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:
___________________
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)=
_____________
2.
Retention volume:
Flow rate= _______________ml/min.
3.
Retention factor:
Retention time of unretained peak (tm)=
_____________
4.
Separation factor (α):
5.
Resolution:
Retention time of unretained peak (tm)=
_____________
6.
Efficiency:
7.
Height equivalent to a theoretical plate (HETP):
Length of column = ________________________
8.
Symmetry factor (tailing factor):
9.
Response factor & Relative response factor:
Conc. (mg/ml)= ___________________
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:
________________________________________________________________________________________________________________________________________________
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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
|