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Printed by: Nguyen Nhan Official Date: Official as of 01-May-2020 Document Type: USP @2021 USPC
1

Acceptance criteria: 97.0%–102.0% on the anhydrous and

Abacavir Sulfate solvent-free basis
IMPURITIES
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 Time Solution A Solution B

2-Cyclopentene-1-methanol, 4-[2-amino-6-(cyclopropyl (min) (%) (%)
amino)-9H-purin-9-yl]-, (1S-cis)-, sulfate (salt) (2:1); 0 95 5
(1S,4R)-4-[2-Amino-6-(cyclopropylamino)-9H-purin-9-yl]-
2-cyclopentene-1-methanol sulfate (salt) (2:1) [188062- 20 70 30
50-2]. 35 10 90
DEFINITION 35.1 95 5
Abacavir Sulfate contains NLT 97.0% and NMT 102.0% of
(C14H18N6O)2 · H2SO4, calculated on the anhydrous and 50 95 5
solvent-free basis.

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System suitability solution: 0.25 mg/mL of USP Abacavir
IDENTIFICATION Related Compounds Mixture RS in water
Change to read: Sample solution: 0.25 mg/mL of Abacavir Sulfate in water
Chromatographic system
• A. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared ci (See Chromatography á621ñ, System Suitability.)
Spectroscopy: 197K▲ (CN 1-May-2020) Mode: LC
• B. The retention time of the major peak of the Sample Detector: UV 254 nm
solution corresponds to that of the System suitability Column: 3.9-mm × 15-cm; 5-µm packing L1
solution, obtained as directed in the test for Organic Column temperature: 30°
Impurities, Procedure 2. Flow rate: 1 mL/min
• C. IDENTIFICATION TESTS—GENERAL, Sulfate á191ñ Injection size: 20 µL
ffi
Sample solution: 5 mg/mL System suitability
Sample: System suitability solution
ASSAY Suitability requirements
• PROCEDURE Resolution: NLT 1.5 between abacavir and trans-abacavir
Mobile phase: Acetonitrile, phosphoric acid, and water Analysis
(20:1:180) Sample: Sample solution
O

Standard solution: 0.04 mg/mL of USP Abacavir Sulfate RS Calculate the percentage of each impurity in the portion of
in water Abacavir Sulfate taken:
Sample solution: 0.04 mg/mL of Abacavir Sulfate in water
Chromatographic system Result = (rU/rT) × 100
(See Chromatography á621ñ, System Suitability.)
Mode: LC rU = peak area of each impurity from the Sample
Detector: UV 254 nm solution
Column: 4.6-mm × 5-cm; 5-µm packing L1 rT = sum of the areas of all the peaks from the Sample
Column temperature: 30° solution
Flow rate: 1 mL/min
Injection size: 20 µL Acceptance criteria
System suitability Individual impurities: See Impurity Table 1.
Sample: Standard solution Total impurities: NMT 0.8%
Suitability requirements
Relative standard deviation: NMT 1.5% Impurity Table 1
Analysis Relative Acceptance
Samples: Standard solution and Sample solution Retention Criteria,
Calculate the percentage of (C14H18N6O)2 · H2SO4 in the Name Time NMT (%)
portion of Abacavir Sulfate taken: Descyclopropyl abacavira 0.65 0.2

Result = (rU/rS) × (CS/CU) × 100 Abacavir 1.00 —

trans-Abacavirb 1.04 0.2

rU = peak area of abacavir from the Sample solution
rS = peak area of abacavir from the Standard solution O-Pyrimidine derivative abacavirc 1.33 0.2
CS = concentration of USP Abacavir Sulfate RS in the t-Butyl derivative abacavird 1.67 0.2
Standard solution (mg/mL)
CU = concentration of Abacavir Sulfate in the Sample Any unspecified impurity — 0.1
solution (mg/mL) a [(1S,4R)-4-(2,6-Diamino-9H-purin-9-yl)cyclopent-2-enyl]methanol.
b {(1R,4R)-4-[2-Amino-6-(cyclopropylamino)-9H-purin-9-yl]-cyclopent-2-enyl}
methanol.

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c N6-Cyclopropyl-9-{(1R,4S)-4-[(2,5-diamino-6-chloropyrimidin-4-yloxy) Column temperature: 30°

methyl]cyclopent-2-enyl}-9H-purine-2,6-diamine. Injection size: 20 µL
d 9-[(1R,4S)-4-(tert-Butoxymethyl)cyclopent-2-enyl]-N6-cyclopropyl-9H-
System suitability
purine-2,6-diamine.
[NOTE—The relative retention times for trans-abacavir,
abacavir enantiomer, and abacavir are 0.8, 0.9, and 1.0,
• Procedure 2: Enantiomeric Purity
respectively.]
Solution A: Heptane, 2-propanol, and diethylamine
Sample: System suitability solution
(850:150:1).
Suitability requirements
Solution B: Heptane and 2-propanol (1:1)
Resolution: NLT 1.0 between trans-abacavir and abacavir
Mobile phase: See the gradient table below.
enantiomer; NLT 1.5 between abacavir enantiomer and
abacavir
Time Solution A Solution B Flow Rate Analysis
(min) (%) (%) (mL/min) Sample: Sample solution
0 100 0 1.0 Calculate the percentage of abacavir enantiomer in the
portion of Abacavir Sulfate taken:
25 100 0 1.0

27 0 100 0.8 Result = (rU/rT) × 100

37 0 100 0.8 rU = peak area of abacavir enantiomer from the Sample
39 100 0 1.0 solution
rT = total peak areas of abacavir and abacavir
55 100 0 1.0 enantiomer from the Sample solution

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Diluent: Methanol and trifluoroacetic acid (200:1) Acceptance criteria
System suitability solution: Transfer a quantity of USP Individual impurities: NMT 0.3% of abacavir enantiomer
Abacavir Stereoisomers Mixture RS to a suitable volumetric
SPECIFIC TESTS
flask, add a volume of Diluent equivalent to 30% of the final
• WATER DETERMINATION, Method Ic á921ñ: NMT 0.5%
volume, and sonicate until the solid is fully dissolved. Add a
volume of 2-propanol equivalent to about 30% of the final
volume, mix, and dilute with heptane to volume to obtain
0.4 mg/mL of USP Abacavir Stereoisomers Mixture RS.
ci ADDITIONAL REQUIREMENTS
• PACKAGING AND STORAGE: Preserve in well-closed
containers. Store at room temperature.
Sample solution: Transfer 4 mg of Abacavir Sulfate to a • USP REFERENCE STANDARDS á11ñ
10-mL volumetric flask. Add 3 mL of Diluent, and sonicate USP Abacavir Sulfate RS
ffi
until the solid is fully dissolved. Add 3 mL of 2-propanol, mix, USP Abacavir Stereoisomers Mixture RS
and dilute with heptane to volume. A mixture of abacavir sulfate, abacavir enantiomer, and
Chromatographic system trans-abacavir.
(See Chromatography á621ñ, System Suitability.) USP Abacavir Related Compounds Mixture RS
Mode: LC A mixture of abacavir glutarate, O-pyrimidine derivative
Detector: UV 286 nm abacavir, descyclopropyl abacavir, trans-abacavir, and
O

Column: 4.6-mm × 25-cm; 10-µm packing L51 t-butyl derivative abacavir.

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