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:
Sr.#
Blank solution
Temperature
Optical rotation
(α)












                                                                                                 Average: _______________
Optical rotation of blank solution: _______________
Sample solution:
Sr.#
Sample solution
Temperature
Optical rotation
(α)












                                                                                                 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: ___________________           
Column size:
Length=
θ=
Stationary phase:

Temperature:

Mobile phase:

Flow rate:

Injection size:

Detector:

Wavelength:
λ=

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)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Height of peak of interest
(h)n
Width of peak of interest
(w)n
Area of peak of interest
A=1/2(h x w)




















2.      Retention volume:
Flow rate= _______________ml/min.
No. of peaks
Retention time of peak of interest
(tr)n
Retention volume = retention time x flow rate












3.      Retention factor:
Retention time of unretained peak (tm)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Retention factor of a component
k= (tr-tm)/tm














4.      Separation factor (α):
No. of peaks
Retention factor of a component
(kn)
Relative retention of two adjacent peaks
α = k2/k1












5.      Resolution:
Retention time of unretained peak (tm)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Width of peak of interest
(w)n
Resolution
Rs = 2 (tr2-tr1)
        (w1-w2)
















6.      Efficiency:
No. of peaks or components
Retention time of peak of interest
(tr)n
Width of peak of interest
(w)n
Efficiency
(No. of theoretical plates)
N= 16 (tr/w)2


















7.      Height equivalent to a theoretical plate (HETP):
Length of column = ________________________
No. of peaks or components
No. of theoretical plates
(N)
Height equivalent to a theoretical plate HETP = L/N












8.      Symmetry factor (tailing factor):
No. of peaks or components
Distance from the peak max. to leading edge of the peak
(f)
Width w
Symmetry factor
At 5%
At 10%
As = w5%
       2f
As = w10%
       2f
























9.      Response factor & Relative response factor:
Conc. (mg/ml)= ___________________
No. of peak
Peak area
Response factor = (peak area/conc.)
Relative response factor = (response factor of impurity/response factor of API)


















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:
________________________________________________________________________________________________________________________________________________






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


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