LIMIT TEST

Dr. MEENU
Assistant Professor,
Shri Guru Ram Rai Institute of Technology & Sciences
2016
 Impurities
Definition: A compound is said
to be impure if it is having
foreign matter, i.e. impurities.
Chemical purity implies
freedom from foreign matter. It
means that a pure chemical
refers to that compound which
is having no foreign matter, i.e.
impurities.

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 :Source of impurities
1. Raw material employed in manufacturing
2. Reagent used in manufacturing process
3. Method or process used in
4. Chemical method used in manufacture
5. Atmospheric contamination during manufacturing
process
6. Defect in manufacturing process
7. Manufacturing hazards
8. Storage conditions
9. Accidental substitution with spurious or useless
materials

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Pharmacopoeia
A description of the article
Tests for identity
Physical constants
Quantitative assay
Limit test Storage conditions
official = obey the requirement

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Pharmacopoeia
British pharmacopoeia (BP)
European pharmacopoeia (Eu.Ph)
United state pharmacopoeia (USP)

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit test

They are quantitative or semi-quantitative

test designed to identify or control small
quantities of impurity ,these test should be
specific and sensitive

Nessler cylinder
Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun
 Limit test
Type:
1. Comparison method
2. Quantitative determination
3. Test in which there is no visible
reaction

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit test
General principle
If the sample is lighter than the
standard solution then it is within the
pharmacopeial limit (accepted)
If the sample is darker/heavier than the
standard solution then it is above the
pharmacopeial limit (rejected)

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit test
NESSLER cylinder
(appendix VII A127)
clear glasses with normal capacity 50ml, the
overall height is about 15cm, the external height
to the 50ml mark 11.0 to 12.4 cm ,the thickness
of the wall 1.0 to 1.5 mm and the thickness of
the base 1.0 to 3.0mm the external heights to
the 50 mark of cylinders used to test must not
differ by more than 1 mm

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit test
General precaution
1. The liquid used must be clean and filtered if
necessary
2. The Nessler cylinder must be made of
colorless glass and of the same inner
diameter
3. Detecting opalescence or color
development must be performed in daylight
4. When comparing turbidity it should be
done against black background
5. When comparing color it should be done
against white background
Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun
Limit test

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit Test for Chloride
:Chemical reaction
Cl- + AgNO3AgCl

1ml AgNO3 1ml AgNO3

Dist. H2O to 50 ml Dist. H2O to 50ml

1ml HNO3
1ml HNO3 10ml St Cl- (5ppm)
15ml sample 5ml H2O+

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

Principle:
This test, which is mainly used to control
chloride impurity in inorganic substance,
depends upon the simple reaction between silver
nitrate and soluble chlorides to give insoluble
silver chloride in the presence of dilute nitric
acid.
The insoluble silver chloride makes the solution
opalescent and the extent of opalescence
depending upon the amount of chloride present
in the substance, is compared with a standard
opalescence produced in a standard solution
having a known amount of chloride by adding
silver nitrate and same volume of dilute nitric
acid as used in the test solution.
Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun
 If the opalescence produced in the test is less
intense than that of standard opalescence, the
sample passes the limit test for chloride and vice
versa.

Note: Dilute nitric acid in used to prevent the

precipitation of other acid radicals with silver
nitrate solution. It acts by providing common ion
i.e. nitrate.

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit Test of Sulphate
:Chemical reaction
SO4-2 + BaCl2 BaSO4

Complete1.5ml
volume
ethanolic
with Dil.water
SO4standard
to 50ml
15ml water
&MIX by rotation
wait
Mixfor
by 1 minute
rotation

1ml 25%BaCl2

Add 0.5ml acetic acid(5M 15ml sample

15ml Standard

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

:Principle
It is based upon the reaction between barium
chloride and soluble sulphate in presence of
dil. Hydrocloric acid.
The comparison of turbidity is done with the
standard turbidity obtained from a known
amount of sulphate and same volume of dilute
HCl in the both solutions.
In I.P. barium chloride solution is replaced with
barium chloride reagent to enhance the
reactivity of test.
Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun
 The opalescence produced in the test is compared
with that of standard opalescence obtained from a
standard sulphate solution containing known amount
of sulphate produced in the same manner.
If the test opalescence is less intense than that of
standard opalescence, the sample passes the limit
test for sulphate and vice versa.
Note: The solubilities of barium sulphate
precipitates are very much affected by the
concentration of the acid. The acidity of the
solution is controlled by using acetic acid.

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit Test for Iron
Chemical reaction:

Procedure:
2 ml 20% citric acid
2 ml 20% citric acid
2 drops thioglycollic acid
H2O to 40ml 2 drops thioglycollic acid
H2O to 40ml
10ml sample
sample 2 ml
Made slution alkaline
Made slution alkaline

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

Principle:
The limit test for iron is based on the formation of
pale pink to deep reddish purple colour by reaction
of iron with thioglycollic acid in the presence of
citric acid in a solution made alkaline with
ammonia solution.
The colour is due to the formation of the co-
ordination compound, ferrous thioglycolate
Fe(HSCH2COO)2.
The original state of oxidation of the iron is
immaterial since thioglycollic acid also reduces
ferric iron to the ferrous state.

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

The colour produced is compared with the standard
colour containing a definite quantity of iron by
viewing transversely through the Nessler cylinders.

NOTE: Citric acid is added in the official process

to prevent the precipitation of iron by ammonia.
Iron gets precipitated in the form of Fe)OH(2 and
Fe)OH(3 depending upon the oxidation state of
the iron in the presence of ammonia.
Citric acid makes soluble complex with iron and
ammonia will not be able precipitate iron and it
will just provide alkaline medium.
Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun
Limit Test of Heavy Metal
Chemical reaction:
Pb+2 +NaS PbS

1.2ml thioacetamide
Sample preparation 1.2ml thioacetamide
mix 2.5g aspirin mix
2ml acetate bufferin 30ml 2ml acetate buffer
dissolve
12ml sample 10ml standard Pb
acetone then
2ml sample
complete to 50
ml with H2O

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

Principle:
Method A, B and C are based upon the reaction
of the heavy metal ion with hydrogen sulphide (in
method A and B) or sodium sulphide (in method
C) leading to the formation of heavy metal
sulphides.
The metal sulphides remain distributed in a
colloidal state and give rise to a brownish
colouration.
The colour produced in the test solution is
compared with that of standard solution
containing a definite amount of the lead.

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Method D is based upon the precipitation of
relatively insoluble and characteristically coloured
sulphides of heavy metals when aqueous solutions
are treated with alkali metal sulphides (NaSH).
The colour is compared by keeping the two Nessler
cylinders side by side and viewing vertically
downwards against a white background. The usual
limit for heavy metals as per I.P. is 20 ppm.

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Limit Test of Lead
Principle: The limit test for lead is based on the
reaction between lead and diphenylthiocarbazone
(dithizone) in an alkaline medium to form lead
dithizonate complex.
The lead present as an impurity in the substance is
separated by extracting an alkaline solution with
dithizone extraction solution.
The interference by other metal ions is eliminated
by adjusting the optimum pH for the extraction by
using reagents like ammonium citrate, potassium
cyanide and hydroxylamine hydrochloride.
Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun
 Chemical reaction:

The original colour of dithizone in chloroform

is green while the lead-dithizonate complex is
violet in colour.
The intensity of the violet colour of the
complex depending upon the quantity of lead
present in the solution is compared with
standard colour produced by standard
solution.
Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun
 Limit Test of Arsenic
Principle: The limit test for arsenic is based on the
reduction of the arsenic in the arsenious state to the
arsine gas (AsH3) with zinc and hydrochloric acid. The
arsine gas stains the mercuric chloride paper yellow.
The sample is dissolved in acid whereby the arsenic
present as impurity in the sample gets converted to
arsenic acid.
The arsenic acid is reduced to arsenious acid by
reducing agents like stannous acid, potassium iodine
etc. The nascent hydrogen formed during the reaction
further reduced arsenious acid to the arsine gas. The
arsine gas reacts with mercuric chloride paper to
produce a yellow stain.

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 The depth of the yellow stain depending upon the amount of
arsenic present in the sample, is compared with that of
standard stain produced from a known amount of arsenic.

Chemical reaction:

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

Non Aqueous Acid-Base Titration
Main reasons for non aqueous titration :
Solubility: many organic compound (acid
or base) sparingly soluble in water but
readily soluble in organic solvent
Too weak acid or base : they didnt give
sharp end point in aqueous titration but
can titrated accurately in suitable non
aqueous solvent

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Non Aqueous Acid-Base Titration
Protic solvent :
Amphiprotic :it self dissociation (autoprotolysis)
which process acidic and basic properties
e,g:H2O,AcOH, ALCOHOL and liquid ammonia
2H2O H3O+ + OH-
2C2H5OH C2H5OH2+ +C2H5O-
Protogenic: they have stronger acidic than basic
properties (more acidic than water)
E,g: CH3COOH & H2SO4
B + CH3COOH BH + + CH3COO-

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Non Aqueous Acid-Base Titration
Protic solvent :
Protophilic : basic substance and react with acidic
solute with the formation of solvated proton and
conjugated base of acid (more basic than water)
E,g :ether ,ketone & amine
HB +S HS+ + B-

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Non Aqueous Acid-Base Titration
Advantage of These Titrant
Excellent potentiometric curve using
ordinary glass or calomel electrode

The salt formed from this titration are

soluble in the solvent commonly used

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

 Safety
Crystal violet
may cause cancer. Severe eye irritant. Harmful by inhalation,
ingestion and through skin contact.

Perchloric acid
It is very corrosive to skin and eyes, It can also ignite or explode upon
contact with organic material such as cloth or wood also Causes
hypothyroidism digestive and respiratory tract burns& it is Corrosive
to metal
glacial acetic acid
This material is strongly corrosive and causes serious
burns. Very harmful if swallowed. Lachrymator

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

References:
1. Indian Pharmacopoeia, 1996, Vol. I & II.
2. United States Pharmacopoeia, USP 29 and National Formulary, NF 24, Asian
ed.,2006 3. A.H. Beckett, & J.B.Stenlake, Practical Pharmaceutical Chemistry,
5th ed., Part One, CBS Publishers and Distributors, 2004
4. L.M.Atherden, Bentley and Drivers Textbook of Pharmaceutical Chemistry,
8th ed., Oxford University Press, 2005
5. Holleman-Wiberg, Inorganic Chemistry, Academic Press, 2001
6. J.D.Lee, Concise Inorganic Chemistry, 5th ed., Blackwell Publishing, 2006
7. Remingtons Pharmaceutical Sciences,Indian ed., 21st ed., Vol. I & Vol. II,
Mack Publishing Co.
8. J.H.Block, E.Roche, T.O.Soine, & C.O.Wilson, Inorganic Medicinal and
Pharmaceutical Chemistry, Lea and Febiger.
9. F.A. Cotton, & G., Wilkinson, Basic Inorganic Chemistry, Wiley
10. L.A. Discher, Modern Inorganic Pharmaceutical Chemistry.
11. Vogel, s qualitative inorganic analysis, revised by G.Svehla, 7th ed.,
Pearson Education Publisher

Dr. MEENU, Asst. Prof., Dept. of Pharmacy, SGRRITS, Dehradun

Thank you