Printed on: Wed Feb 08 2023, 11:18:38 PM(EST) Status: Currently Official on 09-Feb-2023 DocId: GUID-0F0CC075-551C-4DC0-BBDD-D317593E2324_3_en-US

Printed by: Dang Van Vu Official Date: Official as of 01-May-2020 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: iu9go DOI: https://doi.org/10.31003/USPNF_M2653_03_01
1

IMPURITIES
Abacavir Sulfate INORGANIC IMPURITIES
• Residue on Ignition á281ñ: NMT 0.2%
ORGANIC IMPURITIES
• Procedure 1: Related Compounds
Solution A: Trifluoroacetic acid and water (0.05: 99.95)
Solution B: Methanol and water (17:3)
Mobile phase: See the gradient table below.
(C14H18N6O)2 · H2SO4 670.74
2-Cyclopentene-1-methanol, 4-[2-amino-6-(cyclopropyl Time Solution A Solution B
amino)-9H-purin-9-yl]-, (1S-cis)-, sulfate (salt) (2:1); (min) (%) (%)
(1S,4R)-4-[2-Amino-6-(cyclopropylamino)-9H-purin-9-yl]-
2-cyclopentene-1-methanol sulfate (salt) (2:1) CAS RN®: 0 95 5
188062-50-2; UNII: J220T4J9Q2. 20 70 30
DEFINITION 35 10 90
Abacavir Sulfate contains NLT 97.0% and NMT 102.0% of
35.1 95 5
(C14H18N6O)2 · H2SO4, calculated on the anhydrous and
solvent-free basis. 50 95 5

IDENTIFICATION
System suitability solution: 0.25 mg/mL of USP Abacavir
Change to read: Related Compounds Mixture RS in water
Sample solution: 0.25 mg/mL of Abacavir Sulfate in water

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• A. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared Chromatographic system
Spectroscopy: 197K▲ (CN 1-May-2020) (See Chromatography á621ñ, System Suitability.)
• B. The retention time of the major peak of the Sample Mode: LC
solution corresponds to that of the System suitability Detector: UV 254 nm
solution, obtained as directed in the test for Organic Column: 3.9-mm × 15-cm; 5-µm packing L1
Impurities, Procedure 2.
• C. IDENTIFICATION TESTS—GENERAL, Sulfate á191ñ
Sample solution: 5 mg/mL
ci Column temperature: 30°
Flow rate: 1 mL/min
Injection size: 20 µL
ASSAY System suitability
• PROCEDURE Sample: System suitability solution
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Mobile phase: Acetonitrile, phosphoric acid, and water Suitability requirements
(20:1:180) Resolution: NLT 1.5 between abacavir and trans-abacavir
Standard solution: 0.04 mg/mL of USP Abacavir Sulfate RS Analysis
in water Sample: Sample solution
Sample solution: 0.04 mg/mL of Abacavir Sulfate in water Calculate the percentage of each impurity in the portion of
Chromatographic system Abacavir Sulfate taken:
O

(See Chromatography á621ñ, System Suitability.)

Result = (rU/rT) × 100
Mode: LC
Detector: UV 254 nm rU = peak area of each impurity from the Sample
Column: 4.6-mm × 5-cm; 5-µm packing L1 solution
Column temperature: 30° rT = sum of the areas of all the peaks from the Sample
Flow rate: 1 mL/min solution
Injection size: 20 µL
System suitability Acceptance criteria
Sample: Standard solution Individual impurities: See Impurity Table 1.
Suitability requirements Total impurities: NMT 0.8%
Relative standard deviation: NMT 1.5%
Analysis Impurity Table 1
Samples: Standard solution and Sample solution
Relative Acceptance
Calculate the percentage of (C14H18N6O)2 · H2SO4 in the Retention Criteria,
portion of Abacavir Sulfate taken: Name Time NMT (%)

Result = (rU/rS) × (CS/CU) × 100 Descyclopropyl abacavira 0.65 0.2

Abacavir 1.00 —
rU = peak area of abacavir from the Sample solution
trans-Abacavirb 1.04 0.2
rS = peak area of abacavir from the Standard solution
CS = concentration of USP Abacavir Sulfate RS in the O-Pyrimidine derivative abacavirc 1.33 0.2
Standard solution (mg/mL)
t-Butyl derivative abacavird 1.67 0.2
CU = concentration of Abacavir Sulfate in the Sample
solution (mg/mL) Any unspecified impurity — 0.1

a [(1S,4R)-4-(2,6-Diamino-9H-purin-9-yl)cyclopent-2-enyl]methanol.
Acceptance criteria: 97.0%–102.0% on the anhydrous and
b {(1R,4R)-4-[2-Amino-6-(cyclopropylamino)-9H-purin-9-yl]-cyclopent-2-enyl}
solvent-free basis
methanol.
c N6-Cyclopropyl-9-{(1R,4S)-4-[(2,5-diamino-6-chloropyrimidin-4-yloxy)
methyl]cyclopent-2-enyl}-9H-purine-2,6-diamine.

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Printed on: Wed Feb 08 2023, 11:18:38 PM(EST) Status: Currently Official on 09-Feb-2023 DocId: GUID-0F0CC075-551C-4DC0-BBDD-D317593E2324_3_en-US
Printed by: Dang Van Vu Official Date: Official as of 01-May-2020 Document Type: USP @2023 USPC
Do Not Distribute DOI Ref: iu9go DOI: https://doi.org/10.31003/USPNF_M2653_03_01
2

d 9-[(1R,4S)-4-(tert-Butoxymethyl)cyclopent-2-enyl]-N6-cyclopropyl-9H- Injection size: 20 µL

purine-2,6-diamine. System suitability
[NOTE—The relative retention times for trans-abacavir,
• Procedure 2: Enantiomeric Purity abacavir enantiomer, and abacavir are 0.8, 0.9, and 1.0,
Solution A: Heptane, 2-propanol, and diethylamine respectively.]
(850:150:1). Sample: System suitability solution
Solution B: Heptane and 2-propanol (1:1) Suitability requirements
Mobile phase: See the gradient table below. Resolution: NLT 1.0 between trans-abacavir and abacavir
enantiomer; NLT 1.5 between abacavir enantiomer and
Time Solution A Solution B Flow Rate abacavir
(min) (%) (%) (mL/min) Analysis
0 100 0 1.0
Sample: Sample solution
Calculate the percentage of abacavir enantiomer in the
25 100 0 1.0 portion of Abacavir Sulfate taken:
27 0 100 0.8
Result = (rU/rT) × 100
37 0 100 0.8

39 100 0 1.0
rU = peak area of abacavir enantiomer from the Sample
solution
55 100 0 1.0 rT = total peak areas of abacavir and abacavir
enantiomer from the Sample solution
Diluent: Methanol and trifluoroacetic acid (200:1)
Acceptance criteria

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System suitability solution: Transfer a quantity of USP
Abacavir Stereoisomers Mixture RS to a suitable volumetric Individual impurities: NMT 0.3% of abacavir enantiomer
flask, add a volume of Diluent equivalent to 30% of the final SPECIFIC TESTS
volume, and sonicate until the solid is fully dissolved. Add a • WATER DETERMINATION, Method Ic á921ñ: NMT 0.5%
volume of 2-propanol equivalent to about 30% of the final
volume, mix, and dilute with heptane to volume to obtain ADDITIONAL REQUIREMENTS
0.4 mg/mL of USP Abacavir Stereoisomers Mixture RS.
Sample solution: Transfer 4 mg of Abacavir Sulfate to a
10-mL volumetric flask. Add 3 mL of Diluent, and sonicate
ci • PACKAGING AND STORAGE: Preserve in well-closed
containers. Store at room temperature.
• USP REFERENCE STANDARDS á11ñ
until the solid is fully dissolved. Add 3 mL of 2-propanol, mix, USP Abacavir Sulfate RS
and dilute with heptane to volume. USP Abacavir Stereoisomers Mixture RS
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Chromatographic system A mixture of abacavir sulfate, abacavir enantiomer, and
(See Chromatography á621ñ, System Suitability.) trans-abacavir.
Mode: LC USP Abacavir Related Compounds Mixture RS
Detector: UV 286 nm A mixture of abacavir glutarate, O-pyrimidine derivative
Column: 4.6-mm × 25-cm; 10-µm packing L51 abacavir, descyclopropyl abacavir, trans-abacavir, and
Column temperature: 30° t-butyl derivative abacavir.
O

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