LEVOFLOXACIN HEMIHYDRATE SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the
Levofloxacin hemihydrate from the Pharmacopoeial specifications.
2.0 SCOPE:
This
SOP shall be applicable in Q.C laboratory.
3.0 RESPONSIBILITY:
3.1
Q.C Analysts.
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
Glassware
(according to the requirement).
5.1.1.3
Spectrophotometer
(detector).
5.1.2
Material
and reagents:
5.1.2.1
Levofloxacin
hemihydrate sample.
5.1.2.2
8.5g/L of ammonium
acetate.
5.1.2.3
1.25g/L of cupric
sulfate, pentahydrate.
5.1.2.4
1.3g/L of L-isoleucine.
5.1.2.5
Purified water.
5.1.2.6
Methanol.
5.1.3
Requirements:
5.1.3.1
Buffer:
5.1.3.1.1
8.5g/L of ammonium
acetate, 1.25g/L of cupric sulfate, pentahydrate, and 1.3g/L of L-isoleucine
in water.
5.1.3.2
Mobile
phase:
5.1.3.2.1
Methanol and
Buffer (3:7).
5.1.3.3
Standard
solution:
5.1.3.3.1
1mg/ml of USP
Levofloxacin in mobile phase.
5.1.3.4
Sample
solution:
5.1.3.4.1
1mg/ml of
Levofloxacin in mobile phase.
5.1.3.5
Chromatographic
system:
5.1.3.5.1
Mode:
Liquid chromatography.
5.1.3.5.2
Detector:
UV
360nm.
5.1.3.5.3
Column:
4.6mm
x 25cm; 5μm packing L1.
5.1.3.5.4
Column
temperature: 45oC.
5.1.3.5.5
Flow
rate: 0.8ml/min.
5.1.3.5.6
Injection
size: 25μL.
5.1.3.6
System
suitability:
5.1.3.6.1
Sample:
5.1.3.6.1.1 Standard
solution.
5.1.3.6.2
Suitability
requirements:
5.1.3.6.2.1 Tailing factors:
5.1.3.6.2.1.1
0.5-1.5.
5.1.3.6.2.2 Relative standard
deviation:
5.1.3.6.2.2.1
NMT 1.0%.
5.1.3.7
Analysis:
5.1.3.7.1
Sample:
5.1.3.7.1.1 Standard
solution and sample solution.
5.1.3.7.2
Calculate the
percentage of levofloxacin in the portion of Levofloxacin taken.
Result= (rU/rS) x (CS/CU)
x 100.
rU=
peak response of levofloxacin from the sample solution.
rS=
peak response of levofloxacin from the standard solution.
CS=
concentration of USP levofloxacin RS in the standard solution (mg/mL).
CU=
concentration of levofloxacin in the sample solution (mg/mL).
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 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.
5.1.5
Limit:
5.1.5.1 98%
to 102% on anhydrous basis.
6.0 REVISION LOG:
Revision No.
|
Effective Date
|
Reason
|
00
|
New SOP
|
7.0 REFERENCES:
7.1 USP38NF33
Volume-5 Official Monograph/Levofloxacin: 2015, pp.: 4080-4082.
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
spectrum.
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.,
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
|
Q.C
|
Quality
control
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
g/L
|
Gram
per liter
|
mg/ml
|
Milligram
per milliliter
|
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
|
BM
|
Biomark
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|