CLINDAMYCIN HYDROCLORIDE SOP

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