Etoposide Injection
» Etoposide Injection contains not less than 90.0 percent and not more than 110.0 percent of the labeled amount of etoposide (C29H32O13) in a sterile solution in a nonaqueous medium intended for dilution with a suitable parenteral vehicle prior to intravenous infusion.
Packaging and storage— Preserve in single-dose or multiple-dose containers of Type I glass.
Labeling— Label it to indicate that it must be diluted with suitable parenteral vehicle prior to intravenous infusion.
USP Reference standards 11
USP Etoposide RS
.
USP Etoposide Related Compound A RS
.
USP Endotoxin RS
.
Identification—
A: Diluting solution—Prepare a mixture of chloroform and methanol (9:1).
Spray reagent— Add 10 mL of sulfuric acid with cooling and stirring to 70 mL of dehydrated alcohol in a 100-mL volumetric flask. Dilute with dehydrated alcohol to volume, and mix.
Standard solution— Dissolve USP Etoposide RS in Diluting solution to obtain a solution having a known concentration of about 0.8 mg per mL.
Test solution— Transfer a volume of Injection, equivalent to about 20 mg of etoposide, to a 25-mL volumetric flask, dilute with Diluting solution to volume, and mix.
Procedure— Apply separately 10 µL each of the Standard solution and the Test solution to a suitable thin-layer chromatographic plate (see Chromatography 621) coated with a 0.25-mm layer of chromatographic silica gel mixture. Allow the spots to dry, and develop the plate in a suitable chromatographic chamber containing a developing solvent consisting of a mixture of chloroform, acetone, alcohol, and water (80:25:2.5:0.5). Allow the chromatogram to develop until the solvent front has moved about 17 cm from the origin. Remove the plate, and allow it to air-dry in a fume hood for 5 minutes. Replace the plate in the tank and develop again to a distance of about 17 cm from the origin. Remove the plate and air-dry it in a fume hood for about 20 minutes. Spray the plate with the Spray reagent and heat in a forced-air oven at 120 for about 15 minutes: the principal spot from the Test solution corresponds in appearance and RF value to that from the Standard solution.
B: The retention time of the major peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay.
pH 791: between 3.0 and 4.0 in a solution prepared by diluting 5.0 mL of it with 45 mL of water.
Bacterial endotoxins 85 Use a test solution, prepared by diluting the Injection with Sterile Water for Injection to obtain a concentration of 0.31 mg etoposide activity per mL: it contains not more than 2.0 USP Endotoxin Units per mg of etoposide.
Alcohol content, Method II 611 (if present): between 90.0% and 110.0% of the labeled amount of C2H5OH, n-propyl alcohol being used as the internal standard.
Benzyl alcohol content (if present)—
Buffer solution, Mobile phase, System suitability solution, and Chromatographic system— Proceed as directed in the Assay under Etoposide.
Standard preparation— Transfer 0.75 mL of freshly distilled benzyl alcohol, accurately weighed, to a 50-mL volumetric flask, dissolve in and dilute with Mobile phase to volume, and mix. Transfer 1.0 mL of this solution to a 50-mL volumetric flask, dilute with Mobile phase to volume, and mix.
Test preparation— Use the Assay preparation.
Procedure— Separately inject equal volumes (about 20 µL) of the Standard preparation and the Test preparation into the chromatograph, record the chromatograms, and measure the responses for the benzyl alcohol peaks. Calculate the quantity, in mg per mL, of benzyl alcohol in the volume of Injection taken by the formula:
500(C/V)(rU / rS)
in which C is the concentration, in mg per mL, of benzyl alcohol in the Standard preparation; V is the volume of Injection taken; and rU and rS are the peak responses of benzyl alcohol obtained from the Test preparation and the Standard preparation, respectively: between 90.0% and 110.0% of the labeled amount is found.
Related compounds— Proceed as directed in the Related compounds test under Etoposide. Not more than 3.0% total impurities is found.
Other requirements— It meets the requirements under Injections 1.
Assay—
Buffer solution, Mobile phase, Standard preparation, System suitability solution, and Chromatographic system— Proceed as directed in the Assay under Etoposide.
Assay preparation— Transfer an accurately measured volume of Injection, equivalent to about 100 mg of etoposide, to a 50-mL volumetric flask, dilute with Mobile phase to volume, and mix. Pipet 5.0 mL of this solution into another 50-mL volumetric flask, dilute with Mobile phase to volume, and mix.
Procedure— Proceed as directed for Procedure in the Assay under Etoposide. Calculate the quantity, in mg, of etoposide (C29H32O13) in each mL of the Injection taken by the formula:
500(C/V)(rU / rS)
in which C is the concentration, in mg per mL, of USP Etoposide RS in the Standard preparation; V is the volume, in mL, of Injection taken; and rU and rS are the responses of the etoposide peak obtained from the Assay preparation and the Standard preparation, respectively.
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Topic/Question Contact Expert Committee
Monograph Feiwen Mao, M.S.
Scientist
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(MDOOD05) Monograph Development-Ophthalmics Oncologics and Dermatologicals
Reference Standards Lili Wang, Technical Services Scientist
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RSTech@usp.org
85 Radhakrishna S Tirumalai, Ph.D.
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USP32–NF27 Page 2339
Pharmacopeial Forum: Volume No. 29(6) Page 1885