1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Chondroitin Sulfate Sodium from the
Pharmacopoeial specifications.
2.0 SCOPE:
This
SOP shall be applicable in Q.C laboratory.
3.0 RESPONSIBILITY:
3.1 Q.C Analyst.
4.0 ACCOUNTABILITY:
4.1 Q.C Manager.
5.0 PROCEDURE:
5.1 Characters:
5.1.1
Appearance:
5.1.1.1 White or almost white.
5.1.1.2 Hygroscopic powder.
5.1.2
Solubility:
5.1.2.1 Material and equipment:
5.1.2.1.1
Glassware
(2 test tubes, 1 spatula, 1 pipette).
5.1.2.1.2
Ethanol
(96%).
5.1.2.1.3
Acetone.
5.1.2.1.4
Purified
water.
5.1.2.2 Sample:
5.1.2.2.1
Small
quantity.
5.1.2.3 Method:
5.1.2.3.1
Take
a test tube and add small quantity of sample for testing solubility according
to B.P specifications.
5.1.2.3.2
Add
purified water in test tube 1 and observe.
5.1.2.3.3
Add
acetone in test tube 2 and observe.
5.1.2.3.4
Add
Ethanol (96%) in test tube 3 and observe.
5.1.2.4 Observations:
5.1.2.4.1
The
sample in test tube 1 containing with purified water is freely soluble.
5.1.2.4.2
The
sample in test tube 2 & 3 containing with acetone and Ethanol (96%) are
practically insoluble respectively.
5.2 Solution
S1:
5.2.1
Material
and equipment:
5.2.1.1 Glassware (1 50.0ml of beaker, 1 spatula,
1 glass rod, 1 pipette).
5.2.1.2 Analytical weighing balance.
5.2.1.3 Magnetic stirrer.
5.2.1.4 Carbon-dioxide free water.
5.2.2
Sample:
5.2.2.1 2.5g.
5.2.3
Preparation
of solution S:
5.2.3.1 Take a beaker of 50.0ml and add sample 2.5g
in it.
5.2.3.2 Add in it 50.0ml of carbon-dioxide free
water, dissolve by using magnetic stirrer operate.
5.3 Identification
tests:
5.3.1
Sodium test:
5.3.1.1 Material and equipment:
5.3.1.1.1
Glassware
(test tube, spatula, beaker, glass rod).
5.3.1.1.2
Analytical
weighing balance.
5.3.1.1.3
Burner.
5.3.1.1.4
1.0ml
of dilute ammonia.
5.3.1.1.5
Ice-water.
5.3.1.1.6
1.0ml
of ammonium carbonate solution R.
5.3.1.2 Sample:
5.3.1.2.1
0.5ml
of solution S1.
5.3.1.3 Method:
5.3.1.3.1
Take
a test tube and add 0.5ml of solution S1.
5.3.1.3.2
Add
1.5ml of methoxyphenylacetic reagent R.
5.3.1.3.3
Cool
in ice-water for 30min.
5.3.1.3.4
Observe
the changes.
5.3.1.3.5
A
voluminous, white, crystalline ppt is formed.
5.3.1.3.6
Place
in purified water at 20oC and stir for 5minutes.
5.3.1.3.7
The
ppt does not disappear.
5.3.1.3.8
Add
1.0ml of dilute ammonia.
5.3.1.3.9
The
ppt dissolves completely.
5.3.1.3.10 Add 1.0ml of ammonium carbonate solution
R.
5.3.1.3.11 Observe the changes.
5.3.1.4 Observations:
5.3.1.4.1
No
ppt is formed.
5.4 Loss
on drying:
5.4.1
Material
and equipment:
5.4.1.1 Glassware (according to requirement).
5.4.1.2 Analytical weighing balance.
5.4.1.3 Oven.
5.4.2
Sample:
5.4.2.1 1.0g.
5.4.3
Method:
5.4.3.1 Weigh 1.0g of the test sample.
5.4.3.2 Set the oven apparatus.
5.4.3.3 Place the sample into the tray and dry it.
5.4.3.4 Set the temperature of oven at 105oC
for at least 4hour.
5.4.3.5 And wait till the sample loses its moisture.
5.4.3.6 After 4 hours weigh the sample again by
using analytical weighing balance i.e. the final weight.
5.4.3.7 Note down readings on given Annexure-1.
5.4.4
Observation:
5.4.4.1 Maximum 12.0%.
5.5 Assay:
5.5.1
Apparatus:
5.5.1.1 Glassware (according to requirement).
5.5.1.2 Titration apparatus.
5.5.1.3 Magnetic stirrer.
5.5.2
Material and reagents:
5.5.2.1 0.1g of chondroitin sulfate sodium CRS.
5.5.2.2 4.0g cetylpyridinium chloride monohydrate.
5.5.2.3 0.1ml of a 1% solution of methylene blue
R.
5.5.2.4 Purified water.
5.5.3
Sample:
5.5.3.1 0.1g.
5.5.4
Method of analysis:
5.5.4.1 Test
solution (a):
5.5.4.1.1
Take
a 100.0ml of beaker and add weighed 0.1g (m1) of the substance in
it.
5.5.4.1.2
Dissolve
it in sufficient quantity of purified water.
5.5.4.1.3
Finally
dilute it to 100.0ml with the same solvent.
5.5.4.2 Reference
solution (a):
5.5.4.2.1
Take
a 100.0ml of beaker and add weighed 0.1g (m0) of chondroitin sulfate
sodium CRS, previously dried as described in the test for loss on drying in 5.4
point in it.
5.5.4.2.2
Dissolve
it in sufficient quantity of purified water.
5.5.4.2.3
Finally
dilute it to 100.0ml with the same solvent.
5.5.4.3 Titrant
solution (a):
5.5.4.3.1
Take
a 1000.0ml of beaker and add weighed 4.0g cetylpyridinium chloride monohydrate
R in it.
5.5.4.3.2
Dissolve
it in sufficient quantity of purified water.
5.5.4.3.3
Finally
dilute it to 1000.0ml with purified water.
5.5.4.4 Visual
titration:
5.5.4.4.1
Set
titration apparatus.
5.5.4.4.2
Fill
the right hand side burette with titrant solution (a).
5.5.4.4.3
Take
40.0ml of test solution (a) in volumetric flask and titrate it with titrant
solution (a).
5.5.4.4.4
The
solution becomes turbid.
5.5.4.4.5
At
the end point, the liquid appears clear, with an almost-white ppt in
suspension.
5.5.4.4.6
The
ppt is more apparent if 0.1ml of a 1% solution of methylene blue R is added
before starting the titration.
5.5.4.4.7
The
precipitated particles are more apparent against the blue background.
5.5.4.4.8
Similarly,
take 40.0ml of reference solution (a) in volumetric flask and titrate it with
titrant solution (a).
5.5.4.4.9
Note
down the volume used as shown in Annexure-2.
5.5.4.5 Calculate the percentage content of
chondroitin sulfate sodium using the following expression:
u1
x m0
x 100
x Z
u0
x m1 100-h
u0=
volume of appropriate titrant solution when titrating the appropriate reference
solution, in milliliters.
u1=
volume of appropriate titrant solution when titrating the appropriate test
solution, in milliliters.
h=
loss on drying of the substance to be examined, as a percentage.
Z=
percentage content of H2O (C14H19NNa2O14S)x
in chondroitin sulfate sodium CRS.
5.5.5
Limit:
5.5.5.1 95.0% to 105.0% (dried substance).
6.0 REVISION
LOG:
Revision No.
|
Effective Date
|
Reason
|
00
|
New
SOP
|
7.0 REFERENCES:
7.1 The British Pharmacopoeia. Vol I., Official Monograph /Chondroitin sulfate sodium: 2015, pp. 538-541.
7.2 The British Pharmacopoeia. Vol V., Official Monograph /Qualitative
Reactions and Tests: 2015, pp. 266-270.
8.0 ANNEXURES:
Annexure 1: Observations
of percentage loss of drying.
Annexure 2: Observations
and calculations of assay of Chondroitin sulfate sodium.
Annexure:
1
Observations
of percentage loss of drying
Percentage loss
of drying
Apparatus:
____________________
Temperature:
__________________
Pressure:
_____________________
Weight
of Sample = _____________
Time
period = _____________
Average
% Loss of Moisture: _____________
Remarks: ____________________________________________________________
|
||||||||||||||||||||||||
Annexure:
2
Observations
and calculations of assay of Chondroitin sulfate sodium
VISUAL TITRATION
Indicator:____________________________
Weight
of sample( m1):_____________________
Weight
of reference material (m0):____________
Titrant:______________________________
Percentage
content of H2O (C14H19NNa2O14S)x
in chondroitin sulfate sodium CRS (Z):________________________
Sample titration
Average volume (u1):
_________________
Blank titration
Average volume(u0):
_________________
Calculations:
Calculate
the percentage content of chondroitin sulfate sodium using the following
expression:
u1
x m0 x 100
x Z
u0
x m1 100-h
Result: ____________________________________________________________________
|
9.0
ABBREVIATIONS:
Abbreviation
|
Expanded Form
|
SOP
|
Standard
operating procedure
|
&
|
And
|
No.
|
Number
|
Ltd.
|
Limited
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
Q.C
|
Quality
control
|
Vol
|
Volume
|
mg
|
Milligram
|
ml
|
Milliliter
|
ppt
|
Precipitate
|
M
|
Molar
|
g
|
Grams
|
%
|
Percentage
|
R
|
Reagent
|
BP
|
British
Pharmacopoeia
|
vi
|
Initial
volume
|
vf
|
Final
volume
|
h
|
Hour
|
oC
|
Degree
centigrade
|