Zanamivir

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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C₁₂H₂₀N₄O₇      332.31

D-glycero-d-galacto-Non-2-enonic acid, 5-(acetylamino)-4-[(aminoiminomethyl)amino]-2,6-anhydro-3,4,5-trideoxy-;

5-Acetamido-2,6-anhydro-3,4,5-trideoxy-4-guanidino-d-glycero-d-galacto-non-2-enonic acid CAS RN: 139110-80-8; UNII: L6O3XI777I.

1 DEFINITION

Zanamivir contains NLT 98.0% and NMT 102.0% of zanamivir (C₁₂H₂₀N₄O₇), calculated on the anhydrous and solvent-free basis.

2 IDENTIFICATION

Change to read:

A. Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197K or 197M _{(CN 1-May-2020)}

Wavenumber range: 4000 cm−1 to 400 cm−1

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

Mobile phase: Acetonitrile and 7.5 mM sulfuric acid (60:40). Adjust with ammonia TS to a pH of 6.2.

Resolution solution: Prepare 2.5 µg/mL of talo-zanamivir and 0.05 mg/mL of zanamivir from USP Zanamivir Resolution Mixture RS in Mobile phase.

Standard solution: 0.045 mg/mL of USP Zanamivir RS in Mobile phase

Sample solution: 0.045 mg/mL of Zanamivir in Mobile phase

Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 234 nm

Column: 4.6-mm × 25-cm; 5-µm packing L82

Column temperature: 30°

Flow rate: 1.5 mL/min

Injection volume: 20 µL

System suitability

Samples: Resolution solution and Standard solution

Suitability requirements

Resolution: NLT 1.5 between talo-zanamivir and zanamivir, Resolution solution

Relative standard deviation: NMT 1.5%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of zanamivir (C₁₂H₂₀N₄O₇) in the portion of Zanamivir 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 Zanamivir RS in the Standard solution (mg/mL)

C_u = concentration of Zanamivir in the Sample solution (mg/mL)

Acceptance criteria: 98.0%–102.0% on the anhydrous and solvent-free basis

4 IMPURITIES

ORGANIC IMPURITIES

Mobile phase: Proceed as directed in the Assay.

System suitability solution: Prepare 0.45 mg/mL of zanamivir and 9 µg/mL each of imidazole, imidazole carboximidamide, zanamivir Urea analog, 4-amino zanamivir, 4-biguanide zanamivir, and talo-zanamivir from USP Zanamivir Related Compounds Mixture RS in Mobile phase.

Sample solution: 0.45 mg/mL of Zanamivir prepared as follows. Dissolve the sample using 40% of the final volume with water, and dilute with acetonitrile to volume.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 210 nm and 234 nm

Column: 4.6-mm x 25-cm; 5-µm packing L82

Column temperature: 30^o

Flow rate: 1.5 mL/min

Injection volume: 20 µL

System suitability

Sample: System suitability solution

Suitability requirements

Resolution: NLT 1.5 between the peaks of talo-zanamivir and zanamivir at. 234 nm

Analysis

Sample: Sample solution

Calculate the percentage of imidazole in the portion of Zanamivir taken:

Result = [(r_i /F)/(r_i /F + r_z )] × 100

r_i = peak response of imidazole at 210 nm

F = relative response factor for imidazole (see Table 1)

r_z = peak response of zanamivir at 210 nm

Calculate the percentage of imidazole carboximidamide and 4-biguanide zanamivir in the portion of Zanamivir taken:

Result = [(r_i /F)/(r_i /F + r_z )] × 100

r_i = peak response of imidazole carboximidamide or 4-biguanide zanamivir at 234 nm

F = relative response factor for imidazole carboximidamide or 4-biguanide zanamivir (see Table 1)

r_z = sum of the responses of all the peaks including the zanamivir peak at 234 nm

Calculate the percentage of O-triazinyl zanamivir, zanamivir urea analog, 4-amino zanamivir, talo-zanamivir, zanamivir dimer, and any other unspecied impurity in the portion of Zanamivir taken:

Result = (r_u /r_T ) × 100

r_u = peak response of O-triazinyl zanamivir, zanamivir urea analog, 4-amino zanamivir, talo-zanamivir, zanamivir dimer, or any other unspecied impurity at 234 nm

r_T = sum of the responses of all the peaks including the zanamivir peak at 234 nm

Acceptance criteria: See Table 1.

Table 1

^a 1H-Imidazole (No individual limit. Included in the determination of total impurities.)

^b 1H-Imidazole-1-carboximidamide.

^c 5-Acetamido-9-O-[4-amino-6-(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl]-2,6-anhydro-3,4,5-trideoxy-4-guanidino-d-glycero-d-galacto-non-2-enonic acid.

^d 5-Acetamido-9-O-[4-amino-6-(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl]-2,6-anhydro-3,4,5-trideoxy-4-ureido-d-glycero-d-galacto-non-2-enonic acid.

^e 5-Acetamido-9-O-[4-amino-6-(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl]-2,6-anhydro-3,4,5-trideoxy-4-amino-d-glycero-d-galacto-non-2-enonic acid.

^f 5-Acetamido-9-O-[4-amino-6-(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl]-2,6-anhydro-3,4,5-trideoxy-4-(1-biguanidyl)-d-glycero-d-galacto-non-2-enonic acid.

^g 5-Acetamido-2,6-anhydro-3,4,5-trideoxy-4-guanidino-d-glycero-d-talo-non-2-enonic acid (No individual limit. Included in the determination of total impurities.)

^h 4,4'-(2-Amino-4-oxo-1,3,5-triazapent-2-ene-1,5-diyl) bis(5-acetamido-2,6-anhydro-3,4,5-trideoxy-d-glycero-d-galacto-non-2-enonic acid.)

5 SPECIFIC TESTS

Optical Rotation, Specific Rotation〈781S〉

Sample solution: 10 mg/mL in water

Acceptance criteria: +36.0° to +38.0°, measured at 20°, determined on the anhydrous and solvent-free basis

Water Determination, Method Ic〈921〉: 4.0%–9.0%

6 ADDITIONAL REQUIREMENTS

Packaging and Storage: Store in a well-closed container at controlled room temperature.

USP Reference Standards 〈11〉

USP Zanamivir RS

USP Zanamivir Related Compounds Mixture RS

This Reference Standard is a mixture of zanamivir, imidazole, imidazole carboximidamide, zanamivir urea analog, 4-amino zanamivir, 4- biguanide zanamivir, talo-zanamivir, and zanamivir dimer. The chemical names are given below:

Imidazole: 1H-Imidazole.

Imidazole carboximidamide: 1H-Imidazole-1-carboximidamide.

Zanamivir urea analog: 5-Acetamido-9-O-[4-amino-6-(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl]-2,6-anhydro-3,4,5-trideoxy-4-ureido-d-glycero-d-galactonon-2-enonic acid.

4-Amino zanamivir: 5-Acetamido-9-O-[4-amino-6-(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl]-2,6-anhydro-3,4,5-trideoxy-4-amino-d-glycero-d-galacto-non-2-enonic acid.

4-Biguanide zanamivir: 5-Acetamido-9-O-[4-amino-6-(1H-pyrazol-1-yl)-1,3,5-triazin-2-yl]-2,6-anhydro-3,4,5-trideoxy-4-(1-biguanidyl)-d-glycero-dgalacto-non-2-enonic acid.

talo-Zanamivir: 5-Acetamido-2,6-anhydro-3,4,5-trideoxy-4-guanidino-d-glycero-d-talo-non-2-enonic acid.

Zanamivir dimer: 4,4'-(2-Amino-4-oxo-1,3,5-triazapent-2-ene-1,5-diyl) bis(5-acetamido-2,6-anhydro-3,4,5-trideoxy-d-glycero-d-galacto-non-2-enonic acid.)

USP Zanamivir Resolution Mixture RS

This Reference Standard is a mixture of zanamivir and talo-zanamivir.