Edition: BP 2025 (Ph. Eur. 11.6 update)
Action and use
Progestogen.
DEFINITION
Medroxyprogesterone Injection is a sterile suspension of Medroxyprogesterone Acetate in a suitable vehicle
The injection complies with the requirements stated under Parenteral Preparations and with the following requirements.
Content of medroxyprogesterone acetate, C24H34O4
90.0 to 110.0% of the stated amount.
IDENTIFICATION
Dissolve a volume of the injection containing 50 mg of Medroxyprogesterone Acetate in 8 mL of petroleum spirit (boiling range, 40° to 60°) and extract with three 8-mL quantities of a mixture of 7 volumes of glacial acetic acid and 3 volumes of water. Wash the combined extracts with 10 mL of petroleum spirit (boiling range, 40° to 60°), dilute with water until the solution becomes turbid, allow to stand in ice for 2 hours and filter. Wash the precipitate with water and dry at 105°. The infrared absorption spectrum, Appendix II A, is concordant with the reference spectrum of medroxyprogesterone acetate (RS 421).
TESTS
Impurity F (6a-methyl-3,20-dioxo-5b-pregnan-17-yl acetate)
Carry out the method for thin-layer chromatography, Appendix III A, using the following solutions.
(1) The injection, diluted with dichloromethane if necessary, to contain 2.0% w/v of Medroxyprogesterone Acetate.
(2) 2.0% w/v of medroxyprogesterone acetate for performance test EPCRS (containing 0.5% of impurity F) in dichloromethane.
CHROMATOGRAPHIC CONDITIONS
(a) Use as the coating silica gel (Merck silica gel 60 plates are suitable)
(b) 10 volumes of tetrahydrofuran, 45 volumes of 1,1-dimethylethyl methyl ether and 45 volumes of hexane as the mobile phase.
(c) Apply to the plate 10 µL of each of solutions (1) and (2).
(d) After removal of the plate, allow it to dry in air and carry out a second development in the same direction using a freshly prepared mobile phase.
(e) Dry the plate at 100° to 105° and allow to cool, spray with a 200 g/L solution of toluenesulfonic acid in ethanol (96 %). Heat at 120 °C for 10 min, allow to cool and examine the plate in ultraviolet light 365 nm.
SYSTEM SUITABILITY
The test is not valid unless the chromatogram obtained with solution (2) shows two clearly separated spots.
LIMITS
In the chromatogram obtained with solution (1) any blue fluorescent spot with an Rf value higher than the principal spot is not more intense than the corresponding blue fluorescent spot due to impurity F in the chromatogram obtained with solution (2) (0.5%).
Related substances
Carry out the method for liquid chromatography, Appendix III D, using the following solutions.
(1) Dilute a quantity of the injection containing 40 mg of Medroxyprogesterone Acetate with sufficient of the mobile phase to produce 100 mL.
(2) Dilute 1 volume of solution (1) to 100 volumes with the mobile phase.
(3) 0.002% w/v of medroxyprogesterone acetate BPCRS and 0.005% w/v of megestrol acetate BPCRS in the mobile phase.
CHROMATOGRAPHIC CONDITIONS
(a) Stainless steel column (25 cm × 4.6 mm) packed with base-deactivated end-capped octadecylsilyl silica gel for chromatography (5 µm) (Phenomenex Prodigy ODS3 is suitable).
(b) Isocratic elution using the mobile phase described below.
(c) Flow rate of 2 mL per minute.
(d) Maintain the temperature of the column at 40°.
(e) Detection wavelength of 241 nm.
(f) Injection volume of 20 µL for each solution.
MOBILE PHASE
A mixture of 100 volumes of tetrahydrofuran, 350 volumes of acetonitrile, 500 volumes of water and allowed to equilibrate and the volume adjusted to 1000 volumes with water.
SYSTEM SUITABILITY
The test is not valid unless, in the chromatogram obtained with solution (3):
the resolution factor between the two principal peaks is at least 3.3;
the symmetry factor of the peak due to medroxyprogesterone acetate is not more than 1.3.
LIMITS
In the chromatogram obtained with solution (1):
the area of any secondary peak is not greater than the area of the principal peak in the chromatogram obtained with solution (2) (1%);
the sum of the areas of any such peaks is not greater than 1.5 times the area of the principal peak in the chromatogram obtained with solution (2) (1.5%); disregard any peaks with an area not greater than 0.05 times the area of the principal peak in solution (2) (0.05%).
ASSAY
Carry out the method for liquid chromatography, Appendix III D, using the following solutions.
(1) The injection, diluted if necessary with the mobile phase, to contain 0.004% w/v of Medroxyprogesterone Acetate.
(2) 0.004% w/v of medroxyprogesterone acetate BPCRS in the mobile phase.
(3) 0.002% w/v of medroxyprogesterone acetate BPCRS and 0.005% w/v of megestrol acetate BPCRS in the mobile phase.
CHROMATOGRAPHIC CONDITIONS
The chromatographic procedure described under Related substances may be used.
SYSTEM SUITABILITY
The test is not valid unless, in the chromatogram obtained with solution (3):
the resolution factor between the two principal peaks is at least 3.3;
the symmetry factor of the peak due to medroxyprogesterone acetate is not more than 1.3.
DETERMINATION OF CONTENT
Calculate the content of medroxyprogesterone acetate, C24H34O4, in the injection using the declared content of C24H34O4 in medroxyprogesterone acetate BPCRS.
STORAGE
Medroxyprogesterone Injection should be protected from light. On standing, solid matter may separate; it should be redissolved by heating before use.
IMPURITIES
A. 6-hydroxy-6-methyl-3,20-dioxopregn-4-en-17-yl acetate (6-hydroxymedroxyprogesterone acetate),
B. 17-hydroxy-6-methylpregn-4-ene-3,20-dione (medroxyprogesterone),
C. 6,17a-dimethyl-3,17-dioxo-D-homoandrost-4-en-17a-yl acetate,
D. 6-methyl-3,20-dioxopregn-4-en-17-yl acetate (6-epimedroxyprogesterone acetate),
E. 6-methylidene-3,20-dioxopregn-4-en-17-yl acetate (6-methylenehydroxyprogesterone acetate),
F. 6-methyl-3,20-dioxo-5-pregnan-17-yl acetate (4,5-dihydromedroxyprogesterone acetate),
G. 6-methyl-3,20-dioxopregna-4,6-dien-17-yl acetate (megestrol acetate).
