Mitomycin

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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C₁₅H₁₈N₄O₅                   334.33

Azirino[2′,3′:3,4]pyrrolo[1,2-a]indole-4,7-dione, 6-amino-8-[[(aminocarbonyl)oxy]methyl]-1,1a,2,8,8a,8b-hexahydro-8a-methoxy-5-methyl-, [1aS-(1aα,8β,8aα,8bα)]-; (1aS,8S,8aR,8bS)-(6-Amino-8a-methoxy-5-methyl-4,7-dioxo-1,1a,2,4,7,8,8a,8b-octahydroazirino[2′,3′:3,4]pyrrolo[1,2-a]indol-8-yl)methyl carbamate; Mitomycin C

CAS RN: 50-07-7; UNII: 50SG953SK6.

1 DEFINITION

Mitomycin has a potency of NLT 970 µg/mg of mitomycin (C₁₅H₁₈N₄O₅).

2 IDENTIFICATION

Change to read:

A. SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy:

Analysis: Do not dry the sample and standard.

Acceptance criteria: Meets the requirements 197M _{(CN 1-MAY-2020)}

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: Dissolve 1.54 g of ammonium acetate in 250 mL of methanol. Add 5.0 mL of 0.83 N acetic acid and water to make 1000 mL.

System suitability solution: 0.5 mg/mL of USP Mitomycin RS and 7.5 mg/mL of 3-ethoxy-4-hydroxybenzaldehyde in N,N-dimethylacetamide

Standard solution: 0.5 mg/mL of USP Mitomycin RS in N,N-dimethylacetamide

Sample solution: 0.5 mg/mL of Mitomycin in N,N-dimethylacetamide

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 365 nm

Column: 3.9-mm x 30-cm; 10-µm packing L11

Flow rate: 2 mL/min

Injection volume: 10 µL

System suitability

Samples: System suitability solution and Standard solution

[NOTE-The relative retention times for mitomycin and 3-ethoxy-4-hydroxybenzaldehyde are 1.0 and 1.4, respectively.]

Suitability requirements

Resolution: NLT 1.8 between mitomycin and 3-ethoxy-4-hydroxybenzaldehyde, System suitability solution

Tailing factor: NMT 1.3, Standard solution

Relative standard deviation: NMT 2.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the quantity, in µg/mg, of mitomycin (C₁₅H₁₈N₄O₅) in the portion of Mitomycin taken:

Result = (r_u /r_s ) × (C_s /C_u ) × P

r_u = peak area from the Sample solution

r_s = peak area from the Standard solution

C_s = concentration of USP Mitomycin RS in the Standard solution (mg/mL)

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

P = potency of mitomycin in USP Mitomycin RS (µg/mg)

Acceptance criteria: NLT 970 µg/mg

4 IMPURITIES

ORGANIC IMPURITIES

Solution A: 0.77 g/L of ammonium acetate

Solution B: Methanol and Solution A (20:80)

Solution C: Methanol and Solution A (50:50)

Mobile phase: See Table 1.

Table 1

Standard solution: 0.025 mg/mL of USP Mitomycin RS in methanol

Sensitivity solution: 2.5 µg/mL of USP Mitomycin RS in methanol

Sample solution: 5 mg/mL of Mitomycin in methanol

Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 254 nm

Column: 4.6-mm × 25-cm; 5-µm packing L1

Column temperature: 30°

Flow rate: 1 mL/min

Injection volume: 10 µL

System suitability

Samples: Sensitivity solution and Standard solution

Suitability requirements

Signal-to-noise ratio: NLT 10, Sensitivity solution

Tailing factor: NMT 2.0, Standard 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 Mitomycin taken:

Result = (r_u /r_s ) × (C_s /C_u ) × P × (F₁ /F₂ ) × 100

r_u = peak response of each impurity from the Sample solution

r_s = peak response of mitomycin from the Standard solution

C_s = concentration of USP Mitomycin RS in the Standard solution (mg/mL)

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

P = potency of mitomycin in USP Mitomycin RS (µg/mg)

F₁ = conversion factor, 0.001 mg/µg

F₂ = relative response factor (see Table 2)

Acceptance criteria: See Table 2. The reporting threshold is 0.05%.

Table 2

^a {(1S,2S,4aS,8aR,9S,9aR)-7-Amino-9a-methoxy-6-methyl-5,8-dioxo-1,2,3,5,8,8a,9,9a-octahydro-1,2,4a-metheno-(epinitrilo)pyrrolo[1,2-a]indol9-yl}methyl carbamate.

^b {(1aS,8S,8aR,8bS)-8a-Hydroxy-6-methoxy-1,5-dimethyl-4,7-dioxo-1,1a,2,4,7,8,8a,8b-octahydroazirino[2′,3′:3,4]pyrrolo[1,2-a]indol-8-yl}methyl carbamate.

^c {(1aS,8S,8aR,8bS)-6,8a-Dimethoxy-5-methyl-4,7-dioxo-1,1a,2,4,7,8,8a,8b-octahydroazirino[2′,3′:3,4]pyrrolo[1,2-a]indol-8-yl}methyl carbamate.

5 SPECIFIC TESTS

CRYSTALLINITY (695): Meets the requirements

PH (791)

Sample: 5-mg/mL suspension in water

Acceptance criteria: 6.0-7.5

WATER DETERMINATION, Method (921): NMT 2.5%

STERILITY TESTS (71): Where the label states that Mitomycin is sterile, it meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration.

BACTERIAL ENDOTOXINS TEST (85): Where the label states that Mitomycin is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 10.0 USP Endotoxin Units/mg of mitomycin,

6 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in tight, light-resistant containers. Store at 25°, excursions permitted between 15° and 30°.

LABELING: Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms.

USP REFERENCE STANDARDS (11)

USP Mitomycin RS