DULOXETINE HYDROCHLORIDE SOP


1.0  OBJECTIVE:
To lay down a procedure of analytical report for the active raw material of Duloxetine Hydrochloride 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 test:
5.1.1        Chloride test:
5.1.1.1  Material and equipment:
5.1.1.1.1        Glassware (according to requirement).
5.1.1.1.2        Dilute nitric acid.
5.1.1.1.3        Purified water.
5.1.1.1.4        0.5ml of Silver nitrate solution.
5.1.1.1.5        Ammonia.
5.1.1.2  Sample:
5.1.1.2.1        5mg/ml in methanol.
5.1.1.3  Method:
5.1.1.3.1        Take a test tube prepare in it 5mg/ml in methanol TS.
5.1.1.3.2        Add one drop of diluted nitric acid.
5.1.1.3.3        And add 0.5ml of silver nitrate TS.
5.1.1.3.4        Shake and allow it to stand.
5.1.1.3.5        A curdled, white ppt is formed.
5.1.1.3.6        Centrifuge the mixture without delay in centrifugation machine, operate it according to SOP No. BM/QCEO/SOP010-00.
5.1.1.3.7        Decant the supernatant layer.
5.1.1.3.8        Wash the obtained ppt with three 1.0ml of portions of nitric acid solution (1 in 100), and discard the washing.
5.1.1.3.9        Add ammonia TS drop-wise to this ppt.
5.1.1.3.10    Observe the changes.
5.1.1.4  Observations:
5.1.1.4.1        The precipitate dissolves readily.
5.2  Assay:
5.2.1        Apparatus:
5.2.1.1  Liquid chromatography apparatus.
5.2.1.2  Analytical weighing balance.
5.2.1.3  Glassware (according to the requirement).
5.2.1.4  Magnetic stirrer.
5.2.1.5  UV/VIS Spectrophotometer.
5.2.2        Material and reagents:
5.2.2.1  Phosphoric acid.
5.2.2.2  Sodium hydroxide solution.
5.2.2.3  10.3g of sodium 1-hexanesulfonate monohydrate.
5.2.2.4  n-propanol.
5.2.2.5  Acetonitrile.
5.2.2.6  Purified water.
5.2.2.7  USP Duloxetine hydrochloride RS.
5.2.3        Requirements:
5.2.3.1  Buffer:
5.2.3.1.1        Take a beaker of 1000.ml with purified water.
5.2.3.1.2        Add 2.9g of phosphoric acid in it, dissolve it by using magnetic stirrer operate according to SOP No. BM/QCEO/SOP007-00.
5.2.3.1.3        Adjust with sodium hydroxide solution to a pH of 2.5.
5.2.3.1.4        To each L of this solution add 10.3g of sodium 1-hexanesulfonate monohydrate, and dissolve by using magnetic stirrer.
5.2.3.2  Mobile phase:
5.2.3.2.1        Acetonitrile, n-propanol and buffer (13:17:70).
5.2.3.3  Diluent:
5.2.3.3.1        Acetonitrile and purified water (25:75).
5.2.3.4  System suitability solution:
5.2.3.4.1        0.2mg/ml of USP Duloxetine hydrochloride RS in mobile phase. Heat the solution to at least 40o for a minimum of 1h.
[NOTE___The resulting solution contains Duloxetine impurity B, Duloxetine impurity C, Duloxetine impurity D, Duloxetine impurity E and Duloxetine related compound F.]
5.2.3.5  Standard solution:
5.2.3.5.1        0.1mg/ml of USP Duloxetine hydrochloride RS in diluent.
5.2.3.6  Sample solution:
5.2.3.6.1        0.1mg/ml of Duloxetine hydrochloride in diluent.
5.2.3.7  Chromatographic system:
5.2.3.7.1        Mode: Liquid chromatography.
5.2.3.7.2        Detector: UV 230nm.
5.2.3.7.3        Column: 4.6mm x 15cm; 3.5μm packing L7.
5.2.3.7.4        Column temperature: 40±3o.
5.2.3.7.5        Flow rate: 1.0ml/min.
5.2.3.7.6        Injection size: 10μL.
5.2.3.7.7        Run time: 2 times the retention time of Duloxetine.
5.2.3.8  System suitability:
5.2.3.8.1        Sample: System suitability solution.
[NOTE___ see table in Annexure-1 for relative retention times.]
5.2.3.8.2        Suitability requirements:
5.2.3.8.2.1  Resolution: NLT 1.5 between Duloxetine and Duloxetine related compound F peaks.
5.2.3.8.2.2  Tailing factor: NMT 1.5, for the Duloxetine peak.
5.2.3.8.2.3  Relative standard deviation: NMT 1.0%, for the Duloxetine peak.
5.2.3.9  Analysis:
5.2.3.9.1        Samples:
5.2.3.9.1.1  Standard solution and sample solution.
5.2.3.9.2        Calculate the percentage of Duloxetine hydrochloride (C18H19NOS.HCl) in the portion of sample taken:
Result= (rU/rS) x (CS/CU) x 100.
rU= peak response from the sample solution.
rS= peak response from the standard solution.
CS= concentration of USP Duloxetine hydrochloride RS in the standard solution (mg/mL).
CU= concentration of Duloxetine hydrochloride in the sample solution (mg/mL).
5.2.3.9.3        Acceptance criteria:
5.2.3.9.3.1  97.0%-102.0% on the dried basis.
5.2.4        Procedure:
5.2.4.1  Protect the solution of Duloxetine from light.
5.2.4.2  Equilibrate the column and detector with mobile phase at specified flow rate until a constant signal is received.
5.2.4.3  Inject a sample and standard solution of 10μl through the injector, or use an auto-sampler.
5.2.4.4  Begin the gradient program.
5.2.4.5  Record the spectrum.
5.2.4.6  Analyze as directed in the monograph.
6.0  REVISION LOG:
Revision No.
Effective Date
Reason
00

New SOP

7.0  REFERENCES:
7.1  USP38NF33 Volume-4 Official Monograph/ Duloxetine hydrochloride: 2015, pp.: 3243-3244.
8.0  ANNEXURES:
Annexure 1: Table for Relative retention time.
Annexure 2: Observations and calculations of HPLC method.



Annexure: 1
Table for Relative retention time
Name
Relative retention time
Related response factor
Acceptance criteria NMT (%)
Duloxetine impurity Ba,g
0.15
0.36
-
Duloxetine impurity  Cb,g
0.43
1.0
-
Duloxetine impurity Dc,g
0.48
1.8
-
Duloxetine impurity Ed,g
0.74
1.0
-
Duloxetine
1.0
-
-
Duloxetine related compound Fe
1.1
1.0
0.5
Duloxetine impurity Gi,g
1.4
0.51
-
Any individual unspecified impurity
-
1.0
0.1
Total impurities
-
-
0.6
a 3-(Methylamino)-1-(thiophen-2-yl) propan-1-ol.
b 4-[3-( Methylamino)-1-(thiophen-2-yl) propyl] naphthalen-1-ol.
c Naphthalen-1-ol.
d 1-(3-(Methylamino)-1-(thiophen-2-yl) proply) naphthalen-2-ol.
e (S)-N-Methly-3-(naphthalen-1-yloxy)-3-(thiophen.3-yl) propan-1-amine.
f 1- Fluoronaphthalene.
g controlled at Any individual unspecified impurity level.


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 spectrum.






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
ml
Milliliter
ppt
Precipitate
L
Length
g/L
Gram per liter
mg/ml
Milligram per milliliter
μg/ml
Microgram per milliliter
UV/VIS
Ultra violet/ Visible
μg
Microgram
mg
Milligram
RS
Reference standard
UV
Ultra violet
USP
United states pharmacopoeia
nm
Nanometer
mm
Millimeter
cm
Centimeter
μm
Micron
oC
Degree centigrade
ml/min.
Milliliter per minute
μL
Microliter
NMT
Not more than
%
Percentage
NF
National formulary


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