Etoposide Injection
»Etoposide Injection contains not less than 90.0percent and not more than 110.0percent 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 Iglass.
Labeling— Label it to indicate that it must be diluted with suitable parenteral vehicle prior to intravenous infusion.
USP Reference standards á11ñ— 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 10mLof sulfuric acid with cooling and stirring to 70mLof dehydrated alcohol in a 100-mLvolumetric flask.Dilute with dehydrated alcohol to volume,and mix.
Standard solution— Dissolve USP Etoposide RSin Diluting solutionto obtain a solution having a known concentration of about 0.8mg per mL.
Test solution— Transfer a volume of Injection,equivalent to about 20mg of etoposide,to a 25-mLvolumetric flask,dilute with Diluting solutionto volume,and mix.
Procedure— Apply separately 10µLeach of the Standard solutionand the Test solutionto a suitable thin-layer chromatographic plate (see Chromatography á621ñ)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 17cm from the origin.Remove the plate,and allow it to air-dry in a fume hood for 5minutes.Replace the plate in the tank and develop again to a distance of about 17cm from the origin.Remove the plate and air-dry it in a fume hood for about 20minutes.Spray the plate with the Spray reagentand heat in a forced-air oven at 120for about 15minutes:the principal spot from the Test solutioncorresponds in appearance and RFvalue to that from the Standard solution.
B: The retention time of the major peak in the chromatogram of the Assay preparationcorresponds to that in the chromatogram of the Standard preparation,as obtained in the Assay.
pHá791ñ: between 3.0and 4.0in a solution prepared by diluting 5.0mLof it with 45mLof water.
Bacterial endotoxins á85ñ— Use a test solution,prepared by diluting the Injection with Sterile Water for Injection to obtain a concentration of 0.31mg etoposide activity per mL:it contains not more than 2.0USP Endotoxin Units per mg of etoposide.
Alcohol content,Method IIá611ñ(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 Assayunder Etoposide.
Standard preparation— Transfer 0.75mLof freshly distilled benzyl alcohol,accurately weighed,to a 50-mLvolumetric flask,dissolve in and dilute with Mobile phaseto volume,and mix.Transfer 1.0mLof this solution to a 50-mLvolumetric flask,dilute with Mobile phaseto volume,and mix.
Test preparation— Use the Assay preparation.
Procedure— Separately inject equal volumes (about 20µL)of the Standard preparationand the Test preparationinto 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 Cis the concentration,in mg per mL,of benzyl alcohol in the Standard preparation;Vis the volume of Injection taken;and rUand rSare the peak responses of benzyl alcohol obtained from the Test preparationand the Standard preparation,respectively:between 90.0%and 110.0%of the labeled amount is found.
Related compounds— Proceed as directed in the Related compoundstest under Etoposide.Not more than 3.0%total impurities is found.
Other requirements— It meets the requirements under Injections á1ñ.
Assay—
Buffer solution,Mobile phase,Standard preparation,System suitability solution,and Chromatographic system— Proceed as directed in the Assayunder Etoposide.
Assay preparation— Transfer an accurately measured volume of Injection,equivalent to about 100mg of etoposide,to a 50-mLvolumetric flask,dilute with Mobile phaseto volume,and mix.Pipet 5.0mLof this solution into another 50-mLvolumetric flask,dilute with Mobile phaseto volume,and mix.
Procedure— Proceed as directed for Procedurein the Assayunder Etoposide.Calculate the quantity,in mg,of etoposide (C29H32O13)in each mLof the Injection taken by the formula:
500(C/V)(rU/rS),
in which Cis the concentration,in mg per mL,of USP Etoposide RSin the Standard preparation;Vis the volume,in mL,of Injection taken;and rUand rSare the responses of the etoposide peak obtained from the Assay preparationand the Standard preparation,respectively.
Auxiliary Information— Staff Liaison:Lawrence Evans,III,Ph.D.,Scientist
Expert Committee:(PA6)Pharmaceutical Analysis 6
USP28–NF23Page 803
Pharmacopeial Forum:Volume No.29(6)Page 1885
Phone Number:1-301-816-8389