GLUCOSAMINE SULFATE SODIUM CHLORIDE SOP
1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Glucosamine Sulfate Sodium Chloride 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
Crystalline
powder.
5.1.2
Solubility:
5.1.2.1
Material and equipment:
5.1.2.1.1
Glassware (3 test
tubes, 1 spatula, 1 pipette).
5.1.2.1.2
Methanol.
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 methanol in
test tube 2 and observe.
5.1.2.3.4
Add acetone 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 containing with methanol is sparingly soluble.
5.1.2.4.3
The sample in test
tube 3 containing with acetone is practically insoluble
5.2 Solution
S:
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 sufficient
quantity of carbon-dioxide free water, dissolve by using magnetic stirrer
operate according to SOP.
5.2.3.3
Dilute it to
25.0ml with the same solvent.
5.3 Identification
tests:
5.3.1
Chlorides
test:
5.3.1.1
Material and equipment:
5.3.1.1.1
Glassware
(according to requirement).
5.3.1.1.2
Centrifugation
machine.
5.3.1.1.3
Dilute nitric
acid.
5.3.1.1.4
Purified water
(q.s).
5.3.1.1.5
0.4ml of Silver
nitrate R1.
5.3.1.1.6
1.5ml of ammonia.
5.3.1.2
Sample:
5.3.1.2.1
Quantity of substance
to be examined equivalent to about 2.0mg of chloride.
5.3.1.3
Method:
5.3.1.3.1
Take a test tube
add in it 2.0ml of water with the help of pipette.
5.3.1.3.2
Dissolve in 2.0ml
of water a quantity of the substance to be equivalent to about 2.0mg of
chloride.
5.3.1.3.3
Acidify with
dilute nitric acid.
5.3.1.3.4
And add 0.4ml of
silver nitrate R1.
5.3.1.3.5
Shake and allow it
to stand.
5.3.1.3.6
A curdled, white
ppt is formed.
5.3.1.3.7
Centrifuge it in
centrifugation machine, according to SOP.
5.3.1.3.8
The obtained ppt
is wash with 3 quantities, each of 1ml, of water.
5.3.1.3.9
Carry out this
operation rapidly is subdued light, degrading the fact that the supernatant
solution may not become perfectly clear.
5.3.1.3.10 Suspend
the precipitate in 2.0ml of water and add 1.5ml of ammonia.
5.3.1.4
Observations:
5.3.1.4.1
The precipitate
dissolves easily with the possible exception of a few large particles which
dissolves slowly.
5.3.2
Sodium
test:
5.3.2.1
Material and equipment:
5.3.2.1.1
Glassware (test
tube, spatula, beaker, glass rod).
5.3.2.1.2
2.0ml of 150g/L
solution of potassium carbonate.
5.3.2.1.3
4.0ml of potassium
pyroantimonate solution.
5.3.2.1.4
Iced water.
5.3.2.1.5
Purified water.
5.3.2.2
Sample:
5.3.2.2.1
1.0ml of solution
S.
5.3.2.3
Method:
5.3.2.3.1
Take a test tube
and add 1.0ml of solution S.
5.3.2.3.2
Add 2.0ml of
150g/L solution of potassium carbonate and heat it to boiling on burner.
5.3.2.3.3
Observe the
changes.
5.3.2.3.4
No ppt is formed.
5.3.2.3.5
Add 4.0ml of potassium
pyroantimonate solution again heat it to boiling on burner.
5.3.2.3.6
And then take a
beaker of filled with iced water, put test tube in it and allow it to cool.
5.3.2.3.7
If necessary rub
the inside of the test tube with a glass rod.
5.3.2.3.8
Observe the
changes.
5.3.2.4
Observations:
5.3.2.4.1
A dense white ppt
is formed.
5.3.3
Sulfates
test:
5.3.3.1
Material and equipment:
5.3.3.1.1
Glassware (test
tube, spatula, beaker, glass rod).
5.3.3.1.2
1.0ml of dilute
hydrochloric acid.
5.3.3.1.3
1.0ml of barium
chloride solution.
5.3.3.1.4
5.0ml of purified
water.
5.3.3.2
Sample:
5.3.3.2.1
45mg of the substance
to be examined.
5.3.3.3
Method:
5.3.3.3.1
Take a test tube
and add 45mg of the substance to be examined and 5.0ml of purified water.
Dissolve it.
5.3.3.3.2
Add 1.0ml of
dilute hydrochloric acid and 1.0ml of barium chloride solution R1.
5.3.3.3.3
Observe the
changes.
5.3.3.4
Observations:
5.3.3.4.1
A white ppt is
formed.
5.3.4
Specific
optical rotation:
5.3.4.1
Material and equipment:
5.3.4.1.1
Polarimeter.
5.3.4.1.2
Analytical
weighing balance.
5.3.4.1.3
Glassware (1
beaker of 50.0ml, 1 stirrer, 1 spatula).
5.3.4.2
Sample:
5.3.4.2.1
Solution S.
5.3.4.3
Method:
5.3.4.3.1
Perform the test
on the solution S.
5.3.4.3.2
Examine 3hours
after preparation of solution S.
5.3.4.3.3
Firstly clean the
Polarimeter with clean dry cloth, according to.
5.3.4.3.4
Operate the
Polarimeter according to SOP.
5.3.4.3.5
Fill the
Polarimeter tube with blank solution and determine the observed optical
rotation.
5.3.4.3.6
Similarly, fill
the Polarimeter tube with sample solution (i.e solution S) and determine the
observed optical rotation.
5.3.4.3.7
Note down the
values in annexure-1.
5.3.4.3.8
Calculate the
specific optical rotation by using formula:
[α]λ T = α/lc
5.3.4.4
Observations:
5.3.4.4.1
+50.0 to +55.0
(dried substance).
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. Operate it according to the SOP.
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-2.
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
Potentiometer.
5.5.1.3
Magnetic stirrer.
5.5.2
Material
and reagents:
5.5.2.1
0.1ml of 0.1M
hydrochloric acid.
5.5.2.2
0.1M sodium
hydroxide.
5.5.2.3
50.0ml of Purified
water.
5.5.3
Sample:
5.5.3.1
0.250g.
5.5.4
Method
of analysis:
5.5.4.1 Take
a 100.0ml of volumetric flask and add 0.250g of sample in it.
5.5.4.2 Add
50.0ml of purified water and 0.1ml of 0.1M hydrochloric acid in it and dissolve
it by using magnetic stirrer.
5.5.4.3 Fill
the right hand side burette with titrant 0.1M sodium hydroxide.
5.5.4.4 Carry
out a Potentiometric titration.
5.5.4.5
Operate potentiometer
according to SOP.
5.5.4.6 To
neutralize analyte add titrant fixed volume (1ml, 0.5ml or 0.1ml) from burette
every time note the reading of change in potential difference (millivolts) for
each addition in given annexure-3.
5.5.4.7 Plot
a graph, volume used v/s millivolts.
5.5.4.8 Find
out the END POINT. Read the volume added between the 2 points of inflexion.
5.5.4.9 Peak
of graph indicates END POINT i.e. the point at which maximum millivolts. Note
down volume used at that point.
5.5.4.10 Perform
blank titration without using sample. Similarly, as sample titration performed.
Record observations in annexure-3.
5.5.4.11 Calculate
volume used by substance by using formula:
Volume
used by substance = Blank titration - Sample titration.
5.5.4.12 Calculate
percentage purity of the sample by using formula:
%age
purity = volume used by substance x factor x 100
Weight of sample
5.5.5
Factor:
5.5.5.1 1ml
of 0.1M sodium hydroxide is equivalent to 28.67mg of glucosamine sulfate sodium
chloride C12H28Cl2N2Na2O14S.
5.5.6
Limit:
5.5.6.1 98.0%
to 102.0% (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 / Glucosamine sulfate sodium chloride: 2015, pp. 1080-1081.
7.2
The British
Pharmacopoeia. Vol V.,
Official Monograph /Qualitative Reactions and Tests: 2015, pp. 266-270.
8.0 ANNEXURES:
Annexure 1: Specific
optical rotation observations and calculations.
Annexure 2: Observations
of percentage loss of drying.
Annexure 3: Assay
observations and calculations (Potentiometric titration).
Annexure: 1
Specific
optical rotation observations and calculations
|
Specific
optical rotation
Instrument:
___________________
Date: _______________
Model:
_______________________ Length
of Polarimeter tube: ________________
Sample:
________________________________g.
Solvent:
________________________________ml.
Concentration
of sample solution: ____________g/ml.
Blank solution:
Average: _______________
Optical
rotation of blank solution: _______________
Sample solution:
Average: _______________
Optical
rotation of sample solution: ______________
Optical rotation of substance = Blank solution - Sample
solution.
Specific
optical rotation of sample solution by using formula:
[α]λ T = α/lc
Result: ________________
Remarks:
___________________________________________________________
|
Annexure:
2
Observations
of percentage loss of drying
|
Percentage
loss of drying
Apparatus:
____________________
Temperature:
__________________
Pressure:
_____________________
Weight
of Sample = _____________
Time
period = _____________
Average % Loss of Moisture: _____________
Remarks:
____________________________________________________________
|
||||||||||||||||||||||||
Annexure:
3
Assay
observations and calculations (Potentiometric titration)
|
Potentiometric titration
Reference
electrode: ___________________
Indicator
electrode: ____________________
Speed
of magnetic stirrer: _______________
Titrant
used: __________________________
Indicator:
____________________________
Blank
titration:
Plot a graph, volume used v/s millivolts and find out peak
of graph i.e. END POINT of blank titration.
Sample
titration:
Plot a graph, volume used v/s millivolts and find out peak
of graph i.e. END POINT of sample titration:
Volume used by Blank titration: __________________
Volume used by Sample titration: _________________
Volume used by substance = Blank titration - Sample
titration.
mV used by Blank titration: __________________
mV used by Sample titration: _________________
mV used by substance = Blank
titration - Sample titration.
Volume
used by substance: _______________________
Voltmeter
(mV) used by substance: _________________
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
|
|
h
|
Hour
|
|
oC
|
Degree
centigrade
|
|
v/s
|
Verses
|
|
mV
|
Millivolts
|
|
o
|
Degree
(angle)
|
|
l
|
Length
|
|
c
|
Concentration
(g/ml)
|
|
g/ml
|
Gram
per milliliter
|
|
α
|
Alpha
|
|
λ
|
Lambda
|
|
g/L
|
Grams
per liter
|
|
Temp.
|
Temperature
|