Nevirapine

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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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 CAS RN^®: 129618-40-2; UNII: 99DK7FVK1H.

Hemihydrate             275.31

1 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. 

2 IDENTIFICATION

Change to read:

A. ^▲SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197K._{▲(CN 1-MAY-2020)} 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.

3 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

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 220 nm

Column: 4.6-mm x 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 = (r_U/r_S) × (C_S/C_U) × 100

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

C_S = concentration of USP Nevirapine Anhydrous RS in the Standard solution (mg/mL) 

C_U = concentration of Nevirapine in the Sample solution (mg/mL)

Acceptance criteria: 98.0%-102.0% on the anhydrous basis

4 IMPURITIES

4.1 RESIDUE ON IGNITION (281)

NMT 0.1%

4.2 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 = (r_U/r_S) × (C_S/C_U) × (1/F) × 100

r_U = peak response of each impurity from the Sample solution

r_S = peak response of nevirapine from the Standard solution 

C_S = concentration of USP Nevirapine Anhydrous RS in the Standard solution (µg/mL)

C_U = 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

5 SPECIFIC TESTS

WATER DETERMINATION, Method I(921).

For Nevirapine anhydrous: NMT 0.2%

For Nevirapine hemihydrate: 3.1%-3.9%

6 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)

USP Neviragine Anhydrous RS

USP Neviranine Hemihydrate RS

USP Neviranine Related Compound ARS

5,11-Dihydro-6^H-11-ethyl-4-methyl- dipyrido[3,2-b:2',3'-e][1,4]diazepin-6-one.

C₁₄H₁₄N₄O           254.29

USP Neviragine 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