Progesterone Injectable Suspension
» Progesterone Injectable Suspension is a sterile suspension of Progesterone in Water for Injection. It contains not less than 93.0 percent and not more than 107.0 percent of the labeled amount of C21H30O2.
Packaging and storage Preserve in single-dose or in multiple-dose containers, preferably of Type I glass.
Identification Filter a volume of well-shaken Injectable Suspension, equivalent to not less than 100 mg of progesterone, through a medium-porosity, sintered-glass crucible, filtering again through the same crucible if the fluid is not clear. Wash with several 5-mL portions of water until 2 mL of the last washing, evaporated on a steam bath, leaves no weighable residue: the washed solid, dried at 105 to constant weight, melts between 126 and 131, and responds to Identification test A under Progesterone.
pH 791: between 4.0 and 7.5.
Other requirements It meets the requirements under Injections 1.
Mobile phase Prepare a filtered and degassed mixture of water and isopropyl alcohol (72:28). Make adjustments if necessary (see System Suitability under Chromatography 621.)
Diluent Prepare a mixture of alcohol and water (85 in 100).
Internal standard solution Dissolve an accurately weighed quantity of USP Methyltestosterone RS in Diluent to obtain a solution containing about 6.6 mg per mL.
Standard preparation Dissolve an accurately weighed quantity of USP Progesterone RS in Diluent, and dilute quantitatively, and stepwise if necessary, with Diluent to obtain a solution having a known concentration of about 2.5 mg per mL.
Standard curve To four separate polytef-lined, screw-capped centrifuge tubes, pipet 3-, 4-, 5-, and 6-mL portions, respectively, of the Standard preparation. Add Diluent to each tube to make about 8 mL of solution. Transfer 1.0 mL of Internal standard solution to each tube, and mix.
Assay preparation Transfer an accurately measured volume of Injectable Suspension, equivalent to about 25 mg progesterone, to a polytef-lined, screw-capped, 25 mL test tube. Add 16 mL of Diluent, and shake until clear. Add 2.0 mL of Internal standard solution, and mix.
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 254-nm detector and a 2.1-mm × 1-m column that contains packing L2. The chromatographic conditions are such that retention times of the internal standard and progesterone are between 3 and 6 minutes and 7 and 11 minutes, respectively. Chromatograph any solution of the Standard curve, and record the peak areas as directed for Procedure: the resolution, R, between progesterone and methyltestosterone is not less than 3.5; and the relative standard deviation for replicate injections is not more than 1.5%.
Procedure Separately inject equal volumes (5.0 µL) of each solution of the Standard curve and the Assay preparation into the chromatograph, record the chromatograms, and measure the peak areas. Plot the progesterone to methyltestosterone peak ratio versus the concentration, in mg per mL, of progesterone in the Standard curve solutions. Extrapolate the progesterone to methyltestosterone peak ratio of the Assay preparation in the Standard curve so obtained, and determine the progesterone concentration, C, in mg per mL, in the Assay preparation. Calculate the amount, in mg, of progesterone (C21H30O2) in each mL of the Injectable Suspension taken by the formula:
V/A(C)in which V is the total volume of the Assay preparation; A is the volume in mL of Injectable Suspension taken; and C is the concentration of progesterone in the Assay preparation calculated above.
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USP32NF27 Page 3401Pharmacopeial Forum: Volume No. 28(5) Page 1442