MOXIFLOXACIN HYDROCHLORIDE SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the
Moxifloxacin HCl 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 Characters:
5.1.1
Appearance:
5.1.1.1
Light yellow or
yellow powder.
5.1.1.2
Crystalline.
5.1.1.3
Slightly
hygroscopic.
5.1.2
Solubility:
5.1.2.1
Material and equipment:
5.1.2.1.1
Glassware (test
tubes, spatula).
5.1.2.1.2
Purified water.
5.1.2.1.3
Ethanol (96%).
5.1.2.1.4
Acetone.
5.1.2.2
Sample:
5.1.2.2.1
Small quantity.
5.1.2.3
Method:
5.1.2.3.1
Take 3 test tubes
and add small quantity of sample for testing solubility according to B.P
specifications.
5.1.2.3.2
Add purified
water, ethanol (96%) and acetone in each test tube separately in a small volume
and observe the solubility of the sample.
5.1.2.4
Observations:
5.1.2.4.1
The sample in test
tube 1 containing with water is sparingly soluble.
5.1.2.4.2
The sample in test
tube 2 containing with ethanol (96%) is slightly soluble.
5.1.2.4.3
The sample in test
tube 3 containing with acetone is practically soluble.
5.2 Identification
tests:
5.2.1
Specific
optical rotation:
5.2.1.1
Material and equipment:
5.2.1.1.1
Glassware (according
to requirement).
5.2.1.1.2
Polarimeter.
5.2.1.1.3
Acetonitrile.
5.2.1.1.4
Purified water.
5.2.1.2
Sample:
5.2.1.2.1
0.2g.
5.2.1.3
Method:
5.2.1.3.1
Take a beaker and
dissolve 0.2g in 20ml of a mixture of equal volumes of acetonitrile and water.
5.2.1.3.2
Determine the zero
of the Polarimeter and the angle of rotation of polarized light at the
wavelength of the D-line of sodium (λ=589nm) and at 20±0.5oC.
5.2.1.3.3
Determine the zero
of the apparatus with the tube closed, for solids filled with prescribed
solvent.
5.2.1.3.4
Calculate the
specific optical rotation by using formula according to B.P monograph.
5.2.1.4
Observations:
5.2.1.4.1
-125 to -138
(anhydrous substance).
5.2.2
5.2.2.1
Material and equipment:
5.2.2.1.1
Glassware
(according to requirement).
5.2.2.1.2
Dilute nitric
acid.
5.2.2.1.3
Purified water.
5.2.2.1.4
Silver nitrate.
5.2.2.1.5
Ammonia.
5.2.2.2
Sample:
5.2.2.2.1
50mg.
5.2.2.3
Method:
5.2.2.3.1
Dissolve 50mg in
5ml of water, add 1ml of dilute nitric acid, mix, allow it to stand for 5 min
and filter. The filtrate gives chlorides reaction according to B.P
specifications.
5.2.2.3.2
Use 2ml of the
above solution.
5.2.2.3.3
Acidify with
dilute nitric acid.
5.2.2.3.4
And add 0.4ml of
silver nitrate.
5.2.2.3.5
Shake and allow it
to stand.
5.2.2.3.6
A curdled, white
ppt is formed.
5.2.2.3.7
Centrifuge it, by
using centrifuge machine i.e. SOP
5.2.2.3.8
Wash the ppt with
3 quantities, each of 1ml, of water.
5.2.2.3.9
Carry out this
operation rapidly is subdued light, degrading the fact that the supernatant
solution may not become perfectly clear.
5.2.2.3.10 Suspend
the precipitate in 2ml of water and add 1.5ml of ammonia.
5.2.2.4
Observations:
5.2.2.4.1
The precipitate
dissolves easily with the possible exception of a few large particles which
dissolves slowly.
5.3 Other
tests:
5.3.1
pH:
5.3.1.1
Material and equipment:
5.3.1.1.1
Glassware
(according to the requirement).
5.3.1.1.2
pH meter.
5.3.1.1.3
Carbon
dioxide-free water.
5.3.1.2
Sample:
5.3.1.2.1
0.10g
5.3.1.3
Method:
5.3.1.3.1
Dissolve 0.10g in
50ml of carbon dioxide-free water.
5.3.1.3.2
Use pH meter for
determining pH of the sample as per SOP of operation of pH meter i.e. SOP
5.3.1.3.3
Calibrate the
instrument before use according to SOP No
5.3.1.3.4
Measurements are
made at the same temperature 25-35oC.
5.3.1.3.5
Immerse the
electrodes in the solutions to be examined and take reading.
5.3.1.3.6
And follow the B.P
conditions accordingly.
5.3.1.4
Observation:
5.3.1.4.1
3.9 to 4.6.
5.4 Assay:
5.4.1
Apparatus:
5.4.1.1
HPLC apparatus.
5.4.1.2
Glassware
(according to the requirement).
5.4.2
Material
and reagents:
5.4.2.1
Moxifloxacin
hydrochloride sample.
5.4.2.2
End-capped
phenylsilyl silica gel for chromatography (5μm).
5.4.2.3
Tetrabutylammonium
hydrogen sulfate.
5.4.2.4
Potassium
dihydrogen phosphate.
5.4.2.5
Phosphoric acid.
5.4.2.6
Anhydrous sodium
sulfite.
5.4.2.7
Methanol.
5.4.2.8
0.5g/L of tetrabutylammonium
hydrogen sulfate.
5.4.2.9
1.0g/L of
potassium dihydrogen phosphate.
5.4.2.10 3.4g/L
of phosphoric acid.
5.4.2.11 Purified
water.
5.4.3
Requirements:
5.4.3.1
Solution
A:
5.4.3.1.1
Take 1000ml of
beaker and dissolve 0.5g of tetrabutylammonium hydrogen sulfate and 0.1g of
potassium dihydrogen phosphate in about 500ml of water.
5.4.3.1.2
Add 2ml of
phosphoric acid and 0.50g of anhydrous sodium sulfite.
5.4.3.1.3
And then dilute to
1000ml with water.
5.4.3.2
Sample:
5.4.3.2.1
50.0mg (sample to
be examined).
5.4.3.2.2
50.0mg
(Moxifloxacin hydrochloride CRS) for reference solution.
5.4.3.3
Test
solution:
5.4.3.3.1
Test solution (a):
5.4.3.3.1.1 Take
100ml of beaker and dissolve 50.0mg of the substance to be examined in solution
A.
5.4.3.3.1.2 And
dilute to 50.0ml with the same solution.
5.4.3.3.2
Test solution (b):
5.4.3.3.2.1 Take
a beaker and dilute 2.0ml of test solution (a) to 20.0ml with solution A.
5.4.3.4
Reference
solutions:
5.4.3.4.1
Reference solution
(a):
5.4.3.4.1.1 Take
100ml beaker and dissolve 50.0mg of Moxifloxacin hydrochloride CRS in solution
A and dilute to 50.0ml with the same solution.
5.4.3.4.1.2 Dilute
2.0ml of this solution to 20.0ml with solution A.
5.4.3.5
Column:
5.4.3.5.1
Size:
5.4.3.5.1.1 Length=0.25m,
5.4.3.5.1.2 θ=4.6mm.
5.4.3.5.2
Stationary
phase:
5.4.3.5.2.1 End-capped
phenylsilyl silica gel for chromatography (5μm).
5.4.3.5.3
Temperature:
5.4.3.5.3.1 45oC.
5.4.3.6
Mobile
phase:
5.4.3.6.1
Mix 28 volumes of
methanol and 72 volumes of solution containing 0.5g/L of tetrabutylammonium
hydrogen sulfate, 1.0g/L of potassium dihydrogen phosphate and 3.4g/L of
phosphoric acid.
5.4.3.7
Flow
rate:
5.4.3.7.1
1.3ml/min.
5.4.3.8
Detection:
5.4.3.8.1
Spectrophotometer
at 293nm.
5.4.3.9
Injection:
5.4.3.9.1
10μL of the test
solution (b) and reference solution (a).
5.4.3.10 Run time:
5.4.3.10.1 2.5
times the retention time of Moxifloxacin.
5.4.4
Method
of analysis:
5.4.4.1 Firstly
prepare the test solution, reference solution and mobile phase according to the
requirements.
5.4.4.2 The
solutions must be free from solid particles.
5.4.4.3 Prepare
the apparatus.
5.4.4.4 The
mobile phase solvent mixtures must be deaerated prior to use either by boiling
or by applying a partial vacuum to the solvent reservoir.
5.4.4.5 Equilibrate
the column with the prescribed mobile phase, flow rate and at temperature
specified until a suitable baseline is achieved.
5.4.4.6 Test
solution of the mixture to be separated is now introduced into the mobile phase
with the help of an injector just before entering the separating column.
5.4.4.7 As
the eluate leaves the column it enters a detector, where it is continuously
monitored at the specified λ.
5.4.4.8 The
electrical signal obtained from detector is amplified and routes to recorder
which record the developed chromatogram.
5.4.4.9 Calculate
the percentage content of Moxifloxacin HCl (C21H25ClFN3O4).
5.4.5
Limit:
5.4.5.1 98%
to 102% (anhydrous substance).
6.0 REVISION LOG:
|
Revision No.
|
Effective Date
|
Reason
|
|
00
|
|
New SOP
|
7.0 REFERENCES:
7.1
The British
Pharmacopoeia. Vol II.,
Official Monograph /Moxifloxacin hydrochloride: 2015, pp. 318-319.
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
|
|
HCl
|
Hydrochloric
acid
|
|
Q.C
|
Quality
control
|
|
%
|
Percentage
|
|
B.P
|
British
pharmacopoeia
|
|
g
|
Grams
|
|
ml
|
Milliliter
|
|
λ
|
Lamda
|
|
oC
|
Degree
Celsius
|
|
mg
|
Milligram
|
|
Min
|
Minute
|
|
ppt
|
Precipitate
|
|
μm
|
Micron/
micrometer
|
|
g/L
|
Gram
per liter
|
|
CRS
|
Chemical
reference solution
|
|
m
|
Meter
|
|
θ
|
Theta
|
|
mm
|
Millimeter
|
|
ml/min
|
Milliliter
per minute
|
|
nm
|
Nanometer
|
|
μL
|
Microliter
|
|
Vol
|
Volume
|
|
QCA
|
Quality
control active ingredient
|
|
F
|
Format
|
|
mg/ml
|
Milligram
per milliliter
|
|
UV
|
Ultra
violet
|