Revision Bulletin
Official April 1, 2010
Add the following:
■
Azithromycin Tablets
» Azithromycin Tablets contain not less than 90.0 per-
cent and not more than 110.0 percent of the labeled
amount of azithromycin (C38H72N2O12).
Packaging and storage—Preserve in tight containers, and store at
controlled room temperature.
USP Reference standards 〈11〉—USP Azaerythromycin A RS. USP
Azithromycin RS.
Identification—The retention time of the major peak in the chro-
matogram of the Assay preparation corresponds to that in the chro-
matogram of the Standard preparation, as obtained in the Assay.
Dissolution 〈711〉—
Medium: pH 6.0 phosphate buffer; 900 mL.
Apparatus 2: 75 rpm.
Time: 30 minutes.
Octanesulfonate buffer and Mobile phase—Prepare as directed in
the Assay.
Diluent—Transfer about 17.5 g of dibasic potassium phosphate to
a 1000-mL volumetric flask, and dilute with water to volume. Ad-
just with phosphoric acid to a pH of 8.00 ± 0.05. Prepare a mixture
of this solution and acetonitrile (80 : 20).
Standard solution—Transfer an accurately weighed quantity of
USP Azithromycin RS, and dissolve in and dilute with Medium to
obtain a solution having a known concentration of about L/1000 mg
per mL, where L is the Tablet label claim, in mg. Dilute this solu-
tion with Diluent to obtain a solution having a known concentration
of about L/2000 mg per mL, where L is the Tablet label claim, in
mg.
Test solution—Pass a portion of the solution under test through a
suitable 0.45-µm filter. Dilute a portion of the filtrate with Diluent
to obtain a solution having a nominal concentration of about L/
2000 mg per mL, where L is the Tablet label claim, in mg, assum-
ing complete dissolution.
Chromatographic system (see Chromatography 〈621〉)—The liq-
uid chromatograph is equipped with a 210-nm detector and a 4.6-
mm ×15-cm column that contains 5-µm packing L1. The flow rate
is about 1.5 mL per minute. The column is maintained at a constant
temperature of about 50°. Chromatograph the Standard solution,
and record the peak responses as directed for Procedure: the tailing
factor of the azithromycin peak is not more than 2.0; the column
efficiency for the azithromycin peak is not less than 1000 theoreti-
cal plates; and the relative standard deviation of the azithromycin
peak for five replicate injections is not more than 2.0%.
Procedure—Separately inject equal volumes (about 50 µL) of the
Standard solution and the Test solution into the chromatograph, and
record the chromatograms and the peak responses. Calculate the
percentage of C38H72N2O12 dissolved by the formula:
in which rU and rS are the peak responses for the Test solution and
the Standard solution, respectively; CS is the concentration, in mg
per mL, of the Standard solution; 900 is the volume, in mL, of the
Medium; 100 is the conversion factor to percentage; and L is the
Tablet label claim, in mg.
 2010 The United States Pharmacopeial Convention
Date: 26-JAN-2010 Time: 11:26
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Azithromycin 1
Tolerances—Not less than 80% (Q) of the labeled amount of
azithromycin is dissolved in 30 minutes.
Uniformity of dosage units 〈905〉: meets the requirements.
Change to read:
Related compounds—[NOTE—Use low-actinic glassware. Refrig-
erate the Standard solution and the Test solution immediately after
preparation and during analysis, using a refrigerated autosampler
set at 4°. The solutions must be analyzed within 24 hours of
preparation.]
Ammonium phosphate buffer pH 10, Diluent A, and Resolution
solution—Prepare as directed in the Assay.
Solution A—Transfer about 1.8 g of dibasic sodium phosphate to
a 1000-mL volumetric flask, and dilute with water to volume. Pass
through a filter having a porosity of 0.45-µm, and degas.
Solution B—A filtered and degassed mixture of acetonitrile and
methanol (75 : 25).
Mobile phase—Use variable mixtures of Solution A and Solution
B as directed for Chromatographic system. Make adjustments if
necessary (see System Suitability under Chromatography 〈621〉).
Diluent B—A mixture of Ammonium phosphate buffer pH 10 and
methanol (1 : 1).
Blank—Use Diluent A.
Standard stock solution—Prepare as directed for Standard prepa-
ration in the Assay.
Standard solution—Dilute the Standard stock solution quantita-
tively with Diluent A to obtain a solution having a known concen-
tration of about 0.02 mg of azithromycin per mL.
System sensitivity solution—Dilute the Standard solution quan-
titatively with Diluent A, and mix to obtain a solution having a
known concentration of about 0.004 mg of azithromycin per mL.
Test solution—Weigh and finely powder not fewer than 20 Tab-
lets. Transfer an accurately weighed portion of the powder,
equivalent to about 1335 mg of azithromycin, to a 100-mL volu-
metric flask. Add about 75 mL of acetonitrile, and sonicate for not
less than 15 minutes. Shake by mechanical means for not less than
15 minutes. Allow the solution to equilibrate to room temperature,
dilute with acetonitrile to volume, and mix. Centrifuge an aliquot
for 15 minutes. Transfer 3.0 mL of the supernatant to a 10-mL volu-
metric flask, dilute with Diluent B to volume, and mix to obtain a
solution having a nominal concentration of about 4 mg of azithro-
mycin per mL. Pass through a filter having a porosity of 0.45-µm.
Chromatographic system (see Chromatography 〈621〉)—The liq-
uid chromatograph is equipped with a 210-nm detector and a 4.6-
mm × 25-cm column that contains 5-µm packing L1. The flow rate
is about 0.8 mL per minute. The column is maintained at a constant
temperature of about 60°. The autosampler temperature is main-
tained at 4°. The chromatograph is programmed as follows:
Time Solution A Solution B
(minutes) (%) (%) Elution
0–25 50 50 isocratic
25–30 50→45 50→55 linear gradient
30–40 45→40 55→60 linear gradient
40–55 40→35 60→65 linear gradient
55–60 35 65 isocratic
60–61 35→50 65→50 linear gradient
61–70 50 50 re-equilibration
Chromatograph the System sensitivity solution, and record the peak
responses as directed for Procedure: the signal-to-noise ratio for the
azithromycin peak is not less than 10. Chromatograph the Resolu-
tion solution, and record the peak responses as directed for Proce-
All Rights Reserved.
 Revision Bulletin
2 Azithromycin Official April 1, 2010

dure: the resolution, R, between the azaerythromycin A and azithro- Mobile phase—Prepare a filtered and degassed mixture of
mycin peaks is not less than 2.5. Chromatograph the Standard acetonitrile, Octanesulfonate buffer, and methanol (45 : 40 : 15).
solution, and record the peak responses as directed for Procedure: Make adjustments if necessary (see System Suitability under Chro-
the relative standard deviation of the azithromycin peak for repli- matography 〈621〉).
cate injections is not more than 10.0%. Ammonium phosphate buffer pH 10—Transfer about 1.7 g of
Procedure—Separately inject equal volumes (about 100 µL) of monobasic ammonium phosphate to a 1-L volumetric flask, and
the Blank and the Test solution into the chromatograph, and meas- dilute with water to volume. Adjust with ammonium hydroxide to a
ure the peak area responses for all the peaks. Calculate the percent- pH of 10.00 ± 0.05, and mix.
age of each related compound in the portion of Tablets taken by the Diluent A—A mixture of Ammonium phosphate buffer pH 10,
formula: methanol, and acetonitrile (35 : 35 : 30).
(CS / CU)(ri / rS)(P/1000)(1/F)(100) Standard preparation—Transfer an accurately weighed quantity
of USP Azithromycin RS to a suitable volumetric flask, add Diluent
in which CS is the concentration, in mg per mL, of azithromycin in A using about 75% of the final volume, sonicate to dissolve, dilute
the Standard solution; CU is the nominal concentration, in mg per with Diluent A to volume, and mix to obtain a solution having a
mL, of azithromycin in the Test solution; ri is the peak area for any known concentration of about 0.4 mg of azithromycin per mL.
impurity obtained from the Test solution; rS is the peak area for Stock azaerythromycin A solution—Dissolve an accurately
azithromycin obtained from the Standard solution; (P/1000) is the weighed quantity of USP Azaerythromycin A RS in acetonitrile
potency of azithromycin, converted from µg per mg to mg per mg, with the aid of sonication to obtain a solution having a known con-
of USP Azithromycin RS; and F is the relative response factor as centration of about 0.2 mg of azaerythromycin A per mL.
listed in Table 1. The specified and unspecified impurities meet the
limits listed in Table 1. The reporting level for impurities is 0.1%. Resolution solution—Transfer suitable volumes of the Stock
Disregard any peaks in the chromatogram for the Test solution that azaerythromycin A solution and the Standard preparation into a
correspond to peaks in the chromatogram of the Blank. volumetric flask of the appropriate size, and dilute quantitatively
with Diluent A to obtain a solution having a known concentration of
about 0.02 mg each of azaerythromycin A and azithromycin per
Table 1 mL.
Relative Relative Assay preparation—Weigh and finely powder not fewer than 20
Retention Response Limit Tablets. Transfer an accurately weighed portion of the powder,
Peak Identification Time Factor (%) equivalent to about 667 mg of azithromycin, to a 200-mL volumet-
Azithromycin 3′-N-oxide 0.28 0.45 •1.0 (RB ric flask. Add about 75 mL of Diluent A, and sonicate for not less
• than 15 minutes. Shake by mechanical means for not less than 15
1-Apr-2010)
minutes. Allow the solution to equilibrate to room temperature, di-
3′-(N,N-Didemethyl)-3′-N- 0.38 1.9 1.0 lute with Diluent A to volume, and mix. Transfer 6.0 mL of the so-
formylazithromycin lution to a 50-mL volumetric flask, dilute with Diluent A to volume,
3′-(N,N-Didemethyl)azithro- 0.40 0.52 0.5 and mix to obtain a solution having a nominal concentration of
mycin (aminoazithro- about 0.4 mg of azithromycin per mL. Pass through a filter having a
mycin) porosity of 0.45-µm.
Desosaminylazithromycin 0.47 1.1 0.5 Chromatographic system (see Chromatography 〈621〉)—The liq-
Azithromycin related com- 0.53 4.8 •1.0 uid chromatograph is equipped with a 210-nm detector and a 4.6-
•(RB mm × 25-cm column that contains 5-µm packing L1. The flow rate
pound Fa 1-Apr-2010)
3′-N-Demethylazithromycin 0.57 0.53 0.7 is about 1.5 mL per minute. The column is maintained at a constant
•1.0 temperature of about 50°. Chromatograph the Resolution solution,
3′-De(dimethylamino)-3′- 0.78 1.6 •(RB and record the peak responses as directed for Procedure: identify
oxoazithromycin 1-Apr-2010) the peaks by their relative retention times, which are 0.64 and 1.0
6-Demethylazithromycin 0.82 — — for azaerythromycin and azithromycin, respectively; the resolution,
(azaerythromycin A)b R, between the azaerythromycin A and azithromycin peaks is not
Azithromycin 1.0 — — less than 2.5. Chromatograph the Standard preparation, and record
3-Deoxyazithromycin 1.3 — — the peak responses as directed for Procedure: the tailing factor is
not more than 2.0; the column efficiency is not less than 1000 theo-
(azithromycin B)b retical plates; and the relative standard deviation for five replicate
3′-N-Demethyl-3′-N-[(4- 1.4 — — injections is not more than 2.0%.
methylphenyl)sulfon- Procedure—Separately inject equal volumes (about 50 µL) of the
yl]azithromycinb Standard preparation and the Assay preparation into the chromato-
Any other unspecified im- 1.0 0.2 graph, and measure the peak area responses for all the peaks. Calcu-
—
purity late the percent label claim of C38H72N2O12 in the portion of Tablets
Total impuritiesb — — •5.0 taken by the formula:
•(RB
1-Apr-2010)
a
3′-(N-Demethyl)-3′-N-formylazithromycin. (P/1000)(CS / CU)(rU / rS)(100)
b
These compounds are synthetic process impurities of azithromycin. They are con- in which (P/1000) is the potency, converted from µg per mg to mg
trolled in the drug substance and are listed here for information only. The total per mg, of C38H72N2O12 in the USP Azithromycin RS; CS is the con-
impurities specification does not include these impurities. centration, in mg per mL, of azithromycin in the Standard prepara-
tion; CU is the nominal concentration, in mg per mL, of azithro-
Assay— mycin in the Assay preparation; and rU and rS are the peak
Octanesulfonate buffer—Transfer about 4.4 g of dibasic potas- responses of the azithromycin peaks obtained from the Assay prep-
sium phosphate and 500 mg of sodium 1-octanesulfonate to a 1000- aration and the Standard preparation, respectively.■2S (USP32)
mL volumetric flask, and dilute with water to volume. Adjust with
phosphoric acid to a pH of 8.20 ± 0.05.

 2010 The United States Pharmacopeial Convention All Rights Reserved.

Date: 26-JAN-2010 Time: 11:26
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