GENERAL

TEST
PROCEDURE
BY PRASHANSA SHRESTHA
DGM (TECH)
HEAD OF QUALITY
GENERAL TEST FOR RAW
MATERIAL ANALYSIS
The general test performed in analysis of raw material is as follows:

1. Color and clarity of solution

2. Limit tests
3. Arsenic test
4. Heavy metal
5. Loss on drying
6. Determination of water
7. Determination of pH
8. Sulphated Ash
9. Specific gravity, relative density and weight per ml
10. Optical rotation
11. Potentiometric titration
12. Acid base titration
13. Melting point
14. Conductivity
LIMIT TESTS
Limit tests are quantitiative or semi quantitative tests designed to
identify and control small quantities of impurity which are likely to be
present in the substance
They usually involve simple comparison of opalescence, colour or
turbidity with that of standard sample prescribed in pharmacopoeia
In these tests, concentration of impurity is expressed in ppm or as
percentage.
GENERAL LIMIT TEST FOR
NON SPECIFIC IMPURITY
Clarity of solution
Insoluble matter
Colour of solution
Moisture, volatile matter and residual solvents
Non volatile matter
Residue on ignition(loss on ignition)
Ash value
Acid insoluble ash
Water soluble ash and water soluble extractive
Sulphated ash
CLARITY OF SOLUTION
The degree of clarity or opalescence of solution is defined as clear, very
slightly opalescent, slightly opalescent, opalescent and very opalescent.
Measurement is made by direct comparison with reference solution, either
with the aid of a beam of light of standard luminosity or by viewing through
the solution in diffused light against black background
Solution are clear if their clarity is the same as that of water or solvent used
CLARITY OR OPALESCENCE AS PER IP/BP/USP
IP BP USP

Express the degree of Clarity of solution in BP is done Do not have the

by visual method and chapter and
opalescence standard in instrumental method. follow EP
terms of the opalescence
standard. A liquid is
considered clear if its
clarity is the same as
that of water or if its
opalescence is not more
than that of opalescence
standard OS.
CLARITY OR OPALESCENCE AS PER IP
Opalescence standard Preparation:
Standard preparation: Dissolve 1.0g of hydrazine sulphate in sufficient water to
produce 100ml and set aside for about 6 hours. To 25ml of this solution add 25ml of
a 10% w/v of hexamine, mix well and allow to stand for 24 hours. Keep in a glass
container with a smooth internal surface in which the suspension does not adhere to
the glass. Store in this manner, the solution is stable for 2 months.
Prepare standard suspension by diluting 15 ml of well mixed suspension to 1000ml
with water. The standard solution should be used within 24 hours of preparation.
Opalescence Standards: Prepare opalescence standards by mixing aliquots of the
standard suspension with water as indicated in table. Each opalescence standard
should be shaken well before use.
Opalescence standard Standard suspension Water (ml)
(ml) (ml)
OS1 5.0 95.0
OS2 10.0 90.0
OS3 30.0 70.0
OS 4 50.0 50.0
CLARITY OR OPALESCENCE AS PER IP
Method: transfer to a flat bottom test tube of neutral glass 15 to 25mm in a
diameter, suitable volume of the solution under examination such that the
test tubes is filled to a depth of 40mm. Into another matched test tube add the
same volume of freshly prepared Opalescence standard. After 5 minutes,
compare the contents of the test tube against a black background by viewing
under diffused light down the vertical axis of tubes
CLARITY OR OPALESCENCE AS PER BP
Using identical test-tubes of colourless, transparent, neutral glass with a
flat base and an internal diameter of 15-25 mm, compare the liquid to be
examined with a reference suspension freshly prepared as described below,
the depth of the layer being 40 mm. Compare the solutions in diffused
daylight 5 min after preparation of the reference suspension, viewing
vertically against a black background. The diffusion of light must be such
that reference suspension I can readily be distinguished from water R, and
that reference suspension II can readily be distinguished from reference
suspension I.
A liquid is considered clear if its clarity is the same as that of water R or of
the solvent used when examined under the conditions described above, or if
its opalescence is not more pronounced than that of reference suspension I
Hydrazine sulfate solution  Dissolve 1.0 g of hydrazine sulfate R in water R
and dilute to 100.0 mL with the same solvent. Allow to stand for 4-6 h.
Hexamethylenetetramine solution  In a 100 mL ground-glass-stoppered
flask, dissolve 2.5 g of hexamethylenetetramine R in 25.0 mL of water R.
CLARITY OR OPALESCENCE AS PER BP
Primary opalescent suspension (formazin suspension): To the
hexamethylenetetramine solution in the flask add 25.0 mL of the hydrazine sulfate
solution. Mix and allow to stand for 24 h. This suspension is stable for 2 months,
provided it is stored in a glass container free from surface defects. The suspension
must not adhere to the glass and must be well mixed before use.
Standard of opalescence  Dilute 15.0 mL of the primary opalescent suspension to
1000.0 mL with water R. This suspension is freshly prepared and may be stored for
up to 24 h.
Reference suspensions  Prepare the reference suspensions according to Table 2.2.1.-
1. Mix and shake before use.
CLARITY OR OPALESCENCE AS PER BP
The degree of opalescence may also be determined by instrumental
measurement of the light absorbed or scattered on account of submicroscopic
optical density in homogeneities of opalescent solutions and suspensions. 2
such techniques are nephelometry and turbidimetry. For turbidity
measurement of coloured samples, ratio turbidimetry and nephelometry with
ratio selection are used.
The light scattering effect of suspended particles can be measured by
observation of either the transmitted light (turbidimetry) or the scattered light
(nephelometry). Ratio turbidimetry combines the principles of both
nephelometry and turbidimetry. Turbidimetry and nephelometry are useful for
the measurement of slightly opalescent suspensions. Reference suspensions
produced under well-defined conditions must be used.
For quantitative measurements, the construction of calibration curves is
essential, since the relationship between the optical properties of the suspension
and the concentration of the dispersed phase is at best semi-empirical.
COLOUR OF SOLUTION
A solution may be described as colourless if it has appearance of water
which examined under specified condition. Varying degrees of colour are
permissible and controlled by direct comparison of solutions with specified
standard colour solution.
COLOUR OF SOLUTION AS PER IP
The colour of the solution is compared with standard reference solution.
Special reagents:
Ferric chloride colorimetric solution (FCS):
Dissolve about 46g of ferric chloride hexahydrate in enough of a mixture of
25ml of hydrochloric acid and 975ml of water to produce 1000ml. Pippette
10 ml of this solution into 250 ml iodine flask, add 15ml of water,3g of
potassium iodide and 5 ml of hydrochloric acid and allow the mixture to
stand for 15 minutes. Dilute to 100 ml with water and titrate the liberated
iodine with 0.1M sodium thiosulphate using 0.5 ml of starch solution, added
towards the end of the titration, as indicator. Carry out a blank titration.
1ml of 0.1M sodium thiosulphate is equivalent to 0.02703g of FeCl3.6H2O.
Adjust the final volume of the solution by the addition of enough of the
mixture of hydrochloric acid and water so that each ml contains 0.045g of
Fecl3.6H2O
Cobaltous chloride Colourimetric Solution (CCS):
Dissolve about 65 g of cobaltous chloride in enough of a mixture of 25ml of
hydrochloric acid and 975ml of water to produce 1000ml. Pipette 5ml of
this solution into a 250ml iodine flask, add 5ml of hydrogen peroxide
solution (10 volume) and 15 ml of sodium hydroxide solution, boil for 10
minutes, cool and add 2g of potassium iodide and 60ml of dilute sulphuric
acid. Dissolve the precipitate by gentle shaking, if necessary, and
titrate the liberated iodine with 0.1m sodium thiosulphate using 0.5ml of
starch solution, added towards the pink end-point, as indicator. Perform a
blank determination and make any necessary correction. Each ml of 0.1m
sodium thiosulphate is equivalent to 0.02379g of CoCl2,6H2O. Adjust the
final volume of the solution by the addition of enough of the mixture of
hydrochloric acid and water so that each ml contains 0.0595g of
CoCl2,6H2O
Cupric Sulphate Colorimetric Solution (CSS):
Dissolve about 65g of cupric sulphate in enough of a mixture of
25 ml of hydrochloric acid and 975 ml of water to produce
1000ml. Pipette 10ml of this solution into a 250ml iodine flask ,
add 40 ml of water ,4 ml of acetic acid, 3g of potassium iodide ,
and 5ml of hydrochloric acid and titrate the liberated iodine
with 0.1m sodium thiosulphate using 0.5ml of starch solution,
added towards the pale brown end-point, as indicator. Perform
a blank determination and make any necessary correction. Each
ml of 0.1M sodium thiosulphate is equivalent to 0.02497g of
CuSO4,5H2O. Adjust the final volume of the solution by the
addition of enough of the mixture hydrochloric acid and water
so that each ml contains 0.0624 g of CuSO4,5H2
Reference Solution: Prepare by mixing the volumes of colorimetric
solutions and hydrochloric acid (1% w/v HCL) as indicated in Table
METHOD
Transfer to a flat bottomed test tube of neutral glass 15 to 25mm in
diameter, a suitable volume of a liquid been examined such that the
test tube is filled to a depth of 40mm. Into another match test tube
add the same volume of water or of the solvent used for preparing
the solution being examined or of the reference solution stated in
the individual monograph. Examined the column of liquid in
diffused light by viewing down the vertical axis of the tubes against
a white background.
Colourless Solution: A solution is considered colouorless if it has
the same appearance as water or the solvent used for preparing the
solution or is not more intensely coloured than reference solution
BS8.
COLOUR OF SOLUTION AS
PER BP
The examination of the degree of coloration of liquids in the
range brown-yellow-red is carried out by one of the 2
methods below, as prescribed in the monograph.

A solution is colourless if it has the appearance of water R

or the solvent or is not more intensely coloured than
reference solution B9.
Method I
Using identical tubes of colourless, transparent, neutral glass of
12 mm external diameter, compare 2.0 mL of the liquid to be
examined with 2.0 mL of water R or of the solvent or of the
reference solution (see Tables of reference solutions) prescribed in
the monograph. Compare the colours in diffused daylight, viewing
horizontally against a white background.
Method II
Using identical tubes of colourless, transparent, neutral glass with
a flat base and an internal diameter of 15 mm to 25 mm, compare
the liquid to be examined with water R or the solvent or the
reference solution (see Tables of reference solutions) prescribed in
the monograph, the depth of the layer being 40 mm. Compare the
colours in diffused daylight, viewing vertically against a white
background.
Primary solutions
Yellow solution:  Dissolve 46 g of ferric chloride R in about 900
mL of a mixture of 25 mL of hydrochloric acid R and 975 mL of
water R and dilute to 1000.0 mL with the same mixture. Titrate and
adjust the solution to contain 45.0 mg of FeCl3,6H2O per millilitre
by adding the same acidic mixture. Protect the solution from light.
Titration. Place in a 250 mL conical flask fitted with a ground-glass
stopper, 10.0 mL of the solution, 15 mL of water R, 5 mL of
hydrochloric acid R and 4 g of potassium iodide R, close the flask,
allow to stand in the dark for 15 min and add 100 mL of water R.
Titrate the liberated iodine with 0.1 M sodium thiosulfate, using 0.5
mL of starch solution R, added towards the end of the titration, as
indicator.
1 mL of 0.1 M sodium thiosulfate is equivalent to 27.03 mg of
FeCl3,6H2O.
Red solution:  Dissolve 60 g of cobalt chloride R in about 900 mL of
a mixture of 25 mL of hydrochloric acid R and 975 mL of water R and
dilute to 1000.0 mL with the same mixture. Titrate and adjust the
solution to contain 59.5 mg of CoCl2,6H2O per millilitre by adding the
same acidic mixture.
Titration. Place in a 250 mL conical flask fitted with a ground-glass
stopper, 5.0 mL of the solution, 5 mL of dilute hydrogen peroxide
solution R and 10 mL of a 300 g/L solution of sodium hydroxide R.
Boil gently for 10 min, allow to cool and add 60 mL of dilute sulfuric
acid R and 2 g of potassium iodide R. Close the flask and dissolve
the precipitate by shaking gently. Titrate the liberated iodine with 0.1
M sodium thiosulfate, using 0.5 mL of starch solution R, added
towards the end of the titration, as indicator. The end-point is
reached when the solution turns pink.
1 mL of 0.1 M sodium thiosulfate is equivalent to 23.79 mg of
CoCl2,6H2O.
Blue primary solution:  Dissolve 63 g of copper sulfate R in about
900 mL of a mixture of 25 mL of hydrochloric acid R and 975 mL of
water R and dilute to 1000.0 mL with the same mixture. Titrate and
adjust the solution to contain 62.4 mg of CuSO4,5H2O per millilitre
by adding the same acidic mixture.

Titration. Place in a 250 mL conical flask fitted with a ground-glass

stopper, 10.0 mL of the solution, 50 mL of water R, 12 mL of dilute
acetic acid R and 3 g of potassium iodide R. Titrate the liberated
iodine with 0.1 M sodium thiosulfate, using 0.5 mL of starch solution
R, added towards the end of the titration, as indicator. The end-point
is reached when the solution shows a slight pale brown colour.

1 mL of 0.1 M sodium thiosulfate is equivalent to 24.97 mg of

CuSO4,5H2O.
STANDARD SOLUTION
Using the 3 primary solutions, prepare the 5 standard solutions as
follows:
Reference solutions for Methods I and II
Using the 5 standard solutions, prepare the following
reference solutions.
Storage

For Method I, the reference solutions may be stored in

sealed tubes of colourless, transparent, neutral glass of 12
mm external diameter, protected from light.

For Method II, prepare the reference solutions immediately

before use from the standard solutions.