Printed on: Tue Jan 04 2022, 07:59:15 AM(EST) Official Status: Currently Official on 04-Jan-2022 DocId: 1_GUID-80BCB1B5-31D4-416A-A492-8DDD07C8F7F9_6_en-US

Printed by: Steven Alvarez Aguilar Official Date: Official as of 01-May-2020 Document Type: USP @2022 USPC
DOI Ref: 7xyz9 DOI: https://doi.org/10.31003/USPNF_M362_06_01
1

PERFORMANCE TESTS
Azithromycin Tablets
Change to read:
DEFINITION
Azithromycin Tablets contain NLT 90.0% and NMT 110.0% • DISSOLUTION á711ñ
of the labeled amount of azithromycin (C38H72N2O12). Medium: pH 6.0 phosphate buffer; 900 mL
Apparatus 2: 75 rpm
IDENTIFICATION Time: 30 min
• A. The retention time of the major peak of the Sample Solution A: 4.4 mg/mL of dibasic potassium phosphate and
solution corresponds to that of the Standard solution, as 0.5 mg/mL of sodium 1-octanesulfonate; adjusted with
obtained in the Assay. phosphoric acid to a pH of 8.20 ± 0.05
Mobile phase: Acetonitrile, methanol, and Solution A
Change to read: (9:3:8)
Diluent: 17.5 mg/mL of dibasic potassium phosphate.
▲
• B. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared Adjust with phosphoric acid to a pH of 8.00 ± 0.05.
Spectroscopy: 197A▲ (CN 1-May-2020) Prepare a mixture of this solution and acetonitrile (80:20).
Standard solution: 25 mg/mL of USP Azithromycin RS in Standard stock solution: Dissolve USP Azithromycin RS in
acetonitrile. Pass the solution through a suitable filter, and Medium to obtain a solution with a known concentration of
remove the solvent by natural evaporation. about (L/1000) mg/mL, where L is the label claim in mg/
Sample solution: Equivalent to 25 mg/mL of azithromycin Tablet.
from Tablets in acetonitrile. Pass the solution through a Standard solution: Dilute the Standard stock solution with
suitable filter, and remove the solvent by natural Diluent to obtain a solution with a known concentration of
evaporation. about (L/2000) mg/mL, where L is the label claim in mg/

al
Acceptance criteria: Meet the requirements▲ (USP 1-Dec-2019) Tablet.
ASSAY Sample solution: Pass a portion of the solution under test
• PROCEDURE through a suitable filter of 0.45-µm pore size. Dilute a
Buffer: Dissolve 4.6 g of monobasic potassium phosphate portion of the filtrate with Diluent to obtain a solution with a
anhydrous in 900 mL of water. Adjust with 1 N sodium ci theoretical concentration of about (L/2000) mg/mL, where
hydroxide to a pH of 7.5, and dilute with water to 1 L. L is the label claim in mg/Tablet, assuming complete
Mobile phase: Acetonitrile and Buffer (65:35) dissolution.
Standard solution: 1 mg/mL of USP Azithromycin RS in Chromatographic system
Mobile phase. Sonicate and shake as needed to dissolve. (See Chromatography á621ñ, System Suitability.)
Sample solution: Nominally 1 mg/mL of azithromycin in Mode: LC
Detector: UV 210 nm
ffi
Mobile phase from NLT 20 Tablets, finely powdered.
Sonicate and shake as needed to dissolve. Column: 4.6-mm × 15-cm; 5-µm packing L1
Chromatographic system Column temperature: 50°
(See Chromatography á621ñ, System Suitability.) Flow rate: 1.5 mL/min
Mode: LC Injection volume: 50 µL
Detector: UV 210 nm System suitability
Sample: Standard solution
O

Column: 4.6-mm × 25-cm; 5-µm packing L1

Column temperature: 50° Suitability requirements
Flow rate: 2 mL/min Tailing factor: NMT 2.0
Injection volume: 100 µL Relative standard deviation: NMT 2.0%
System suitability Analysis
Sample: Standard solution Samples: Standard solution and Sample solution
Suitability requirements Calculate the percentage of the labeled amount of
Tailing factor: NMT 2.0 azithromycin (C38H72N2O12) dissolved:
Relative standard deviation: NMT 2.0%
Analysis Result = (rU/rS) × (CS/L) × V × ▲D▲ (USP 1-Dec-2019) × 100
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of rU = peak response of azithromycin from the Sample
azithromycin (C38H72N2O12) in the portion of Tablets solution
taken: rS = peak response of azithromycin from the Standard
solution
Result = (rU/rS) × (CS/CU) × P × F × 100 CS = concentration of USP Azithromycin RS in the
Standard solution (mg/mL)
rU = peak response of azithromycin from the Sample L = label claim (mg/Tablet)
solution V = volume of Medium, 900 mL
rS = peak response of azithromycin from the Standard ▲
D = dilution factor for the Sample solution, if
solution necessary▲ (USP 1-Dec-2019)
CS = concentration of USP Azithromycin RS in the
Standard solution (mg/mL) Tolerances: NLT 80% (Q) of the labeled amount of
CU = nominal concentration of azithromycin in the azithromycin (C38H72N2O12) is dissolved.
Sample solution (mg/mL) • UNIFORMITY OF DOSAGE UNITS á905ñ: Meet the
P = potency of USP Azithromycin RS (µg/mg) requirements
F = conversion factor, 0.001 mg/µg

Acceptance criteria: 90.0%–110.0%

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Printed on: Tue Jan 04 2022, 07:59:15 AM(EST) Official Status: Currently Official on 04-Jan-2022 DocId: 1_GUID-80BCB1B5-31D4-416A-A492-8DDD07C8F7F9_6_en-US
Printed by: Steven Alvarez Aguilar Official Date: Official as of 01-May-2020 Document Type: USP @2022 USPC
DOI Ref: 7xyz9 DOI: https://doi.org/10.31003/USPNF_M362_06_01
2

IMPURITIES Chromatographic system

(See Chromatography á621ñ, System Suitability.)
Change to read: Mode: LC
• ORGANIC IMPURITIES Detector: UV 210 nm
Protect all solutions containing azithromycin from light. Column: 4.6-mm × 15-cm; 3.5-µm packing L1
Refrigerate the Standard solution and the Sample solution Temperatures
after preparation and during analysis, using a refrigerated Autosampler: 4°
autosampler set at 4°. The solutions must be analyzed Column: 50°
within 24 h of preparation. Flow rate: 1.2 mL/min
Solution A: Water and ammonium hydroxide (2000: 1.2). Injection volume: 100 µL
The pH of this solution is about 10.5. System suitability
Solution B: Acetonitrile, methanol, and ammonium Samples: System suitability solution, Standard solution, and
hydroxide (1800: 200: 1.2) Sensitivity solution
Mobile phase: See Table 1. Suitability requirements
Resolution: NLT 1.0 between desosaminylazithromycin
Table 1 and azithromycin related compound F, System suitability
solution
Time Solution A Solution B Relative standard deviation: NMT 2.0%, Standard
(min) (%) (%)
solution
0 54 46 Signal-to-noise ratio: NLT 10, Sensitivity solution
20 54 46
Analysis
Samples: ▲Standard solution,▲ (USP 1-Dec-2019)Sample

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35 10 90 solution, and Blank
35.1 54 46 Calculate the percentage of each impurity in the portion of
Tablets taken:
▲
50.1▲ (USP 1-Dec-2019) 54 46
Result = (rU/rS) × (CS/CU) × P × F1 × (1/F2) × 100
Buffer: 1.7 g/L of monobasic ammonium phosphate in
water. Adjust with ammonium hydroxide to a pH of 10
▲
± 0.05.▲ (USP 1-Dec-2019)
Diluent A: Methanol, acetonitrile, and Buffer
ci rU

rS
= peak response of each impurity from the Sample
solution
= peak response of azithromycin from the Standard
(350:300:350). ▲ [NOTE—Diluent A is stable for 24 h after solution
mixing the organic and buffer phases.]▲ (USP 1-Dec-2019) CS = concentration of USP Azithromycin RS in the
ffi
Diluent B: Methanol and Buffer (1:1) Standard solution (mg/mL)
System suitability stock solution: 0.1 mg/mL each of USP CU = nominal concentration of azithromycin in the
Desosaminylazithromycin RS, USP Azithromycin Related Sample solution (mg/mL)
P = potency of USP Azithromycin RS (µg/mg)
Compound F RS, and ▲USP
F1 = conversion factor, 0.001 mg/µg
N-Demethylazithromycin RS▲ (USP 1-Dec-2019) in acetonitrile
System suitability solution: 0.028 mg/mL each of USP F2 = relative response factor (see Table 2)
O

Desosaminylazithromycin RS, USP Azithromycin Related

Compound F RS, and ▲USP Acceptance criteria: See Table 2. The reporting threshold is
N-Demethylazithromycin RS▲ (USP 1-Dec-2019) from the System 0.1%. Disregard any peaks in the Sample solution that
suitability stock solution in Diluent A correspond to peaks in the Blank.
▲
Peak identification solution: 0.004 mg/mL each of USP
Table 2
Desosaminylazithromycin RS, USP Azithromycin Related
Compound F RS, and USP N-Demethylazithromycin RS Accept-
Relative Relative ance
from the System suitability solution in Diluent A▲ (USP 1-Dec-2019) Retention Response Criteria,
Standard stock solution: 0.4 mg/mL of USP Name Time Factor NMT (%)
Azithromycin RS in acetonitrile. Sonicate and shake as
Azithromycin
needed to dissolve. N-oxidea 0.20 ▲
0.46▲ (USP 1-Dec-2019) 1.0
Standard solution: 0.02 mg/mL of azithromycin from the
Standard stock solution in Diluent A 3′-(N,N-Didemeth- 0.29
Sensitivity solution: 0.004 mg/mL of azithromycin from yl)-3′-N-formylazi-
thromycinb,▲c▲
the Standard solution in Diluent A (USP 1-Dec-2019) 0.30 1.7 1.0
Sample stock solution: Nominally 14.3 mg/mL of
azithromycin prepared as follows. Transfer nominally 3′-(N,N-
Didemethyl)azi-
1430 mg of azithromycin, from finely powdered Tablets thromycin (amino-
(NLT 20), to a 100-mL volumetric flask. Add 75 mL of azithromycin)d 0.34 ▲
0.44▲ (USP 1-Dec-2019) 0.5
acetonitrile, and sonicate for NLT 15 min. Shake by
mechanical means for NLT 15 min. Allow the solution to
▲
0.40▲
Azithromycin relat-
equilibrate to room temperature, dilute with acetonitrile to ed com-
(USP 1-Dec-2019)

volume, and mix. pound F▲c,▲ ▲

0.46▲
Sample solution: Nominally 4 mg/mL of azithromycin (USP 1-Dec-2019)
e
(USP 1-Dec-2019)
▲
5.5▲ (USP 1-Dec-2019) 1.0
prepared as follows. Centrifuge an aliquot of the Sample ▲
0.47▲
Desosaminylazithro-
stock solution for NLT 15 min. Transfer 7.0 mL of the mycinf (USP 1-Dec-2019) 1.1 0.5
supernatant to a 25-mL volumetric flask, and dilute with
Diluent B to volume. N-Demethylazithro-
mycing 0.50 ▲
0.47▲ (USP 1-Dec-2019) 0.7
Blank: Diluent A

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Printed on: Tue Jan 04 2022, 07:59:15 AM(EST) Official Status: Currently Official on 04-Jan-2022 DocId: 1_GUID-80BCB1B5-31D4-416A-A492-8DDD07C8F7F9_6_en-US
Printed by: Steven Alvarez Aguilar Official Date: Official as of 01-May-2020 Document Type: USP @2022 USPC
DOI Ref: 7xyz9 DOI: https://doi.org/10.31003/USPNF_M362_06_01
3

Table 2 (continued) h (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3,3-dimethyl-α-L-

ribo-hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-
Accept- heptamethyl-11-[[3,4,6-trideoxy-3-oxo-β-D-xylo-hexopyranosyl]oxy]-1-oxa-6-
Relative Relative ance azacyclopentadecan-15-one.
Retention Response Criteria, i Process impurities that are controlled in the drug substance are not to be
Name Time Factor NMT (%)
reported. They are listed here for information only. The unspecified impurities
3′-De(dimethylami- and total impurities limits do not include these impurities.
no)-3′-oxoazithro- j 9-Deoxo-9a-aza-9a-homoerythromycin A.
mycinh 0.87 ▲
1.7▲ (USP 1-Dec-2019) 1.0 k (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O-
methyl-α-L-ribo-hexopyranosyl)oxy]-2-propyl-3,4,10-trihydroxy-
Azaerythromy- 3,5,6,8,10,12,14-heptamethyl-11-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-
— —
cin Ai, j 0.94 xylo-hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one dihydrate.
l (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O-
Azithromycin 1.0 — —
methyl-α-L-ribo-hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy-
2-Desethyl-2- 3,5,6,8,10,12,14-heptamethyl-11-[[3-[N-(4-methylphenylsulfonyl)-N-
propylazithro methylamino]-3,4,6-trideoxy-β-D-xylo-hexopyranosyl]oxy]-1-oxa-6-
— — azacyclopentadecan-15-one.
-
m (2R,3R,4S,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O-
mycini, k 1.10
methyl-α-L-ribo-hexopyranosyl)oxy]-2-ethyl-4,10-dihydroxy-3,5,6,8,10,12,14-
3′-N-Demethyl- heptamethyl-11-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-xylo-hexopyranosyl]
3′-N-[(4-methyl- oxy]-1-oxa-6-azacyclopentadecan-15-one.
phenyl)sulfonyl]
— —
azithro
- ADDITIONAL REQUIREMENTS
mycini, l 1.11 • PACKAGING AND STORAGE: Preserve in tight containers.
Store at controlled room temperature.
3-Deoxyazithromy-

al
cin (azithromycin — —
B)i, m 1.14 Change to read:
Any individual • USP REFERENCE STANDARDS á11ñ
unspecified — USP Azithromycin RS
impurityi 1.0 0.2
ci USP Azithromycin Related Compound F RS
Total impuritiesi — — 5.0 3′-(N-Demethyl)-3′-N-formylazithromycin;
(2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-
a (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O-
Dideoxy-3-C-methyl-3-O-methyl-α-L-ribo-
methyl-α-L-ribo-hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy-
3,5,6,8,10,12,14-heptamethyl-11-[[3,4,6-trideoxy-3-(dimethylazinoyl)-β-D-
hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy-
xylo-hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one. 3,5,6,8,10,12,14-heptamethyl-11-[[3-(N-methyl)
formamido-3,4,6-trideoxy-β-D-xylo-hexopyranosyl]
ffi
b (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O-
methyl-α-L-ribo-hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy- oxy]-1-oxa-6-azacyclopentadecan-15-one.
3,5,6,8,10,12,14-heptamethyl-11-[[3-formamido-3,4,6-trideoxy-β-D-xylo- C38H70N2O13 762.97
hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one.
c The system may resolve two rotamers. The limit is for the sum of the two
▲
USP N-Demethylazithromycin RS
rotamers. (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-
d (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O- Dideoxy-3-C-methyl-3-O-methyl-α-L-ribo-
O

methyl-α-L-ribo-hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy- hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy-
3,5,6,8,10,12,14-heptamethyl-11-[[3-amino-3,4,6-trideoxy-β-D-xylo-
hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one.
3,5,6,8,10,12,14-heptamethyl-11-[[3,4,6-trideoxy-3-
e 3′-(N-Demethyl)-3′-N-formylazithromycin; (2R,3S,4R,5R,8R,10R,11R,12S, methylamino-β-D-xylo-hexopyranosyl]oxy]-1-oxa-6-
13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O-methyl-α-L-ribo-hexopyranosyl) azacyclopentadecan-15-one.
oxy]-2-ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-heptamethyl-11-[[3-(N- C37H70N2O12 734.96▲ (USP 1-Dec-2019)
methyl)formamido-3,4,6-trideoxy-β-D-xylo-hexopyranosyl]oxy]-1-oxa-6-
azacyclopentadecan-15-one.
USP Desosaminylazithromycin RS
f (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2-Ethyl-3,4,10,13-tetrahydroxy- (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2-Ethyl-
3,5,6,8,10,12,14-heptamethyl-11-[[3,4,6-trideoxy-3-dimethylamino-β-D-xylo- 3,4,10,13-tetrahydroxy-3,5,6,8,10,12,14-heptamethyl-
hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one. 11-[[3,4,6-trideoxy-3-dimethylamino-β-D-xylo-
g (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-13-[(2,6-Dideoxy-3-C-methyl-3-O-
hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-
methyl-α-L-ribo-hexopyranosyl)oxy]-2-ethyl-3,4,10-trihydroxy- one.
3,5,6,8,10,12,14-heptamethyl-11-[[3,4,6-trideoxy-3-methylamino-β-D-xylo-
hexopyranosyl]oxy]-1-oxa-6-azacyclopentadecan-15-one. C30H58N2O9 590.79

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