FENOFIBRATE SOP


1.0  OBJECTIVE:
To lay down a procedure of analytical report for the active raw material of Fenofibrate 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  Assay:
5.1.1        Apparatus:
5.1.1.1  Liquid chromatography apparatus.
5.1.1.2  Analytical weighing balance.
5.1.1.3  Glassware (according to the requirement).
5.1.1.4  Magnetic stirrer.
5.1.1.5  UV/VIS Spectrophotometer.
5.1.2        Material and reagents:
5.1.2.1  Acetonitrile.
5.1.2.2  Purified water.
5.1.2.3  Phosphoric acid.
5.1.2.4  USP Fenofibrate RS.
5.1.3        Requirements:
5.1.3.1  Mobile phase:
5.1.3.1.1        Acetonitrile and purified water acidified with phosphoric acid to a pH of 2.5 (7:3).
5.1.3.2  Standard solution:
5.1.3.2.1        1mg/ml of USP Fenofibrate RS in mobile phase.
5.1.3.3  Sample solution:
5.1.3.3.1        1mg/ml of Fenofibrate in mobile phase.
5.1.3.4  Chromatographic system:
5.1.3.4.1        Mode: Liquid chromatography.
5.1.3.4.2        Detector: UV 286nm.
5.1.3.4.3        Column: 4.0mm x 25cm; packing L1.
5.1.3.4.4        Flow rate: 1.0ml/min.
5.1.3.4.5        Injection size: 5μL.
5.1.3.5  System suitability:
5.1.3.5.1        Sample: Standard solution.
5.1.3.5.2        Suitability requirements:
5.1.3.5.2.1  Relative standard deviation: NMT 1.0%, for six replicate injections.
5.1.3.6  Analysis:
5.1.3.6.1        Samples: Standard solution and sample solution.
5.1.3.6.2        Calculate the percentage of Fenofibrate (C20H21ClO4) in the portion of Fenofibrate 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 Fenofibrate RS in the standard solution (mg/mL).
CU= concentration of Fenofibrate in the sample solution (mg/mL).
5.1.3.6.3        Acceptance criteria:
5.1.3.6.3.1  98.0%-102.0% on the dried basis.
5.1.4        Procedure:
5.1.4.1  Equilibrate the column and detector with mobile phase at specified flow rate until a constant signal is received.
5.1.4.2  Inject a sample and standard solution of 5μl through the injector, or use an auto-sampler.
5.1.4.3  Begin the gradient program.
5.1.4.4  Record the spectrum.
5.1.4.5  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/ Fenofibrate: 2015, pp.: 3454-3455.
8.0  ANNEXURES:
Annexure 1: Observations and calculations of HPLC method.

Annexure: 1
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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