TIZANIDINE HYDROCHLORIDE METHOD OF ANALYSIS
SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for raw active material of the Tizanidine hydrochloride from
the Pharmacopoeial specification.
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 Identification
tests:
5.1.1
Chlorides
test:
5.1.1.1
Material and equipment:
5.1.1.1.1
Centrifuge
machine.
5.1.1.1.2
Diluted nitric
acid.
5.1.1.1.3
Purified water.
5.1.1.1.4
Silver nitrate.
5.1.1.1.5
Ammonia.
5.1.1.1.6
Excess of 6N
ammonium hydroxide.
5.1.1.2
Sample:
5.1.1.2.1
10mg/ml.
5.1.1.3
Method:
5.1.1.3.1
Take a test tube
add in it 10mg/ml with the help of pipette.
5.1.1.3.2
Add silver
nitrate. Observe the changes.
5.1.1.3.3
A white, curdy ppt
is formed that is insoluble in nitric acid but is soluble in a slight excess of
6N ammonium hydroxide.
5.1.1.3.4
Observe the
changes.
5.1.1.4
Observations:
5.1.1.4.1
A white, curdy ppt
is soluble in a slight excess of 6N ammonium hydroxide.
5.2 pH determination
test:
5.2.1
Material and equipment:
5.2.1.1
Glassware
(according to the requirement).
5.2.1.2
pH meter.
5.2.2
Sample:
5.2.2.1
10mg/ml.
5.2.3
Method:
5.2.3.1
Firstly clean the
pH meter with clean dry cloth, according to SOP
5.2.3.2
Operate the pH
meter according to SOP for operation of pH meter i.e. SOP
5.2.3.3
Rejuvenate the
electrode before use according to SOP instructions, if there is any need of.
5.2.3.4
Calibrate the
electrode of the pH meter according to SOP
5.2.3.5
Perform the test
on 10mg/ml solution.
5.2.3.6
Take a beaker of
100.0ml and add 10mg/ml solution in it. Such that it immersed electrodes in it
completely.
5.2.3.7
Maintain the
temperature of sample at 25oC±2oC.
5.2.3.8
Dip the electrode
along with temperature sensor into the sample.
5.2.3.9
When dipping
electrode into sample, it must be completely immersed in it.
5.2.3.10 Stir
the probe gently in the sample to create a homogeneous sample.
5.2.3.11 Allow
the reading to stabilize for a time.
5.2.3.12 Record
the observed values of pH & temperature in the respective Annexure-1.
5.2.3.13 Wash
the electrodes again after use and store the electrode in storage solution as
recommended in SOP of cleaning i.e. SOP. Calibrate the instrument before use according to SOP.
5.2.4
Observation:
5.2.4.1
4.3-5.3.
5.3 Loss on drying:
5.3.1
Material and equipment:
5.3.1.1
Glassware
(according to requirement).
5.3.1.2
Analytical
weighing balance.
5.3.1.3
Oven.
5.3.2
Sample:
5.3.2.1
0.5g.
5.3.3
Method:
5.3.3.1
Weigh 0.5g of the
test sample.
5.3.3.2
Set the oven
apparatus. Operate it according to the SOP
5.3.3.3
Place the sample
into the tray and dry it.
5.3.3.4
Set the
temperature of oven at 105oC for at least 3 hours.
5.3.3.5
And wait till the
sample loses its moisture.
5.3.3.6
After 3 hours
weigh the sample again by using analytical weighing balance i.e. the final
weight.
5.3.3.7
Note down readings
on given Annexure-2
5.3.4
Observation:
5.3.4.1
NMT 0.5%.
5.4 Assay:
5.4.1
Buffer:
5.4.1.1 Take
beaker of 1000.0ml add 6.8g/L of monobasic potassium phosphate.
5.4.1.2 Adjust
pH with5.3N potassium hydroxide to a pH of 7.5±0.05.
5.4.2
Mobile
phase:
5.4.2.1
Take a beaker of 100.0ml.
5.4.2.2
Add acetonitrile
and buffer (20:80), stir by using magnetic stirrer.
5.4.3
System
suitability solution:
5.4.3.1 46μg/ml
of USP Tizanidine hydrochloride RS and 0.12μg/ml of USP Tizanidine related
compound C RS in mobile phase.
5.4.4
Standard
solution:
5.4.4.1 0.046mg/ml
of USP Tizanidine hydrochloride RS in mobile phase.
5.4.5
Sample
solution:
5.4.5.1 0.046mg/ml
of Tizanidine hydrochloride in mobile phase.
5.4.6
Chromatographic
system:
5.4.6.1
Mode:
Liquid chromatography.
5.4.6.2
Detector:
UV
230nm.
5.4.6.3
Column:
4.6mm
x 15cm; packing L7.
5.4.6.4
Column
temperature: 35o.
5.4.6.5
Flow
rate: 1.0ml/min.
5.4.6.6
Injection
size: 20μL.
5.4.7
System
suitability:
5.4.7.1 Samples: System
suitability solution and Standard solution.
[NOTE:
The relative retention times for Tizanidine related compound C and Tizanidine
are 0.5 and 0.1, respectively.]
5.4.7.2 Suitability
requirements:
5.4.7.2.1
Resolution:
NLT
6 between Tizanidine and Tizanidine related compound C, System suitability
solution.
5.4.7.2.2
Tailing
factor: NMT 2.0 for Tizanidine, standard solution.
5.4.7.2.3
Relative
standard deviation: NMT 2.0%, standard solution.
5.4.8
Analysis:
5.4.8.1 Samples:
Standard
solution and sample solution.
5.4.9
Calculate the
percentage of Tizanidine hydrochloride (C9H8ClN5S.HCl)
in the portion of Tizanidine
hydrochloride taken:
Result: (rU/rS) x (CS/CU)
x 100
rU=
peak area of Tizanidine from the sample solution.
rS=
peak area of Tizanidine from the standard solution.
CS=
concentration of USP of Tizanidine hydrochloride RS in the standard solution
(mg/ml).
CU=
concentration of the sample solution (mg/ml).
5.4.10 Equilibrate
the column and detector with mobile phase at specified flow rate until a
constant signal is received.
5.4.11 Separately
inject equal volumes about 20μL of the standard solution and sample solution.
5.4.12 Record
the chromatogram.
5.4.13 Measure
the responses for the major peaks.
5.4.14 Analyze
as directed in the monograph.
5.5 Limit:
5.5.1
98.0 to 102.0% on
the dried basis.
6.0 REVISION LOG:
|
Revision No.
|
Effective Date
|
Reason
|
|
00
|
|
New SOP
|
7.0 REFERENCES:
7.1 USP38NF33
Volume-6 Official Monograph/ Tizanidine hydrochloride: 2015, pp.: 5590-5591.
8.0 ANNEXURES:
Annexure 1: pH
measurement.
Annexure 2: Observations
of Percentage Loss of drying by using oven.
Annexure 3: Observations
and calculations of HPLC method.
Annexure:
1
pH
measurement
|
Sr.#
|
Temperature
|
pH
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Result: _________________________________________________________________
Annexure: 2
Observations
of percentage loss of drying by using Oven
|
Percentage
loss of drying by using Oven
Weight
of Sample = _____________
Time
period = _____________
Pressure=
_________________
Average % Loss of Moisture: _____________
Remarks: _______________________________________________________________
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Annexure:
3
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
|
|
Sr.#
|
Serial
number
|
|
Q.C
|
Quality
control
|
|
Vol
|
Volume
|
|
g
|
Grams
|
|
ml
|
Milliliters
|
|
ppt
|
Precipitates
|
|
mg
|
Milligrams
|
|
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
|
|
QCA
|
Quality
control active ingredient
|
|
F
|
Format
|
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