Ribavirin Tablets
DEFINITION
Ribavirin Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of ribavirin (C8H12N4O5).
IDENTIFICATION
• The retention time of the ribavirin peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
ASSAY
• Procedure
Buffer:
4.0 g/L of sodium dihydrogen phosphate dihydrate in water. Adjust with 5% sodium hydroxide solution to a pH of 5.0 ± 0.05. Pass the solution through a suitable filter of 0.45-µm pore size.
Mobile phase:
Acetonitrile and Buffer (1:49)
Diluent:
Acetonitrile and water (3:7)
Standard stock solution:
0.6 mg/mL of USP Ribavirin RS in Diluent
Standard solution:
0.03 mg/mL of USP Ribavirin RS in Mobile phase from the Standard stock solution
Sample stock solution:
Transfer a portion of ribavirin, equivalent to 1000 mg of ribavirin from finely powdered Tablets (NLT 10), to a 1000-mL volumetric flask. Add about 750 mL of Diluent, and sonicate with occasional shaking for 30 min. Cool to room temperature, dilute with Diluent to volume, and mix. Centrifuge and decant the supernatant.
Sample solution:
0.03 mg/mL of ribavirin in Mobile phase from the Sample stock solution. Pass the solution through a suitable filter of 0.45-µm pore size.
Chromatographic system
Mode:
LC
Detector:
UV 207 nm
Column:
4.6-mm × 25-cm; 5-µm packing L1
Flow rate:
1 mL/min
Injection size:
20 µL
Run time:
10 min
System suitability
Sample:
Standard solution
Suitability requirements
Column efficiency:
NLT 2000 theoretical plates
Tailing factor:
NLT 2.0
Relative standard deviation:
NMT 2.0%
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of C8H12N4O5 in the portion of Tablets taken:
Result = (rU/rS) × (CS/CU) × 100
| rU | = | = peak response of ribavirin from the Sample solution |
| rS | = | = peak response of ribavirin from the Standard solution |
| CS | = | = concentration of USP Ribavirin RS in the Standard solution (mg/mL) |
| CU | = | = nominal concentration of ribavirin in the Sample solution (mg/mL) |
Acceptance criteria:
90.0%–110.0%
PERFORMANCE TESTS
• Dissolution 
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711
Medium:
Water; 900 mL
Apparatus 2:
50 rpm
Time:
30 min
Buffer and Mobile phase:
Proceed as directed in the Assay.
Standard solution:
0.22 mg/mL of USP Ribavirin RS in Medium
Sample solution:
Pass the solution through a suitable filter of 0.45-µm pore size.
Chromatographic system
Mode:
LC
Detector:
UV 225 nm
Column:
4.6-mm × 25-cm; 5-µm packing L1
Flow rate:
1 mL/min
Injection size:
10 µL
System suitability
Sample:
Standard solution
Suitability requirements
Column efficiency:
NLT 2000 theoretical plates
Tailing factor:
NMT 2.0
Relative standard deviation:
NMT 2.0%
Analysis
Calculate the percentage of C8H12N4O5 dissolved:
Result = (rU/rS) × (CS/L) × V × 100
| rU | = | = peak response of ribavirin from the Sample solution |
| rS | = | = peak response of ribavirin from the Standard solution |
| CS | = | = concentration of USP Ribavirin RS in the Standard solution (mg/mL) |
| L | = | = label claim (mg/Tablet) |
| V | = | = volume of Medium, 900 mL |
Tolerances:
NLT 80% (Q) of the labeled amount of ribavirin is dissolved.
• Uniformity of Dosage Units 905:
Meet the requirements
905:
Meet the requirements
IMPURITIES
Organic Impurities
[Note—If uracil and/or uridine are known impurities, Procedure 2 is recommended. ]
• Procedure 1
Solution A:
3.4 g/L of potassium dihydrogen phosphate in water. Adjust with 5% potassium hydroxide solution to a pH of 5.00 ± 0.05. Pass the solution through a suitable filter of 0.45-µm pore size.
Solution B:
Acetonitrile
Mobile phase:
See the gradient table below.
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 100 | 0 |
| 30 | 90 | 10 |
| 40 | 75 | 25 |
| 50 | 50 | 50 |
| 55 | 50 | 50 |
| 56 | 100 | 0 |
| 70 | 100 | 0 |
Standard stock solution:
0.4 mg/mL of USP Ribavirin RS in Solution A
Standard solution:
1 µg/mL of USP Ribavirin RS in Solution A from the Standard stock solution
Sample solution:
Transfer a portion of ribavirin, equivalent to 100 mg of ribavirin from finely powdered Tablets (NLT 20), to a 200-mL volumetric flask. Add about 150 mL of Solution A, and sonicate with occasional shaking for 15 min. Cool to room temperature, dilute with Solution A to volume, and mix. Pass the solution through a suitable filter of 0.45-µm pore size.
Chromatographic system
Mode:
LC
Detector:
UV 220 nm
Column:
4.6-mm × 25-cm; 5-µm packing L1
Flow rate:
1 mL/min
Injection size:
20 µL
Run time:
70 min. [Note—Data collection is only for the first 55 min. The remaining gradient steps re-equilibrate the column. ]
System suitability
Sample:
Standard solution
Suitability requirements
Relative standard deviation:
NMT 5.0%
Analysis
Samples:
Standard solution and Sample solution
[Note—Impurities are listed in Impurity Table 1. ]
Calculate the percentage of any unknown impurity in the portion of Tablets taken:
Result = (rU/rS) × (CS/CU) × 100
| rU | = | = peak response of any unknown impurity from the Sample solution |
| rS | = | = peak response of ribavirin from the Standard solution |
| CS | = | = concentration of USP Ribavirin RS in the Standard solution (mg/mL) |
| CU | = | = nominal concentration of ribavirin in the Sample solution (mg/mL) |
Acceptance criteria
Individual unknown impurity:
NMT 0.10%. [Note—Disregard any peak area less than 0.05%. ]
Total impurities:
NMT 0.30%
Impurity Table 1
| Name | Relative Retention Time |
|---|---|
| Triazole acida | 0.35 |
| Ribavirin acidb | 0.40 |
| Triazole amidec | 0.64 |
| Ribavirin | 1.0 |
| Ribavirin 5-isomerd | 1.37 |
| Ribavirin methyl estere | 2.09 |
| Ribavirin 5'-acetylf | 2.43 |
| Ribavirin 5'-benzoylg | 4.83 |
|
[Note—These are process impurities listed for information only. ]
|
|
|
a
1H-1,2,4-Triazole-3-carboxylic acid.
b
1-
 -d-Ribofuranosyl-1H-1,2,4-triazole-3-carboxylic acid.
c
1H-1,2,4-Triazole-3-carboxamide.
d
1-
-d-Ribofuranosyl-1H-1,2,4-triazole-3-carboxamide.
e
Methyl 1-
-d-Ribofuranosyl-1H-1,2,4-triazole-3-carboxylate.
f
1-(5-O-Acetyl-
-d-ribofuranosyl)-1H-1,2,4-triazole-3-carboxamide.
g
1-(5-O-Benzoyl-
-d-ribofuranosyl)-1H-1,2,4-triazole-3-carboxamide.
|
|
• Procedure 2
Buffer:
3.0 g/L of dibasic potassium phosphate in water. Adjust with phosphoric acid to a pH of 6.0 ± 0.1. Pass the solution through a suitable filter of 0.45-µm pore size.
Mobile phase:
Methanol and Buffer (1:39)
Standard stock solution:
1 mg/mL of USP Ribavirin RS in water. [Note—Sonicate with occasional shaking to dissolve the solids. ]
Standard solution:
0.01 mg/mL of USP Ribavirin RS in water from the Standard stock solution
Sensitivity solution:
0.5 µg/mL of USP Ribavirin RS from the Standard solution in water
Sample solution:
1.0 mg/mL. Transfer a portion of ribavirin, equivalent to 1000 mg of ribavirin from finely powdered Tablets (NLT 20), to a 1000-mL volumetric flask. Add about 500 mL of water, and sonicate with occasional shaking for 15 min. Shake the solution for 15 min, and cool to room temperature. Dilute with water to volume, and centrifuge the solution for 10 min.
Chromatographic system
Mode:
LC
Detector:
UV 207 nm
Column:
4.6-mm × 25-cm; 5-µm packing L1
Column temperature:
30
Flow rate:
1 mL/min
Injection size:
10 µL
Run time:
NLT 4.3 times the retention time of the ribavirin peak
System suitability
Samples:
Standard solution and Sensitivity solution
Suitability requirements
Signal-to-noise ratio:
NLT 10, Sensitivity solution
Relative standard deviation:
NMT 2.0%, Standard solution
Analysis
Samples:
Standard solution and Sample solution
[Note—Impurities are listed in Impurity Table 2. ]
Calculate the percentage of any impurity in the portion of Tablets taken:
Result = (rU/rS) × (CS/CU) × (1/F) × 100
| rU | = | = peak response of any impurity from the Sample solution |
| rS | = | = peak response of ribavirin from the Standard solution |
| CS | = | = concentration of USP Ribavirin RS in the Standard solution (mg/mL) |
| CU | = | = nominal concentration of ribavirin in the Sample solution (mg/mL) |
| F | = | = relative response factor (see Impurity Table 2) |
Acceptance criteria
Individual impurities:
See Impurity Table 2 below.
Total impurities:
NMT 1.0%
Impurity Table 2
| Name | Relative Retention Time |
Relative Response Factor |
Acceptance Criteria, NMT (%) |
|---|---|---|---|
| Ribavirin acida | 0.55 | 0.98 | 0.25 |
| Triazole amideb | 0.73 | 1.1 | 0.25 |
| Uracilc | 0.89 | 1.6 | 0.25 |
| Ribavirin | 1.00 | — | — |
| Uridined | 1.71 | 1.0 | 0.25 |
| Any other individual impuritye | — | 1.0 | 0.17 |
|
a
1-
-d-Ribofuranosyl-1H-1,2,4-triazole-3-carboxylic acid.
b
1H-1,2,4-Triazole-3-carboxamide.
c
Pyrimidine-2,4(1H,3H)-dione.
d
1-
-d-Ribofuranosylpyrimidine-2,4(1H,3H)-dione.
e
[Note—Disregard any peak area less than 0.05%. ]
|
|||
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight containers. Store between 15 and 30.
and 30.
• Labeling:
If a test for Organic Impurities other than Procedure 1 is used, the labeling states with which Organic Impurities test the article complies.
• USP Reference Standards 11
11
USP Ribavirin RS 
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Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Leonel M. Santos, Ph.D.
Senior Scientific Liaison 1-301-816-8168 |
(SM12010) Monographs - Small Molecules 1 |
| 711 |
Margareth R.C. Marques, Ph.D.
Senior Scientific Liaison 1-301-816-8106 |
(GCDF2010) General Chapters - Dosage Forms |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 4544