BRIEFING

Nitazoxanide. Because there is no existing USP monograph for this drug substance, a new monograph
based on validated methods of analysis is proposed. The liquid chromatographic procedures in the Assay
and the test for Organic Impurities are based on analyses performed with the Waters µBondapak C18
brand of column with L1 packing. The typical retention time for the nitazoxanide peak is about 9 min.

This monograph is contingent on FDA approval of a product that meets the proposed monograph.

(CHM1: S. Shivaprasad.)
Correspondence Number—C186508

Comment deadline: September 30, 2018

Add the following:

Nitazoxanide

C12H9N3O5S 307.28

2-(Acetyloxy)-N-(5-nitro-2-thiazolyl) benzamide;
N-(5-Nitrothiazol-2-yl)salicylamide acetate ester [55981-09-4].

DEFINITION

Nitazoxanide contains NLT 98.0% and NMT 102.0% of nitazoxanide (C12H9N3O5S), calculated on the
dried basis.

IDENTIFICATION
• A. I A 〈197K〉
• 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
• P
Buffer: 1.8 g/L of potassium phosphate, monobasic. Adjust with phosphoric acid to a pH of 2.5.
Mobile phase: Acetonitrile and Buffer (450:550). To this solution add 1.83 g of hexane sulfonic acid
sodium salt.
Standard stock solution: 1 mg/mL of USP Nitazoxanide RS prepared as follows. Transfer a suitable
amount of USP Nitazoxanide RS to a suitable volumetric flask. Add 10% of the flask volume of
dimethyl sulfoxide and dilute with acetonitrile to volume.
Standard solution: 0.2 mg/mL of USP Nitazoxanide RS in Mobile phase from Standard stock solution
Sample stock solution: 1 mg/mL of Nitazoxanide prepared as follows. Transfer a suitable amount of
Nitazoxanide to a suitable volumetric flask. Add 10% of the flask volume of dimethyl sulfoxide and
 dilute with acetonitrile to volume.
Sample solution: 0.2 mg/mL of Nitazoxanide in Mobile phase from Sample stock solution
Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC
Detector: UV 360 nm
Column: 3.9-mm × 30-cm; 10-µm packing L1
Autosampler temperature: 4°
Flow rate: 1 mL/min
Injection volume: 20 µL
Run time: NLT 2 times the retention time of nitazoxanide
System suitability
Sample: Standard solution
Suitability requirements
Tailing factor: NMT 2.0
Relative standard deviation: NMT 1.0%
Analysis
Samples: Standard solution and Sample solution

Calculate the percentage of nitazoxanide (C12H9N3O5S) in the portion of Nitazoxanide taken:

Result = (rU/rS) × (CS/CU) × 100

rU = peak response of nitazoxanide from the Sample solution

rS = peak response of nitazoxanide from the Standard solution
CS = concentration of USP Nitazoxanide RS in the Standard solution (mg/mL)
CU = concentration of Nitazoxanide in the Sample solution (mg/mL)

Acceptance criteria: 98.0%–102.0% on the dried basis

IMPURITIES
• O I
Buffer and Mobile phase: Prepare as directed in the Assay.
System suitability stock solution: 0.6 mg/mL each of USP Nitazoxanide RS, USP Nitazoxanide
Related Compound A RS, USP Nitazoxanide Related Compound B RS, and USP Aspirin RS prepared as
follows. Transfer suitable amounts of USP Nitazoxanide RS, USP Nitazoxanide Related Compound A
RS, USP Nitazoxanide Related Compound B RS, and USP Aspirin RS to a suitable volumetric flask. Add
5% of the flask volume of dimethyl sulfoxide and dilute with acetonitrile to volume.
System suitability solution: 0.06 mg/mL each of USP Nitazoxanide RS, USP Nitazoxanide Related
Compound A RS, USP Nitazoxanide Related Compound B RS, and USP Aspirin RS in acetonitrile from
System suitability stock solution. [N —This solution is stable for about 6 h at 4°.]
Standard stock solution: 0.6 mg/mL of USP Nitazoxanide RS prepared as follows. Transfer a suitable
amount of USP Nitazoxanide RS to a suitable volumetric flask. Add 5% of the flask volume of dimethyl
sulfoxide and dilute with acetonitrile to volume.
Standard solution: 2.5 µg/mL of USP Nitazoxanide RS in Mobile phase from Standard stock solution
Sample stock solution: 6 mg/mL of Nitazoxanide prepared as follows. Transfer a suitable amount of
Nitazoxanide to a suitable volumetric flask. Add 5% of the flask volume of dimethyl sulfoxide and
dilute with acetonitrile to volume.
Sample solution: 1.2 mg/mL of Nitazoxanide in Mobile phase from Sample stock solution. [N —
This solution is stable for about 6 h at 4°.]
Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC
Detector: UV 237 nm
Column: 3.9-mm × 30-cm; 10-µm packing L1
Autosampler temperature: 4°
Flow rate: 1 mL/min
Injection volume: 20 µL
System suitability
Samples: System suitability solution and Standard solution
Suitability requirements
Resolution: NLT 1.2 between nitazoxanide related compound A and aspirin; NLT 1.5 between
nitazoxanide and nitazoxanide related compound B, System suitability solution
Tailing factor: NMT 2.0 for the nitazoxanide peak, Standard solution
Relative standard deviation: NMT 5.0% for the nitazoxanide peak, Standard solution
Analysis
Samples: Standard solution and Sample solution

Calculate the percentage of any individual impurity in the portion of Nitazoxanide taken:

Result = (rU/rS) × (CS/CU) × (1/F) × 100

rU = peak response of any individual impurity from the Sample solution

rS = peak response of nitazoxanide from the Standard solution
CS = concentration of USP Nitazoxanide RS in the Standard solution (mg/mL)
CU = concentration of Nitazoxanide in the Sample solution (mg/mL)
F = relative response factor (see Table 1)

Acceptance criteria: See Table 1. The reporting threshold is 0.02%.

 Table 1
Relative Relative Acceptance
Retention Response Criteria,
Name Time Factor NMT (%)
Nitazoxanide related compound A 0.40 1.9 0.10
Aspirina 0.48 1.2 0.10
Salicylic acidb 0.61 1.8 0.10
Nitazoxanide 1.0 — —
Nitazoxanide related compound B 1.2 1.0 1.0
Any individual unspecified impurity — 1.0 0.10
Total impurities — — 1.0
a 2-Acetoxybenzoic acid.
b 2-Hydroxybenzoic acid.

SPECIFIC TESTS
• L D 〈731〉
Analysis: Dry at 60° for 3 h.
Acceptance criteria: NMT 0.50%

ADDITIONAL REQUIREMENTS
• P S : Preserve in well-closed containers.
• USP R S 〈11〉
USP Aspirin RS
2-Acetoxybenzoic acid.
C9H8O4 180.16
USP Nitazoxanide RS
USP Nitazoxanide Related Compound A RS
5-Nitrothiazole-2-amine.
C3H3N3O2S 145.14
USP Nitazoxanide Related Compound B RS
Tizoxanide;
2-Hydroxy-N-(5-nitrothiazol-2-yl)benzamide.
C10H7N3O4S 265.25

(TBD)

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