of Titration 8

Titrant

 The key point of a good titration is the accurate determination of the volume
of titrant used until the equivalence point.

 For this reason the following two requirements have to be fulfilled:

- A titrant addition in small quantities must be possible.
- An accurate reading for the volume used is needed.

 A manual burette fulfills these requirements

- Stopcock: add small volumes
- Grading: accurate volume reading

Basic of Titration 9

Indication

 The equivalence point (when all analyte has reacted with the titrant) of a titration should be
easily visible or measurable.
 The two basic indication principles are:
- Indicator
- Electrode

 Indicator:
- An indicator is a substance which changes its color at the equivalence point.
- Small amounts of an indicator are added to the solution prior to the analysis.

- Example:
Phenolphthalein is an indicator which is pink under basic conditions and colorless
under acidic conditions. At the equivalence point of an acetic acid titration by sodium
hydroxide, the color will immediately change from colorless to pink.
Basic of Titration 10

Indication

 Electrode:
- An electrode is an instrument that measures a specific property of a solution
electrochemically.
- A suitable electrode has to be chosen for each type of reaction. The electrode must
measure a property which is related to the titration reaction.
- The whole titration procedure can be followed with an electrode  titration curve

- Example:
For the titration of acetic acid by sodium hydroxide a pH electrode is used. The pH
electrode measures the H+ concentration related to acetic acid that hasn't reacted.
The equivalence point is reached when the pH value changes suddenly from acidic to
basic.

Basic of Titration 11

Titration Curve
 If an electrode is used for a titration the signal vs. titrant volume can be plotted.
 Such a plot is called a titration curve.
 The equivalence point can be determined directly from a titration curve and is the
inflection point of the curve.

 Example: Titration curve (pH vs. volume) of acetic acid titration with sodium hydroxide.
The equivalence point is clearly visible.

pH

Equivalence point

Volume

Basic of Titration 12

Titration Modes

 Two different titration modes are used:

- Equivalence point titration (EQP)
- Endpoint titration (EP)
Signal
 EQP:
- The EQP is reached as soon as all analyte has EQP
reacted with the titrant.
- In a titration curve the EQP is the inflection point of the
curve.
- The titration is carried out over the EQP and evaluated
afterwards. Volume
Signal

 EP:
- The EP is reached as soon as the signal reaches a
predefined value.
- The titration is usually stopped at the EP EP

Volume
Basic of Titration 13

Titration Types and Calculation – Direct Titration

 The most common and basic titration type is: direct titration.
 In a direct titration the analyte reacts directly with the titrant.
 Example: In the titration of acetic acid with sodium hydroxide the acetic acid reacts directly
with sodium hydroxide by exchanging a proton. The content of acetic acid can be
calculated directly from the used sodium hydroxide (titrant) volume (consumption).

CH3COOH + NaOH CH3COONa + H2O

(analyte) (titrant) (reaction products)

Basic of Titration 14

Titration Types and Calculation – Direct Titration

 The analyte content in a direct titration is calculated from the titrant consumption at the
equivalence or end point using the following formula:
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅=
𝑚

R: Result
VEQ: Titrant consumption at the equivalence or end point (in mL)
c: Titrant concentration (in mol/L)
t: Titrant titer (no unit)
C: Constant for unit conversion (unit part, unit is dependent on the result
needed)
m: Sample size (in g or mL)
 The constant C is dependent on the result unit wanted and on the unit of the sample size.

Basic of Titration 15

Titration Types and Calculation – Direct Titration

 Examples of constants:
Sample size
g mL
entry unit
𝑀
% 𝐶=
10 ∙ 𝑧
𝑀
mg/g g/L 𝐶=
Result unit

𝑧
𝑀 ∙ 1000
ppm mg/L 𝐶=
𝑧
1000
mmol/kg mmol/L 𝐶=
𝑧
1
mol/kg mol/L 𝐶=
𝑧

M: Molar mass of the analyte (in g/mol)

z: Equivalent number (no unit)
Basic of Titration 16

Titration Types and Calculation – Direct Titration

 Example: For the titration of the acetic acid (M = 60.04 g/mol) in a 1 g sample of vinegar, 5
mL sodium hydroxide (c = 0.1 mol/L, titer: 0.9900) were consumed until the equivalence
point. To calculate the acetic acid content in % you have to use the following formula:

𝑀 60.04 g/mol g∙%

𝐶= = mg = 6.004
10 ∙ 𝑧 10 g ∙ % ∙ 1 mmol

mol g∙%
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 5 mL ∙ 0.1 L ∙ 0.9900 ∙ 6.004 mmol
𝑅= = = 2.97 %
𝑚 1g

Basic of Titration 17

Titration Types and Calculation – Titer

 The titer is usually determined by a direct titration, but the calculation is a little different.
 The titer (t) is defined as the actual concentration (cact) divided by the nominal
concentration (cnom) of a titrant:

𝑐act
𝑡=
𝑐nom
 The following formula can be used to calculate the titer of a titrant, if this titrant was used
to titrate a titer standard by a direct titration:

𝑚
𝑡=
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝐶
t: Titrant titer (no unit)
m: Titer standard sample size (in g or mL)
VEQ: Titrant consumption at the equivalence or end point (in mL)
c: (Nominal) titrant concentration (in mol/L)
C: Constant for unit conversion (unit part, unit is dependent on the titer
substance form)

Basic of Titration 18

Titration Types and Calculation – Titer

 If the titer standard substance is a solid, use the following formula for the constant C:

𝑀
𝐶=
10 ∙ 𝑝 ∙ 𝑧
 For liquid titer standards the following constant is used:

1
𝐶=
𝑐𝑠𝑡 ∙ 𝑧
M: Molar mass of the analyte (in g/mol)
p: Purity of the solid titer standard (in %)
z: Equivalent number of the titer standard (no unit)
cst: Concentration of the liquid titer standard (in mol/L)
Basic of Titration 19

Titration Types and Calculation – Titer

 Example: Potassium hydrogen phthalate (KHP, M= 204.23 g/mol) was used as a solid
titer standard to determine the titer of sodium hydroxide (c = 0.1 mol/L). 0.0931 g of
KHP (purity: 99 %) was weighed into a titration beaker and diluted with deionized water.
This solution was titrated with sodium hydroxide. The equivalence point was detected at
a titrant consumption of 4.5238 mL. For this measurement the titer can be calculated as
follows:

g
𝑀 204.23 mol g
𝐶= = = 0.206
10 ∙ 𝑝 ∙ 𝑧 10 mg ∙ 99 % ∙ 1 mmol
g∙%
𝑚 0.0931 g
𝑅= = = 0.9990
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝐶 4.5238 mL ∙ 0.1 mol ∙ 0.206 g
L mmol

Basic of Titration 20

Titration Types and Calculation – Blank Titration

 In some titrations it may happen that the solvent itself also reacts with the titrant. The
amount of titrant used for the solvent is called the blank value.
 The blank value has to be compensated to get the correct result.
 To determine the blank value a titration of the solvent without any sample has to be
performed. The blank value is the volume of the titrant used until the equivalence or
endpoint is reached.
 For a titration where a blank value is used, take care to always use the same solvent
volume!

Basic of Titration 21

Titration Types and Calculation – Blank Titration

 Example: The acid number of motor oil is determined by a non-aqueous acid/base

titration with potassium hydroxide (in 2-propanol) as titrant. As solvent a mixture of
toluene, 2-propanol and water is used. Before titrating the sample, the solvent mixture
without any sample is titrated. The titrant consumption of this blank measurement is
compensated in the calculation of the sample measurement.
Basic of Titration 22

Titration Types and Calculation – Blank Titration

 The calculation of a blank value compensated titration is similar to the direct titration:

(𝑉𝐸𝑄 − 𝐵) ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅=
𝑚
R: Result
VEQ: Used titrant volume until the equivalence or end point (in mL)
B: Blank value, titrant volume used for the titration of the solvent (in mL)
c: Titrant concentration (in mol/L)
t: Titrant titer (no unit)
C: Constant for unit conversion (unit part, unit is depending on the result
needed)
m: Sample size (in g or mL)
 The constant C is the same as for the direct titration

Basic of Titration 23

Titration Types and Calculation – Back Titration

 In a back titration a known excess amount of reagent (titrant A with known concentration)
is added to the sample. This titrant reacts with the analyte. Afterwards the amount of non-
reacted titrant A will be titrated with a second titrant (titrant B).
 The added volume of titrant A has to be known very precisely.
 For accurate measurements a so called back value is determined by titrating the added
volume of titrant A without any sample. The result is based on the used volume of titrant B
without any sample.
 Such titrations are often used when the reaction between the analyte and the first titrant is
slow.

Basic of Titration 24

Titration Types and Calculation – Back Titration

 Example: For the titration of nitrite in soya sauce a known volume of potassium
permanganate (titrant A) is added to the sample. Potassium permanganate will react with
the nitrite and the excess of the permanganate is titrated with ammonium ferrous sulfate(II)
(titrant B).

5 NO2- + 2 MnO4- + 6 H+ 2 Mn2+ + 5 NO3- + 3 H2O

(analyte) (titrant A)
MnO4- + 5 Fe2+ + 8 H+ Mn2+ + 5 Fe3+ + 4 H2O
(titrant A) (titrant B)
Basic of Titration 25

Titration Types and Calculation – Back Titration

 The analyte content in a back titration is calculated using the following formula:
(𝐵𝑘 − 𝑉𝐸𝑄) ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅=
𝑝∙𝑚

R: Result
Bk: Back value, volume of titrant B used for the titration of titrant A without sample (in mL)
VEQ: Volume of titrant B used to back titrate the excess of titrant A after
sample addition (in mL)
c: Concentration of titrant B (in mol/L)
t: Titer of titrant B (no unit)
C: Constant for unit conversion (unit part, unit is depending on the result
needed)
p: Stoichiometric proportion between titrant A and titrant B of the second
reaction, stoichiometric factor of titrant B divided by factor of titrant A (no
unit)
m: Sample size (in g or mL)
 The constant C is the same as for the direct titration

Agenda 26

1 Basic of Titration

2 Reaction types in Titration

3 Manual VS Automatic Titration

4 Titrator Instrument

5 Aplication in F&B

6 Tips and Hints

Reaction types in Titration 27

1. Acid / Base
 Reaction:
- This type of titration is based on the reaction between an acid (AH) and a base (B),
where a proton (H+) is exchanged.
AH + B A- + BH+
- If the analyte is an acid the titrant will be a base and vice versa.
 Analytes:
- Weak or strong acid / base
- Examples: Hydrochloric acid, sulphuric acid, acetic acid, sodium hydroxide, ammonia, …
 Titrants:
- Strong base / acid solution
- Examples: Sodium hydroxide 0.1 mol/L, hydrochloric acid 0.1 mol/L
 Indication:
- pH indicators: pH dependent dyes (for manual titration), examples: phenolphthalein,
methyl orange, …
- pH sensor (for automated titration)
Reaction types in Titration 28

1. Acid / Base

 The parameter which changes during a acid / base titration is the pH value.
 The pH value is defined as the negative decadic logarithm of the H+ (or H3O+)
concentration in a solution:

𝑝𝐻 = − log 𝑐 H + = − log 𝑐 H3 O+

 Even in pure deionized or distilled water a small amount of ions is present due to the
amphoteric properties of water (it can react as a acid or a base). Water molecules react
with each other to build H3O+ and OH-:

H2O + H2O H3O+ + OH-

 In pure deionized water the concentration of H3O+ and OH- ions are both 10-7 mol/L. The
product of these two concentrations is called the ionic product of water (Kw) and this value
is a constant:

mol mol 𝑚𝑜𝑙 2

𝐾w = 𝑐 H3 O+ ∙ 𝑐 OH − = 10−7 ∙ 10 −7 = 10−14 2
L L 𝐿

Reaction types in Titration 29

1. Acid / Base

 If we calculate the pH value for pure water (c(H3O+) = 10-7 mol/L) it is 7.

A pH value of 7 is neutral.
 If an acid (AH) is added to an aqueous solution, it will react (dissociation) with water to
form more H3O+ ions (and less OH- ions due to Kw):

AH + H2O A- + H3O+

The H3O+ concentration will increase and the pH value will decrease.
A pH value below 7 is acidic.
 If a base (B) is added to an aqueous solution, it will react with water to form more OH- ions
(and less H3O+ ions due to Kw):

B + H2O BH+ + OH-

The H3O+ concentration will decrease and the pH value will increase.
A pH value above 7 is alkaline.

Reaction types in Titration 30

1. Acid / Base

 For a manual titration various pH indicators exist. The selection must be based on the pH
range where the equivalence point occurs: the indicator has to be chosen such that its
color change occurs at the equivalence point.
 The following graph shows common pH indicators and their color change range:
Thymol blue

Congo red

Methyl orange

Bromcresol green

Methyl red

Bromthymol blue

Phenol red

Phenolphthalein

Thymolphthalein

0 2 4 6 8 10 12
pH
 Reaction types in Titration 31

1. Acid / Base

 For automated titration the pH value is measured by a pH sensor (usually combined

sensor), which consists of a reference system (usually Ag/AgCl) and a glass membrane
which is sensitive to H+ (or H3O+).
 Since the measured signal is a potential (mV) and not pH, the sensor has to be calibrated
(correlate the mV signal to a pH value).
 The calibration is done by measuring the potential of various calibration buffers with
defined pH value. From these measurements (calibration points) a calibration line is
calculated.
 Theoretically (at 25°C) the slope of the calibration line is -59.16 mV/pH and the zero point
is pH 7.00.
E [mV]
zero point
hydrated layer membrane glass

0 pH
1 14
slope

Reaction types in Titration 32

1. Acid / Base

Example 1: Titration of acetic acid in vinegar with sodium hydroxide

 The acetic acid concentration of a vinegar sample should be determined. Since acetic acid
is an acid this will be done by titrating with sodium hydroxide. The reaction for this titration
is
CH3COOH + NaOH CH3COONa + H2O

 For this sample an acetic acid content of 5 % is expected. From this value one can
calculate the sample size by re-arranging the calculation formula so that the sample size
can be calculated

𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶
𝑅= → 𝑚=
𝑚 𝑅
 If we use a 20 mL burette, a titrant consumption of about 10 mL (half of burette volume)
would be perfect. We use a sodium hydroxide solution with c = 0.1 mol/L and assume a
titer of 1. Form these values the approximate sample size can be calculated.

Reaction types in Titration 33

1. Acid / Base
𝑀 60.04 g/mol g∙%
𝐶= = mg = 6.004
10 ∙ 𝑧 10 g ∙ % ∙ 1 mmol

mol g∙%
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 10 mL ∙ 0.1 L ∙ 1 ∙ 6.004 mmol
𝑚= = = 1.2 g
𝑅 5%

 Based on this calculation 1.3134 g of vinegar was weighed on an analytical balance into a
titration beaker and diluted in 50 mL deionized water. This solution was titrated with sodium
hydroxide (c = 0.1 mol/L and t = 0.9877) on an automated titrator equipped with a pH
sensor until the equivalence point was recognized.
Reaction types in Titration 34

1. Acid / Base
Titration Curve
12

10

8
pH

2
0 2 4 6 8 10
V [mL]
VEQ
 From the titrant consumption (8.3784 mL) the content of acetic acid in the sample was
calculated:

mol g∙%
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 8.3784 mL ∙ 0.1 L ∙ 0.9877 ∙ 6.004 mmol
𝑅= = = 3.78 %
𝑚 1.3134 g

Reaction types in Titration 35

1. Acid / Base

Example 2: Simultaneous titration of sodium hydroxide and carbonate

 If a sodium hydroxide solution is not closed and not protected against carbon dioxide
intake, carbon dioxide will be dissolved and will react to carbonate. If one performs a
titration, this carbonate will also be titrated.
 During the titration three different bases are present: sodium hydroxide (NaOH), sodium
carbonate (Na2CO3) and sodium hydrogen carbonate (NaHCO3). Since NaOH is the
strongest base this species will react first with the titrant (hydrochloric acid, HCl).
Afterwards Na2CO3 will react to NaHCO3 and this base will react at the end to CO2.

1. NaOH + HCl NaCl + H2O

2. Na2CO3 + HCl NaHCO3 + NaCl
3. NaHCO3 + HCl CO2 + NaCl + H2O

Reaction types in Titration 36

1. Acid / Base

 Theoretically, we should observe three equivalence points but the first point will be very flat
and only two points will be clearly visible:
- 1. EQP: Reaction of NaOH and Na2CO3
- 2. EQP: Reaction of NaHCO3

 From these two EQP's the NaOH and Na2CO3 concentrations can be
determined.

 To perform this titration 7 mL of sodium hydroxide sample is added with a

accurate pipette into a titration beaker and the solution is diluted with 50 mL of
deionized water. This solution is titrated with hydrochloric acid (c = 0.1 mol/L
and t = 0.9932) until two equivalence points are recognized (automatic titrator
with pH sensor).
Reaction types in Titration 37

1. Acid / Base

Titration Curve

12

10

8
pH

2
0 2 4 6 8 10 12 14 16
V [mL] VEQ1 VEQ2

Reaction types in Titration 38

2. Precipitation

 Reaction:
- The analyte (A) and the titrant (T) react and form a compound which precipitates.
A + T AT

- Usually silver nitrate (AgNO3) is used as a titrant, which forms hardly soluble silver salts
(e.g. AgCl)
 Analytes:
- Ions which form hardly soluble silver salts
- Examples: chloride, cyanide, thiocyanate
 Titrants:
- Silver nitrate
 Indication:
- Indicators: Potassium dichromate (for manual titration)
- Silver ring electrode (for automated titration)

Reaction types in Titration 39

2. Precipitation

Example: Titer determination of silver nitrate

 The titer of silver nitrate (AgNO3) is determined by sodium chloride (NaCl) as standard
substance.
NaCl + AgNO3 AgCl + NaNO3

 To determine the titer, 43.2 mg of NaCl standard (purity: 99.5 %) is weighed into a titration
beaker and dissolved in 50 mL nitric acid (c = 0.02 mol/L). The nitric acid is needed to
keep a pH value of 4.5 in the sample solution, since the precipitation reaction takes place
under slightly acidic conditions.
 The sample solution is titrated with AgNO3 (c = 0.1 mol/L) while the signal is measured
with a silver ring sensor (measures Ag+ concentration). The titration is stopped as soon as
the equivalence point is recognized.
 During the titration the precipitation of AgCl is clearly visible.
Reaction types in Titration 40

2. Precipitation

Titration Curve
200

150

100
E [mV]

50

-50

-100
0 2 4 6 8 10
V [mL] VEQ

Reaction types in Titration 41

3. Redox
 Reaction:
- The analyte and the titrant exchange electrons during this reaction and the oxidation
state of both changes.
- If the analyte is an oxidizing agent (O) the titrant is a reducing agent (R) and vice versa.
Oxidation: R R+ + e-
Reduction: O + e- O-

 Analytes:
Redox reaction: R + O O- + R+
- Oxidizing and reducing agents
- Examples: Sulphurdioxide, Nitrite, Iron, …
 Titrants:
- Oxidizing and reducing agents
- Examples: Iodine, Sodium thiosulfate, potassium permanganate, …
 Indication:
- Color change due to colored titrant, starch solution (for Iodine)
- Platinum sensor (for automated titration)

Reaction types in Titration 42

3. Redox
Example 1 : Titer determination of potassium permanganate
 The titer of potassium permanganate (KMnO4) is determined by using Di-sodium oxalate
(Na2C2O4) as the standard. During this titration Na2C2O4 is oxidized to CO2 and KMnO4 is
reduced to Mn2+:

Oxidation: Na2C2O4 2 CO2 + 2 Na+ + 2 e-

Reduction: KMnO4 + 8 H+ + 5 e- Mn2+ + 4 H2O + K+

Redox reaction:
5 Na2C2O4 + 2 KMnO4 + 16 H+ 10 CO2 + 10 Na+ + 2 Mn2+ + 8 H2O + 2 K+

 Since two electrons are released by the oxidation of Na2C2O4 the equivalent number of
this substance is z = 2.
 Since 5 electrons are needed for the reduction of the titrant KMnO4 the concentration of
the titrant will be given as c(1/5 KMnO4) which is five times higher than the concentration
c(KMnO4).

𝑐 = 𝑐 1/5 KMnO4 = 𝑐 KMnO4 ∙ 5

Reaction types in Titration 43

3. Redox

Example 2: Iodometric determination of Cr6+

 Cr6+ should be determined in an electroplating bath. This is done in a two step reaction.
First Cr6+ is reduced to Cr3+ using iodide (I-) which is oxidized to iodine (I2). The iodine
produced is then titrated with sodium thiosulfate (Na2S2O3):

1. 2 Cr6+ + 6 I- 2 Cr3+ + 2 I2
2. 2 Na2S2O3 + I2 Na2S4O6 + 2 I-
 Since three electrons are needed to reduce Cr6+ to Cr3+ the equivalent number is z = 3.
 5 mL of a 1:100 diluted bath sample (m = 0.05 mL) is added into a titration beaker. The
sample is diluted in
- 5 mL of 50 % H2SO4 solution to acidify the solution
- 40 mL deionised water.
 To have an excess of iodide in the solution 5 mL of 10 % potassium iodide (KI) solution
was added. This iodide reacts with the Cr6+ and a corresponding amount of iodine is
produced.

Reaction types in Titration 44

4. Complexometric

 Reaction:
- Analyte (A) and titrant (T) react together to build a complex ([AT]).
A + T [AT]

 Analytes:
- Metal ions with a charge > 1
- Examples: Zinc ion, magnesium ion, calcium ion, …
 Titrants:
- Ligands which build complexes with the analyte
- Example: Ethylenediaminetetraacetic acid (EDTA)
 Indication:
- Indicators (also ligands) that change color if they are not bound to the analyte, examples:
Murexide, Eriochrome Black T
- Indicators and a photometric sensor (for automated titration)

Reaction types in Titration 45

4. Complexometric

 In a manual titration the color change of the indicator is observed visually with the naked
eye.
 In an automated titration an indicator is also added, but the color change is observed by a
photometric sensor.
Reaction types in Titration 46

4. Complexometric

Example: Total hardness of water

 Total hardness of water is expressed as the CaCO3 (M = 100.09 g/mol, z = 1) content in
water. It is a typical analysis done as a complexometric titration of calcium and magnesium
at pH 10.
 50 mL of tap water, 10 mL of 5 % ammonia solution (for pH 10), and 1 mL of 1 %
Eriochrome Black T indicator solution were added into a titration beaker. This solution was
titrated by EDTA (c = 0.1 mol/L, t = 0.9763) and the color change, from violet to blue, was
observed with a photometric sensor at pH 10. The equivalence point was found at a titrant
consumption of 0.7392 mL.
 The hardness (as CaCO3 content) can be calculated as follows:

g mg
𝑀 ∙ 1000 100.09 mol ∙ 1000 g mg
𝐶= = = 100090
𝑧 1 mol
mol mg
𝑉𝐸𝑄 ∙ 𝑐 ∙ 𝑡 ∙ 𝐶 0.7392 mL ∙ 0.1 L ∙ 0.9763 ∙ 100090 mol
𝑅= = = 144.47 ppm
𝑚 50 mL

Reaction types in Titration 47

5. Karl Fischer
 Reaction:
- Karl Fischer titration is a special type of redox titration for water determination.

ROH + SO2 + 3 R'N + I2 + H2O (R'NH)·SO4R + 2 (R'NH)I

- The Karl Fischer reaction needs an alcohol (ROH, usually methanol), sulfur dioxide, a
base (imidazole or pyridine), iodine and the analyte water.
 Analyte: Water
 Titrants and Solvents:
- One-component reagent: contains iodine, sulfur dioxide and a base dissolved in
methanol. The solvent is methanol.
- Two-component reagent: Contains iodine dissolved in methanol. The solvent contains
sulfur dioxide, a base and methanol.
 Indication:
- Polarized double pin platinum electrode (voltammetry)

Reaction types in Titration 48

5. Karl Fischer

 The Karl Fischer reaction is a two step reaction:

1. ROH + SO2 + R'N (R'NH)·SO3R
2. (R'NH)·SO3R + I2 + H2O + 2 R'N (R'NH)·SO4R + 2 (R'NH)I

 From this reaction it can be seen that water reacts with iodine attaining 1:1
stoichiometry.
 The solvent ROH (usually methanol) is also involved in the reaction, this means that at
least 50 % of the solvent should always be methanol.
 A suitable base (R'N, usually imidazole, in the past: pyridine) keeps the pH value between
5 and 7. This is the optimal pH range so that the reaction is fast and yet no side reactions
occur.
 Reaction types in Titration 49

5. Karl Fischer

 Karl Fischer titration needs a special setup: The titration vessel has to be protected from
humidity as it has an influence on the result. For this reason Karl Fischer titrations are
almost always done using an automatic titrator.
 A typical setup for a Karl Fischer titration is shown below:

Dosing tube for titrant Sensor

Molecular sieve

Glass vessel
Burette filled with titrant

Solvent
Magnetic stirrer

Reaction types in Titration 50

5. Karl Fischer

 Karl Fischer titration is an endpoint titration.

 The endpoint is detected with a polarized double platinum pin sensor. A fixed polarization
current (Ipol) is applied to this sensor and the voltage (U in mV), which is used to maintain
this current is measured.

 During the titration:

The iodine from the titrant reacts directly with
the water
 No free iodine in the solution
 High voltage is needed to maintain the
polarization current

Reaction types in Titration 51

5. Karl Fischer

 At the end of the titration:

No water is available for the reaction
with iodine
 Free iodine in the solution causes
ionic conduction
 Low voltage is needed, signal drop

E
 As soon as the voltage signaldrops below a
defined endpoint the titration stops

t
Reaction types in Titration 52

5. Karl Fischer

Example: Water content determination in ethanol

 The titration is started. Once the pretitration is done the titrator goes into standby mode. A
stable and low drift value (typically < 25 µg/min) is awaited.
 The drift value read just before the sample determination starts is noted.
 Since ethanol is completely soluble in methanol, the sample can be directly inserted into
the titration vessel. For this purpose about 5 mL of ethanol is drawn up into a syringe,
placed on a balance and tared.
 The sample determination is started (drift value: d = 11.3 µg/min, titrant concentration: c =
4.8976 mg/mL). About 1 mL of ethanol is added with the syringe into the titration beaker.
The syringe is once again placed on the balance. The sample size is the difference
between the weight of the syringe before and after the sample addition (back weighing):
1.0348 g

Reaction types in Titration 53

5. Karl Fischer

 The titration ends (endpoint was set to 100 mV) after 2.1865 mL and a titration time of
2.293 min.

Titration Curve

600

400
E [mV]

EP
200

0
0 50 100 150
t [s]

Agenda 54

1 Basic of Titration

2 Reaction types in Titration

3 Manual VS Automatic Titration

4 Titrator Instrument

5 Aplication in F&B

6 Tips and Hints

Manual vs Automatic Titration 55

Dosing

Evaluation
indication
Calculation

Documentation

Manual vs Automatic Titration 56

Dosing
Manual Titration Automatic Titration
 Automatic dosing via precision glass
 Open and close tap of manual burettes and high-resolution motor
burette driven piston
 Wait in between to allow reaction to  High dosing accuracy
take place
 Fully controlled by titrator via method
 Check indication during complete parameters
titration
 Automatic preparation and refill
 Burette volume and piston stroke
can be calibrated with traceable
standards

Manual vs Automatic Titration 57

Indication

Manual Titration Automatic Titration

 Automatic indication via sensors of
 Color indicator different kind
 Color change indicates  High measurement accuracy
Endpoint of titration
 Measured value recorded after every
volume increment added
 Fully controlled by titrator via method
parameters
 Big application range (not just color-
indicated EP titrations)
 Independent from operator
 Sensor inputs as well as sensors can
be calibrated with traceable standards
Before End After
point
 Manual vs Automatic Titration 58

Evaluation
Manual Titration Automatic Titration
 Automatic evaluation of endpoint and
 Read off dispensed volume equivalence point by optimized
when endpoint is reached algorithms
 Only for endpoint titration  Mathematically, based on measured
 Equivalence point titration value table
not possible  High analysis accuracy and
repeatability
 Big application range (not just color-
indicated EP titrations)
 Independent from operator

Manual vs Automatic Titration 59

Calculation
Manual Titration Automatic Titration
 Calculate content or concentration  Automatic calculation of result
 Inputs  Various results simultaneously
- Dispensed volume to endpoint available
- Concentration of titrant  Predefined or user defined
- Sample size (weight or volume) calculation formulas
- Molecular weight and equivalence  Automatic calculation of statistics,
number of analyzed substance outlier test etc.
 Calculate by hand, calculator, PC etc  No typing or calculation mistakes
 Independent from operator

Manual vs Automatic Titration 60

Documentation
Automatic Titration
Manual Titration  Automatic print-out
 Write calculated result on paper
 Slip or A4 printer
 Archive paper
 Contains all data, not just result
 Type result into electronic file
- Who did what when and how
 Archive files
- Fully GLP compliant
- Guarantees traceability
 Archive paper still manually
 Type result into electronic file
 Archive files
 Transfer data to LIMS
Manual vs Automatic Titration 61

Automatic Titration Benefit

 Accurate and precise results - due to the automatic titrant addition and the possibility of
adding very small amounts of titrant (lower than 0.001 mL).
 Fast but still precise titrations - due to the controlled titrant addition determined by an
algorithm (small increments near the equivalence point).
 Highly reproducible results - since no operator interaction is needed the results are
highly reproducible even if another operator performs the analysis.
 Full titration curve is recorded, which allows a titration to the equivalence point and post-
processing.
 The results can be calculated and printed automatically by the titrator.
 Traceability - the titration curve and the results can be stored (e.g. on a computer if the
titrator is connected) or printed directly.
 Operator has minimal contact with chemicals since the titrant and the reagents can be
added directly from the bottle.
 Depending on the titrator, the analysis can be fully automated thus saving time

Agenda 62

1 Basic of Titration

2 Reaction types in Titration

3 Manual VS Automatic Titration

4 Titrator Instrument

5 Aplication in F&B

6 Tips and Hints

Titration Innovations at Mettler-Toledo 63

DV/DK
The first METTLER TOLEDO titrator

 Analogue titrator
 Modular system

1973  Up to 16 modules for

EP, EQP, KF etc.
 Innovative, especially for
photometric titration
Titration Innovations at Mettler-Toledo 64

SR10 1st computerised system

 Controlled by microprocessor
(PDP 9-bit)
 Largest in the world

1974  Sample preparation included

 100-position sample changer
 10 automated burettes
 25 systems sold worldwide
 Price then: 90‘000 CHF for the basic unit, 330‘000 CHF for the
complete system
 Displayed in the Technical Museum in Munich

Light-years ahead of the times

Titration Innovations at Mettler-Toledo 65

DL40 MemoTitrator
1st compact general and KF titrator

 Z8 processor
 1st titrator with memory
 40 programmable methods

1979  Data storage

 Processing software
 Propeller stirrer
 Automatic titration
 MemoCard

The first modern microprocessor titrator

Titration Innovations at Mettler-Toledo 66

DL25 Titrator
Introduces dynamic addition of titrant

 50 programmable methods
 EQP detection with non-linear

1985 regression

Simple, quick and flexible

Titration Innovations at Mettler-Toledo 67

DL70 Line Autotitrators

 Online titration curve

 Flexible method creation
 Logic conditions

1989  4 automated burettes

 4 processing criteria
 Titrant database
 Automatic titrations
 2 titrations in parallel

Great application flexibility. The first for safety and GLP functions

Titration Innovations at Mettler-Toledo 68

DL50 Family of titrators

 1st titrator with memory card

(results, methods, config.)
 Expandable
 Online titration curve

1996  Automated 2-phase titration

 Titrant expiry verification
 Routine/Expert mode
 Intuitive and easy to use

The most modern operator interface. The first with online help.

MT Automatic Titrator Today...

FAMILY
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Titration Portfolio
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Titration Excellence Karl Fischer Compact Potentiometric

T5 T7 T9 V10S V20S V30S C10S C20S C30S G10S G20S

MODEL
PAGES

T5 T7 T9

+
One Click Titration
Official URL links:

[Link]/Titration
[Link]/Titration-Excellence
Sodium Determination [Link]/Karl-Fischer
[Link]/Compact-Titration
The New Compact 10S [Link]/One-Click-Titration
[Link]/Sodium-Determination
[Link]/Titration-10S
69
Introduction Titration Excellence

Introduction Titration Excellence

One Click® Titration Simple

Methods Manual Series

One Click® offers easy to understand simplicity

Internal usage only

Introduction Titration Excellence

One Click® Titration Excellence

Efficient
Efficient
Secure
Secure

Modular

Internal usage only

Introduction Titration Excellence

Titration Excellence approach to modularity:

1. Tailored Exactly to Your Needs.

 with segment specific applications
 with a huge variety of accessories
2. Get all You Need in One Box.
 expand efficiency all in one device
3. Be Future Ready.
 simple addition of accessories
 upgradability (T7 to T9)

Internal usage only

New Titration Excellence Line Highlights

 Multiple Standard Addition  SmartSample™ reader for
efficient and secure data transfer
for completely automated
sodium determination

 Coulometric Option  PDF writer and

expanding analysis range new data export for
with KF coul titration more flexibility

 Touchscreen terminal and new  StatusLight™ for

One Click® interface for increased efficiency
increased efficiency

New features emphasize Titration Excellence utmost modularity

New Titration Excellence Line Highlights

 Touchscreen terminal with a new look and

increased efficiency.
- Large 7 inch WVGA color TFT screen
- USB connection on right hand side
- Built in StatusLight™

 New One Click® User Interface

- Save up to 24 shortcuts
- User management with color
skinning
- Scrolling and Swiping

Modern appearance, more comfort and attractive for demos!

New Titration Excellence Line Highlights

 Coulometric Option enables Titration

Excellence to perform potentiometric, KF vol
and KF coul analyses.
- All-in-one system integrated
- Improved accuracy of Bromine
Index determination
- Simultaneous KF Vol and KF
Coul analysis on T9
- Overflow protection for KF Vol

Coulometer Board No Y cable

Always in the 4th slot anymore!

Potentiometric and Karl Fischer titration in one device

New Titration Excellence Line Highlights

 SmartSample™ reader to offer efficient and secure

data transfer for single samples

- Installed SmartSample reader on the titrator as standard

- Enables wireless data transfer from balance to titrator
- Elimination of transcription errors

Intelligent choice for single- and automated multi-sample titration

New Titration Excellence Line Highlights

The integration of the Sodium

Analyzer functionalities on Titration
Excellence offers:
- Ion determination based on multiple
standard addition
- Completely automated sodium analysis
- Accurate ion specific analysis

Strong feature for Food and Beverage industry

New Titration Excellence Line Highlights

 StatusLight™ supports you in routine operations and daily work by visual and acoustic
signaling.

- StatusLight™ on titrator and terminal

- StatusLight™ concept aligned with InMotion™, UV/VIS and
Excellence Balances
- Built-in loudspeaker for acoustic signals

Stay in touch with your titration

New Titration Excellence Line Highlights

 Flexible Data Export

- CSV
- PDF
- XML

More flexibility in data export

Data Export
Ethernet

RS232
RS-P26
USB
Excellence
XML Stream

USB-P25

USB Stick
Network Printer
PC
XML Stream
Local Printer

USB
Data Export Box
Ethernet
Compact Server
LabX

LabX connectivity:

 New LabX Version 2016 with launch

 New titrators can be connected

 New features not yet compatible with Version 2016

 Compatibility will be assured with Version 2017!
 Launch Version 2017: JULY 2016

Agenda 83

1 Basic of Titration

2 Reaction types in Titration

3 Manual VS Automatic Titration

4 Titrator Instrument

5 Aplication in F&B

6 Tips and Hints

Salt Content 84

As chloride or sodium chloride

 Titrant: 0.1 mol/L AgNO3
 Solvent: water
 Auxiliary reagent: HNO3
 Sensor: DMi141-SC or DMi145-SC or DMi148

NaCl + AgNO3 AgCl + Na+ + NO3-

Tips & Hints

- pH must be below pH 6 (ideal pH 4.5) to prevent AgOH formation.
- pH should not be below pH 2 - it dissolves the silver ring of the
electrode.
- For high chloride content take care not to titrate too fast:
co-precipitation
- Can add PVA to prevent co-precipitation
- To clean sensor, stirrer and burette tip, dip in ammonium
hydroxide (Automation – use conditioning beaker)
Salt Content 85

Sodium, Na+
 Sodium has been linked to high blood pressure.
 High blood pressure can increase the risk of
heart disease, kidney disease and stroke.

 Na-content is relevant
 Determination of Na-content by standard addition technique

Salt Content

 Examples
- Ketchup
- Dairy product (cheese, butter)
- Vegetable juice
(carrots, tomato, etc.)
- Soup, meat and
vegetable bouillon
- Sauces, meat extract
- Bread and pastries
- Snack food (potato chip)
- Breakfast cereals
- Seasoning powders
- Spices
- Bean paste sauce
- Pasta sauces
86

Acid Content 87

Acid content as:

 Acetic acid, Tartaric acid, Citric acid, Malic acid, Free fatty acid, etc.

Acetic acid M = 60.01z = 1

Tartaric acid M = 150.09 z =2
Citric acid M = 192.13 z =3
Malic acid M = 134.09 z =3

CH3COOH + NaOH CH3COONa + H2O

 Titrant: 0.1 mol/L NaOH

 Solvent: water
 Sensor: DGi115 or DGi114 or DGi111
 Acid Content

 Examples
- Vinegar
- Juices (apple, orange)
- Wine
- Coca Cola
- Yoghurt drink
- Milk
- Sour cream
- Honey
- Edible oil

88

Calcium content 89

Calcium
 Titrant:0.1 mol/L EDTA
 Solvent: water
 Sensor: DP5 Phototrode™
or
DX240 Ca-ISE
Ca2+ + EDTA  [Ca-EDTA]2+

Tips & Hints

- pH must be adjusted to pH 10
with ammonia or borate buffer

- Not suitable to use the DP5 Phototrode™

for beverage samples due to opacity,
natural colors and protein suspensions

- Alternative sensor – Ca ISE

- Indicator not necessary when Ca ISE sensor is used

Calcium content

 Examples
- Milk
- Soy Milk
- Cheese
- Yoghurts
- Juices

90
Vitamin C determinations 91

 Three ways to titrate

- Potentiometric
- Titrant: Iodine (direct titration) or sodium thiosulfate (back-titration)
- Reaction: Redox reaction
- Sensor: DMi140-SC or DM147i-SC

- Voltametric
- Titrant: DPI
- Reaction: Redox reaction
- Sensor: DM143-SC Polarized sensor

- Amperometric
- Titrant: DPI
- Reaction: Redox reaction
- Sensor: DM143-SC Polarized sensor

Vitamin C determinations 92

Potentiometric Indication
 Titrant: 0.05 mol/L I2
 Solvent: water
 Auxiliary reagent: 5% H2SO4
 Sensor: DMi140-SC or DMi147-SC
C6H8O6 (aq) + I2 (aq) C6H6O6 (aq) + 2 HI

- Direct titration with Iodine (Iodimetric titration)

 Quantitative oxidation of ascorbic acid to
dehydroascorbic acid by iodine in acid solution.
- Back titration with sodium thiosulfate
 Adding KI and KIO3 to generate I2
 Excess iodine is titrated with Na2S2O3
 Results more reliable than direct titration (less I2 is lost to the environment).

 Disadvantages:
Iodine is not selective to Vitamin C; other components will be oxidized, e.g. SO 2.

Vitamin C determinations 93

 Voltametric or amperometric Indication

 Titrant: 0.01 mol/L 1/2 DPI (2,6-Dichlorophenolindophenol)

 Solvent: water
 Auxiliary reagent: Oxalic acid or NaOH to adjust pH to 3
 Sensor: DM143-SC

C6H8O6 + DPI  C6H6O6 + H2-DPI

Advantages:
 DPI is more selective for Vitamin C than iodine
 DPI gives a higher jump at the EQP compared to the titration with iodine.
Vitamin C determinations 94

Voltametric Indication
 Titration curve: “sudden” jump
- Accurate results requires incremental addition.
Fixed increment = 0.03 – 0.1 mL
(In same cases also dynamic titrant addition works)
- Evaluation: Standard or asymmetric

700 mV

Tx
Ipol = 12.0 µA

Standard curve
100 mV

Vitamin C determinations 95

Amperometric Indication
 Segmented titration curve:
- Evaluation: segmented
- Segmented evaluation allows (requires) the use of larger increments.
Fixed increments 0.1 – 0.2 mL
 faster titration; titration time 1 to 3 minutes

4 µA
Tx
Upol = 200 mV

0 µA

Vitamin C determinations

 Examples
- Juices

- Canned fruits
- Dried fruits

96
Vitamin C determinations 97

Tips & Hints I

 Titrant DPI preparation.

- Use DPI sodium salt monohydrate, which dissolves good in water.
DPI sodium salt dihydrate does not dissolve very good in water (Filtration needed.)
 Titrant DPI is not stable.
- Titrant DPI is unstable and should be prepared freshly, at least every 2 nd day, and should
be stored in the dark when not used.
- Run a titer determination every day.
 Titer determination (Standard: Ascorbic acid).
- Since the sample size is very small (20 mg) use a stock solution.
- Make a fresh stock solution for every titer determination, since ascorbic acid is not stable.
 pH adjustment to pH 3 is very important.
- For the titer determination, the pH should also be adjusted to pH 3, because deionized
water has a pH between 6 and 7, which is too high.

Vitamin C determinations 98

Tips & Hints II

 Clean sensor between samples for better precision.

 Vitamin C is not stable against light and oxygen.

- For accurate work bubble nitrogen through sample during titration and titrate with red
titration beakers to protect against light.

 Iodine and DPI are not selective for Vitamin C

- Iodine is less selective than DPI.
If orange juice contains also SO2,
- with Iodine: Vitamin C and SO2 will be titrated.
- with DPI: Vitamin C and approx. 20 % SO2 will be titrated.

- It is possible to selectively titrate Vitamin C in a sample which also contains SO 2 by

masking SO2 with glyoxal (ethane-1,2-dione, C2H2O2).

Free and Total SO2 99

Free sulphur dioxide, SO2

 Titrant: 0.1 mol/L ½ I2
 Auxiliary reagent: 5 mL KI 10 % and 5mL 25% H2SO4
 Sensor: DM143-SC

SO2 + I2 + 2 H2O H2SO4 + 2 HI

Total sulphur dioxide, SO2

- Add 5mL 5 M NaOH and allow to react for 15 minutes to release
bound SO2 (Swiss Food Manual)
- Acidify with 7mL 25% H2SO4 and add 5 mL KI 10 %.
(KI addition to delay reduction of I2 by other components in sample)
- Titrate with I2 as for free SO2
Free and Total SO2

 Voltametric titration to endpoint EP=100 mV (similar to KF)

100

Free and Total SO2

Tips & Hints

 SO2 is volatile
- In order to avoid loss of SO2, the samples must be taken from freshly
opened bottle.
- Working with sample changer leads to small SO 2 losses from the samples
already prepared.
 Iodine is not selective for SO2
- These competing reactions can partially be delayed by addition of KI.
(not ascorbic acid).
 SO2 Standard
- Sodium metabisulfite, Na2S2O5 , can be used as a SO2 Standard
(Na2S2O5  2 SO2)
 Very low content of SO2 in wine (< 20-50 ppm)
- Titrate with EQP und Iodine solution (for 3 ppm VEQ ca. 0.2 mL)
101

Free and Total SO2

Examples
- Wine
- Cider
- Fruit based drinks
- Dried fruit
- Flour
- Sausages

102
Free and Total SO2 103

 Use of SO2
 Used as a preservative in wine for its antibacterial properties
- Sorbic acid in wine is reduced by bacteria to foul smelling ether

 Bleaching agent in flour

 Fumigate fruit and vegetables to extend their shelf life

Kjeldahl Nitrogen 104

Principle of Nitrogen Determination according to Kjeldahl

 Digestion
Sample strongly heated for 2 – 3 hours in digestion flask containing sulfuric acid 98 %.
 Distillation of ammonia
NaOH solution 30 % is added. Ammonia is generated according:
(NH4)2SO4 + 2 NaOH  2 NH3 + Na2SO4 + H2O
Ammonia is distilled using water steam and collect in 1 – 2 % boric acid solution
2 NH3 + 4 H3BO3  (NH4)2B4O7 + 5 H2O
 Titration
Ammonium tetraborate, (NH4)2B4O7, can be titrated with HCl.
(NH4)2B4O7 + 2 HCl + 5 H2O  2 NH4Cl + 4 H3BO3

 Titrant: 0.1 mol/L HCl

 Solvent: Water
 Sensor: DGi111 or DGi115
Johan Gustav Christoffer Thorsager Kjeldahl
(Danish chemist, 1849 - 1900 )

Kjeldahl Nitrogen 105

TTL I/O (RS232) for automated Kjeldahl analysis

Büchi (KjelFlex K-360)

Communication between Distillation unit

• Compact Titrator G20

• Titration Excellence T50 T70 and T90

via TTL I/O (and RS232 for Txx)

Büchi K-360 with T50

 Kjeldahl Nitrogen

 Examples

- Milk
- Cheese
- Yogurt
- Butter

106

Additional Applications 107

 Reducing sugar acc. Rebelein (M496* – furthermore M568, M341, M342)

 Peroxide Value (M624* - M369)

 Iodine Value (M617*)

 Saponification Value (M618* - M090*)

 Formol Value (M493* - M061, M240)

* with Titration Excellence

Saponification value

Agenda 108

1 Basic of Titration

2 Reaction types in Titration

3 Manual VS Automatic Titration

4 Titrator Instrument

5 Aplication in F&B

6 Tips and Hints

Tips and Hints 109

 Titration beakers
- Use absolutely clean titration beakers.
- Clean titration beakers in a dishwasher and rinse at least 2 times with
deionised water.
- The material of the beakers (plastic or glass) should be resistant to all
chemicals used for the titration.
- If the analytes are sensitive to light use special beakers which protect
the sample from UV radiation.

 Burette
- Choose the appropriate burette size. The titrant consumption should
be within 20 % to 90 % of the burette volume for an accurate
measurement
- Estimate the burette size by calculating the titrant consumption from
the expected value or perform a fast test titration.
- Avoid bubbles in the burette or remove them before starting the
titration (rinsing) as they will produce results that are too high. De-gas
the titrant or reduce the titration speed if bubbles start coming from the
titrant.

Tips and Hints 110

 Tubes (automated titration)

- Remove all bubbles from the tubes (or the results will be too high) by tapping gently
during rinsing.
- Use a siphon to avoid diffusion of titrant into the titration solution.

 Indicator (manual titration)

- Choose an indicator with a color change range within the range of the equivalence
jump.
- Don't add too much indicator; the indicator may influence the analyte/titrant
reaction. Just a few drops of indicator solution usually suffice.

Tips and Hints 111

 Sensor (automated titration)

- Store and maintain the sensor as described in the operating
instructions.
- Ensure that the electrolyte level is high enough, namely higher
than internal elements and titration solution
- Poor sensor response results in titrant consumption and results
that are too high.
- Perform a sensor test periodically to check slope, offset and drift
value of sensors which need calibrating (pH sensor).
- Perform a sensor calibration, especially before endpoint
titrations and after every maintenance action.
- Some sensors need conditioning (see operating instructions)
before they can be used or after they have not been used for a
long time.
- Reference electrodes (which are also part of a combined
electrode) need to be in contact with the sample solution by a
junction (diaphragm). Avoid junction blockage for correct and
stable results. When required clean the junction using an
appropriate solution (see operating instructions).
Tips and Hints 112

 Sample solution
- The overall sample solution (sample, reagents and solvent) should be enough to cover
the sensor, typically 50 mL. Add solvent (water for aqueous titrations) if the volume is
too small.
- The amount of sample should be chosen such that the titrant consumption is within 20
% to 90 % of the burette volume.
- Use accurate measuring tools (analytical balances, pipettes, …) to measure the
sample size.
- Perform a blank value determination if the solvent isn't pure.
- Depending on the titration reaction (e.g. redox and precipitation titration) make sure that
the pH value of the sample solution is within the requested range.

Tips and Hints 113

 Titrant
- Perform a titer determination to know the titrant concentration accurately.
- Use high purity, stable, non-hygroscopic substances with a high molecular weight (for a
high sample size) as the titer standard substance.
- A titer determination interval (usable life) and a lifespan should be defined depending
on the stability of the titrant.
- Protect titrant from CO2, UV radiation, humidity or other factors which may have an
influence on the concentration.

114
Getting the best results possible

1 Internal usage only

Agenda

Sources of errors

Using the right titration method

Reagent handling

pH calibration and sensor care

The effects of temperature

Sampling & weighing the right way

Conclusion

2
Sources of Error

Systematic error:
a generally unknown component of the error which remains constant or
drifts slightly

Examples:
- Different analytical method
Inaccurate result
- Wrong sample size
- Titrant concentration
- Blank value
- Sensor adjustment/calibration
- Sensor response
- Sensor conditioning

3
Sources of Error

Random error:
a component of the error which varies in an unpredictable way

Examples:
- Inadequate equipment
- Method parameters Imprecise result
- Bubbles
- Sample handling
- Sample
- Rinsing
- Operator

4
Agenda

Sources of errors

Using the right titration method

Reagent handling

pH calibration and sensor care

The effects of temperature

Sampling & weighing the right way

Conclusion

5
Using the right method for the task

The end point titration

pH

Fast reaction, slow electrode

Slow reaction, fast electrode

pH 8.2

mL

6
Using the right method for the task

The end point titration

- Often faster
- Approximation
- Electrode calibration
- Temperature
- Reaction speed
- Electrode response

7
Using the right method for the task

The equivalence point titration

pH

Equivalence point

mL

8
Using the right method for the task

The equivalence point titration

- ‘True’ result
- Curve shape
- Increment size
- Time between increments
- Recognition

Whenever possible
use an equivalence point titration!

9
Agenda

Sources of errors

Using the right titration method

Reagent handling

pH calibration and sensor care

The effects of temperature

Sampling & weighing the right way

Conclusion

10
Reagent handling

Preparation and storage of titrants

- CO2-free water
- Protected from CO2
- Freshly prepared e.g. DPI for vitamin C
- Light-sensitivity
- Temperature
- Standardization

11
Titrant Standardization: Why?

To know the exact concentration of the titrant

To get accurate and precise results

To use it as a test titration

To test the whole measuring system

- Titrator
- Burette drive
- Burette
- Sensor

12
Titrant Standardization: Definition

Titration of a pure primary or secondary standard of exactly known

content to determine the true concentration of the titrant.

Actual concentration
Titer =
Nominal concentration

Example:
Nominal conc.= 0.1 mol/L Actual conc. = 0.09899 mol/L

Titer = 0.9899

13
Examples of Primary Standards

Acid-base reaction:
KHP, Tris(hydroxymethyl)aminomethane (THAM),
sodium carbonate, benzoic acid (non-aq)

Redox reaction:
Potassium dichromate, sodium oxalate, ascorbic acid,
potassium iodate

Complexation reaction:
Calcium carbonate, zinc sulfate, Na-EDTA

Precipitation reaction:
Sodium chloride, sodium sulfate, sodium dodecylsulfate (SDS)

14
Titrant Standardization: Primary Standards

Known composition and high purity

Stable at room temperature, easily dried:
- Not hygroscopic
- Do not react with oxygen and moisture in the air
- Not light sensitive

Reaction with compounds goes to completion (100%)

Equivalence mass should generally be high to keep weighing error
small

Easily soluble in the solvent:
Ensure sufficient stir time, e.g. 180 s for Na-tartrate

Primary standard solutions do not require other standardization if prepared

by accurately weighing and dissolving the standard in the solvent.

15
 Standardization of 0.1 M HCl

Primary standard:
50-80 mg THAM

Glass titration beaker

50 ml deion. water

Results:

Series start time 01.06.2010 [Link]

No. Comment / ID Start time Rx Result Unit
Name
1/3 TRIS 01.06.2010 [Link] R1 = 0.99604 -- Titer
2/3 TRIS 01.06.2010 [Link] R1 = 0.99958 -- Titer
3/3 TRIS 01.06.2010 [Link] R1 = 1.00074 -- Titer

Rx Name n Mean value Unit s srel [%]

R1 Titer 3 0.99879 -- 0.002447 0.245

16
Standardization of 0.1 M NaOH

Primary standard: 70-120 mg KHP

Glass titration beaker, 50 ml deion. water

Results:

Rx R1
Name Titer
n 6
Mean value 0.99527
Unit --
S 0.00234
srel [%] 0.235

(Method: CSTiter435,
21.7.2010)

17
Standardization of 0.1 NaOH: Uptake of CO2

Alkaline titrants (alkalimetry): 30000

Conc.
CO2 + H2O = H2CO3 = CO32- + 2 H+ [mg/L]

20000

CO32- : Exp. air Result

days [mg/L]
1 2526 10000
2 5026
3 8793
4 14422 0

6 20684 0 2 4 6 8

7 24568 days

NaOH: Exp. air Result Theor. Conc. Dev. RSD

days [g/L] [g/L] [%] [%]
1 4.837 4.9017 -1.31 0.051
2 4.586 4.9017 -6.44 0.139
3 4.308 4.9017 -12.11 0.178
4 4.152 4.9017 -15.20 0.108
6 3.906 4.9017 -20.35 0.162

18
Effect of Carbon Dioxide

mV

With carbon dioxide

Without carbon dioxide

mL Acid
19
Standardization of 0.1 M HClO4 (non-aq)

Primary standard: 50-90 mg THAM

Glass titration beaker with 50 ml 100% acetic acid

Results:
Rx Name n Mean value Unit s srel [%]
R1 Titer 8 1.007340 -- 0.002122 0.211

Movable
(Method: CDCTiterHClO4, 15.6.2010, sample 1) sleeve

20
Standardization procedures

Frequency depends on titrant stability

- Karl Fischer Reagents: daily
- Basic titrants: daily to weekly
- Acids / Metal salts: weekly or less frequently

Check each titrant individually

Lower concentrated titrants e.g. 0.01 M NaOH
require more frequent standardization

Use same conditions (i.e. temperature, stir speed,..) for standardization

as for sample titration

21
Titrant Standardization: Tips and hints I

Protect titrants against:
- Humidity (Silica gel, molecular sieve)
- Carbon dioxide
8Drying tube ME-23961 and holder ME-23915
8CO2 absorbing material NaOH on support

Follow appropriate sample preparation conditions

- E.g. use glass beaker with KHP to avoid problems caused by
electrostatic effects
- Use correct balance and check for suitable sample handling

Run standardizations more than once
- Recommended: at least 3 x
- Set upper and lower acceptance limits for srel values

Order small quantities of primary standards
22
Titrant Standardization: Tips and hints II

Date chemicals on delivery

Only use deionized water or better

Glass beakers reduce disturbances due to electrostatic effects

If using the same titrants on two different instruments:
- Standardization on both instruments is required.

When using PP beakers:
- Leave to stand for a few minutes
to release electrostatic build-up
and then weigh out the sample

23
Tips and Hints III: Effect of Water on HClO4

Too much water

mV

More water

Little water

mL Perchloric Acid

24
Volumetric Buret

Sample size, buret size and titrant concentration

shall be designed to use 30 to 80% of the buret volume
to reach the endpoint.

Refilling a buret during a titration should be avoided since it can lead to

signal instability or irregularities in the curve profile.

25
Buret maintenance

Cleaning
• Empty and clean buret, tubes and
tips at least once every 3 months

Flushing
• Flush buret whenever bubbles are
present, but at least once every day

Replacement of glass cylinder
• Have buret checked annually,
especially when bases are used

Regular servicing
• Have instrument serviced once
per year, depending on usage

26
Agenda

Sources of errors

Using the right titration method

Reagent handling

pH calibration and sensor care

The effects of temperature

Sampling & weighing the right way

Conclusion

27
Electrode calibration, adjustment and care
What is a calibration?
- Establishing the relationship between the
measured signal and pH or pM/pX by
measuring solutions of known and traceable values
mV

4 7 10 pH

28
Electrode calibration, adjustment and care

When is a sensor adjustment required?

- If sample pH is measured as a result
- If a pH endpoint titration is performed
- If a calibration indicates that one is required

How to calibrate and adjust and how often?

- Measure the potential of at least 2 known buffers e.g. pH 4 and
pH 7
- Calibrate at least once per day (depending on temperature
fluctuations)
- Adjust when necessary based on the calibration

29
Troubleshooting of electrodes

If an electrode is not performing well check for

- Membrane dehydrated, contaminated or damaged
- Electrolyte contaminated or missing
- Air bubbles behind the ceramic junction or membrane
- Junction contaminated
- Crystallized KCl blocking outlet of electrolyte
- Electrode worn out (out of specifications)

30
Electrode maintenance

Cleaning the electrode

After each measurement rinse the electrode with distilled water

Never wipe the membrane with a paper towel!

31
Re-conditioning of electrodes

Dry electrode

Dry membrane:
Re-conditioning in 0.1 M HCl for 12 hours

Electrode out of specifications

Regeneration of the membrane:
Place electrode in re-conditioning solution (NH4HF2) for 1 - 2
minutes. Caution: harmful etching agent!

It is easier to keep an electrode working than to recover a non-working

one

32
Electrode maintenance in non-aqueous media

Electrode cleaning
- Rinse electrode in pure solvent followed by a water rinse
- If non-polar solvents are used for titration, increase polarity of
rinse solution stepwise, i.e. start with toluene or xylene, followed
by 2-propanol and finally water

Electrode conditioning in water
- After 1-3 samples condition electrode about 5 minutes. If
necessary, condition after each sample.
- Before the next titration, rinse off the conditioning solution with
solvent

After daily use:
- Store electrode over night in diluted acid or 3M KCl for hydration.

The electrodes must be adjusted after regeneration

33
Conditioning of electrode

In non-aqueous medium the electrode loses the water

layer on the membrane.
Without water layer the signal of the electrode is
incorrect and instable.

Solution:

After every sample conditioning in water
(or buffer solution) at pH 4.5 - 5.5
for minimum 3-5 minutes to rebuild the water layer of
the pH sensitive glass membrane

Remove the excess of water by rinsing with solvent
prior to next determination.

34
Example: Missing electrode conditioning

Customer analysis
- Assay of Metformine-HCl with HClO4 (USP 32, 2010) in acetic acid
as a titrant, and formic acid/acetic acid anhydride as solvent.

Problem:
- After few samples within tolerances (recovery: 98-101%), the
variation becomes too large and the recovery is outside the specs.
- This can be hardly seen from the titration curve since it looks great.
Variation starts appearing after 2-3 days of continuous titration

Solution:
- Before the subsequent titration, the sensor is conditioned in
aqueous 0,02 M HCl during 180 s.
- At the end of a sample series, the sensor is parked in aq. 0.02 M HCl.
- Optional: Additional conditioning in acetic acid anhydride during 10 s.
35
Example: Missing electrode conditioning

Experimental setup

36
Example: Missing electrode conditioning

Results

Method:
CDC001Sample
21.06.2010 09:16

Rx Name n Mean value Unit s srel [%]

R1 Content 5 100.0140 % 0.2562 0.256

Hint:
- Always run a dummy titration before starting each analysis, e.g.
before a blank value determination, a titer determination and a
sample determination. Discard the result of this titration since it is
used to condition the electrode 37to the acetic anhydride solvent.
Reference electrolytes

Exchange electrolyte at least once every 3 months
- Do not simply top up
- Drain electrode and refill with fresh electrolyte
- Ensure that the correct electrolyte is used (with or without AgCl)

Electrolyte level in the electrode higher than in the sample solution
- If the level of the electrolyte is lower than the sample solution, then the
latter flows into the reference system of the electrode; the
consequences are wrong results

No air bubbles behind the junction

- Vertical shaking of the electrode
Which electrolyte?
- Aqueous solutions: 3 M KCl, 3 M KCl saturated with AgCl, 1 M KNO3,..
- Non aqueous solutions: 1 M LiCl in ethanol, 1 M LiClO4 in acetic acid,
…
38
Cleaning of junction

- Blocked with AgCl: with concentrated ammonia

- Blocked with AgS: with 8 % thiourea in 0.1 M HCl

- Blocked with protein: with 5 % pepsin in 0.1 M HCl

- Other contaminations:
in ultrasonic bath with water or 0.1 M HCl

Treat one hour, rinse with distilled water and re-calibrate

39
Aging of electrodes

Symptoms of an old electrode

- Sluggish response
- Small slope
- Offset out of specifications
- Unstable signal, drifting pH value
Tests
• Calibration with buffer 4 and 7:
Offset: 0 mV ± 30 mV;
slope -55 mV/pH to -59,2 mV/pH

• Determination response time:

30 seconds after change from buffer 4 to buffer 7 the potential
should not change more than 2 mV.

40
 Electrode test and adjustment

Sensor test and calibration with shortcuts on the

touch screen of the automated titrator

Simple test
and sensor adjustment.

Message and warning,

if sensor out of specs.

Message, when
sensor adjustment
is needed.

41
Aging of electrodes

Lifetime
The approximate lifetime of a pH electrode with good storage,
maintenance and measurement in aqueous solutions in the pH range
of pH 1 to pH 12:

at ambient temperature 1 - 3 years

at 90°C 3 - 9 month
at 120 °C few weeks only

42
Electrode Storage: Combined pH Glass Electrodes

Always store in one of the following

- 3 M KCl
- Dilute HCl diluted (approx. 0.1 M)
- Buffer solutions pH 4 or 7

Never store glass electrode in distilled water or dry

- Shortens lifetime
- Electrode needs to be re-conditioned before use

43
Electrode Storage: Metal Ring Electrodes

Redox electrodes - Storage

- Combined: stored in the reference electrolyte (3 M KCl)

Redox electrodes - Passivation (thin oxide coating)
- Immerse the electrode in a solution of 0.5 g quinhydrone in
50 mL pH 4 buffer, rinse with water

Ag metal electrodes - Storage
- Combined electrodes: in the reference electrolyte - saturated
KNO3

Ag metal electrodes - Refilling
- Only with KNO3 solution; never use KCl
- Crystals of KNO3: rinse with water and add fresh reference
electrolyte

44
Agenda

Sources of errors

Using the right titration method

Reagent handling

pH calibration and sensor care

The effects of temperature

Sampling & weighing the right way

Conclusion

45
The effect on temperature on pH
Electrode slope and temperature compensation

mV By measuring the buffer and

sample temperatures, the titrator
(or pH meter) can compensate for
the change in slope

Caution:
pH no instrument can compensate for
real changes in pH as a result of
temperature change!

T1 Conclusion
T2 The only really accurate result is
T3 when the buffers and samples are
at the same temperature!

46
Agenda

Sources of errors

Using the right titration method

Reagent handling

pH calibration and electrode care

The effects of temperature

Sampling & weighing the right way

Conclusion

47
Requirements of the sample

Sample taken should be representative of the overall

material
- Mix sample thoroughly before sampling
- Make sure sample is large enough to be
representative

Must be stable during weighing
(methods other than weighing possible)
- Take care of hygroscopicity of the sample
with Karl Fischer samples

Corrections for water or LOD, if desired, are possible

It has to react stoichiometrically with the titrant

No secondary/side reactions

48
Balance

Every balance has precision and

accuracy specifications

THE AMOUNT OF SAMPLE HAS

TO BE ACCURATELY KNOWN

The balance should be selected

based on the sample size to
provide at least 0.1% accuracy
and precision
(For 100 mg sample, need to
weigh to 0.1 mg or less)

49
Agenda

Sources of errors

Using the right titration method

Reagent handling

pH calibration and electrode care

The effects of temperature

Sampling & weighing the right way

Conclusion

50
Conclusion

For accurate and precise results you need….

Suitable and Optimized Titration Method

Calibrated and Working Instrument

Calibrated and Working Balance

Correctly Prepared and Stored Titrant

Proper Reagents and Solvents

Appropriate and Adjusted Electrode

Maintenance and Care

51
Good Titration Practice
in the Pharmaceutical Industry
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrationstitration)
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
1
The strength of acids and bases

Examples of acids Examples of bases

Acid pKa Base pKb

Perchloric acid -9 Guanidine 0.5

Formic acid 3.75 Methylamine 3.36
Acetic acid 4.73 Ammonia 4.75
Lactic acid 3.86 Diethanolamine 5.12
Benzoic acid 4.20 Picoline 7.52
Phenol 9.95 Quinoline 8.94
Hydroquinone 10.0 Pyrazol 11.51
Caffeine 13.39

The weaker an acid (base) the higher the pKa (pkb) value

Note: the pK values are defined in aqueous systems

2
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrationstitration)
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
3
How to titrate a weak acid or base?

In aqueous media the H3O+ or the OH- ions are the strongest
acids or bases

The solvent water shows a levelling effect on the acid or
base strength

Very strong acids or bases [pKa(b) <0] or very weak acids or
bases [pKa(b)>9] cannot be titrated in water

In non-aqueous media there is no competition from the auto
dissociation of water
H2O + H2O H3O+ + OH-

In non-aqueous solvents the dissociation of an acid or a
base is totally different compared to water

Example: A solution of picrinic acid in acetone is more
acidic than a solution of HCl in water
4
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrationstitration)
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
5
Advantages of non-aqueous titrations

Expansion of the solubility range: many substances that are
insoluble in water can be titrated in non aqueous solvents.

Expansion of the application range: weak bases and weak acids

that cannot be analyzed in water can be easily titrated in non-
aqueous solvents. Example: Pyrazole (pKb = 11.51), 2-
Aminobenzoicacidmethylester (pKb = 11.82), Phenol (pKa = 9.95)

Titration of substance mixtures that cannot be separated in

aqueous media. Example: Isopropylamine (pKb = 3.37) and
Aniline (pKb = 9.42) mixture

Undesirable side reactions of titrant and sample with water are

avoided
6
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrationstitration)
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
7
Non-aqueous base titration

Determination of amines

R-NH2 + HClO4 R-NH3+ + ClO4-

Examples for direct titration of amines

- Benzylnicotinate 1 (Rheumatism) 1
- Dequalinium chloride 2 (Anti-bacterial)

Examples of protonated amine salts titrations
2
- Diphenylpyraline Hydrochloride 3 (Anti-Histamine)
- Tramadol Hydrochloride 4 (Analgesic, “pain killer”)

Note:
Mercuric acetate is used to facilitate titrations by 3
Releasing free amine from the protonated amine salts:

2R-NH3+Cl- + Hg(OAc)2 2R-NH2 + HgCl2 + 2HOAc

4
8
Titration of bases – levelling/differentiating

In aqueous systems only weak bases with a pKb value of 7 can be

titrated, e.g. aliphatic amines

Bases with a pKb value from 7 – 12 (aromatic amines or

heterocycles) can be titrated in organic solvents, mostly glacial
acetic acid.

Perchloric acid in glacial acetic acid is used as the titrant

9
Solvents for titration of bases

Crucial for the success of a titration

Must have a high dielectric constant in order to
solubilize the analyte and to enable a sharper
potential jump

The nature of the solvent determines the maximum
permissible base strength

Must be available in sufficient purity and dryness

10
Solvents for titration of bases

Acetic acid O
- Very good solvent
- Enables titration of weak bases up to pKb values of 12 H3C OH
- Analysis of bases mixtures is not possible – levelling effect
- Rarely acetylation is a side reaction

Acetic acid anhydride O O
- Very good solvent, similar to acetic acid
- Added as auxiliary reagent to maintain anhydrous conditions
H3C O CH3
- Preferred solvent for titration of very weak bases up to pKb
values of 12

Methanol
- Very good solvent, similar to water H3 C OH
- Enables titration of bases up to a pKb value of 9

2- Propanol (Isopropanol)
- Good differentiation of base mixtures H3C CH CH3

- Enables titration of bases up to a pKb value of 10 OH

Acetone
- Excellent differentiation of base mixtures O

- Very good solvent

H3C CH3
11
Titrants for the titration of bases

Perchloric acid can be purchased as a standard stock solution (0.1M in
70% aqueous solution) for dilution with the appropriate solvent

Perchloric acid in glacial acetic acid

- Glacial acetic acid (99.8%) as the solvent
- This solution contains approx. 0.5% water
- If non aqueous conditions are required, acetic anhydride must be added. Tip:
0.5% water require 28 g acetic anhydride

Perchloric acid in acetic anhydride
- Dilution of the standard stock solution with acetic anhydride

Perchloric acid in 2-Propanol (Isopropanol)
- The standard stock solution is mixed with 2-Propanol into 500 mL of 2-
Propanol
- This titrant is stable

12
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrations
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
13
Non-aqueous acid titration

Determination of aromatic alcohols

R-OH + (R‘)4NOH R-O - + N(CH3)4+ + H2O

R‘ = Methyl, Butyl
OH

Examples for direct titration of phenol

and derivatives OH

- Phenol HO O
- Rutoside OH
OH

HO OH
O
O
O
OH O

H3C O
HO

HO
OH
14
 Solvents for titration of acids

1,2 - Diaminoethane H2
- Determination of acids with pKa values up to 11 C NH2
H2N CH2
- Increasing levelling effect with increasing acid strength, at pKa<6 it is
not possible to distinguish between acids in a mixture
- Must be protected against CO2 contamination

Dimethylformamide O
- Excellent solvent, high dielectric constant
- Highly differentiating solvent, only strong acids with pKa<0 are levelled, H N(CH3)2
i.e. mixtures of strong acids cannot be determined
- Steep inflections at the equivalence point
- Must be protected against moisture to avoid hydrolysis

2-Propanol (Isopropanol)
- Good solvent H3C CH CH3

- Highly differentiating solvent, only strong acids with pKa<0 are levelled, OH
i.e mixtures of strong acids cannot be titrated

Acetone O

- Excellent solvent
- Even very strong acids can be differentiated from each other H3C CH3

15
Titrants for titration of acids

Almost exclusively Tetramethylammoniumhydroxide (TMAH) or
Tetrabutylammoniumhydroxide (TBAH)

TMAH solutions in 2-Propanol yield maximum specifity but are not
stable

TMAH solutions in methanol yield medium to low specificity but
higher stability

Mixtures of 2-propanol and methanol and a small portion of water
(1%) increase the stability: titrants may be stored for 1 year

TBAH has the same basicity as TMAH

The TBA salts generated during the titration are more soluble than
the TMA salts

Some TMA salts are poorly soluble and precipitate at the electrode
surface
16
Titrants for titration of acids

0.1 M TMAH or 0.1 M TBAH can be purchased as a standard stock

solution in 2-propanol/methanol

TMAH can be stored for 1 year, however slight decomposition to
TMA can be observed

TBAH is less stable, shall be consumed within 6 months

If other solvents are required the respective standard stock solution
is diluted with the required solvent.

Protect titrant from atmospheric CO2!

Recommendation: Use NaOH coated, activated Charcoal

17
Tips for non-aqueous acid titrations

TMAH and TBAH are susceptible to

- Decomposition to the free amines TMA and TBA
- CO2 contamination from ambient atmosphere

The free amine causes an additional inflection in the titration curve
before the actual equivalence point of the analyte

The CO2 causes an additional inflection in the titration curve after
the actual equivalence of the analyte

The CO2 inflection can be surpressed by saturation of the solution
with nitrogen

18
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrations
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
19
Standardization of titrants

Standardization of perchloric acid

- Potassium hydrogenphtalate (KHP)
8Stable
8Non-hygroscopic
8Easy to dry (105 – 135°C)
8Must be dissolved in hot glacial acetic acid
8Disadvantage: precipitation of KClO4
- 2-Methylimidazole
8Stable
8Good solubility in organic solvents

Standardization of TMAH or TBAH

- Benzoic acid
- Perchloric acid in 2-Propanol (not in acetic acid!)

20
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrations
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
21
Sensors for non-aqueous titrations

Potentiometric indication – requirements for electrode

Large pH glass membrane to achieve fast signal

response and equilibration

Electrostatic shielding in order to minimize interferences
that contribute to noisy curves

Large (moveable) junction to provide sufficiently large
electrolyte flow for signal stabilization

LiCl in Ethanol recommended as electrolyte

22
Sensor maintenance

Cleaning
- Rinse sensor in pure solvent followed by a water rinse

Conditioning in water.
- After 5 - 10 samples condition electrode about 5 minutes.
- Before the next titration, rinse off the conditioning solution with
solvent

After daily use:

- Store electrode over night in diluted acid or KCl 3 mol/L for
regeneration and hydration.

The sensors must be calibrated after regeneration

23
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrations
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
24
Blank determination
In non-aqueous media it is highly recommended to perform a blank determination

Application:
Solvent: DMA with 10 % water (DMA = N,N-Dimethylacetamid)
Titrant: HClO4 0.1 mol/L in ethanol; Potentiometric indication
Blank determination with 60 mL solvent.

The solvent is without ions!

Titration curve: Add inert ions: KCl or LiCl

E [mV] E [mV]

0.0 0.1 0.2 0.3 0.4 0.5 0.0 0.1 0.2 0.3 0.4 0.5
V [mL] V [mL]
25
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrations
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
26
Effect of temperature

Problem
The thermal expansion coefficient of the non-aqueous solvents is 5
to 10 times higher than that of water

A temperature increase will significantly decrease the density of
the solvent and consequently reduce the titer of the titrant

A temperature difference of 1 causes an error of 0.3%

27
Effect of temperature

Solution

Maintain constant temperature conditions, i.e. titrant and sample are

at the same temperature

If a temperature change has occured between the standardization
and sample titration the standardization of the titrant must be
repeated

Measure the temperature during standardization and titration and
calculate a correction factor. In the case of perchloric acid the
correction factor is in the range from 0.9988 – 1.0020

28
Agenda

The strength of acids and bases

How to titrate a weak acid or base?

Advantages of non-aqueous titrations

Non-aqueous base titrations
- Titration of bases – levelling/differentiating
- Solvents for titration of bases
- Titrants for titration of bases

Non-aqueous acid titrations
- Solvents for titration of acids
- Titrants for analysis of acids
- Tips for non-aqueous acid titrations

Standardization of non-aqueous titrants

Sensors for non-aqueous titrations

Blank determination

Effect of temperature

Application examples
29
Assay Benzylnicotinate

Titrant: 0.1 M HCLO4

Solvent: Acetic anhydride, 50 mL

Sample: 0.17 – 0.19 g Benzylnicotinate

Sensor: pH glass electrode with 1M LiCl in ethanol
(DGi113-SC or DGi116-Solvent)

Blank determination on 50 mL solvent (triple determination)

Correction factor CF of perchloric acid solution: temperature measurement
during standardization and titration (0.999644)

Reaction
- R-C5H4N + HClO4 → R-C5H4NH++ClO4-

Result: % Benzylnicotinate = (Consumption [mmol] – Blank mean value
[mmol])x CF x Molar Mass Benzyl nicotinate x 10/Sample weight [g]

Result: 97.921%, RSD (6 samples): 0.047%

30
Assay Sodium Citrate, tribasic, anhydrous

Titrant: 0.1 M HCLO4

Solvent: Glacial acetic acid, 50 mL

Sample: 0.05 g Sodium citrate

Sensor: pH glass electrode with 1M LiCl in ethanol
(DGi113-SC or DGi116-Solvent)

Blank determination on 50 mL solvent (triple determination)

Correction factor (CF) of perchloric acid solution: temperature
measurement during standardization and titration (0.99734)

31
Assay Sodium Citrate, tribasic, anhydrous

Reaction
C6H5Na3O7 + 3HClO4 → C6H5O4(OH)3+NaClO4

Result: % Benzylnicotinate = (VEQ*c*t [mmol] – Blank
[mmol]) x CF x Molar Mass Benzyl nicotinate x
10/Sample weight [g]*3
VEQ: Volume to equivalence point [mL]
c: Nominal volume of titrant mol/L
t: Titer of titrant
3: Equivalent number, since 3 moles of perchloric
acid are required for the reaction

Result: 100.017%, RSD (6 samples): 0.312%

32
Assay Rutoside

Application description

Titrant: 0.1 M TBAH

Solvent: Dimethylformamide

Sample: 0.19 – 0.21 g Rutoside

Sensor: pH glass electrode with 1M LiCl in ethanol

Reaction
- R-(OH)2 + TBAH → R-O2TBA2+2H2O

Result: % Rutoside = Consumption [mmol] x 2 x Molar Mass
Rutoside x 10/Sample weight [g]

Result: 99.93%, RSD (6 samples): 0.399%

33