Progesterone Injection
» Progesterone Injection is a sterile solution of Progesterone in a suitable solvent. It contains not less than 90.0 percent and not more than 110.0 percent of the labeled amount of C21H30O2.
Packaging and storage— Preserve in single-dose or in multiple-dose containers, preferably of Type I or Type III glass.
USP Reference standards 11
USP Progesterone RS
Identification— Insert a pledget of fine glass wool into the base of a chromatographic tube of about 200 × 25 mm. Mix 8.0 mL of nitromethane with 7.0 g of purified siliceous earth in a 150-mL beaker 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 1 mL of the Injection with n-heptane to obtain a solution having a concentration of about 1 mg of progesterone per mL. Transfer 4.0 mL of this solution to the prepared column. Pass 300 mL of n-heptane through the column, discarding the first 120 mL of the eluate. Collect the subsequent eluate in a 250-mL beaker. Evaporate the solution under a stream of nitrogen on a steam bath to about 50 mL, transfer to a 100-mL beaker, and evaporate to dryness. Remove the last traces of n-heptane by adding 1 mL of methanol and again drying. Dry the specimen over silica gel for 4 hours: the IR absorption 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 under Injections 1.
Mobile phase— Prepare a degassed mixture of alcohol and water (11:9). Make adjustments if necessary (see System Suitability under Chromatography 621).
Standard preparation— Dissolve an accurately weighed quantity of USP Progesterone RS in 20 mL of tetrahydrofuran, and dilute quantitatively, and stepwise if necessary, with alcohol to obtain a solution having a known concentration of about 0.08 mg per mL.
Assay preparation— Transfer an accurately measured volume of Injection, equivalent to about 100 mg of progesterone, to a 100-mL volumetric flask, add 20 mL of tetrahydrofuran to dissolve, and dilute with alcohol to volume. Transfer 8 mL of this solution to a 100-mL volumetric flask, dilute with alcohol to volume, and mix.
Chromatographic system (see Chromatography 621)— 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 1 mL per minute. The column temperature is maintained at about 40. Chromatograph a sample of dimethyl sulfoxide, and identify the retention time, t, of this nonretarded compound to calculate the capacity factor, k¢. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: 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 10 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. [note—The run time for the Assay preparation must be at least twice that of the Standard preparation.] Calculate the quantity, in mg, of progesterone (C21H30O2) in each mL of Injection taken by the formula:
1250(C/V)(rU / rS)
in which C is the concentration, in mg per mL, of USP Progesterone RS in the Standard preparation; V is the volume, in mL, of Injection taken; and rU and rS are the peak responses for progesterone obtained from the Assay preparation and the Standard preparation, respectively.
Auxiliary Information— Please check for your question in the FAQs before contacting USP.
Topic/Question Contact Expert Committee
Monograph Daniel K. Bempong, Ph.D.
Senior Scientist
(MDPS05) Monograph Development-Pulmonary and Steroids
Reference Standards Lili Wang, Technical Services Scientist
USP32–NF27 Page 3399
Pharmacopeial Forum: Volume No. 27(5) Page 3038
Chromatographic Column—
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.