1.0 OBJECTIVE:
To
lay down a procedure for the active raw material of the Buspirone Hydrochloride USP 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 Identification
tests:
5.1.1
Chlorides test:
5.1.1.1 Material and equipment:
5.1.1.1.1
Glassware
(according to requirement).
5.1.1.1.2
Analytical
weighing balance.
5.1.1.1.3
Centrifuge
machine.
5.1.1.1.4
Dilute
nitric acid.
5.1.1.1.5
0.5ml
of Silver nitrate TS.
5.1.1.1.6
Nitric
acid solution.
5.1.1.1.7
Ammonia.
5.1.1.1.8
Purified water.
5.1.1.2 Sample:
5.1.1.2.1
10.0mg/ml
in purified water.
5.1.1.3 Method:
5.1.1.3.1
Take
a test tube add in it 10.0mg/ml in purified water of test solution with the
help of pipette.
5.1.1.3.2
Add
one drop of diluted nitric acid and 0.5ml of silver nitrate TS to a solution of
the substance being examined containing.
5.1.1.3.3
Centrifuge
the mixture without delay, in centrifugation machine,
5.1.1.3.4
And
decant the supernatant layer.
5.1.1.3.5
The
obtained ppt is wash with three 1ml, of portions of nitric acid solution (1 in
100).
5.1.1.3.6
And
discard the washing.
5.1.1.3.7
Add
ammonia TS dropwise to this precipitate.
5.1.1.3.8
Observe
the changes.
5.1.1.4 Observations:
5.1.1.4.1
It
dissolves readily.
5.2 Assay:
5.2.1
Apparatus:
5.2.1.1 Glassware (according to requirement).
5.2.1.2 Magnetic stirrer.
5.2.1.3 HPLC apparatus.
5.2.1.4 Analytical weighing balance.
5.2.1.5 Monobasic potassium phosphate.
5.2.1.6 Sodium hydroxide.
5.2.1.7 Acetonitrile.
5.2.1.8 Propylparaben.
5.2.1.9 Methanol.
5.2.1.10 1N hydrochloric acid.
5.2.1.11 Purified water.
5.2.2
Buffer:
5.2.2.1 Prepare 1.36 mg/ml of monobasic potassium
phosphate in water.
5.2.2.2 Adjust the solution with 10% sodium
hydroxide (w/v) to a pH of 7.5.
5.2.2.3 And then filter it by using filtration
apparatus.
5.2.3
Mobile phase:
5.2.3.1 Prepare mobile phase Acetonitrile and
Buffer (2:3) according to the need.
5.2.4
Internal standard stock solution:
5.2.4.1 Prepare internal standard stock solution
2.5mg/ml of propylparaben in methanol.
5.2.5
Internal standard solution:
5.2.5.1 Prepare internal standard solution
0.125mg/ml of propylparaben from the internal standard stock solution in
purified water.
5.2.6
Standard stock solution:
5.2.6.1 Take a 100.0ml of volumetric flask and add
50.0mg of USP Buspirone Hydrochloride RS.
5.2.6.2 Dissolve it in 25.0ml of 1N hydrochloric
acid by using magnetic stirrer.
5.2.6.3 Dilute it with purified water up to the
volume.
5.2.7
Standard solution:
5.2.7.1 Take a 50.0ml of volumetric flask and add
10.0 ml of standard stock solution and 10.0 ml of internal standard solution in
it.
5.2.7.2 Dilute it with purified water up to the
volume.
5.2.8
Sample stock solution:
5.2.8.1 Take a 100.0ml of volumetric flask and add
50.0mg of buspirone hydrochloride in it.
5.2.8.2 Add 25.0ml of 1N hydrochloric acid. Dissolve
it by using magnetic stirrer
5.2.8.3 Dilute it with purified water up to the
volume.
5.2.9
Sample solution:
5.2.9.1 Prepare 0.1mg/ml of buspirone
hydrochloride from the sample stock solution and 0.025mg/ml of propylparaben
from the internal standard solution in purified water.
5.3
Chromatographic system:
5.3.1
Mode: Liquid
chromatography.
5.3.2
Detector: UV
254 nm.
5.3.3
Column: 3.9-mm × 30-cm;
packing L1.
5.3.4
Flow rate: 2.0mL/min.
5.3.5
Injection size: 25μL.
5.4
System suitability:
5.4.1
Samples:
Standard solution.
[NOTE-----The
relative retention times for propylparaben and buspirone hydrochloride are 0.55
and 1.0, respectively.]
5.4.2
Suitability
requirements:
5.4.2.1 Resolution:
NLT 4 between buspirone hydrochloride and
the internal standard.
5.4.2.2 Relative
standard deviation: NMT 2.0%.
5.5
Analysis:
5.5.1
Samples:
Standard solution and sample solution.
5.5.2
Calculate
the percentage of buspirone hydrochloride (C21H31N5O2.HCl)
in the portion of buspirone hydrochloride taken:
Result
= (RU/RS) x (CS/CU) x 100
RU
= peak response ratio of Buspirone hydrochloride to propylparaben from the
sample solution.
RS
= peak response ratio of Buspirone hydrochloride to propylparaben from the standard
solution.
CS
= concentration of USP Buspirone Hydrochloride RS in the standard solution
(mg/ml).
CU
= nominal concentration of Buspirone Hydrochloride in the standard solution
(mg/ml).
5.6 Procedure:
5.6.1
Equilibrate
the column and detector with mobile phase at specified flow rate until a
constant signal is received.
5.6.2
Separately
inject equal volumes about 25μL of the standard solution and sample solution.
5.6.3
Record
the chromatogram.
5.6.4
Measure
the responses for the major peaks.
5.6.5
Analyze
as directed in the monograph.
5.7
Limit:
5.7.1
97.5%-102.5%.
6.0 REVISION
LOG:
Revision No.
|
Effective Date
|
Reason
|
00
|
New
SOP
|
7.0 REFERENCES:
7.1 USP38NF33
Volume-3 Official Monograph/ Buspirone Hydrochloride: 2015, pp.: 2495-2496.
7.2 The British Pharmacopoeia. Vol V., Official Monograph /Qualitative
Reactions and Tests: 2015, pp. 266-270.
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
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|
Q.C
|
Quality
control
|
Vol
|
Volume
|
g
|
Grams
|
ml
|
Milliliter
|
Min
|
Minutes
|
oC
|
Degree
centigrade
|
mg
|
Milligram
|
M
|
Molar
|
%
|
Percentage
|
TS
|
Test
solution
|
R
|
Reagent
|
g/L
|
Gram
per liter
|
ppt
|
Precipitates
|
N
|
Normal
|
USP
|
United
states pharmacopoeia
|
NF
|
National
formulary
|
cm
|
Centimeter
|
nm
|
Nanometer
|
mm
|
Millimeter
|
Approx.
|
Approximately
|
%
|
Percentage
|
μL
|
Microliter
|
θ
|
Theta
|
λ
|
Lambda
|
w/v
|
Weight
by volume
|
mg/ml
|
Milligram
per milliliter
|
UV
|
Ultraviolet
|
ml/min
|
Milliliter
per minute
|