Packaging and storage—
Preserve in tight, light-resistant containers. Store at 25
, excursion permitted between 15
and 30
.
Other requirements—
Where the label states that Mitomycin is sterile, it meets the requirements of the tests for
Sterility and
Bacterial endotoxins under
Mitomycin for Injection. Where the label states that Mitomycin must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements of the test for
Bacterial endotoxins under
Mitomycin for Injection.
Assay—
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, and mix. Pass through a filter having a 0.5-µm or finer porosity, and degas. Make adjustments if necessary (see
System Suitability under
Chromatography 
621
).
Standard preparation—
Quantitatively dissolve an accurately weighed quantity of
USP Mitomycin RS in N,N-dimethylacetamide to obtain a solution having a known concentration of about 0.5 mg per mL.
Resolution solution—
Dissolve suitable quantities of
USP Mitomycin RS and 3-ethoxy-4-hydroxybenzaldehyde in N,N-dimethylacetamide to obtain a solution containing about 0.5 mg and 7.5 mg per mL, respectively.
Assay preparation—
Transfer about 25 mg of Mitomycin, accurately weighed, to a 50-mL volumetric flask, add N,N-dimethylacetamide to volume, and mix to dissolve.
Chromatographic system (see Chromatography 621)—
The liquid chromatograph is equipped with a 365-nm detector and a 4-mm × 30-cm column that contains packing L11. The flow rate is about 2 mL per minute. Chromatograph the
Resolution solution, and record the peak responses as directed under
Procedure: the resolution,
R, between the mitomycin and the 3-ethoxy-4-hydroxybenzaldehyde peaks is not less than 1.8. The relative retention times are about 1.0 for mitomycin and 1.4 for 3-ethoxy-4-hydroxybenzaldehyde. Chromatograph the
Standard preparation, and record the peak responses as directed for
Procedure: the tailing factor for the mitomycin peak is not more than 1.3; and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure—
Separately inject equal volumes (about 10 µL) of the
Standard preparation and the
Assay preparation into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in µg, of C
15H
18N
4O
5 in each mg of the Mitomycin taken by the formula:
50(CP/W)(rU / rS),
in which
C is the concentration, in mg per mL, of
USP Mitomycin RS in the
Standard preparation; P is the stated potency, in µg per mg, of the
USP Mitomycin RS;
W is the weight, in mg, of Mitomycin taken to prepare the
Assay preparation; and
rU and
rS are the peak areas obtained from the
Assay preparation and the
Standard preparation, respectively.
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