DUTASTERIDE USP
SOP
1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Dutasteride USP 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 Identification
tests:
5.1.1
Specific
optical rotation:
5.1.1.1
Material and equipment:
5.1.1.1.1
Polarimeter.
5.1.1.1.2
Analytical
weighing balance.
5.1.1.1.3
Magnetic stirrer.
5.1.1.1.4
Glassware (1
beaker of 50.0ml, 1 stirrer, 1 spatula).
5.1.1.1.5
Purified water.
5.1.1.2
Sample:
5.1.1.2.1
10.0mg/ml.
5.1.1.3
Method:
5.1.1.3.1
Take a beaker and
prepare 10.0mg/ml in chloroform and alcohol (98:2).
5.1.1.3.2
Firstly clean the
Polarimeter with clean dry cloth, according to SOP No. BM/QCEC/SOP031-00.
5.1.1.3.3
Operate the
Polarimeter according to SOP.
5.1.1.3.4
Fill the
Polarimeter tube with blank solution and determine the observed optical
rotation.
5.1.1.3.5
Similarly, fill
the Polarimeter tube with sample solution and determine the observed optical
rotation.
5.1.1.3.6
Note down the
values in annexure-1.
5.1.1.3.7
Calculate the
specific optical rotation with reference to anhydrous substance by using
formula:
[α]λ T = α/lc
5.1.1.4
Observations:
5.1.1.4.1
+15.0o
to 25.0o.
5.2 Assay:
5.2.1
Apparatus:
5.2.1.1
Glassware
(according to requirement).
5.2.1.2
Magnetic stirrer.
5.2.1.3
HPLC apparatus.
5.2.1.4
Analytical
weighing balance.
5.2.1.5
Acetonitrile.
5.2.1.6
Trifluoroacetic
acid.
5.2.1.7
0.5mg/mL of USP
Dutasteride Resolution Mixture RS.
5.2.1.8
0.5mg/mL of USP
Dutasteride RS.
5.2.1.9
0.5mg/mL of USP
Dutasteride.
5.2.1.10 Purified
water.
5.2.2
Diluent:
5.2.2.1 Take
a beaker and prepare 60 volumes of acetonitrile and 40 volumes of purified
water.
5.2.3
Mobile
phase:
5.2.3.1
Take a beaker and
prepare 52 volumes of acetonitrile, 48 volumes of purified water and 0.025
volumes of trifluoroacetic acid.
5.2.4
System
suitability solution:
5.2.4.1
Prepare 0.5mg/mL
of USP Dutasteride Resolution Mixture RS in Diluent.
5.2.4.2
Sonicate to
dissolve by using Sonicator operate according to SOP
5.2.5
Standard
solution:
5.2.5.1
Prepare 0.5mg/mL
of USP Dutasteride RS in Diluent.
5.2.5.2 Sonicate
to dissolve by using Sonicator operate according to SOP
5.2.6
Sample
solution:
5.2.6.1
Prepare 0.5mg/mL
of Dutasteride in Diluent.
5.2.6.2 Sonicate
to dissolve by using Sonicator operate according to
5.2.7
Chromatographic
system:
5.2.7.1
Mode:
Liquid chromatography.
5.2.7.2
Detector:
UV
220nm.
5.2.7.3
Column:
4.6-mm
× 25-cm; 5-µm packing L1.
5.2.7.4
Column
temperature: 35o.
5.2.7.5
Flow
rate: 1.0mL/min.
5.2.7.6
Injection
size: 10µL.
5.2.7.7
Run
time: 1.5 times the retention time of Dutasteride.
5.2.8
System
suitability:
5.2.8.1 Sample: System
suitability solution and standard solution.
[NOTE---- See Table for the relative
retention times.]
|
Name
|
Relative retention time
|
Relative response factor
|
Acceptance criteria NMT (%)
|
|
Dutasteride acida
|
0.10
|
1.0
|
0.2
|
|
Dutasteride dimethylamideb
|
0.11
|
1.4
|
0.2
|
|
Dutasteride methyl esterc
|
0.28
|
1.0
|
0.15
|
|
Dutasteride ethyl esterd
|
0.39
|
1.0
|
0.2
|
|
Dutasteride 17α-5-enee
|
0.90
|
1.0
|
0.2
|
|
Dutasteride 17α-epimerf
|
0.93
|
1.0
|
0.3
|
|
Dutasteride
|
1.00
|
-
|
-
|
|
Chlorodutasterideg
|
1.15
|
0.33
|
0.4
|
|
Dihydrodutasterideh
|
1.19
|
1.0
|
0.15
|
|
Dutasteride 5-ene
|
1.20
|
1.0
|
0.3
|
|
Any other individual impurity
|
-
|
-
|
0.1
|
5.2.9
Suitability
requirements:
5.2.9.1 Resolution: NLT 1.5 between
Dutasteride 17α-epimer and Dutasteride, system suitability solution
5.2.9.2 Relative
standard deviation: NMT
1.5%, Standard solution.
5.2.10 Analysis:
5.2.10.1 Samples:
Standard
solution and sample solution.
5.2.10.2 Calculate
the percentage of dutasteride (C27H30F6N2O2)
in the portion of Dutasteride taken:
Result=
(rU/rS) × (CS/CU) × 100
rU=
peak response from the Sample solution.
rS=
peak response from the Standard solution.
CS=
concentration of USP Dutasteride RS in the Standard solution (mg/mL).
CU=
concentration of Dutasteride in the Sample solution (mg/mL).
5.3 Procedure:
5.3.1
Equilibrate the
column and detector with mobile phase at specified flow rate until a constant
signal is received.
5.3.2
Separately inject
equal volumes about 10μL of the standard solution and sample solution.
5.3.3
Record the spectrum
.
5.3.4
Measure the
responses for the major peaks.
5.3.5
Analyze as
directed in the monograph.
5.4 Acceptance
criteria:
5.4.1
97.0% - 102.0% on
the anhydrous and solvent free basis.
6.0 REVISION LOG:
|
Revision No.
|
Effective Date
|
Reason
|
|
00
|
|
New SOP
|
7.0 REFERENCES:
7.1
USP38NF33
Volume-4 Official Monograph/ Dutasteride: 2015, pp.: 3247-3250.
7.2
USP38NF33
Volume-1 Official Monograph/ Chromatography: 2015, pp.: 424-434.
8.0 ANNEXURES:
Annexure 1: Specific
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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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
|
|
g
|
Grams
|
|
ml
|
Milliliter
|
|
Min
|
Minutes
|
|
oC
|
Degree
centigrade
|
|
mg
|
Milligram
|
|
N
|
Normal
|
|
USP
|
United
states pharmacopoeia
|
|
NF
|
National
formulary
|
|
cm
|
Centimeter
|
|
nm
|
Nanometer
|
|
mm
|
Millimeter
|
|
Approx.
|
Approximately
|
|
%
|
Percentage
|
|
μL
|
Microliter
|
|
λ
|
Lambda
|
|
UV
|
Ultraviolet
|
|
ml/min
|
Milliliter
per minute
|
|
o
|
Degree
(angle)
|
|
l
|
Length
|
|
c
|
Concentration
(g/ml)
|
|
g/ml
|
Gram
per milliliter
|
|
α
|
Alpha
|
|
T
|
Temperature
|