Pharmaceutical Analysis

Laida Neti Mulyani, M.Si

Kontrak Kuliah

Presensi
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Tugas
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Total

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(100%)

Analysis Method

Chemical Methods
Electrochemical Methods
Optical Methods
Assay Methods Based on Separation Technique
Miscellenous Assay Methods

ELECTROCHEMICAL
METHODS

Potensiometri
Introduction
16.2 Theory
16.2.1 General considerations
16.2.2 End-point determination
16.3. Electrodes
16.4 Applications of potentiometric titrations in
pharmaceutical analysis
Amperometri

INTRODUCTION

Titrasi potensiometri merupakan salah satu metode analisis

berdasarkan pengukuran beda potensial yang dihasilkan oleh suatu
sel elektrokimia
Memanfaatkan hubungan antara konsentrasi dengan beda potensial
berdasarkan persamaan nernst

THEORY
Titrasi potensiometri bermanfaat untuk
menentukan titik akhir titrasi yang tidak dapat
ditentukan menggunakan suatu indikator
Memiliki akurasi dan presisi yang tinggi
In a situation, where a metal M is placed in a
solution containing its own ions Mn+, an
electrode potential is established across the
two electrodes, whose actual value is
provided by the Nernst equation as shown
below :

THEORY

GENERAL
CONSIDERATIONS
The potentiometric titrations invariably cover a broadspectrum of chemical reactions that may be classified as
follows :
Neutralization reactions,
Redox reactions,
Precipitation reactions,
Complexation reactions, and
Potentiometric titrations in non-aqueous solvents

Neutralization Reactions
Advantages
It is found to be useful to titrate a mixture of acids having a significant
difference in their strengths, for instance : HCl and CH3COOH
(alcoholic). In this case, the first-break in the titration curve signifies that
the stronger of the two acids i.e., HCl, gets neutralized ; whereas, the
secondbreak represents the entire completion (i.e., HCl + CH3COOH)
In order to get fruitful and reproducible results it is quite necessary that
the strengths between either
the two acids or bases in question must vary by at least 105 to 1.
Demerits of the Method
The neutralization reactions often found to be giving unsatisfactory results
in the following two instances. They are :
(a) when both the acid and the base are appreciably weak, and
(b) when either the acid or the base is very weak (i.e., K < 108) and also
the prevailing solutions are dilute.

Neutralization Reactions

Note : In (a) above, an accuracy up to 1% is achievable in

0.1 M solution.
Choice of Electrodes :
Indicator Electrodes : Hydrogen, Glass or Antimony
electrodes ;
Reference Electrode : Calomel electrode.

Redox Reactions
Titrasi Potensiometri melibatkan reaksi redox
Nilai potensil di pengaruhi oleh perbandingan konsentrasi zat
tereduksi dan teroksidasi dari spesi ioniknya

Contoh
Titrasi antara Fe2+ dengan kalium permanganat atau potasium
dikromat
Elektroda yang digunakan : Pt
Oksidator biasanya ada didalam buret

Precipitation Reactions
Titrasi potensiometri dimana konsentrasi ioniknya ditentukan oleh
penurunan kelarutan spesi yangmenghasilkan suatu bentuk tidak
terlarut melaui suatu reaksi presipitasi.
Contoh
Titrasi Ag+ dengan senyawa halida seperti Cl-,, Br-, atau I- atau dengan
senyawa SCN Pemilihan elektrode
Elektrode referensi :SCE, Silver-silver chloride elektroda
Elektroda indikator: Peratk Pt,
( elektrode harus mudah berkeseimbangan dengan ion presipitat)
Jembatan garam
Untuk penentuan suatu Halida jembatan garam harus dijenukan
dengan potasium nitrat
Note dapat juga dikembangkan ion selektive presipitat

Complexation Reactions
Titrasi potensiometri dimana terjadi interaksi antara suatu presipitat
terlarut dengan suatu perekasi secara berlebih sehingga menghasilkan
suatu senyawa kompleks
Contoh

Pada reaksi tersebut pada tahap satu dihasikan suatu presipitat AgCN,
presipitat ini akan larut dengan penambahan KCN berlebih sehingga
membentuk suatu senyawa kompleks [Ag(CN)2].
Sehingga pada titrasi dibagi menjadi 3 bagian
Up to End-point (titik ahir awal titrasi )
CN- akan dirubah menjadi suatu sennyawa kompleks dengan Ag+
At the end point (titik ahir titrasi)
Dilihat dari visuallisai melalui perbedaan dan kenaikan potensial sel

Complexation Reactions

Beyond end point (titik ahir bawah titra

Penambahan AgNO3 akan menyebabkan AgCN mengalami
presipitasii. Peruban ke dua pada nilai potensial dapat diamati
dengan adanya sebagian besar CN- yang akan mengendap
sebagai AgCN-

Choice of Electrodes :
Indicator Electrode : Silver electrode ;
Reference Electrodes : Colomel electrode ; Mercury-mercury (I)
sulphate electrode.
Salt-Bridge : A saturated solution of KNO3 or K2SO4 isolated from
the reference electrode

Potentiometric Titration in
Non-Aqueous Solvents
Metode ini digunakan untuk menentukan titik ahir titrasi pada media non
air. Sekala Mv lebih digunakan daripada sekala pH, karena
Skala pH berdasarkan bufer tidak ada memiliki makna yang logis
pada media non air
Nilai potensial pada media non air melampaui nilai skala pH
Kurva hasil titrasi umumnya lebih atau kurang empiris dan menghasilkan
deteksi titik ahir yang dependable dan reprodusibilitas yang lebih
bermakna.
Choice of Electrodes :
Indicator Electrodes : Glass electrode ;
Reference Electrode : Calomel electrode ;
Salt-Bridge : A saturated solution of KCl

END POINT
DETERMINATION
Table 16.1 : Data of Potentiometric Titration of 25.0 ml 0.01 M NaF
Against 0.01 M La (NO3)3

END POINT
DETERMINATION
Sigmoid-curve

Kurva turunan pertama

Titik akhir titrasi

Titik akhir titrasi

Kurva turunan kedua

Titik akhir titrasi

INSTRUMENTATION

Jenis- Jenis Elektroda

Function

Type

First type

Reference

Second type

1-Normal
hydrogen

1- Redox
reaction

4- Acid base
reaction

2-Ag/AgCl

3Preceptimetric
reaction

2Complexometric
reaction

Third type

3-Calomel

Indicator

Mode of action
Redox type
Electron transfer

Ion- Selective

Accordingto the Type

1) First type (kind or order):
Potensial tergantung pada keseimbangan
antara unsur dan ionnya.

(Mo / Mn+)

Mo Mn+ + ne E
E25oC = Eo - 0.059/n log 1 / [Mn+]
From Nernest equation E [Mn+]

E.p.

mls of titrant
e.g. Titration of Cu2+ with EDTA
Keadaan awal menujukkan konsentrasi ion Cu2+ sangat tinggi sehingga
potensial selnya sangat tinggi, sebagai fugsi E [Cu2+], kemudian terjadi
penurunan potensial ketika Cu2+ bereaksi dengan EDTA
It is used for any metal except Fe, Cr, Co, Ni WHY?
Because:
1- mudah teroksidasi oleh udara sehingga nilai potensial tidak stabil
2- dapat mengalami deformasi kristal dan menimbulkan laisan oksida

2) Electrode of Second type:

In this type equilibrium is established between three phases.

The electrode is a metal electrode which is covered with a layer of its
sparingly soluble salt immersed in a solution of electrolyte.
This electrode is used for determination of anions (non metal ions).
The electrode potential is indirectly affected by the analyte.
e.g. Ag0 electrode: (Ag0 / AgCl / Cl-)
EAg0 1/ [Cl-]
i.e. Ag0 is coated with a layer of silver chloride and dipped in chloride
solution.
Calomel electrode: (Hg0 / Hg2Cl2 / Cl-)
EHg0 1/ [Cl-]
Mercury electrode: (Hg0 / Hg2y2- / y4-)
EHg0 1/ [Y4-]
Similar electrodes for bromide, iodidecan be used.

In Ag electrode; the potential depends on the chloride

concentration in the solution and this is used for its determination.
The electrode potential is:
E25 = E0 - 0.059 log [Cl-]
i.e Potential is inversely proportional to non metal ion concentration.
e.g. titration of Cl- with Ag+
Before titration:
Conc of Cl- is high so E is low
During titration:
E
Conc of Cl decreases so E increases
At e.p.
conc of Cl- is zero so sharp increase in E
mls of titrant

3) Electrode of Third type:

In this type equilibrium is established between four phases.
E.g. We want to determine Ca 2+ ions in CaCl2 but we have Pb rod
E.g. Pbo / PbC2O4 / CaC2O4 / CaCl2
Represented by the following equations:
PbC2O4
Pb2+ + C2O42Lead acetae and calcium acetate
2+
2 CaC2O4
Ca + C2O4
supresses the ionization of each
other by common ion
CaCl2
Ca2+ + 2Cl Increase in [Ca2+] will supress dissociation of CaC2O4, leading to lower
availability of C2O42- .
In this case stress of C2O42- on PbC2O4 decreased leading to more
dissociation of PbC2O4 producing more Pb2+ and consequently potential
developed on Pbo is increased.
EPbo [Ca2+]

4) Redox electrode:

Pada tipe ini kesetimbangan dicapai antara oxidan dan reduktan pada
larutan yang sama denganadanya elektroda innert e.g pt wire (i.e it is one
phase system electrode).

e.g. Fe3+ / Fe2+ in presence of pt

Ce4+/Ce3+

E25oC = Eo - 0.059/n log Fe2+ /Fe3+

E25oC 1/ [red] / [oxid]

E
Fe3+/Fe2+

E.g. Determination of Fe2+ with Ce4+

E.p.
mls of titrant

Accordingto the Funtion

Refference electrode
In this type electrode must have known and constant potential,
therefore used as a reference to measure the potential of indicator
electrode through galvanic cell.
e.g

a)Normal hydrogen electrode (N.H.E.),

b) calomel electrode,
c) silver, silver Chloride electrode.

Standard Hydrogen Electrode (SHE)

Half cell presentation:

Pt, H2 / H+ (1.0 N)//
Type :1st type
Electrode reaction: 2H+ + 2e H2
Nernest equation:
E25oC = Eo - 0.059/n log H2 / [H+]2
E25oC = Eo + 0.059 log [H+]
E [H+] in N.H.E [H+] = 1 , log 1 is zero
So E25oC = Eo = zero.
Advantage:
It is a primary reference electrode as its potential is zero.
Disadvantages:
It is difficult to keep H2 gas at 1 atm during all determinations.
it needs periodical re-plating of pt sheet with pt blac

The Secondary Reference

Electrodes:
1- The Calomel Electrode:
Half cell presentation

Type :2nd type

Electrode reaction:
Hg22+ + 2e 2Hg0

Nernst Equation
E25oC = Eo - 0.059/2 log Hg0 / [Hg22+]2

 From the Nernst equation it is clear that the electrode potential

depends on the [Hg22+]; which is produced from the dissociation of the

very small amount of the sparingly soluble Hg2Cl2.

Hg2Cl2 2Hg22+ + 2 Cl The mercurous ion concentration is controlled by the solubility product

of the salt KspHg2Cl2

KspHg2Cl2 = [Hg22+]2 [Cl-]2
[Hg22+]2 = KspHg2Cl2 / [Cl-]2
E25oC = Eo - 0.059/2 log [Cl-]2 / KspHg2Cl2

E 1/ [Cl-] electrode potential depends on the [Cl-] **

Although maximum potential is obtained on using 0.1N KCl; however,

we usually use Sat. KCl WHY?

Silver Electrode:
Half cell presentation:
Ag0/AgCl /,KCl (saturated, 1 N or 0.1N) //
Type :2nd type
Electrode reaction: Ag+ + e Ag0
Nernst Equation :
E25oC = Eo - 0.059/1 log Ag0 / [Ag+]

As explained under calomel electrode, the Ag+ ion concentration is

controlled the solubility product of the salt KspAgCl
Ksp AgCl = [Ag+] [Cl-]
[Ag+] = Ksp AgCl / [Cl-]
E25oC = Eo - 0.059/1 log [Cl-] / Ksp AgCl
E 1/ [Cl-]
electrode potential depends on the [Cl-] **

Indicator electrodes
An indicator electrode is that which its potential is sensitive to the
concentration of one of the participant or products of reaction.
Its potential changes rapidly with the change of the particular ion.,It must:
Give rapid response.
Its response is reproducible.
Now when it is connected with a reference electrode, the change in e.m.f. of
the formed cell will be due to the change of indicator electrode potential,
which in terns reflects the change of the ion under investigation.
Indicator electrodes are classified into two classes:
Electrodes where electron transfer takes place at the electrode surface.
i.e. redox reaction occurs (metallic electrodes).
Electrodes where (charge) ion exchange takes place at a specific
membrane surface.
i.e. ion selective electrodes or specific ion electrodes.

N.B:
We must notice that electrodes potential are related to activity.
The most important application of this method is the determination of pH.
The observed potential of the cell employed during the measurement is:
Eobs = Eref + Ej Eind
where Eref , Ej and
Eind are the potentials of reference electrode, liquid
junction and indicator electrode.

Liquid junction potential

Liquid junction - interface between two solutions containing different

electrolytes or different concentrations of the same electrolyte
A junction potential occurs at every liquid junction.
Caused by unequal mobilities of the + and - ions.

The disadvantage of the cell with liquid junction is there is a potential

associated with the liquid junction, called the liquid junction
potential.
A potential is developed at both boundaries of the junction
This liquid junction potential arises from difference in rate of migration
of anion and cations of the bridge salt and the electrolytes in the
electrodes solutions. This difference results in unequal charge
distribution at the boundaries, thus a potential is developed

Liquid junction potential

It depends upon:
Charge mobilities
Concentration of the ionic species
Nature of the solvent on each side of the boundary
pH dependent

The presence of liquid junction potential limit the accuracy of

potentiometric measurements.
How to minimize liq.junct. Pot.?
By using a high concentration of a salt, its ions has nearly equal mobility
e.g. KCl or KNO3 (K+ = 74, Cl- = 73, NO3- = 76)
Use high concentration of salt for preparation of the bridge to reduce the
effect of difference in migration rates of other ions in the electrode solution.

Ecell = Eind - Eref + Ej

Electrodes for Redox

Reactions
:
These are inert electrodes which acquire the potential of the system in
which they are dipped.
They are made of platinum or gold, in the form of foils, coils or plates
WHY?
to expose a large surface area to the solution thus attain the
equilibrium potential rapidly.
It is one phase system electrode and is connected to Ag/AgCl or
calomel electrode to measure the e.m.f. produced.

Electrodes for complexometric

Reactions:

We may use:
A- Electrodes of the First type:
For the determination of a cation (metal ion); where the best
electrode used fora cation is its elemental form.
e.g. in determination of Cu2+ with EDTA , a copper rod is the
indicator electrode, its potential is directly affected by the analyte
concentration as indicated from:
Nernest eq:
E25oC = Eo - 0.059/n log 1 / [Cu2+] so E [Cu2+]

E.p.

B- Electrodes of the Second type:

For the determination of a mixture of metals

(Cu2+,Cd2+, Bi3+ by EDTA)
We can not dip three different metal rods,
So we use an electrode that is affected by
the titrant instead of the sample.
We use:
(Hgo / Hg2y2- / y4-)
EHgo 1/ [Y4-]

mls of titrant

Electrodes for precipitemetric

Reactions:

We may use:
A- Electrodes of the First type:
For the determination of a cation (metal ion); where the best electrode
used for a cation is its elemental form.

in determination of Ag+ with chloride , a silver

rod is the indicator electrode, its potential is
directly affected by the analyte concentration as
E
indicated from:
Nernest eq: E25oC = Eo - 0.059/n log 1/ [Ag+] so
E [Ag+]
mls of titrant

In determination of mix of Cl- & Br- & I- with AgNO3,

So better to choose an indicator electrode which
responds to the titrant , i.e. silver rod.
E

mls of titrant

Before titration: there is no Ag+ ions so E is low

At e.p. :conc of Ag+ ions increases
So E increases sharply

B Electrodes of the Second type:

For the determination of Cl- or Br- or I- with AgNO3
E
Det. of anion so use
Ag0 electrode: (Ag0 / AgCl / Cl-)
EAg0 1/ [Cl-]
mls of titrant

Electrodes for Neutralization

Reactions:
The Hydrogen gas Electrode
Electrode reaction: H+ + e

Pt black
Catalyst

H2

Mode of action: electron transfer electrode

Nernst equation:
E25 = Eo - 0.059 log 1/ [H+]
E25oC = zero - 0.059 log 1/ [H+]
E25oC = - 0.059 pH
(E depends on [H+] or pH)
When connected to a reference electrode the
e.m.f. of the cell
Ecell = Ereference - Eindicator
Ecell = Ereference - (-0.059 pH)
Ecell = Ereference + 0.059 pH
pH = Ecell - Ereference / 0.059

Salt
bridge

platinum
coated with Pt
black

[H+] = X M

Disadvantages:
As N.H.E. in addition:
It can not be used, presence of oxidant or reductant will interfere with the
equilibrium and function of the electrode.
It can not be used in presence of catalytic poisons as S2- which interfere
with the catalytic activity of pt black).
It can not be used in reactions involving volatile constituents e.g. H2CO3,
H2S because bubbling of H2 gas causes their volatilization.

Antimony electrode Sb0/ Sb2O3

Electrode reaction:Sb2O3 + 6H+ + 6e
2Sb0 + 3H2O
Mode of action:
electron transfer electrode
It is prepared by allowing a rod of antimony to cast in air for two or three weeks.
Pt wire

Nernst equation:
E Sb0/ Sb2O3 = E0 - 0.059/6 log [Sb0]2 / [Sb2O3] [H+]6
E 25oC= E0 - 0.059/6 log [Sb0]2 / [Sb2O3] - 0.059/6 log 1/ [H+]6
[Sb0] / [Sb2O3] is considered as unity, as log 1= zero
E 25oC= E0 - 0.059/6 log 1/ [H+]6 , E 25oC= E0 + 0.059 log [H+]

E25oC = E0 - 0.059 pH

Sb2O3
Sb0

Advantages:
Easy to be used cheep and durable.
It can be used for determination of pH of volatile acids (H2CO3)
Disadvantages
Can only be used within pH range 2-8, at lower pH Sb2O3 dissolves and at
higher pH Sb0 dissolves.
It can not be used in presence of oxidizing, reducing agents.
It can not be used in presence of complexing agents e.g. taratric acid because
it gives antimony tartarate complex.
It can not be used in presence of noble metals which will be displaced by
antimony.

Quinhydrone Electrode
:
Is formed by the addition of quinone and hydroquinone in equimolar
proportion to the solution to be analyzed and an inert electrode (pt) is
immersed in the solution.
Electrode reaction: Q + 2H+ + 2e
H 2Q
Mode of action: electron transfer electrode
Nernst equation:
E Q/ H2Q = E0 - 0.059/2 log [H2Q] / [Q] [H+]2
E 25oC= E0 - 0.059/2 log [H2Q] / [Q] - 0.059/2 log 1/ [H+]2
[Q] / [H2Q] =1,as they are in equimolar proportions.
As log 1= zero
E 25oC= E0 - 0.059/2 log 1/ [H+]2 , E 25oC= E0 + 0.059 log [H+]
E25oC = E0 - 0.059 pH

Quinhydrone Electrode
Advantages:
Easily prepared and used.
Not affected by catalytic poisoning.
It comes to equilibrium rapidly.
It can be used for determination of pH of volatile acids (CO2).

Disadvantages
Can not be used at pH > 8 because H2Q is dissociated, altering the pH
of the solution.
It can not be used in presence of oxidizing and reducing agents.
Atmospheric oxygen slowly oxidizes H2Q, therefore, it must be freshly
prepared.

Glass Electrode:

This the mostly widely used electrode for measuring the pH.
It is ion selective electrodes which is specific for hydrogen ion.
Ion selective electrodes respond to activity more than concentration of ions:
a = [Mn+] fa
where: a is the activity ,
[Mn+] is the molar concentration of ion and
fa is the activity coefficient.
fa 1/ ionic strength of the solution.

As the ionic strength the activity coefficient activity E

How to measure E:
The glass electrode is immersed in the unknown solu and is coupled to
saturated calomel electrode then connect both electrodes with a
potentiometer.
Advantages:
It can be used in presence of oxidizing, reducing and complexing agents,
catalytic poisons and noble metals.
used in solutions containing volatile constituents.
Disadvantages:
It is fragile.
Can not be used in presence of dehydrating agents. e.g. concentrated
sulphuric acid, ethyl alcohol.
Can not be used above pH 12 (alkaline error) as Interference from sodium
ions occurs i.e. sodium ion exchange together with H+ (glass membrane
becomes permeable to sodium).It takes certain time to reach equilibrium
due to glass resistance.

Special Applications:

Potentiometric pH Measurement using Glass electrode

One drop of solution

Tooth cavity
Sweat on skin
pH inside a living cell
Flowing liquid stream
Acidity of stomach

APPLICATIONS OF POTENTIOMETRIC
TITRATIONS IN PHARMACEUTICAL
ANALYSIS
A. Assay of Nitrazepam
Materials Required
Nitrazepam : 0.25 g ; acetic anhydride : 25.0 ml ; perchloric acid (0.1 M) : 250 ml
a Potentiometer ; a Magnetic Stirrer ; Burette (50 ml) ;
Theory
Nitrazepam is a weakly basic compound and hence, it may be titrated
conveniently by means of a non-aqueous titration technique and determining the
end-point potentiometrically.
Procedure
Weigh accurately 0.25 g of nitrazepam and
dissolve in 25.0 ml of acetic
anhydride.Titrate with 0.1 M perchloric acid
placed in a burette and adding it carefully
into the beaker kept on a magnetic stirrer
potentiometrically. Each ml of 0.1 M
perchloric acid is equivalent to 28.13 mg of
C15H11N3O3.

Clonidine Hydrochloride :
Materials Required
Clonidine hydrochloride : 0.2 g ; ethanol (96%) : 100 ml ; 0.1 M ethanolic
sodium hydroxide Vs : 1 L (Add 3.3 g of 10 M sodium hydroxide solution to
250 ml of absolute ethanol).
Standardization of 0.1 M Ethanolic Sodium Hydroxide Solution Vs
Dissolve 0.2 g of benzoic acid in a mixture of 10 ml of ethanol (96%) and 2
ml of water and titrate with the ethanolic sodium hydroxide solution using
0.2 ml of thymolphthalein solution (a 0.1 % w/v solution of thymolphthalein
in ethanol (96%) as indicator. Each ml of 0.1 M ethanolic sodium hydroxide
Vs is equivalent to 12.21 mg of C7H6O2.

Procedure
Dissolve 0.2 g of clonidine hydrochloride in 70 ml of ethanol (96%) and
titrate with 0.1M ethanolic sodium hydroxide Vs determining the end-point
potentiometrically. Each ml of 0.1 M ethanolic sodium hydroxide Vs is
equivalent to 26.66 mg of C9H9Cl2,N3, HCl.