Epirubicin Injection

Epirubicin Hydrochloride Injection

Epirubicin Injection is a sterile solution of Epirubicin Hydrochloride in Water for Injections.

Epirubicin Injection contains not less than 95.0 per cent and not more than 110.0 per cent of the stated amount of epirubicinhydrochloride,C₂₇H₃₀ClNO₁₁.

Usual strengths. 2 mg per ml.

Identification

1. A solution containing 0.001 per cent w/v of epirubicin hydrochloride inwater. When examined in the range 220 nm to 350 nm (2.4.7), the solution shows an absorption maximum at about 233, 253 and 292 nm.
2. In the Assay, the principal peak in the chromatogram obtained with the test solution corresponds to the peak in the chromatogram obtained with the reference solution.

Tests

pH(2.4.24). 2.5 to 4.0.

Related substances. Determine byliquid chromatography (2.4.14).

Note-Allow the solutions to stand for 3 hours before use.

Test solution. Dilute a volume of the injection with sufficient mobile phase to produce a solution containing 0.1 per cent w/v of Epirubicin Hydrochloride.

Reference solution(a). Dilute 1.0 ml of the test solution to 100.0 ml with the mobile phase.

Reference solution (b).A 0.01 per cent w/v each of epirubicin hydrochloride RS and doxorubicin hydrochloride RS in the mobile phase.

Reference solution (c). Dissolve 10 mg of doxorubicin hydrochloride RS 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 8 per cent w/v solution of sodium hydroxide, add 15 ml of acetonitrile and 10 ml of methanol and mix (generation of impurity A).

Chromatographic system

     –   a stainless steel column 25 cm × 4.6 mm, packed with trimethylsilane bonded to porous silica (6 µm) (Such as Zorbax TMS),

     –   column temperature: 35°,

     –   mobile phase: a mixture of 54 volumes of a buffer solution prepared by dissolving 3.7 g of sidium lauryl sphate and 28 ml of 1M orthophosphoric acid in 1000 ml ofwater, 17 volumes of methanol and 29 volumes of acetonitrile,

     –   flow rate: 2.5 ml per minute,

     –   spectrophotometer set at 254 nm,

     –   injection volume: 10 µl.

Name                                              Relative           Correction

                                                    retention time          factor

Epirubicin impurity A1                     0.3                      0.7

Epirubicin impurity B2                     0.4                       ---

Epirubicin impurity C3                     0.8                       ---

Epirubicin (Retention time:

about 9.5 minutes)                           1.0                       ---

Epirubicin impurity D4                     1.5                       ---

Epirubicin impurity E5                      1.1                       ---

Epirubicin impurity F6                      1.7                       ---

Epirubicin impurity G7                     2.1                       ---

¹ R = OH: (8S,10S)-6,8,10,11-tetrahydroxy-8-(hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione (doxorubicin one),

² R = H: (8 B.S,10S)-8-acetyl-6,8,10,11-tetrahydroxy-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione (daunorubicin one),

³ (8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-6,8,11-trihydroxy-8-(hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetracene-   5,12-dione (doxorubicin),

⁴ (8S,10S)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-6,8,11-trihydroxy-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione (daunorubicin),

⁵ (8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexopyranosyl)oxy]-6,8,11-trihydroxy-8-[(1RS)-1-hydroxyethyl]-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione (dihydrodaunorubicin),

⁶ (8S,10S)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-arabino-hexopyranosyl)oxy]-6,8,11-trihydroxy-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione (epi-daunorubicin),

⁷ 8,8′-[(2R,4R)-4-hydroxy-2-(hydroxymethyl)-1,3-dioxolan-2,4-diyl]bis[(8S,10S)-10-[(3-amino-2,3,6-trideoxy-α-L-arabino-hexopyranosyl)oxy]-6,8,11-trihydroxy-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione] (epirubicin dimmer).

 

Identification of impurities. Identify any peak in the chromatogram obtained with test solution corresponding to epirubicin impurity A using the second most abundant peak in the chromatogram obtained with reference solution (c) and multiply the area of this peak by the corresponding correction factor of  0.7.

Inject reference solutions (b), (c) and the test solution.

Run the chromatogram 3.5 times the retention time of epirubicin.

Inject reference solution (b). The test is not valid unless the resolution between the peaks corresponding to epirubicin and doxorubicin is not less than 2.0.

Inject reference solution (a) and the test solution. In the chromatogram obtained with the test solution, the area of peak corresponding to epirubicin impurities A is not more than twice the area of the principal peak in the chromatogram obtained with reference solution (a) (2.0 per cent). The area of peak corresponding to epirubicin impurities C is not more than the area of the principal peak in the chromatogram obtained with reference solution (a) (1.0 per cent)  The area of any secondary peak is not more than 0.5 times the area of the principal peak in the chromatogram obtained with reference solution (a) (0.5 per cent).  The sum of areas of all the secondary peaks is not more than four times the area of the principal peak in the chromatogram obtained with reference solution(a) (4.0 per cent). Ignore any peak with an area less than 0.05 times the area of the principal peak in the chromatogram obtained with reference solution (a) (0.05 per cent).

Other tests. Comply with the tests stated under Parenteral Preparations (Injections).

Bacterial endotoxins(2.2.3). 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. Determine by liquid chromatography (2.4.14) as described under related substances with the following modification.

Note: Allow the solutions to stand for 3 hours before use.

Test solution. Dilute a volume of the injection with sufficient mobile phase to produce a solution containing 0.1 per cent  w/v of Epirubicin Hydrochloride.

Reference solution (a). A solution containing 0.1 per cent w/v epirubicin hydrochloride RS in the mobile phase.

Reference solution (b).A 0.01 per cent w/v each of epirubicin hydrochloride RS and doxorubicin hydrochloride RS in the mobile phase.

Inject reference solution (b). The test is not valid unless in the chromatogram obtained with reference solution (b) the resolution between the peaks due to epirubicin and doxorubicin is not less than 2.0.

Inject reference solution (a) and the test solution.

Calculate the content of C₂₇H₃₀ClNO₁₁ in the injection.

Storage.Epirubicin Injection should be stored at a temperature of 2° to 8°.