Non-aqueous Titration

Marjana Khalil
 Non-aqueous Titration
• Here the titration is carried out in a non-aqueous
medium (other than water)
• When applied:
• If the compound to be analyzed is poorly soluble
in water
• If the compound is degraded by water
• To compare the strengths of strong acids or bases
When weak acid and bases are titrated
• Poor end point in water medium
 Principle of non-aqueous 1tra1on

• The theory is based on Brønsted-Lowry

Theory. Thus, an acid is a proton donor and a
base is a proton acceptor. Therefore, when an
acid HA undergoes dissocia@on it gives rise to
a proton and the conjugate base A of the acid.
 Principle of non-aqueous titration

• Hence, from the above definitions it may be

implied that:
• an acid: could be either an electrically neutral
molecule e.g., HNO3; or a negatively charged
anion e.g., HSO4– ; or a positively charged
cation e.g., C6H5NH3+ , H3O+ ;
• a base: could be either an electrically neutral
molecule e.g., C6H5NH2 ; or an anion; e.g. Cl–,
NO3–.
 Different definitions of Base
• Arrhenius base: Any substance that, when
dissolved in water, increases the concentration of
hydroxide ion (OH-)
• Bronsted-Lowery base: A proton acceptor
conjugate acid
• Lewis base: An electron donor
 Types of solvent
• Based on proton interaction solvents are of 4
types:
• Protophilic solvents: Solvents that have
greater tendency to accept protons, i.e., water
(H2O), alcohol (C2H5OH), ammonia
(NH3)/liquid ammonia, etc. Weak acids are
normally used in the presence of strongly
protophilic solvents as their acidic strengths
are thus enhanced and then become
comparable to these of strong acids; this is
known as the levelling effect.
• Protogenic solvents: Solvents that have the
tendency to produce protons, i.e., water
(H2O), liquid hydrogen chloride (HCl), glacial
ace@c acid (CH3COOH), etc. they enhance the
strength of weak bases.
• Amphipro7c solvents: Solvents that act as
both protophilic and protogenic, e.g., water
(H2O), ammonia (NH3)/ammonium (NH4+)
glacial ace@c acid (CH3COOH), ethyl alcohol,
etc. They are all capable of slight dissocia@on.
• Apro7c solvents: These solvents neither
donate nor accept protons. They possess low
dielectric constants, do not cause ioniza@on of
solutes and do not undergo reac@ons with
acids and bases, e.g., benzene (C6H6), Toluene
(C6H5CH3), carbon tetrachloride (CCl4), Carbon
disulphide (CS2), etc.
• Example: HCl acts as an acid in H2O, a
stronger acid in NH3, a weak acid in
CH3COOH, neutral in C6H6 and a weak base in
HClO4.
 What is levelling effect?
• Leveling effect or solvent leveling refers to the effect
of solvent on the properties of acids and bases. The
strength of a strong acid is limited ("leveled") by the
basicity of the solvent. Similarly the strength of a
strong base is leveled by the acidity of the solvent.
• For eg: when ammonia is the solvent, which is a strong
proton acceptor, the strongest acid is ammonium
(NH4+), thus HCl and a super acid exert the same
acidifying effect. If two weak acids are put into a
solvent, such as ammonia, it is likewise hard to tell the
difference between the two acids, since they will both
act like a mol of ammonium ions. So, the leveling effect
changes with the solvent.
• When a strong acid is dissolved in water, it
reacts with it to form hydronium ion (H3O+).
An example of this would be the following
reaction, where "HA" is the strong acid:
• HA + H2O → A− + H3O+
• Any acid that is stronger than H2O reacts with
H2O to form H3O+.
• The same argument applies to bases. In water,
OH− is the strongest base. Thus, even though
sodium amide (NaNH2) is an exceptional base
(pKa of NH3 ~ 33), in water it is only as good as
sodium hydroxide. On the other hand, NaNH2
is a far more basic reagent in ammonia than is
NaOH.
 What is differentiating effect?
• It is the effect of solvent on the acidity/
basicity of solute by which two or more acids
or bases dissolved in the solvent can be
distinguished (showing different initial
strength).
• The solvent is called differentiating solvent.
 Advantages of nonaqueous titration
• 1) A sharp end point is achieved which otherwise
cannot be obtained when the titration is performed in
an aqueous medium:
• Perchloric Acid is a very strong acid and when it is made to
dissolve in acetic acid, the latter can behave as a base and
forms an ‘onium ion’ after combining with protons donated by
the perchloric acid.
• Thus, we have :

• As the CH3COOH2+ ion can instantly donate its proton to a base, therefore,
a solution of perchloric acid in glacial acetic acid, behaves as a strongly
acidic solution.
• the basic characteristics of Pyridine, a weak base is
increased when dissolved in acetic acid. Therefore,
it is practically feasible to titrate a solution of a weak
base in acetic acid against a mixture of perchloric
acid in acetic acid. Thus, a sharp end point is
achieved which otherwise cannot be obtained when
the titration is performed in an aqueous medium.
2) Quan7ta7ve analysis of weakly basic drugs:
In glacial ace@c acid (an acidic solvent) and in
dioxane (C4H802, a neutral solvent), the
perchloric acid (HClO4) behaves as more acidic
(i.e., less protophilic) than HCl; and, therefore,
many base-hydrochlorides (i.e., chlorides) may
be @trated with standard HClO4.
 Advantages of Nonaqueous titration
(Continued):
3. Titration of substances with poor aqueous
solubility: Organic acids and bases that are
insoluble in water are readily soluble in non-
aqueous solvent.

4. Titration of substances with poor aqueous

reactivity

5. Titration of very weak acids or bases: Weak

acids or bases can be directly titrated in
nonaqueous medium using a leveling solvent.
 Disadvantages of nonaqueous
titration
• Expensive,
• Volatile,
• Toxic,
• Removal of water is necessary.
 Examples of some nonaqueous solvents
selection:
• Glacial acetic acid: It is the most frequently used non-
aqueous solvent. Before it is used, it is advisable to
check the water content. This may be between 0.1%
and 1.0%. used for titration of weak bases, Nitrogen
containing compounds.

• Acetonitrile: It is frequently used with other solvents

such as chloroform and phenol and especially with
ethanoic acid. It enables very sharp end points to be
obtained in the titration of metal ethanoates when
titrated with perchloric acid.
• Alcohols (isopropyl alcohol, n-butyl alcohol): Salt of
organic acids can be determined in mixtures of glycols
and alcohols or mixtures of glycols and hydrocarbons.
The most common combinations are ethylene glycol
(dihydroxyethane) with propan-2-ol or butan-1-ol.

• Dimethyl formamide (DMF): It is a protophilic solvent,

employed for benzoic acid, amides etc,

• Dioxane: It is often used in place of glacial ethanoic

acid when mixtures of substances are to be quantified.
Unlike ethanoic acid, dioxane is not a levelling solvent
and produces separate end points, corresponding to
the individual components in the mixtures.
 Common 5trants of nonaqueous 5tra5on:

Acidic solutions:
• HClO4 in glacial acetic acid
• HCl in propylene-glycol /chloroform mixture

Application:
•

• For titration of weak bases with Kb = 10-7−10-12.

E.g. aromatic amines, amides, alkaloids etc.
• For titration of high molecular weight organic
bases that have limited solubility in water.
 Common titrants of nonaqueous
titration:
Basic solutions:
• Tetra butyl ammonium hydroxide in pyridine
• KOH in ethanol

Application:
• For titration of weak acids with Ka = 10-7−10-12.
E.g. carboxylic acids, phenols, enols etc.
• For titration of high molecular weight organic
acids that have limited solubility in water.
 METHODOLOGY
For non-aqueous titrations, the following four
steps are usually taken into consideration,
namely :

(i) Preparation of 0.1 N Perchloric acid, (1L)

(ii) Standardization of 0.1 N Perchloric Acid,
(iii) Choice of Indicators, and
(iv) Effect of Temperature on Assays.
• Materials Required : 8.5 ml of perchloric acid
(70.0 to 72.0%) ; 1 Litre of glacial aceWc acid ; 30
ml of aceWc anhydride.
[1. How can you calculate the molarity of 70%
HClO4? (12.33 M)
Hint: [(% x d) / MW] x 10 = Molarity]
=(70x1.77)/100.5x10 =12.33 M =12.33 N (S2)
Density of HClO4= 1.77 g/cm3
2. How can you calculate the volume required to
prepare 0.1 N (S1) 1L (V1) HClO4 soluWon? (8.5 mL)
] =8.11 ml
 Practice
• How can you prepare 0.1 N 1 L HClO4 solution
from 75% HClO4 solution with desity 1.6
g/cm3?
• [(% x d) / MW] x 10 = Molarity]
=11.94 M = 11.94 N (S2)
• V1x S1 = V2 x S2
• To prepare 0.1 N 1 L Perchloric acid we need
8.4 ml HClO4 from 75% HClO4 Solution. (Ans)
• Procedure : Gradually mix 8.5 ml of perchloric acid
to 900 ml of glacial acetic acid with vigorous and
continuous stirring. Now add 30 ml acetic anhydride
and make up the volume to 1 litre with glacial acetic
acid and allow to stand for 24 hours before use.

• The acetic anhydride reacts with the water (approx.

30%) in perchloric acid and some traces in glacial
acetic acid thereby making the resulting mixture
practically anhydrous. Thus, we have:
• Calculate the amount of 72% HClO4 (in grams)
present in 1000 ml (d= 1.6 g/cm3)
• Calculate the moles. [11.46 mol]
• Calculated the concentration.
• Calculate the concentration when 8.5 ml of
that HClO4 is diluted to 1 L.
• Calculate the amount of 72% HClO4 (in grams)
present in 8.5 ml
• Perchloric acid has a molecular weight of
100.46 and 1 L of 0.1 N solution shall contain
1 /10th the equivalent weight or 10.046 g. To
prepare 1 L of standard perchloric acid
solution, it requires 8.5 ml (sp. gr. 1.6) volume
and a purity of 72% which will calculate out as
9.792 g of HClO4.
STANDARDIZATION OF 0.1 N PERCHLORIC ACID

• In usual practice, potassium hydrogen phthalate (or potassium

biphthalate, KHC8H4O4) is employed as a standardizing agent
for acetous perchloric acid.
Procedure: Weigh accurately about 0.5 g of
potassium hydrogen phthalate in a 100 ml conical
flask.
• Add 25 ml of glacial acetic acid and attach a reflux
condenser fitted with a silica-gel drying tube.
Warm until the salt gets dissolved completely.
Cool and titrate with 0.1 N perchloric acid by
making use of either of the following two
indicators :
(a) acetous crystal violet-2 drops, end point Blue to
Blue-Green (0.5% w/v)
(b) acetous oracet blue B-2 drops, end point Blue to
Pink.
 CHOICE OF INDICATORS
A number of indicators stated below are commonly used in non-aqueous titrations.
It is, however, necessary to mention here that the same indicator must be used
throughout for carrying out the standardization, titration and neutralization of
mercuric acetate solution.
EFFECT OF TEMPERATURE ON ASSAYS
ASSAY BY NON-AQUEOUS TITRATIONS
Assays of various pharmaceutical substances either in
pure form or in dosage form may be assayed successfully
by non-aqueous titrations. For the sake of convenience
these typical titrations can be categorized into two broad
groups, namely :
(a) Acidimetry in Non-aqueous Titrations—It can be
further sub-divided into two heads, namely :
(i) Titration of primary, secondary and tertiary
amines, and
(ii) Titration of halogen acid salts of bases.
(b) Alkalimetry in Non-aqueous Titrations—i.e., titration
of acidic substances.
 Methlyldopa
• In general, the reaction taking place between a primary amine and
perchloric acid may be expressed as follows :
• R.NH2 + HClO4 → [R.NH3]+ + ClO4–
• Materials Required: Methyldopa 0.2 g ;
anhydrous formic acid : 15 ml ; glacial ace@c
acid : 30 ml ; dioxane : 30 ml ; 0.1 N perchloric
acid and crystal violet solu@on.
• Procedure: Weigh accurately about 0.2 g and
dissolve in 15 ml of anhydrous formic acid, 30
ml of glacial ace@c acid and 30 ml of dioxane.
Add 0.1 ml of crystal violet solu@on and @trate
with 0.1 N perchloric acid.
• Each ml of 0.1 N perchloric acid is equivalent
to 0.02112 g of C10H13NO4.
 Titration of Halogen Acid Salts of Bases
• In general, the halide ions, namely : chloride, bromide and
iodide are very weakly basic in character so much so that they
cannot react quantitatively with acetous perchloric acid.
• In order to overcome this problem, mercuric acetate is usually
added to a halide salt thereby causing the replacement of
halide ion by an equivalent amount of acetate ion, which
serves as a strong base in acetic acid as shown below:
 Amitriptyline Hydrochloride
• Materials Required: Amitriptyline hydrochloride:
1.0 g ; mercuric acetate ; crystal violet ; 0.1 N
perchloric acid ; glacial acetic acid.
• Procedure: Weigh accurately about 1.0 g and
dissolve in 50 ml of glacial acetic acid, warming
slightly, if necessary, to affect the solution.
• Cool, add 10 ml of mercuric acetate solution, two
drops of crystal violet solution and titrate with
0.1 N perchloric acid to a green end-point.
• Each ml of 0.1 N perchloric acid is equivalent to
0.03139 g of C20H23N. HCl.
• % amitriptyline???
 ALKALIMETRY IN NON-AQUEOUS
TITRATIONS
• A plethora of weakly acidic pharmaceutical substances
may be titrated effectively by making use of a suitable
non-aqueous solvent with a sharp end-point. The wide
spectrum of such organic compounds include:
anhydrides, acids, amino acids, acid halides, enols (viz.,
barbiturates), xanthines, sulphonamides, phenols,
imides and lastly the organic salts of inorganic acids.
• However, a weak inorganic acid e.g., boric acid, can be
estimated conveniently employing ethylenediamine as
the non-aqueous solvent.
 Preparation of 0.1 N Potassium
Methoxide in Toluene-Methanol
• Materials Required: Absolute methanol: 40 ml ; dry
toluene : 50 ml ; potassium metal : 4 g.
• Procedure: Add into a dry flask, a mixture of methanol
(40 ml) and dry toluene (50 ml) and cover it loosely.
Carefully add freshly cut pieces of potassium metal to
the above mixture gradually with constant shaking.
After complete dissolution of potassium metal, add
enough absolute methanol to yield a clear solution.
• Toluene 50 ml is added with constant shaking until the
mixture turns hazy in appearance. The process is
repeated until 1 litre of solution is obtained.
Preparation of 0.1 N Sodium Methoxide
• It is prepared exactly in a similar manner as for
0.1 N Potassium Methoxide, using 2.3 g of
freshly-cut sodium in place of potassium.
Preparation of 0.1 N Lithium Methoxide
• It is prepared as for 0.1 N Potassium
Methoxide, but using 0.7 g of lithium in place
of potassium.
 Standardization of 0.1 N Methoxide
Solution
• Materials Required : Dimethylformamide (DMF) : 10 ml ;
• thymol blue (0.3% in MeOH) ; 0.1 N lithium methoxide in toluenemethanol
; benzoic acid : 0.6 g.
• Procedure : The apparatus shown in Figure is employed
• for the standardization of 0.1 N methoxide
solution. Transfer 10 ml of DMF in a conical
flask and add to it 3 to 4 drops of thymol blue.
• Quickly introduce 0.6 g of benzoic acid and
titrate immediately with methoxide in
toluenemethanol.
• Caution: Care must be taken to avoid
contamination of neutralized liquid with
atmospheric carbon dioxide.
Precaution contd….
• Reactions for standardization:
 Ethosuximide
• Materials Required : Ethosuximide : 0.2 g ;
dimethylformamide : 50 ml ; azo-violet (0.1% w/v in
• DMF) : 2 drops ; sodium methoxide 0.1 N.
• Procedure : Weigh accurately about 0.2 g, dissolve in
50 ml of dimethylformamide, add 2 drops of azo-
violet soluNon and Ntrate with 0.1 N sodium methoxide
to a deep blue end point, taking precauNons to prevent
absorpNon of atmospheric carbon dioxide.
• Each ml of 0.1 N sodium methoxide is equivalent to
0.01412 g of C7H11NO2.
• %Ethosuximide???
• Home Work:
• Preparation of 0.1 N Tetrabutylammonium
Hydroxide in Toluene-Methanol
• Standardization of 0.1 N
Tetrabutylammonium Hydroxide
• Assay of Chlorthalidone