LUMEFANTRINE SOP


LUMEFANTRINE SOP

1.0  OBJECTIVE:
To lay down a procedure of analytical report for the active raw material of Lumefantrine 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  HPLC 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  5.65g of sodium 1-hexanesulfonate.
5.1.2.2  2.75g of monobasic sodium phosphate.
5.1.2.3  Phosphoric acid.
5.1.2.4  Acetonitrile.
5.1.2.5  2-propanol.
5.1.2.6  USP Lumefantrine related Compound A RS.
5.1.2.7  10% volume of dichloromethane.
5.1.2.8  USP Lumefantrine RS.
5.1.2.9  1.0mg of Lumefantrine (sample).
5.1.2.10 Purified water.
5.1.3        Requirements:
5.1.3.1  Buffer:
5.1.3.1.1        Take a beaker of 1000.0ml.
5.1.3.1.2        Add 5.65g of sodium 1-hexanesulfonate and 2.75g of monobasic sodium phosphate in beaker with 900.0ml of purified water, stir it by using magnetic stirrer operate according to SOP
5.1.3.1.3        Adjust with phosphoric acid to a pH of 2.3 before dilution with purified water.
5.1.3.1.4        And finally dilute it with purified water to a final volume of 1000.0ml.
5.1.3.2  Solution A:  
5.1.3.2.1        Acetonitrile and Buffer (300:700).
5.1.3.3  Solution B:
5.1.3.3.1        Acetonitrile and 2-propanol (540:460).
5.1.3.4  Mobile phase:
Time
(min)
Solution A
(%)
Solution B
(%)
0
65
35
1.2
65
35
6.0
50
50
6.4
30
70
10.0
25
75
15.0
10
90
15.1
65
35
20.0
65
35

5.1.3.5  System suitability stock solution: 10.0μg/ml of USP Lumefantrine related Compound A RS prepared as follow.
5.1.3.5.1        Take a volumetric flask and transfer a suitable quantity of USP Lumefantrine related Compound A RS.
5.1.3.5.2        Dissolve it in 10% volume of dichloromethane.
5.1.3.5.3        Dilute it with acetonitrile up to the volume.
5.1.3.6  System suitability solution: 1mg/ml of USP Lumefantrine RS and 1μg/ml of USP Lumefantrine related Compound A RS prepared as follow.
5.1.3.6.1        Take a 10.0ml of volumetric flask and transfer a 10mg of USP Lumefantrine RS.
5.1.3.6.2        Dissolve it in 1.0ml of dichloromethane.
5.1.3.6.3        Add 1.0ml of the system suitability stock solution.
5.1.3.6.4        Dilute it with acetonitrile to the volume.
5.1.3.7  Standard solution: 1.0mg/ml of USP Lumefantrine RS prepared as follows.
5.1.3.7.1        Take a volumetric flask and transfer a suitable quantity of USP Lumefantrine RS to it.
5.1.3.7.2        Dissolve it in 10% volume of dichloromethane.
5.1.3.7.3        Dilute it with acetonitrile to the volume.
5.1.3.8  Sample solution: 1.0mg/ml of Lumefantrine prepared as follows.
5.1.3.8.1        Take a volumetric flask and transfer a suitable quantity of Lumefantrine to it.
5.1.3.8.2        Dissolve it in 10% volume of dichloromethane.
5.1.3.8.3        Dilute it with acetonitrile to the volume.
5.1.3.9  Chromatographic system:
5.1.3.9.1        Mode: Liquid chromatography.
5.1.3.9.2        Detector: UV 265nm.
5.1.3.9.3        Column: 4.6mm x 50mm; 1.8μm packing L1.
5.1.3.9.4        Column temperature: Beginning of column, 50o; end of column, 35o.
5.1.3.9.5        Flow rate: 2.5ml/min.
5.1.3.9.6        Injection size: 2.5μL.
5.1.3.9.7        Run time: 20min.
5.1.3.10    System suitability:
5.1.3.10.1    Samples:
5.1.3.10.1.1    System suitability solution and standard solution.
[NOTE___ The relative retention times for lumefantrine related compound A and Lumefantrine are 0.9 and 1.0, respectively.]
5.1.3.10.2    Suitability requirements:
5.1.3.10.2.1    Resolution: NLT 1.3 between lumefantrine and lumefantrine related compound A, system suitability solution.
5.1.3.10.2.2    Tailing factor: NMT 2.1, Standard solution.
5.1.3.10.2.3    Relative standard deviation: NMT 1.0%, standard solution.
5.1.3.11    Analysis:
5.1.3.11.1    Samples:
5.1.3.11.1.1    Standard solution and sample solution.
5.1.3.11.2    Calculate the percentage of Lumefantrine (C10H32Cl3NO) in the portion of the Lumefantrine 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 the standard solution (mg/mL).
CU= concentration of the sample solution (mg/mL).
5.1.3.11.3    Acceptance criteria:
5.1.3.11.3.1    98.0%-102.0%.
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 2.5μl 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-5 Official Monograph/Lumefantrine: 2015, pp.: 4162-4164.
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 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.,
PIC
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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