Identification—
A:
The UV absorption spectrum of a methanol solution containing about 25 µg of dactinomycin per mL exhibits maxima and minima at the same wavelengths as that of a similar solution of
USP Dactinomycin RS, concomitantly measured, and the ratio
A240 /
A445 is between 1.30 and 1.50.
B:
The chromatogram obtained from the
Assay preparation in the
Assay exhibits a major peak for dactinomycin, the retention time of which corresponds to that exhibited by the
Standard preparation, and the chromatogram compares qualitatively to that obtained from the
Standard preparation.
Sterility 
71
—
It meets the requirements when tested as directed for
Membrane Filtration under
Test for Sterility of the Product to be Examined, each container being constituted aseptically by injecting Sterile Water for Injection through the stopper, and the entire contents of all the containers being collected aseptically with the aid of 200 mL of
Fluid A before filtering.
Assay—
[NOTE—In this procedure, use freshly prepared
Standard preparation and
Assay preparation, protected from light.
]
Mobile phase—
Mix 6 volumes of acetonitrile and 4 volumes of water, filter through a membrane filter (1-µm or finer porosity), and degas. [NOTE—The acetonitrile concentration may be varied to provide appropriate Chromatographic system performance and to provide a suitable elution time.]
Standard preparation—
Dissolve an accurately weighed quantity of
USP Dactinomycin RS in
Mobile phase, and dilute quantitatively with
Mobile phase to obtain a solution having a known concentration of about 250 µg of dactinomycin per mL.
Assay preparation—
Add an accurately measured volume of Mobile phase to 1 container of Dactinomycin for Injection to obtain a solution containing about 250 µg of dactinomycin per mL, and filter, if necessary, to obtain a clear solution.
Chromatographic system
(see
Chromatography 621)—The liquid chromatograph is equipped with a 254-nm detector and a 3.9-mm × 30-cm column that contains packing L1. The flow rate is about 2.5 mL per minute. Chromatograph the
Standard preparation, and record the peak response as directed under
Procedure: the column efficiency is not less than 1200 theoretical plates, the tailing factor is not more than 2, and the relative standard deviation for replicate injections is not more than 3.0%.
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 responses for the major peaks. The retention time is about 6 minutes for dactinomycin. Calculate the quantity, in mg, of C
62H
86N
12O
16 in the container of Dactinomycin for Injection taken by the formula:
(CV / 1000)(rU / rS),
in which
C is the concentration, in µg, of dactinomycin in each mL of the
Standard preparation, V is the volume, in mL, of the
Assay preparation, and
rU and
rS are the peak responses of the
Assay preparation and the
Standard preparation, respectively.
for 3 hours: it loses not more than 4.0% of its weight.