Nevirapine
C15H14N4O 266.30
6H-Dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one, 11-cyclopropyl-5,11-dihydro-4-methyl-;
11-Cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one
[129618-40-2].
Hemihydrate 275.31
(ne vir' a peen).
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C15H14N4O 266.30
6H-Dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one, 11-cyclopropyl-5,11-dihydro-4-methyl-;
11-Cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one
[129618-40-2].Hemihydrate 275.31
DEFINITION
Nevirapine is anhydrous or contains one-half molecule of water of hydration. It contains NLT 98.0% and NMT 102.0% of C15H14N4O, calculated on the anhydrous basis.
IDENTIFICATION
• A. Infrared Absorption 
197K
:
Do not dry the specimens.
197K:
Do not dry the specimens.
• B.
The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
ASSAY
• Procedure
Buffer:
0.025 M of monobasic ammonium phosphate in water prepared as follows. Dissolve 2.9 g of monobasic ammonium phosphate in 800 mL of water. Adjust with 1 N sodium hydroxide to a pH of 5.0, and dilute with water to 1000 mL.
Mobile phase:
Acetonitrile and Buffer (1:4)
Standard stock solution A:
0.24 mg/mL of USP Nevirapine Anhydrous RS prepared as follows. Dissolve a quantity of USP Nevirapine Anhydrous RS in Acetonitrile and Mobile phase (1:20). Sonicate for at least 15 min, allow to cool to room temperature, and dilute with Mobile phase to volume. [Note—Do not use after 78 h. ]
Standard stock solution B:
0.24 mg/mL of USP Nevirapine Related Compound A RS prepared as follows. Dissolve a quantity of USP Nevirapine Related Compound A RS in a volume of a mixture of acetonitrile and Mobile phase (1:3). Sonicate for at least 15 min, allow to cool to room temperature, and dilute with Mobile phase to volume.
Standard stock solution C:
0.06 mg/mL of USP Nevirapine Related Compound B RS prepared as follows. Dissolve a quantity of USP Nevirapine Related Compound B RS in a volume of a mixture of acetonitrile and Mobile phase (10:22). Sonicate for at least 30 min, allow to cool to room temperature, and dilute with Mobile phase to volume.
System suitability solution:
0.03 mg/mL each of USP Nevirapine Anhydrous RS and USP Nevirapine Related Compound A RS and 0.015 mg/mL of USP Nevirapine Related Compound B RS from suitable volumes of Standard stock solution A, Standard stock solution B, and Standard stock solution C, respectively, in Mobile phase. [Note—Do not use after 78 h. ]
Standard solution:
0.03 mg/mL of USP Nevirapine Anhydrous RS in Mobile phase from Standard stock solution A. [Note—Do not use after 78 h. ]
Sample stock solution:
0.24 mg/mL of Nevirapine in Mobile phase prepared as follows. Transfer the required amount of Nevirapine to a suitable volumetric flask, and add 4% of the final volume with acetonitrile and 80% of the final volume with Mobile phase. Sonicate for at least 15 min, allow to cool to room temperature, and dilute with Mobile phase to volume.
Sample solution:
0.03 mg/mL of nevirapine anhydrous in Mobile phase from the Sample stock solution
Chromatographic system
Mode:
LC
Detector:
UV 220 nm
Column:
4.6-mm × 15-cm; 5-µm packing L60
Column temperature:
35
Flow rate:
1 mL/min
Injection size:
25 µL
System suitability
Samples:
System suitability solution and Standard solution
[Note—The relative retention times are shown in Table 1. ]
Suitability requirements
Resolution:
NLT 5.0 between nevirapine related compound B and nevirapine, and NLT 7.4 between nevirapine and nevirapine related compound A; System suitability solution
Relative standard deviation:
NMT 2.0%, Standard solution
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of nevirapine (C15H14N4O) in the portion of Nevirapine taken:
Result = (rU/rS) × (CS/CU) × 100
| rU | = | = peak response from the Sample solution |
| rS | = | = peak response from the Standard solution |
| CS | = | = concentration of USP Nevirapine Anhydrous RS in the Standard solution (mg/mL) |
| CU | = | = concentration of Nevirapine in the Sample solution (mg/mL) |
Acceptance criteria:
98.0%–102.0% on the anhydrous basis
IMPURITIES
• Residue on Ignition 281:
NMT 0.1%
281:
NMT 0.1%
• Heavy Metals, Method II 231:
NMT 10 ppm
231:
NMT 10 ppm
• Organic Impurities
Buffer, Mobile phase, System suitability solution, and Standard stock solution A:
Proceed as directed in the Assay.
Standard solution:
0.2 µg/mL of USP Nevirapine Anhydrous RS in Mobile phase from Standard stock solution A
Sample solution:
0.24 mg/mL of Nevirapine in Mobile phase prepared as follows. Transfer the required amount of Nevirapine to a suitable volumetric flask, and add 4% of the final volume with acetonitrile and 80% of the final volume with Mobile phase. Sonicate for at least 15 min, allow to cool to room temperature, and dilute with Mobile phase to volume.
Chromatographic system:
Proceed as directed in the Assay, except for Run time and Injection size.
Run time:
At least 80 min
Injection size:
25 µL for System suitability; 50 µL for Analysis
System suitability
Samples:
System suitability solution and Standard solution
[Note—The relative retention times are shown in Table 1. ]
Suitability requirements
Resolution:
NLT 5.0 between nevirapine related compound B and nevirapine, and NLT 7.4 between nevirapine and nevirapine related compound A; System suitability solution
Relative standard deviation:
NMT 5.0%, Standard solution
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of each impurity in the portion of Nevirapine taken:
Result = (rU/rS) × (CS/CU) × (1/F) × 100
| rU | = | = peak response of each impurity from the Sample solution |
| rS | = | = peak response of nevirapine from the Standard solution |
| CS | = | = concentration of USP Nevirapine Anhydrous RS in the Standard solution (µg/mL) |
| CU | = | = concentration of nevirapine anhydrous in the Sample solution (µg/mL) |
| F | = | = relative response factor for each impurity (see Table 1) |
Acceptance criteria
See Table 1.
Table 1
| Name | Relative Retention Time |
Relative Response Factor |
Acceptance Criteria, NMT (%) |
|---|---|---|---|
| Nevirapine related compound B | 0.7 | 1.3 | 0.2 |
| Nevirapine | 1.0 | 1.0 | — |
| Nevirapine related compound A | 1.5 | 1.0 | 0.2 |
| Nevirapine impurity C | 2.8 | 1.0 | 0.2 |
| Any other individual unspecified impurity | — | 1.0 | 0.1 |
| Total impurities | — | — | 0.6 |
SPECIFIC TESTS
• Water Determination, Method I 921
921
For Nevirapine anhydrous:
NMT 0.2%
For Nevirapine hemihydrate:
3.1%–3.9%
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight containers, and store at controlled room temperature.
• Labeling:
Label to indicate whether it is anhydrous or the hemihydrate.
• USP Reference Standards 11
11
USP Nevirapine Anhydrous RS 
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USP Nevirapine Hemihydrate RS
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USP Nevirapine Related Compound A RS
5,11-Dihydro-6H-11-ethyl-4-methyl-dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one.
C14H14N4O 254.29
5,11-Dihydro-6H-11-ethyl-4-methyl-dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one.
C14H14N4O 254.29
USP Nevirapine Related Compound B RS
5,11-Dihydro-4-methyl-6H-dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one.
C12H10N4O 226.23
5,11-Dihydro-4-methyl-6H-dipyrido[3,2-b:2¢,3¢-e][1,4]diazepin-6-one.
C12H10N4O 226.23
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 |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 4037
Pharmacopeial Forum: Volume No. 30(1) Page 136