Document No.

HP/QC/RM/SAP-0009
Hygeia Pharmaceuticals Revision No. 01
Quality Control Department
Effective Date 16-01-2019
Standard Analytical Procedure
Next Review 16-01-2021
Clarithromycin

Prepared by Reviewed by Approved by Authorized by

QA Inspector Assistant Manager QA Manager Quality Assurance Plant Manager
*Not valid without signatures

1.0 Title:
Standard Analytical Procedure for Clarithromycin.
2.0 Objective:
The purpose of this SAP is to describe a procedure for analysis of Clarithromycin.
3.0 Scope:
This procedure applies to all qualified users of Hygeia Pharmaceuticals.
4.0 Responsibility:
Each Qualified user is responsible for following this SOP and department head is responsible for
overall compliance to this procedure.

5.0 Abbreviations:
SOP Standard Operating Procedure
SOR Standard Operating Records
SAP Standard Analytical Procedure
D/water Distilled water

6.0 Reference:
USP 40/Hygeia Specs.
7.0 Procedure:
7.1 Description:
White to off white crystalline powder.

7.2 Solubility:
Soluble in acetone, slightly soluble in dehydrated alcohol, practically insoluble in water.

7.3 Identification:
Infrared Absorption 197K.

7.4 Water:
Not more than 2.0%.
7.5 Residue on ignition:
Not more than 0.2%, 0.5 g of it being taken, the charred residue being moistened with 1 ml
of sulfuric acid.
7.6 Heavy metals:
Not more than 0.002%.

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 Document No. HP/QC/RM/SAP-0009
Hygeia Pharmaceuticals Revision No. 01
Quality Control Department
Effective Date 16-01-2019
Standard Analytical Procedure
Next Review 16-01-2021
Clarithromycin

7.7 pH:
Between 8.0 and 10.0, determined in a 1 in 500 suspension of it in a mixture of water and
methanol (19:1).
7.8 Related substances:
Solution A:
Prepare a solution containing 4.76 g of monobasic potassium phosphate per L. Adjust with
dilute phosphoric acid (l in 10) or potassium hydroxide (45% w/v) to a pH of 4.4. Pass this
solution through a C18 filtration kit.

Solution:
Use acetonitrile.

Mobile phase:
Use variable mixtures of Solution A and Solution B as directed under Chromatographic
system. Make adjustments if necessary (see System Suitability under

Chromatography:
Diluting solution: prepare a mixture of acetonitrile and water (50:50).

Standard solution A:
Transfer about 75 mg of USP Clarithromycin RS, accurately weighed, to a 50-ml volumetric
flask, and dissolve in 25 ml of acetonitrile. Dilute with water to volume, and mix.

Standard solution B:
Transfer 5.0 ml of Standard solution A to a 100-ml volumetric flask, dilute with diluting
solution to volume, and mix.

Standard solution C:
Transfer 1.0 ml of Standard solution B to a 10-ml volumetric flask, dilute with diluting
solution to volume, and mix. This solution contains about 0.0075 mg of USP Clarithromycin
RS per ml.
Standard solution D: Transfer about 15 mg of USP Clarithromycin Identity RS, accurately
weighed, to a 10-ml volumetric flask, dissolve in 5.0 ml of acetonitrile, dilute with water to
volume, and mix.

Test solution:
Transfer about 75 mg of Clarithromycin, accurately weighed, to a 50-ml volumetric flask,
dissolve in 25 ml of acetonitrile, dilute with water to volume, and mix.

Chromatographic system:
The liquid chromatograph is equipped with a 205-nm detector and a 4.6-mm × 10-cm
column that contains packing L1 and is maintained at a constant temperature of about 40.
The flow rate is about 1.1 ml per minute.

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 Document No. HP/QC/RM/SAP-0009
Hygeia Pharmaceuticals Revision No. 01
Quality Control Department
Effective Date 16-01-2019
Standard Analytical Procedure
Next Review 16-01-2021
Clarithromycin
Relative retention times with reference to Clarithromycin (retention time = about 11
minutes) include the following: impurity I = about 0.38; impurity C = about 0.89; impurity F
= about 1.33; impurity A = about 0.42; impurity D = about 0.96; impurity P = about 1.35;
impurity J = about 0.63; impurity N = about 1.15; impurity K = about 1.59; impurity L =
about 0.74; impurity E = about 1.27; impurity G = about 1.72; impurity B = about 0.79;
impurity 0 = about 1.38; impurity H = about 1.82; and impurity M = about 0.81.

System suitability:
Chromatograph Standard solution B, and record the responses as directed for Procedure: the
tailing factor for the main Clarithromycin peak is not more than 1.7. Chromatograph
Standard solution D, and record the responses as directed for Procedure: the peak-to-valley
ratio (HP / HV) of impurity D and Clarithromycin is not less than 3.0, where HP is the
height above the baseline of the peak due to impurity D; and HV is the height above the
baseline of the lowest point of the curve separating this peak from the peak due to
Clarithromycin.

Procedure:
Separately inject equal volumes (about 10 µL) of the Diluting solution, Standard solution B,
Standard solution D, Standard solution C, and the Test solution into the chromatograph,
record the chromatograms, and measure the peak area responses. Calculate the percentage of
each impurity in the Clarithromycin taken by the formula:

50(CC / W) (riF /rC) P

in which CC is the concentration, in mg per ml, of USP Clarithromycin RS in Standard

solution C; W is the weight, in mg, of Clarithromycin taken to prepare the Test solution; ri
is the peak area response for any individual impurity observed in the chromatogram
obtained from the Test solution; F is 1.0, or the correction factor of 0.27, and 0.15 applied to
the responses for peaks at relative retention times in relation to that of Clarithromycin of
about 1.72, and 1.82, corresponding to related compound G and related compound H,
respectively; rC is the peak area response of the main Clarithromycin peak in the
chromatogram obtained from Standard solution C; and P is the purity of USP
Clarithromycin RS taken to prepare Standard solution A. Not more than 1.0% of any single
related compound is found, not more than four related compounds exceed the limit of 0.4%,
and the total of all related compounds is not more than 3.5%.

7.9 Assay:
Solution A, Solution B, Diluting solution, and Standard solution D: Proceed as directed in
the test for related substances.

Standard preparation:
Use Standard solution A, prepared as directed in the test for Related substances.

Assay preparation:
Use the Test solution, prepared as directed in the test for related substances.

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 Document No. HP/QC/RM/SAP-0009
Hygeia Pharmaceuticals Revision No. 01
Quality Control Department
Effective Date 16-01-2019
Standard Analytical Procedure
Next Review 16-01-2021
Clarithromycin

Chromatographic system:
Proceed as directed in the test for related substances. In addition, the relative standard
deviation for replicate injections of the Standard preparation is not more than 1.5%.

Procedure:
Separately injects equal volumes (about 10 µL) of the Standard preparation and the Assay
preparation into the chromatograph, records the chromatograms, and measure the peak area
responses for the major peaks. Calculate the percentage of C38H69NO13 in the portion of
Clarithromycin taken by the formula:
50(CS / W)(rU / rS)P,
in which CS is the concentration, in mg per ml, of USP Clarithromycin RS in the Standard
preparation; W is the weight, in mg, of Clarithromycin taken to prepare the Assay
preparation; rU and rS are the Clarithromycin peak area responses obtained from the
chromatograms of the Assay preparation and the Standard preparation, respectively; and P is
the purity of USP Clarithromycin RS taken to prepare the Standard preparation.

Limit: 96-102%

Internal Method:
Particle Size through 60 mesh:
Take 5gm of sample and pass through 60 mesh. Calculate the percentage of the of sample
passed or retained.
Limits: Not Less Than 90% Must pass.

Bulk Density:
Take 5 gm of sample in graduated cylinder, note the volume and calculate the Bulk Density
by using formula.
Density = Mass / Volume
Limits:0.500gm/ml--------------0.630gm/ml

Tapped Density:
Take 5 gm of sample in graduated cylinder and tap 100 times and note the volume and
calculate the Tapped Density by using formula.

Density = Mass / Volume

Limits: 0.620gm/ml--------------0.700gm/ml

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 Document No. HP/QC/RM/SAP-0009
Hygeia Pharmaceuticals Revision No. 01
Quality Control Department
Effective Date 16-01-2019
Standard Analytical Procedure
Next Review 16-01-2021
Clarithromycin

1.0 Records:
Required Record Quality Record No.
Raw material entry log Book HP/QC/QF-006
Analytical work sheet HP/QC/QF-009
Analysis report Raw material HP/QC/QF-003
Instrument Usage Log Book HP/QC/QF-002

2.0 Revision history:

Revision No. Change Description
01 Header footer changed

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