Etoposide Capsules
»Etoposide Capsules contain not less than 90.0percent and not more than 110.0percent of the labeled amount of etoposide (C29H32O13).
Caution—Etoposide is potentially cytotoxic.Great care should be taken to prevent inhaling particles of Etoposide and exposing the skin to it.
Packaging and storage— Preserve in tight containers in a cold place.Do not freeze.
USP Reference standards á11ñ— USP Etoposide RS.USP Etoposide Related Compound A RS.
Identification—
A:Infrared Absorption á197Kñ—
Test specimen— Transfer a suitable quantity of the contents of Capsules,equivalent to about 100mg of etoposide,to a separator containing 100mLof water.Extract twice with 20-mLportions of chloroform,separate and combine the organic layer,dry over anhydrous sodium sulfate,and filter.Transfer the dried filtrate to a second separator,extract with 30mLof water,and allow the layers to separate.Drain the chloroform layer through a bed of anhydrous sodium sulfate contained in a filter funnel into a round bottom flask,and evaporate the chloroform at a temperature of 30±5using a rotary evaporator.Dissolve the oily residue obtained in 5mLof water,shake gently,and allow to stand for 30minutes.Filter,collecting the precipitate formed on a glass filter funnel,wash the precipitate with three 20-mLportions of water,and allow the precipitate to dry on the filter for about 90minutes in a vacuum oven at 40.Prepare a dispersion of the precipitate in KBr at a ratio 1in 100.
B: The retention time of the major peak in the chromatogram of theAssay preparation corresponds to that in the chromatogram of theStandard preparation,as obtained in theAssay.
Dissolution á711ñ—
Medium: pH4.5acetate buffer;900mL.
Apparatus 2: 50rpm.
Time: 30minutes.
Determine the amount of C29H32O13dissolved by employing the following method.
Buffer solution— Dissolve 5.44g of sodium acetate in 2000mLof water,adjust with acetic acid to a pHof 4.0,and filter.
Mobile phase— Prepare a filtered and degassed mixture ofBuffer solution and acetonitrile (74:26).Make adjustments if necessary (seeSystem Suitability underChromatography á621ñ).
Standard solution— Dissolve,with sonication,an accurately weighed quantity of USP Etoposide RSin a volume of methanol equivalent to 2%of the final solution volume.Dilute withDissolution Medium to obtain a solution having a known concentration of about 55µg per mL,and mix.
Test solution— At 30minutes withdraw a 10-mLaliquot from the dissolution vessel.
Chromatographic system— The liquid chromatograph is equipped with a 240-nm detector and a 3.9-mm ×30-cm column that contains packing L11.The flow rate is about 2mLper minute.Chromatograph theStandard solution,and record the peak responses as directed forProcedure:the relative standard deviation for replicate injections is not more than 2.0%.
Procedure— Separately inject equal volumes (about 50µL)of the filtered portions of theStandard solution and theTest solution into the chromatograph,record the chromatograms,and measure the responses for the major peaks.Determine the amount of C29H32O13dissolved.
Tolerances— Not less than 80%(Q)of the labeled amount of C29H32O13is dissolved in 30minutes.
Uniformity of dosage units á905ñ: meet the requirements.
Related compounds— Proceed as directed in the test forRelated compounds underEtoposide:not more than 2.0%of picroetoposide is found;and not more than 3.0%of total impurities is found.
Assay—
Buffer solution,Mobile phase,Standard preparation,System suitability solution,and Chromatographic system— Proceed as directed in theAssay underEtoposide.
Assay preparation— Transfer an accurately counted number of Capsules,equivalent to about 500mg of etoposide,to a 500-mLvolumetric flask,add about 400mLofMobile phase,and stir using a magnetic bar for about 15minutes,followed by sonication for about 1hour with occasional shaking.Cool,dilute withMobile phase to volume,stir for an additional 5minutes,and filter.Transfer 10.0mLof this solution to a 50-mLvolumetric flask,dilute withMobile phase to volume,and mix.
Procedure— Proceed as directed in theAssay underEtoposide.Calculate the quantity,in mg,of etoposide (C29H32O13)per Capsule by the formula:
2500(C/N)(rU/rS),
in whichCis the concentration,in mg per mL,of USP Etoposide RSin theStandard preparation;Nis the number of Capsules taken;and rUand rSare the etoposide peak responses obtained from theAssay preparation and theStandard preparation,respectively.
Auxiliary Information— Staff Liaison:Lawrence Evans,III,Ph.D.,Scientist
Expert Committee:(PA6)Pharmaceutical Analysis 6
USP28–NF23Page 802
Pharmacopeial Forum:Volume No.27(5)Page 3004
Phone Number:1-301-816-8389