Edition: BP 2025 (Ph. Eur. 11.6 update)
General Notices
Action and use
Anthracycline antibacterial; cytostatic.
DEFINITION
Epirubicin Injection is a sterile solution of Epirubicin Hydrochloride in Water for Injections.
The injection complies with the requirements stated under Parenteral Preparations and with the following requirements.
Content of epirubicin hydrochloride, C27H29NO11,HCl
95.0 to 110.0% of the stated amount.
IDENTIFICATION
A. Dilute a volume of the injection containing 10 mg of Epirubicin Hydrochloride to 100 mL with water and further dilute 5 mL to 50 mL with water. The light absorption of the resulting solution, Appendix II B, in the range 220 to 350 nm exhibits three maxima at 233, 253 and 292 nm.
B. In the Assay, the retention time of the principal peak in the chromatogram obtained with solution (1) is similar to that of the principal peak in the chromatogram obtained with solution (2).
TESTS
Acidity
pH, 2.5 to 4.0, Appendix V L.
Related substances
Carry out the method for liquid chromatography, Appendix III D, using the following solutions. Allow the solutions to stand for 3 hours before use.
(1) Dilute a volume of the injection with sufficient of the mobile phase to produce a solution containing 0.1% w/v of Epirubicin Hydrochloride.
(2) Dilute 1 volume of solution (1) to 100 volumes with the mobile phase.
(3) 0.01% w/v of each of doxorubicin hydrochloride BPCRS and epirubicin hydrochloride BPCRS in the mobile phase.
(4) Dissolve 10 mg of doxorubicin hydrochloride BPCRS in a mixture of 5 mL of water and 5 mL of orthophosphoric acid and allow to stand for 30 minutes. Adjust the pH of the solution to 2.6 with an 8% w/v solution of sodium hydroxide, add 15 mL of acetonitrile and 10 mL of methanol and mix (generation of impurity A).
CHROMATOGRAPHIC CONDITIONS
(a) Use a stainless steel column (25 cm × 4.6 mm) packed with trimethylsilyl silica gel for chromatography (6 µm) (Zorbax TMS is suitable).
(b) Use isocratic elution and the mobile phase described below.
(c) Use a flow rate of 2.5 mL per minute.
(d) Use a column temperature of 35°.
(e) Use a detection wavelength of 254 nm.
(f) Inject 10 µL of each solution.
(g) For solution (1), allow the chromatography to proceed for 3.5 times the retention time of epirubicin.
MOBILE PHASE
17 volumes of methanol, 29 volumes of acetonitrile and 54 volumes of a solution containing 0.37% w/v of sodium dodecyl sulfate and 2.8% v/v of 1M orthophosphoric acid.
When the chromatograms are recorded under the prescribed conditions, the retention times relative to epirubicin (retention time about 9.5 minutes) are:
impurity A, about 0.3; impurity B, about 0.4; impurity C, about 0.8; impurity E, about 1.1; impurity D, about 1.5; impurity F, about 1.7; impurity G, about 2.1.
SYSTEM SUITABILITY
The test is not valid unless, in the chromatogram obtained with solution (3), the resolution factor between the peaks due to doxorubicin and epirubicin is at least 2.0.
LIMITS
Identify any peak in the chromatogram obtained with solution (1) corresponding to impurity A using the second most abundant peak in the chromatogram obtained with solution (4) and multiply the area of this peak by the corresponding correction factor: 0.7.
In the chromatogram obtained with solution (1):
the area of any peak corresponding to impurity A (doxorubicin aglycone, doxorubicinone) is not greater than twice the area of the principal peak in the chromatogram obtained with solution (2) (2%);
the area of any peak corresponding to impurity C (doxorubicin) is not greater than the area of the principal peak in the chromatogram obtained with solution (2) (1%);
the area of any other secondary peak is not greater than 0.5 times the area of the principal peak in the chromatogram obtained with solution (2) (0.5%);
the sum of the areas of all the secondary peaks is not greater than four times the area of the principal peak in the chromatogram obtained with solution (2) (4%).
Disregard any peak with an area less than 0.05 times the area of the principal peak in the chromatogram obtained with solution (2) (0.05%).
Bacterial endotoxins
Carry out the test for bacterial endotoxins, Appendix XIV C. Dilute the injection, if necessary, with water BET to give a solution containing 2 mg per mL of Epirubicin Hydrochloride (solution A). The endotoxin limit concentration of solution A is not more than 2.2 IU per mL.
ASSAY
Carry out the method for liquid chromatography, Appendix III D, using the following solutions. Allow the solutions to stand for 3 hours before use.
(1) Dilute a volume of the injection with sufficient of the mobile phase to produce a solution containing 0.1% w/v of Epirubicin Hydrochloride.
(2) 0.1% w/v of epirubicin hydrochloride BPCRS in the mobile phase.
(3) 0.01% w/v of each of doxorubicin hydrochloride BPCRS and epirubicin hydrochloride BPCRS in the mobile phase.
CHROMATOGRAPHIC CONDITIONS
(a) Use a stainless steel column (25 cm × 4.6 mm) packed with trimethylsilyl silica gel for chromatography (6 µm) (Zorbax TMS is suitable).
(b) Use isocratic elution and the mobile phase described below.
(c) Use a flow rate of 2.5 mL per minute.
(d) Use a column temperature of 35°.
(e) Use a detection wavelength of 254 nm.
(f) Inject 10 μL of each solution.
MOBILE PHASE
17 volumes of methanol, 29 volumes of acetonitrile and 54 volumes of a solution containing 0.37% w/v of sodium dodecyl sulfate and 2.8% v/v of 1M orthophosphoric acid.
SYSTEM SUITABILITY
The test is not valid unless, in the chromatogram obtained with solution (3), the resolution factor between the peaks due to doxorubicin and epirubicin is at least 2.0.
DETERMINATION OF CONTENT
Calculate the content of C27H29NO11,HCl in the injection using the declared content of C27H29NO11,HCl in epirubicin hydrochloride BPCRS.
STORAGE
Epirubicin Injection should be stored at a temperature of 2° to 8°.
IMPURITIES
The impurities limited by the requirements of this monograph include those listed under Epirubicin Hydrochloride.
