VORICONAZOLE METHOD OF ANALYSIS SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of Voriconazole 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 Characters:
5.1.1
Appearance:
5.1.1.1
White or almost
white powder.
5.1.2
Solubility:
5.1.2.1
Material and equipment:
5.1.2.1.1
Glassware (3 test
tubes, spatula).
5.1.2.1.2
Acetone.
5.1.2.1.3
Methylene
chloride.
5.1.2.1.4
Purified water.
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
in test tube 1, acetone in test tube 2 and methylene chloride in test tube 3 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 purified water is very slightly soluble.
5.1.2.4.2
The sample in test
tube 2 and 3 containing with acetone methylene chloride are freely soluble.
5.2 Assay:
5.2.1
Apparatus:
5.2.1.1
Liquid
chromatography apparatus.
5.2.1.2
Glassware
(according to the requirement).
5.2.1.3
Spectrophotometer
(detector).
5.2.2
Material
and reagents:
5.2.2.1
50.0mg of voriconazole
CRS.
5.2.2.2
Acetonitrile R.
5.2.2.3
Methanol R.
5.2.2.4
1.90g/L solution
of ammonium formate R.
5.2.2.5
Anhydrous formic
acid R.
5.2.3
Sample:
5.2.3.1
50.0mg.
5.2.4
Test
solution:
5.2.4.1
Test solution (a):
5.2.4.1.1
Take 100ml of
beaker and dissolve 50.0mg of the substance to be examined in the mobile phase.
5.2.4.1.2
Sonicate it, if
necessary.
5.2.4.1.3
And dilute to
100.0ml with the mobile phase.
5.2.4.1.4
Mix well to ensure
complete dissolution.
5.2.4.2
Test solution (b):
5.2.4.2.1
Take 100ml of
beaker and add 5.0ml of the test solution (a).
5.2.4.2.2
Dilute it to
100.0ml with the mobile phase.
5.2.5
Reference
solutions:
5.2.5.1
Reference solution
(a):
5.2.5.1.1
Take 100ml of
beaker and dissolve 50.0mg of voriconazole CRS in the mobile phase.
5.2.5.1.2
Sonicate it, if
necessary.
5.2.5.1.3
And dilute to
100.0ml with the mobile phase.
5.2.5.1.4
Mix well to ensure
complete dissolution.
5.2.5.1.5
Take another
100.0ml of the beaker and add 5.0ml of the above solution in it.
5.2.5.1.6
Dilute it to
100.0ml with the mobile phase.
5.2.6
Column:
5.2.6.1
Size:
5.2.6.1.1
Length=0.15m,
5.2.6.1.2
θ=3.9mm.
5.2.6.2
Stationary
phase:
5.2.6.2.1
End-capped
octadecylsilyl silica gel for chromatography R (4μm).
5.2.6.3
Temperature:
5.2.6.3.1
35oC.
5.2.7
Mobile
phase:
5.2.7.1
Mix 15 volumes of
acetonitrile and 30 volumes of Methanol R and 55 volumes of a 1.90g/L solution
of ammonium formate R previously adjusted to pH 4.0 with anhydrous formic acid
R while stirring continuously.
5.2.8
Flow
rate:
5.2.8.1
1.0ml/min.
5.2.9
Detection:
5.2.9.1
Spectrophotometer
at 256nm.
5.2.10
Injection:
5.2.10.1 20μL
test solution (b) and reference solution (a).
5.2.11 Run time:
5.2.11.1 3
times the retention time of voriconazole.
5.2.12 Relative retention
time:
5.2.12.1 With
reference to voriconazole (retention time=about 8 min).
5.2.13 Method of
analysis:
5.2.13.1 Firstly
prepare the test solution, reference solution and mobile phase according to the
requirements.
5.2.13.2 The
solutions must be free from solid particles.
5.2.13.3 Prepare
the apparatus.
5.2.13.4 The
mobile phase solvent mixtures must be deaerated prior to use either by boiling
or by applying a partial vacuum to the solvent reservoir.
5.2.13.5 Equilibrate
the column with the prescribed mobile phase, flow rate and at temperature
specified until a suitable baseline is achieved.
5.2.13.6 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.
5.2.13.7 As
the eluate leaves the column it enters a detector, where it is continuously
monitored at the specified λ.
5.2.13.8 The
electrical signal obtained from detector is amplified and routes to recorder
which record the developed chromatogram.
5.2.13.9 Calculate
the percentage content of Voriconazole (C16H14F3N5O)
from the declared content of Voriconazole CRS.
5.2.14 Observations:
5.2.14.1 97.5%
to 102.0% (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 /Voriconazole: 2015, pp. 1170-1172.
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.,
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
|
Q.C
|
Quality
control
|
B.P
|
British
pharmacopoeia
|
Vol
|
Volume
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
μm
|
Micron/
micrometer
|
g/L
|
Gram
per liter
|
ml
|
Milliliter
|
mg
|
Milligram
|
CRS
|
Chemical
reference solution
|
m
|
Meter
|
θ
|
Theta
|
mm
|
Millimeter
|
oC
|
Degree
Celsius
|
ml/min
|
Milliliter
per minute
|
nm
|
Nanometer
|
μL
|
Microliter
|
Min
|
Minute
|
λ
|
Lamda
|
%
|
Percentage
|
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