DUTASTERIDE USP SOP

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/QCE_C/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        +15.0^o to 25.0^o.

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: 35^o.

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= (r_U/r_S) × (C_S/C_U) × 100

r_U= peak response from the Sample solution.

r_S= peak response from the Standard solution.

C_S= concentration of USP Dutasteride RS in the Standard solution (mg/mL).

C_U= 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