Dactinomycin
;)
Actinomycin D.
Actinomycin D [50-76-0].
»Dactinomycin contains not less than 950µg and not more than 1030µg of C62H86N12O16per mg,calculated on the dried basis.
Caution—Great care should be taken to prevent inhaling particles of Dactinomycin and exposing the skin to it.
Packaging and storage—
Preserve in tight containers,protected from light and excessive heat.
Identification—
Solution:
25µg per mL.
Medium:
methanol.
Absorptivity at 445nm,calculated on the dried basis,is not less than 95.0%and not more than 103.0%of that of USP Dactinomycin RS,the potency of the Reference Standard being taken into account.
Ratio:
A240/A445between 1.30and 1.50.
B:
The chromatogram obtained from the Assay preparationin the Assayexhibits 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.
Specific rotation á781Sñ:
between -292
and -317(t=20).
and -317(t=20).
Test solution:
1mg per mL,in methanol.
Crystallinity á695ñ:
meets the requirements.
Bacterial endotoxins á85ñ—
It contains not more than 100USP Endotoxin Units per mg.
Loss on drying á731ñ—
Dry it in vacuum at a pressure not exceeding 5mm of mercury at 60for 3hours:it loses not more than 5.0%of its weight.
for 3hours:it loses not more than 5.0%of its weight.
Assay—
[NOTE—In this procedure,use a freshly prepared Standard preparation and Assay preparation,protected from light.]
Mobile phase—
Prepare a suitable mixture of acetonitrile,0.04Msodium acetate,and 0.07Macetic acid (approximately 46:25:25),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 RSin Mobile phase,and dilute quantitatively with Mobile phaseto obtain a solution having a known concentration of about 1200µg of dactinomycin per mL.
Assay preparation—
Dissolve about 30mg of Dactinomycin,accurately weighed,in Mobile phaseto make 25.0mL,and mix.
Chromatographic system
(see Chromatography á621ñ)—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 1.0mLper minute.Chromatograph three replicate injections of the Standard preparation,and record the peak responses as directed under Procedure:the relative standard deviation is not more than 1.0%.
Procedure—
Separately inject equal volumes (about 20µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.The retention time is about 25minutes for Dactinomycin.Calculate the potency,in µg,of C62H86N12O16per mg taken by the formula:
25(C/W)(rU/rS),
in which Cis the concentration,in µg,of dactinomycin in each mLof the Standard preparation;Wis the weight,in mg,of Dactinomycin taken;and rUand rSare the peak responses of the Assay preparationand the Standard preparation,respectively.
Auxiliary Information—
Staff Liaison:William W.Wright,Ph.D.,Scientific Fellow
Expert Committee:(PA7)Pharmaceutical Analysis 7
USP28–NF23Page 570
Phone Number:1-301-816-8335