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Lamivudine and Zidovudine Tablets

DEFINITION
Lamivudine and Zidovudine Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of
lamivudine (C8H11 N3O3S) and zidovudine (C10 H13 N5O4 ).

IDENTIFICATION
• A. The retention times of the lamivudine and zidovudine peaks of the Sample solution correspond
to those of the Standard solution, as obtained in the Assay.

ASSAY
• P
Solution A: 25 mM of ammonium acetate. Adjust with glacial acetic acid to a pH of 4.0.
Solution B: Methanol
Solution C: Acetonitrile
Mobile phase: See Table 1.
Table 1
Time Solution A Solution B Solution C
(min) (%) (%) (%)

0 95 5 0

15.0 95 5 0

30.0 70 30 0

38.0 70 30 0

38.1 0 0 100

45.0 0 0 100

45.1 95 5 0

60.0 95 5 0

Diluent: Solution A and Solution B (19:1)

System suitability solution: 0.17 mg/mL of USP Lamivudine Resolution Mixture B RS in
Diluent
Standard solution: 0.15 mg/mL of USP Lamivudine RS and 0.30 mg/mL of USP Zidovudine RS
in Diluent
Sample stock solution: Transfer a counted number of Tablets, equivalent to 1500 mg of
zidovudine and 750 mg of lamivudine, into a 500-mL volumetric flask. Add 250 mL of water,
and disintegrate completely by shaking for a minimum of 15 min. Dilute with water to volume,
and mix.
Sample solution: Pass a portion of the Sample stock solution through a filter of 0.45-µm pore
size, discarding the first 2–3 mL. Further dilute the filtrate to obtain 0.15 mg/mL of lamivudine
and 0.30 mg/mL of zidovudine in Diluent.
Chromatographic system
(See Chromatography 〈621〉 , System Suitability.)
Mode: LC

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Detector: UV 270 nm
Column: 3-mm × 25-cm; packing L1
Flow rate: 0.5 mL/min
Injection volume: 10 µL
System suitability
Samples: System suitability solution and Standard solution
[N — The relative retention times for lamivudine diastereomer and lamivudine are 0.50 and
0.52, respectively. ]
Suitability requirements
Resolution: NLT 1.5 between lamivudine diastereomer and lamivudine, System suitability
solution
Relative standard deviation: NMT 2.0% for zidovudine and lamivudine, Standard solution
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of lamivudine (C8H11N3O3S) and zidovudine
(C10H13N5O4 ) in the portion of Tablets taken:

Result = ( r U /r S )×(C S /C U ) × 100

r U = peak response of zidovudine or lamivudine from the Sample solution

r S = peak response of zidovudine or lamivudine from the Standard solution
C = concentration of USP Zidovudine RS or USP Lamivudine RS in the Standard solution
S (mg/mL)
C = nominal concentration of zidovudine or lamivudine in the Sample solution (mg/mL)
U

Acceptance criteria: 90.0%–110.0%

PERFORMANCE TESTS
• D 〈711〉
Test 1
Medium: 0.1 N hydrochloric acid; 900 mL, deaerated
Apparatus 2: 75 rpm
Time: 15 min
Lamivudine response factor solutions: 0.167 mg/mL of USP Lamivudine RS in Medium.
[N — Prepare in duplicate. ]
Zidovudine response factor solutions: 0.333 mg/mL of USP Zidovudine RS in Medium.
[N — Prepare in duplicate. ]
Sample solution: Pass a portion of the solution under test through a suitable filter (PTFE,
PVDF, or equivalent) of 0.45-µm pore size.
Detector: UV 240–300 nm
Blank: Medium
Cell length: 0.02-cm flowcell
Analysis: The calculations of the percentages dissolved are done using a multicomponent
analysis software.
Tolerances: NLT 85% (Q) of the labeled amount of zidovudine and lamivudine is dissolved.
Test 2: If the product complies with this test, the labeling indicates that it meets USP
Dissolution Test 2.

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Medium: 0.1 N hydrochloric acid; 900 mL

Apparatus 2: 75 rpm
Time: 30 min
Buffer solution: 7.7 g/L of ammonium acetate in water
Mobile phase: Acetonitrile and Buffer solution (1:9)
Standard stock solution: 1.4 mg/mL of USP Lamivudine RS and 2.8 mg/mL of USP
Zidovudine RS in Medium. A small amount of methanol, NMT 20% of the final volume, can be
used to dissolve both compounds.
Standard solution: 0.168 mg/mL of lamivudine and 0.336 mg/mL of zidovudine in Medium
from the Standard stock solution
Sample solution: Pass a portion of the solution under test through a suitable filter.
Chromatographic system
(See Chromatography 〈621〉 , System Suitability.)
Mode: LC
Detector: UV 270 nm
Column: 4.6-mm × 15-cm; packing L1
Column temperature: 40°
Flow rate: 1.2 mL/min
Injection volume: 10 µL
System suitability
Sample: Standard solution
Suitability requirements
Column efficiency: NLT 1500 theoretical plates for lamivudine and NLT 3000 theoretical
plates for zidovudine
Tailing factor: NMT 2.0 for lamivudine and zidovudine
Relative standard deviation: NMT 2.0% for zidovudine and lamivudine
Calculate the percentages of lamivudine (C8H11N3O3S) and zidovudine (C10H13N5O4 ) dissolved:

Result = ( r U /r S )×(C S /L) × V × 100

r U = peak response of lamivudine or zidovudine from the Sample solution

r S = peak response of lamivudine or zidovudine from the Standard solution
C S = concentration of lamivudine or zidovudine in the Standard solution (mg/mL)
L = label claim for lamivudine or zidovudine (mg/Tablet)
V = volume of Medium, 900 mL
Tolerances: NLT 80% (Q) of the labeled amount of zidovudine and lamivudine is dissolved.
• U D U 〈905〉 : Meet the requirements for zidovudine and lamivudine

IMPURITIES
• O I
Solution A, Solution B, Solution C, Mobile phase, Diluent, System suitability solution,
Sample solution, Chromatographic system, and System suitability: Proceed as directed
in the Assay.
Analysis
Sample: Sample solution
Calculate the percentage of each lamivudine related impurity in the portion of Tablets taken:

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Result = ( r U /r T ) × 100

r = peak response of each individual impurity from the Sample solution

U

r = sum of the peak responses of lamivudine and all lamivudine related impurities from
T the Sample solution
Calculate the percentage of each zidovudine related impurity and unspecified impurity in the
portion of Tablets taken:
Result = ( r U /r T ) × (1/F) × 100

r = peak response of each individual impurity from the Sample solution

U

r = sum of the peak responses of zidovudine, all zidovudine related impurities, and
T unspecified impurities from the Sample solution
F = relative response factor (see Table 2)
Acceptance criteria: See Table 2.
Table 2
Relative Relative Acceptance
Retention Response Criteria,
Name Time Factor NMT (%)

Lamivudine-(cytosine)a 0.11 1.0 — b

Lamivudine-(uracil)c 0.14 1.0 — b

Lamivudine-(carboxylic
acid)d 0.17 1.0 0.3

Lamivudine-(S-sulfoxide)e 0.20 1.0 — b

Lamivudine-(R-sulfoxide)f 0.22 1.0 — b

Zidovudine related
compound Cg 0.27 1.7 1.5

Lamivudine
diastereomerh 0.50 1.0 0.2

Lamivudine 0.52 — —

Zidovudine-(thymidine)i 0.60 1.0 — b

Lamivudine-(uracil b
—
derivative)j 0.70 1.0

Lamivudine-(salicylic acid)k 0.80 1.0 — b

Zidovudine 1.00 — —

Zidovudine related b
—
compound Bl 1.10 1.0

Any individual unspecified

—
impurity 1.0 0.1

Total lamivudine
related impurities
(the limit includes — —
all lamivudine related
impurities) 0.6

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Relative Relative Acceptance

Retention Response Criteria,
Name Time Factor NMT (%)

Total zidovudine
related impurities
(the limit includes — —
individual unspecified
impurities) 2.0

a
4-Aminopyrimidin-2(1H )-one.
bThe individual impurity limit is not included because these are process/other impurities
monitored individually in the drug substances.
c
Pyrimidine-2,4(1H ,3H )-dione.
d
(2R ,5S )-5-(4-Amino-2-oxopyrimidin-1(2H )-yl)-1,3-oxathiolane-2-carboxylic acid (2R
,5S )-5-(cytosine-1-yl)-1,3-oxathiolane-2-carboxylic acid.
e 1-[(2R ,3S ,5SS)-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine S-oxide.
f 1-[(2R ,3R ,5S )-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine S-oxide.
g 5-Methylpyrimidine-2,4(1H ,3H )-dione.
h
1-[(2S ,5S )-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine.
i [1-(2-Deoxy-β- -ribofuranosyl)]thymine.
j
(2RS ,5SR )1-[(2R ,5S )-2-(Hydroxymethyl)-1,3-oxathiolan-5-yl]uracil.
k 2-Hydroxybenzoic acid.
l 3'-Chloro-3'-deoxythymidine.

ADDITIONAL REQUIREMENTS
• P S : Preserve in well-closed containers, protected from light, and store
between 2° and 30°.
• L : When more than one Dissolution test is given, the labeling states the Dissolution test
used only if Test 1 is not used.
• USP R S 〈11〉
USP Lamivudine RS
USP Lamivudine Resolution Mixture B RS
USP Zidovudine RS

Auxiliary Information— Please check for your question in the FAQs before contacting USP.
Topic/Question Contact Expert Committee

Monograph Shankari Shivaprasad, (CHM12015) Chemical Medicines

Ph.D. Monographs 1
Scientific Liaison
(301) 230-7426

〈711〉 Margareth R.C. Marques, (GCDF2015) General Chapters-

Ph.D. Dosage Forms 2015
Principal Scientific Liaison
(301) 816-8106

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Topic/Question Contact Expert Committee

Reference RS Technical Services

Standards 1-301-816-8129
rstech@usp.org
USP39–NF34 Page 4484
Pharmacopeial Forum: Volume No. 38(6)

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