Revision Bulletin Official April 1, 2010

Add the following:

Azithromycin 1
TolerancesNot 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[NOTEUse low-actinic glassware. Refrigerate 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 solutionPrepare as directed in the Assay. Solution ATransfer 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-m, and degas. Solution BA filtered and degassed mixture of acetonitrile and methanol (75 : 25). Mobile phaseUse variable mixtures of Solution A and Solution B as directed for Chromatographic system. Make adjustments if necessary (see System Suitability under Chromatography 621). Diluent BA mixture of Ammonium phosphate buffer pH 10 and methanol (1 : 1). BlankUse Diluent A. Standard stock solutionPrepare as directed for Standard preparation in the Assay. Standard solutionDilute the Standard stock solution quantitatively with Diluent A to obtain a solution having a known concentration of about 0.02 mg of azithromycin per mL. System sensitivity solutionDilute the Standard solution quantitatively with Diluent A, and mix to obtain a solution having a known concentration of about 0.004 mg of azithromycin per mL. Test solutionWeigh and finely powder not fewer than 20 Tablets. Transfer an accurately weighed portion of the powder, equivalent to about 1335 mg of azithromycin, to a 100-mL volumetric 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 volumetric flask, dilute with Diluent B to volume, and mix to obtain a solution having a nominal concentration of about 4 mg of azithromycin per mL. Pass through a filter having a porosity of 0.45-m. Chromatographic system (see Chromatography 621)The liquid chromatograph is equipped with a 210-nm detector and a 4.6mm 25-cm column that contains 5-m 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 maintained at 4. The chromatograph is programmed as follows: Time (minutes) 025 2530 3040 4055 5560 6061 6170 Solution A (%) 50 5045 4540 4035 35 3550 50 Solution B (%) 50 5055 5560 6065 65 6550 50

Azithromycin Tablets

Azithromycin Tablets contain not less than 90.0 percent and not more than 110.0 percent of the labeled amount of azithromycin (C38H72N2O12).
Packaging and storagePreserve in tight containers, and store at controlled room temperature. USP Reference standards 11USP Azaerythromycin A RS. USP Azithromycin RS. IdentificationThe retention time of the major peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram 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 phasePrepare as directed in the Assay. DiluentTransfer about 17.5 g of dibasic potassium phosphate to a 1000-mL volumetric flask, and dilute with water to volume. Adjust with phosphoric acid to a pH of 8.00 0.05. Prepare a mixture of this solution and acetonitrile (80 : 20). Standard solutionTransfer 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 solution 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 solutionPass a portion of the solution under test through a suitable 0.45-m 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, assuming complete dissolution. Chromatographic system (see Chromatography 621)The liquid chromatograph is equipped with a 210-nm detector and a 4.6mm 15-cm column that contains 5-m 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 theoretical plates; and the relative standard deviation of the azithromycin peak for five replicate injections is not more than 2.0%. ProcedureSeparately inject equal volumes (about 50 L) 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.

Elution isocratic linear gradient linear gradient linear gradient isocratic linear gradient 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 Resolution solution, and record the peak responses as directed for Proce-

2010 The United States Pharmacopeial Convention

All Rights Reserved.

Date: 26-JAN-2010 Time: 11:26 \\managewise\share\SHARE\USPNF\PRINTQ\pager\xmlIn\AO_20100126112600_m362.xml

Azithromycin

Revision Bulletin Official April 1, 2010

Mobile phasePrepare a filtered and degassed mixture of acetonitrile, Octanesulfonate buffer, and methanol (45 : 40 : 15). Make adjustments if necessary (see System Suitability under Chromatography 621). Ammonium phosphate buffer pH 10Transfer about 1.7 g of monobasic ammonium phosphate to a 1-L volumetric flask, and dilute with water to volume. Adjust with ammonium hydroxide to a pH of 10.00 0.05, and mix. Diluent AA mixture of Ammonium phosphate buffer pH 10, methanol, and acetonitrile (35 : 35 : 30). Standard preparationTransfer an accurately weighed quantity of USP Azithromycin RS to a suitable volumetric flask, add Diluent A using about 75% of the final volume, sonicate to dissolve, dilute with Diluent A to volume, and mix to obtain a solution having a known concentration of about 0.4 mg of azithromycin per mL. Stock azaerythromycin A solutionDissolve an accurately weighed quantity of USP Azaerythromycin A RS in acetonitrile with the aid of sonication to obtain a solution having a known concentration of about 0.2 mg of azaerythromycin A per mL. Resolution solutionTransfer suitable volumes of the Stock azaerythromycin A solution and the Standard preparation into a 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 mL. Assay preparationWeigh and finely powder not fewer than 20 Tablets. Transfer an accurately weighed portion of the powder, equivalent to about 667 mg of azithromycin, to a 200-mL volumetric 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 minutes. Allow the solution to equilibrate to room temperature, dilute with Diluent A to volume, and mix. Transfer 6.0 mL of the solution to a 50-mL volumetric flask, dilute with Diluent A to volume, and mix to obtain a solution having a nominal concentration of about 0.4 mg of azithromycin per mL. Pass through a filter having a porosity of 0.45-m. Chromatographic system (see Chromatography 621)The liquid chromatograph is equipped with a 210-nm detector and a 4.6mm 25-cm column that contains 5-m 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 Resolution solution, and record the peak responses as directed for Procedure: identify the peaks by their relative retention times, which are 0.64 and 1.0 for azaerythromycin and azithromycin, respectively; the resolution, R, between the azaerythromycin A and azithromycin peaks is not less than 2.5. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the tailing factor is not more than 2.0; the column efficiency is not less than 1000 theoretical plates; and the relative standard deviation for five replicate injections is not more than 2.0%. ProcedureSeparately inject equal volumes (about 50 L) of the Standard preparation and the Assay preparation into the chromatograph, and measure the peak area responses for all the peaks. Calculate the percent label claim of C38H72N2O12 in the portion of Tablets taken by the formula: (P/1000)(CS / CU)(rU / rS)(100) in which (P/1000) is the potency, converted from g per mg to mg per mg, of C38H72N2O12 in the USP Azithromycin RS; CS is the concentration, in mg per mL, of azithromycin in the Standard preparation; CU is the nominal concentration, in mg per mL, of azithromycin in the Assay preparation; and rU and rS are the peak responses of the azithromycin peaks obtained from the Assay preparation and the Standard preparation, respectively.s2S (USP32)

dure: the resolution, R, between the azaerythromycin A and azithromycin peaks is not less than 2.5. Chromatograph the Standard solution, and record the peak responses as directed for Procedure: the relative standard deviation of the azithromycin peak for replicate injections is not more than 10.0%. ProcedureSeparately inject equal volumes (about 100 L) of the Blank and the Test solution into the chromatograph, and measure the peak area responses for all the peaks. Calculate the percentage of each related compound in the portion of Tablets taken by the formula: (CS / CU)(ri / rS)(P/1000)(1/F)(100) in which CS is the concentration, in mg per mL, of azithromycin in the Standard solution; CU is the nominal concentration, in mg per mL, of azithromycin in the Test solution; ri is the peak area for any impurity obtained from the Test solution; rS is the peak area for azithromycin obtained from the Standard solution; (P/1000) is the potency of azithromycin, converted from g per mg to mg per mg, of USP Azithromycin RS; and F is the relative response factor as listed in Table 1. The specified and unspecified impurities meet the limits listed in Table 1. The reporting level for impurities is 0.1%. Disregard any peaks in the chromatogram for the Test solution that correspond to peaks in the chromatogram of the Blank. Table 1 Relative Retention Time 0.28 0.38 0.40 0.47 0.53 0.57 0.78 0.82 1.0 1.3 1.4

Peak Identification Azithromycin 3-N-oxide 3-(N,N-Didemethyl)-3-Nformylazithromycin 3-(N,N-Didemethyl)azithromycin (aminoazithromycin) Desosaminylazithromycin Azithromycin related compound Fa 3-N-Demethylazithromycin 3-De(dimethylamino)-3oxoazithromycin 6-Demethylazithromycin (azaerythromycin A)b Azithromycin 3-Deoxyazithromycin (azithromycin B)b 3-N-Demethyl-3-N-[(4methylphenyl)sulfonyl]azithromycinb Any other unspecified impurity Total impuritiesb
a

Relative Response Factor 0.45 1.9 0.52 1.1 4.8 0.53 1.6 1.0

Limit (%) 1.0 (RB

1-Apr-2010)

1.0 0.5 0.5

1.0 1.0

(RB (RB

1-Apr-2010)

0.7 0.2

1-Apr-2010)

5.0

(RB

1-Apr-2010)

3-(N-Demethyl)-3-N-formylazithromycin. b These compounds are synthetic process impurities of azithromycin. They are controlled in the drug substance and are listed here for information only. The total impurities specification does not include these impurities.

Assay Octanesulfonate bufferTransfer about 4.4 g of dibasic potassium phosphate and 500 mg of sodium 1-octanesulfonate to a 1000mL 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 \\managewise\share\SHARE\USPNF\PRINTQ\pager\xmlIn\AO_20100126112600_m362.xml