Progesterone Injection
»Progesterone Injection is a sterile solution of Progesterone in a suitable solvent.It contains not less than 90.0percent and not more than 110.0percent of the labeled amount of C21H30O2.
Packaging and storage— Preserve in single-dose or in multiple-dose containers,preferably of Type Ior Type IIIglass.
Identification— Insert a pledget of fine glass wool into the base of a chromatographic tube of about 200×25mm.Mix 8.0mLof nitromethane with 7.0g of purified siliceous earth in a 150-mLbeaker until uniform,and transfer to the chromatographic tube,packing lightly with a suitable tamping rod.Pack a pledget of glass wool on the top of the column.Dilute 1mLof the Injection withn-heptane to obtain a solution having a concentration of about 1mg of progesterone per mL.Transfer 4.0mLof this solution to the prepared column.Pass 300mLofn-heptane through the column,discarding the first 120mLof the eluate.Collect the subsequent eluate in a 250-mLbeaker.Evaporate the solution under a stream of nitrogen on a steam bath to about 50mL,transfer to a 100-mLbeaker,and evaporate to dryness.Remove the last traces ofn-heptane by adding 1mLof methanol and again drying.Dry the specimen over silica gel for 4hours:the IRabsorption spectrum of a potassium bromide dispersion of the residue so obtained exhibits maxima only at the same wavelengths as that of a similar preparation of USP Progesterone RS.
Other requirements— It meets the requirements underInjections á1ñ.
Assay—
Mobile phase— Prepare a degassed mixture of alcohol and water (11:9).Make adjustments if necessary (seeSystem Suitability underChromatography á621ñ).
Standard preparation— Dissolve an accurately weighed quantity of USP Progesterone RSin 20mLof tetrahydrofuran,and dilute quantitatively,and stepwise if necessary,with alcohol to obtain a solution having a known concentration of about 0.08mg per mL.
Assay preparation— Transfer an accurately measured volume of Injection,equivalent to about 100mg of progesterone,to a 100-mLvolumetric flask,add 20mLof tetrahydrofuran to dissolve,and dilute with alcohol to volume.Transfer 8mLof this solution to a 100-mLvolumetric flask,dilute with alcohol to volume,and mix.
Chromatographic system(see Chromatography á621ñ)— The liquid chromatograph is equipped with a 254-nm detector and a 4.6-mm ×25-cm column that contains 5-µm packing L1.The flow rate is about 1mLper minute.The column temperature is maintained at about 40.Chromatograph a sample of dimethyl sulfoxide,and identify the retention time,ta,of this nonretarded compound to calculate the capacity factor,k¢.Chromatograph theStandard preparation,and record the peak responses as directed forProcedure:the capacity factor,k¢,for progesterone is not less than 2.0;the tailing factor is not more than 2.0;and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure— Separately inject equal volumes (about 10µL)of theStandard preparation and theAssay preparation into the chromatograph,record the chromatograms,and measure the responses for the major peaks.[NOTE—The run time for theAssay preparation must be at least twice that of theStandard preparation.]Calculate the quantity,in mg,of progesterone (C21H30O2)in each mLof Injection taken by the formula:
1250(C/V)(rU/rS),
in whichCis the concentration,in mg per mL,of USP Progesterone RSin theStandard preparation;Vis the volume,in mL,of Injection taken;andrUandrSare the peak responses for progesterone obtained from theAssay preparation and theStandard preparation,respectively.
Auxiliary Information— Staff Liaison:Clydewyn M.Anthony,Ph.D.,Scientist
Expert Committee:(PA1)Pharmaceutical Analysis 1
USP28–NF23Page 1635
Pharmacopeial Forum:Volume No.27(5)Page 3038
Phone Number:1-301-816-8139