CLINDAMYCIN HYDROCLORIDE SOP
1.0 OBJECTIVE:
To
lay down a procedure of analytical report for the active raw material of the Clindamycin hydrochloride 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 White or almost white powder.
5.1.1.2 Crystalline.
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%).
5.1.2.2 Sample:
5.1.2.2.1
Small
quantity.
5.1.2.3 Method:
5.1.2.3.1
Take
2 test tubes and add small quantity of sample for testing solubility according
to B.P specifications.
5.1.2.3.2
Add
purified water and ethanol (96%) 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 very soluble.
5.1.2.4.2
The
sample in test tube 2 containing with ethanol (96%) is slightly soluble.
5.2 Identification
tests:
5.2.1
5.2.1.1 Material and equipment:
5.2.1.1.1
Glassware
(according to requirement).
5.2.1.1.2
Water-bath.
5.2.1.1.3
Dilute
hydrochloric acid.
5.2.1.1.4
Sodium
carbonate.
5.2.1.1.5
20g/L
solution of sodium nitroprusside.
5.2.1.2 Sample:
5.2.1.2.1
10.0mg.
5.2.1.3 Method:
5.2.1.3.1
Take
a beaker and dissolve about 10.0mg of sample in 2ml of dilute hydrochloric acid.
5.2.1.3.2
Heat
on water-bath for 3 min.
5.2.1.3.3
Add
3ml of sodium carbonate solution and 1ml of a 20g/L solution of sodium
nitroprusside.
5.2.1.4 Observations:
5.2.1.4.1
A
violet-red colour develops.
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
Silver
nitrate.
5.2.2.1.4
Ammonia.
5.2.2.1.5
Purified
water.
5.2.2.2 Sample:
5.2.2.2.1
0.1g.
5.2.2.3 Method:
5.2.2.3.1
Take
a test tube and dissolve 0.1g in water and dilute to 10.0ml with the same
solvent.
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 operate it.
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 Observation:
5.2.2.4.1
The
precipitate dissolves easily with the possible exception of a few large
particles which dissolves slowly.
5.3 Assay:
5.3.1
Apparatus:
5.3.1.1 HPLC apparatus.
5.3.1.2 Glassware (according to the requirement).
5.3.2
Material and reagents:
5.3.2.1 Octadecylsilyl silica gel for
chromatography (5μm).
5.3.2.2 Acetonitrile.
5.3.2.3 6.8g/L solution of potassium dihydrogen
phosphate.
5.3.2.4 250g/L solution of potassium hydroxide.
5.3.2.5 Purified water.
5.3.3
Requirements:
5.3.3.1
Sample:
5.3.3.1.1
50.0mg
(sample to be examined).
5.3.3.1.2
50.0mg
(Clindamycin hydrochloride CRS) for reference solution.
5.3.3.2
Test solution:
5.3.3.2.1
Take
a100ml of beaker and dissolve 50.0mg of the substance to be examined in the
mobile phase.
5.3.3.2.2
And
dilute to 50.0ml with the mobile phase.
5.3.3.3
Reference solutions:
5.3.3.3.1
Reference
solution (a):
5.3.3.3.1.1 Take 100ml beaker and dissolve 50.0mg of Clindamycin
hydrochloride CRS in the mobile phase.
5.3.3.3.1.2 And dilute to 50.0ml with the mobile
phase.
5.3.3.4
Column:
5.3.3.4.1
Size:
5.3.3.4.1.1 Length=0.25m,
5.3.3.4.1.2 θ=4.6mm.
5.3.3.4.2
Stationary phase:
5.3.3.4.2.1 Octadecylsilyl silica gel for
chromatography (5μm).
5.3.3.5
Mobile phase:
5.3.3.5.1
Mix
45 volumes of acetonitrile and 55 volumes of a 6.8g/L solution of potassium
dihydrogen phosphate adjusted to pH 7.5 with a 250g/L solution of potassium hydroxide.
5.3.3.6
Flow rate:
5.3.3.6.1
1ml/min.
5.3.3.7
Detection:
5.3.3.7.1
Spectrophotometer
at 210nm.
5.3.3.8
Injection:
5.3.3.8.1
10μL
of the test solution and reference solution (a).
5.3.3.9
Run time:
5.3.3.9.1
Twice
the retention time of Clindamycin.
5.3.3.10 System
suitability:
5.3.3.10.1 Repeatability:
5.3.3.10.1.1 Maximum relative standard deviation of
0.85 percent after 6 injections of reference solution (a).
5.3.4
Method of analysis:
5.3.4.1 Firstly prepare the test solution,
reference solution and mobile phase according to the requirements.
5.3.4.2 The solutions must be free from solid
particles.
5.3.4.3 Prepare the apparatus.
5.3.4.4 The mobile phase solvent mixtures must be
deaerated prior to use either by boiling or by applying a partial vacuum to the
solvent reservoir.
5.3.4.5 Equilibrate the column with the prescribed
mobile phase, flow rate and at temperature specified until a suitable baseline
is achieved.
5.3.4.6 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.
5.3.4.7 As the eluate leaves the column it enters
a detector, where it is continuously monitored at the specified λ.
5.3.4.8 The electrical signal obtained from
detector is amplified and routes to recorder which record the developed Spectrum.
5.3.4.9 Calculate the percentage content of
Clindamycin HCl (C18H34Cl2FN2O5S).
5.3.5
Observations:
5.3.5.1 91% 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 I., Official Monograph /Clindamycin
hydrochloride: 2015, pp. 580-581.
8.0 ANNEXURES:
8.1 Not Applicable.
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/L
|
Gram
per liter
|
mg
|
Milligram
|
Min
|
Minute
|
g
|
Grams
|
ml
|
Milliliter
|
ppt
|
Precipitate
|
μm
|
Micron/
micrometer
|
CRS
|
Chemical
reference substance
|
m
|
Meter
|
θ
|
Theta
|
mm
|
Millimeter
|
ml/min
|
Milliliter
per minute
|
nm
|
Nanometer
|
μL
|
Microliter
|
λ
|
Lamda
|
Vol
|
Volume
|
QCA
|
Quality
control active ingredient
|
F
|
Format
|