FLUOXETINE HYDROCHLORIDE SOP
1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Fluoxetine hydrochloride 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).
5.1.2.1.2
Methanol.
5.1.2.1.3
Methylene
chloride.
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 3 test tubes
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 methylene
chloride in test tube 3 and observe.
5.1.2.4
Observations:
5.1.2.4.1
The sample in test
tube 1and 3 containing with purified water and methylene chloride are sparingly
soluble, respectively.
5.1.2.4.2
The sample in test
tube 2 containing with methanol is freely soluble.
5.2 Identification
tests:
5.2.1
Chlorides
test:
5.2.1.1
Material and equipment:
5.2.1.1.1
Glassware
(according to requirement).
5.2.1.1.2
Centrifuge
machine.
5.2.1.1.3
Dilute nitric
acid.
5.2.1.1.4
Purified water
(q.s).
5.2.1.1.5
0.4ml of Silver
nitrate R1.
5.2.1.1.6
1.5ml of ammonia.
5.2.1.2
Sample:
5.2.1.2.1
Quantity of
substance to be examined equivalent to about 2.0mg of chloride.
5.2.1.3
Method:
5.2.1.3.1
Take a test tube
add in it 2.0ml of water with the help of pipette.
5.2.1.3.2
Dissolve in 2.0ml
of water a quantity of the substance to be equivalent to about 2.0mg of
chloride.
5.2.1.3.3
Acidify with
dilute nitric acid.
5.2.1.3.4
And add 0.4ml of
silver nitrate R1.
5.2.1.3.5
Shake and allow it
to stand.
5.2.1.3.6
A curdled, white
ppt is formed.
5.2.1.3.7
Centrifuge it in
centrifugation machine, according to SOP.
5.2.1.3.8
The obtained ppt
is wash with 3 quantities, each of 1ml, of water.
5.2.1.3.9
Carry out this
operation rapidly is subdued light, degrading the fact that the supernatant
solution may not become perfectly clear.
5.2.1.3.10 Suspend
the precipitate in 2.0ml of water and add 1.5ml of ammonia.
5.2.1.4
Observations:
5.2.1.4.1
The precipitate
dissolves easily with the possible exception of a few large particles which
dissolves slowly.
5.3 Solution
S:
5.3.1
Material and equipment:
5.3.1.1
Glassware (1
100.0ml of beaker, 1 spatula, 1 glass rod, 1 pipette).
5.3.1.2
Analytical
weighing balance.
5.3.1.3
Magnetic stirrer.
5.3.1.4
85 volumes of
methanol.
5.3.1.5
Purified water.
5.3.2
Sample:
5.3.2.1
2.0g.
5.3.3
Preparation of solution
S:
5.3.3.1
Take a beaker of
100.0ml and add sample 2.0g in it.
5.3.3.2
Dissolve it in a
mixture of 15 volumes of purified water and 85 volumes of methanol.
5.3.3.3
Dilute it to
100.0ml with the same mixture of solvent.
5.4 Optical rotation:
5.4.1
Material and equipment:
5.4.1.1
Polarimeter.
5.4.1.2
Magnetic stirrer.
5.4.1.3
Glassware (1
beaker of 50.0ml, 1 stirrer, 1 spatula).
5.4.2
Sample:
5.4.2.1
Solution S.
5.4.3
Method:
5.4.3.1
Firstly clean the Polarimeter
with clean dry cloth, according to SOP No. BM/QCEC/SOP031-00.
5.4.3.2
Operate the
Polarimeter according to SOP.
5.4.3.3
Fill the
Polarimeter tube with blank solution and determine the observed optical
rotation.
5.4.3.4
Similarly, fill
the Polarimeter tube with sample solution and determine the observed optical
rotation.
5.4.3.5
Note down the
values in annexure-1.
5.4.4
Observations:
5.4.4.1
The optical
rotation is -0.05o to +0.05o.
5.5 Assay:
5.5.1
Apparatus:
5.5.1.1
HPLC apparatus.
5.5.1.2
Glassware
(according to the requirement).
5.5.2
Material
and reagents:
5.5.2.1
55.0mg of
Fluoxetine hydrochloride CRS.
5.5.2.2
Methanol.
5.5.2.3
Tetrahydrofuran.
5.5.2.4
Triethylamine.
5.5.2.5
Purified water.
5.5.2.6
Phosphoric acid.
5.5.3
Sample:
5.5.3.1
55.0mg.
5.5.4
Test
solution:
5.5.4.1
Take a beaker of
50.0ml and add 55.0mg of the substance to be examined in it.
5.5.4.2
Add in sufficient
quantity of mobile phase and dissolved it by using magnetic stirrer operate
according to SOP.
5.5.4.3
And dilute it to
50.0ml with the mobile phase.
5.5.4.4
Take 10.0ml of
this solution in another beaker.
5.5.4.5
And dilute it to
100.0ml with the mobile phase.
5.5.5
Reference
solutions:
5.5.5.1
Take a beaker of
50.0ml and add 55.0mg of Fluoxetine hydrochloride CRS in it.
5.5.5.2
Add in sufficient
quantity of mobile phase and dissolved it by using magnetic stirrer operate
according to SOP.
5.5.5.3
And dilute it to
50.0ml with the mobile phase.
5.5.5.4
Take 10.0ml of
this solution in another beaker.
5.5.5.5
And dilute it to
100.0ml with the mobile phase.
5.5.6
Column:
5.5.6.1
Size:
5.5.6.1.1
Length=0.25m,
5.5.6.1.2
θ=4.6mm.
5.5.6.2
Stationary
phase:
5.5.6.2.1
Octylsilyl silica
gel for chromatography R (5μm).
5.5.7
Mobile
phase:
5.5.7.1
Mix 8 volumes of
methanol R, 30 volumes of tetrahydrofuran R and 62 volumes of a solution of
triethylamine R prepared as follows: to 10.0ml of triethylamine R, and add
980.0ml of purified water R, mix and adjust to pH 6.0 with phosphoric acid R
(about 4.5ml) and dilute to 1000.0ml with purified water R.
5.5.8
Flow
rate:
5.5.8.1
1.0ml/min.
5.5.9
Detection:
5.5.9.1
Spectrophotometer
at 227nm.
5.5.10
Injection:
5.5.10.1 10μL.
5.5.11 Run time:
5.5.11.1 3
times the retention time of fluoxetine.
5.5.12 Retention time:
5.5.12.1 Fluoxetine=10min
to 18min; if necessary, adjust the volumes of methanol and of the solution of
triethylamine in the mobile phase.
5.5.13 System
suitability: reference solution:
5.5.13.1 Symmetry factor: Maximum
2.0 calculated at 10% of the height of the peak due to fluoxetine.
5.5.14 Method of
analysis:
5.5.14.1 Firstly
prepare the test solution, reference solution and mobile phase according to the
requirements.
5.5.14.2 The
solutions must be free from solid particles.
5.5.14.3 Prepare
the apparatus.
5.5.14.4 The
mobile phase solvent mixtures must be de-aerated prior to use either by boiling
or by applying a partial vacuum to the solvent reservoir.
5.5.14.5 Equilibrate
the column with the prescribed mobile phase, flow rate and at temperature
specified until a suitable baseline is achieved.
5.5.14.6 Test
solution of the mixture to be separated is now introduced into the mobile phase
with the help of an injector just before entering the separating column.
5.5.14.7 As
the eluate leaves the column it enters a detector, where it is continuously
monitored at the specified λ.
5.5.14.8 The
electrical signal obtained from detector is amplified and routes to recorder
which record the developed chromatogram.
5.5.14.9 Calculate
the percentage content of Fluoxetine hydrochloride (C17H19ClF3NO)
from the declared content of Fluoxetine hydrochloride CRS.
5.5.15 Observations:
5.5.15.1 98.0%
to 102.0% (anhydrous substance).
6.0 REFERENCES:
6.1
The British
Pharmacopoeia. Vol I.,
Official Monograph / Fluoxetine
hydrochloride: 2015, pp. 995-996.
6.2
The British
Pharmacopoeia. Vol V.,
Official Monograph /Qualitative Reactions and Tests: 2015, pp. 266-270.
7.0 ANNEXURES:
Annexure 1: Optical
rotation observations and calculations.
Annexure 2: Observations
and calculations of HPLC method.
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
and calculations of HPLC method
|
Analysis
on HPLC
Instrument:
___________________
Date: _________________
Model:
___________________
Sample
solution: _______________________
Reference
standard solution: ______________
Impurities:
____________________________
(calculate
each component calculation separately)
OBSERVATIONS:
Attach
chromatogram.
CALCULATIONS:
1.
Retention time:
n= no. of peak
Retention time of unretained peak (tm)=
_____________
2.
Retention volume:
Flow rate= _______________ml/min.
3.
Retention factor:
Retention time of unretained peak (tm)=
_____________
4.
Separation factor (α):
5.
Resolution:
Retention time of unretained peak (tm)=
_____________
6.
Efficiency:
7.
Height equivalent to a theoretical plate (HETP):
Length of column = ________________________
8.
Symmetry factor (tailing factor):
9.
Response factor & Relative response factor:
Conc. (mg/ml)= ___________________
10. Relative standard deviation (%RSD):
Use formula of relative standard deviation where it is
required i.e.,
11. Percentage of content:
Percentage content = (rU/rS) x (CS/CU)
x 100.
rU= peak response of substance from the sample
solution.
rS= peak response of substance from the standard
solution.
CS= concentration of substance in the standard
solution (mg/mL).
CU= concentration of substance in the sample
solution (mg/mL).
RESULTS:
________________________________________________________________________________________________________________________________________________
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8.0 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
|
|
g
|
Grams
|
|
ml
|
Milliliter
|
|
oC
|
Degree
centigrade
|
|
mg
|
Milligram
|
|
M
|
Molar
|
|
%
|
Percentage
|
|
R
|
Reagent
|
|
μm
|
Micron/
micrometer
|
|
g/L
|
Gram
per liter
|
|
CRS
|
Chemical
reference solution
|
|
m
|
Meter
|
|
θ
|
Theta
|
|
mm
|
Millimeter
|
|
ml/min
|
Milliliter
per minute
|
|
nm
|
Nanometer
|
|
μL
|
Microliter
|
|
Min
|
Minute
|
|
λ
|
Lamda
|