Printed on: Thu Feb 09 2023, 01:02:37 AM(EST) Status: Not Yet Official on 09-Feb-2023 DocId: GUID-5C010AF5-72F3-44E0-9EE4-6393A31AE6A2_5_en-US

Printed by: Dang Van Vu Official Date: 01-May-2023 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: y1tw2 DOI: https://doi.org/10.31003/USPNF_M890_05_01
1

Tailing factor: NMT 2▲▲ (RB 1-May-2023)

Acyclovir
To view the Notice from the Expert Committee that posted in
conjunction with this accelerated revision, please click
www.uspnf.com/rb-acyclovir-20220429.
Relative standard deviation: NMT 2.0%▲▲ (RB 1-May-2023)
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of acyclovir (C8H11N5O3) in the
portion of Acyclovir taken:

Result = (rU/rS) × (CS/CU) × 100

rU = peak response of acyclovir from the Sample

solution
rS = peak response of acyclovir from the Standard
solution

al
CS = concentration of USP Acyclovir RS in the Standard
solution (mg/mL)
CU = concentration of Acyclovir in the Sample solution
(mg/mL)
C8H11N5O3 225.20

ci
6H-Purin-6-one, 2-amino-1,9-dihydro-9-[(2-hydroxyethoxy) Acceptance criteria: 98.0%–101.0% on the anhydrous
methyl]-; basis
9-[(2-Hydroxyethoxy)methyl]guanine CAS RN®: 59277-89-
IMPURITIES
3; UNII: X4HES1O11F.
DEFINITION Delete the following:

ffi
Acyclovir contains NLT 98.0% and NMT 101.0% of acyclovir ▲
• PROCEDURE 1: LIMIT OF GUANINE▲ (RB 1-May-2023)
(C8H11N5O3), calculated on the anhydrous basis.
Delete the following:
IDENTIFICATION
▲
• ORDINARY IMPURITIES á466ñ▲ (RB 1-May-2023)
Change to read:
O
• A. SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared Add the following:
Spectroscopy: 197K ▲or 197A▲ (RB 1-May-2023) ▲
• ORGANIC IMPURITIES
Buffer A: 3.48 g/L of dipotassium hydrogen phosphate in
Change to read:
water. Adjust with phosphoric acid to pH of 3.1.
• B. The retention time of the major peak of the Sample Buffer B: 3.48 g/L of dipotassium hydrogen phosphate in
water. Adjust with phosphoric acid to pH of 2.5.
e
solution corresponds to that of the Standard solution, as
obtained in the Assay. ▲▲ (RB 1-May-2023) Solution A: Acetonitrile and Buffer A (1:99)
Solution B: Acetonitrile and Buffer B (50:50)
ASSAY Mobile phase: See Table 1.
B

Change to read: Table 1

• PROCEDURE Time Solution A Solution B
Mobile phase: Glacial acetic acid in water (1 in 1000) (min) (%) (%)
▲
▲ (RB 1-May-2023) 0 100 0
Standard stock solution: 0.5 mg/mL of USP Acyclovir RS
To

prepared as follows. Dissolve about 25 mg of USP Acyclovir
RS in 5 mL of 0.1 N sodium hydroxide in a 50-mL volumetric
flask, and dilute with water to volume.
Standard solution: 0.1 mg/mL of USP Acyclovir RS in
0.01 N sodium hydroxide from Standard stock solution
Sample stock solution: 0.5 mg/mL of Acyclovir prepared as
follows. Dissolve about 100 mg of Acyclovir in 20 mL of
0.1 N sodium hydroxide in a 200-mL volumetric flask, and
dilute with water to volume.
Sample solution: 0.1 mg/mL of Acyclovir in ▲0.01 N
sodium hydroxide▲ (ERR 1-May-2023) from Sample stock solution
Chromatographic system
(See Chromatography á621ñ, System Suitability.)
Mode: LC
Detector: UV 254 nm
Column: 4.6-mm × 25-cm; ▲10-µm▲ (RB 1-May-2023) packing
L1
Flow rate: 3 mL/min
Injection volume: 20 µL
System suitability
Sample: ▲Standard solution▲ (RB 1-May-2023)
Suitability requirements
▲
▲ (RB 1-May-2023)
5 100 0
27 80 20
40 80 20
Diluent: Dimethyl sulfoxide and water (20:80 v/v)
System suitability solution: 0.01 mg/mL of USP Acyclovir
RS and 0.5 μg/mL each of USP Acyclovir Related
Compound A RS and USP Acyclovir Related Compound F
RS prepared as follows. Dissolve a suitable amount of USP
Acyclovir RS, USP Acyclovir Related Compound A RS, and
USP Acyclovir Related Compound F RS in 20% of the final
flask volume of dimethyl sulfoxide and dilute with water to
volume.
Standard stock solution: 0.1 mg/mL of USP Acyclovir RS
prepared as follows. Dissolve a suitable amount of USP
Acyclovir RS in 20% of the final flask volume of dimethyl
sulfoxide and dilute with water to volume.
Standard solution: 1.0 µg/mL of USP Acyclovir RS in Diluent
from Standard stock solution
Sensitivity solution: 0.3 µg/mL of USP Acyclovir RS in
Diluent from Standard stock solution
Sample solution: 1 mg/mL of Acyclovir prepared as follows.
Dissolve a suitable amount of Acyclovir in 20% of the final

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Printed on: Thu Feb 09 2023, 01:02:37 AM(EST) Status: Not Yet Official on 09-Feb-2023 DocId: GUID-5C010AF5-72F3-44E0-9EE4-6393A31AE6A2_5_en-US
Printed by: Dang Van Vu Official Date: 01-May-2023 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: y1tw2 DOI: https://doi.org/10.31003/USPNF_M890_05_01
2

flask volume of dimethyl sulfoxide and dilute with water to Table 2 (continued)
volume. Relative Relative Acceptance
Chromatographic system Retention Response Criteria,
(See Chromatography á621ñ, System Suitability.) Name Time Factor NMT (%)
Mode: LC Diacetyl gua-
Detector: UV 254 nm nined 1.48 0.71 0.1
Column: 4.6-mm × 25-cm; 5-µm packing L1
7,9′-Diguanyl an-
Flow rate: 1 mL/min aloge 1.53 0.66 0.1
Injection volume: 10 µL
System suitability 9,9′-Diguanyl an-
alogf 1.59 1.0 0.2
Samples: System suitability solution, Standard solution, and
Sensitivity solution Acyclovir related
Suitability requirements compound F 1.79 1.0 0.1

al
Resolution: NLT 1.0 between acyclovir related Acyclovir related
compound F and acyclovir related compound A, System compound A 1.83 1.0 0.1
suitability solution g
Relative standard deviation: NMT 5.0%, Standard Bis-acyclovir 2.44 1.0 0.2
solution N,O-Diacetyl

ci
Signal-to-noise ratio: NLT 10, Sensitivity solution acyclovir-N9-iso-
Analysis merh 2.67 1.0 0.1
Samples: Standard solution and Sample solution Any individual
Calculate the percentage of any individual impurity in the unspecified im- —
portion of Acyclovir taken: purity 1.0 0.05

ffi
Total impurities — — 1.0
Result = (rU/rS) × (CS/CU) × (1/F) x 100
a 9-(2-Hydroxyethyl)guanine.
rU = peak response of any individual impurity from the b 2-Amino-7-[(2-hydroxyethoxy)methyl]-1,7-dihydro-6H-purin-6-one.
Sample solution c Unknown structure.
rS = peak response of acyclovir from the Standard d N2,9-Diacetylguanine.
solution
O e 2-Amino-9-({2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethoxy}
CS = concentration of USP Acyclovir RS in the Standard methyl)-1,7-dihydro-6H-purin-6-one.
f 1,2-Bis[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethane.
solution (mg/mL) g Bis({9-[(2-hydroxyethoxy)methyl]guanine}-N2-yl)methane.
CU = concentration of Acyclovir in the Sample solution h 2-{[2-(Acetylamino)-6-oxo-1,6-dihydro-9H-purin-9-yl]methoxy}ethyl
(mg/mL) acetate.▲ (RB 1-May-2023)
F = relative response factor (See Table 2)
SPECIFIC TESTS
e
Acceptance criteria: See Table 2. Reporting threshold is
0.03%. • WATER DETERMINATION á921ñ, Method I: NMT 6.0%
ADDITIONAL REQUIREMENTS
Table 2 • PACKAGING AND STORAGE: Preserve in tight containers, and
B

Relative Relative Acceptance store at room temperature. Protect from light and moisture.
Retention Response Criteria,
Name Time Factor NMT (%)
Change to read:
Guanine 0.40 1.0 0.7
• USP REFERENCE STANDARDS á11ñ
Hydroxyethyl USP Acyclovir RS
To

guaninea 0.74 1.0 0.2 ▲

USP Acyclovir Related Compound A RS
N -Isomer
7 b
0.94 0.43 0.1 2-[(2-Amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]
ethyl acetate.
Acyclovir 1.0 — —
C10H13N5O4 267.24
Specified impuri- USP Acyclovir Related Compound F RS
ty 1c 1.34 1.0 0.1 N-[9-[(2-Hydroxyethoxy)methyl]-6-oxo-6,9-dihydro- 1H-
Specified impuri- purin-2-yl]acetamide.
ty 2c 1.39 1.0 0.3 C10H13N5O4 267.24▲ (RB 1-May-2023)

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