Epirubicin Hydrochloride Injection

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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1 DEFINITION

Epirubicin Hydrochloride Injection contains NLT 90.0% and NMT 110.0% of the labeled amount of epirubicin hydrochloride (C₂₇H₂₉NO₁₁ · HCl).

2 IDENTIFICATION

A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

B. The UV absorption spectrum of the major peak of the Sample solution and the Standard solution as obtained in the Assay exhibit maxima and minima at the same wavelengths.

3 ASSAY

Procedure

Solution A: Dissolve 2.9 g of sodium lauryl sulfate in 950 mL of water. To the resulting solution add 1.4 mL of phosphoric acid and dilute with water to 1 L.

Mobile phase: Acetonitrile and Solution A (50:50)

Standard solution: 1 mg/mL of USP Epirubicin Hydrochloride RS in Mobile phase

Sample solution: Nominally 1 mg/mL of epirubicin hydrochloride from Injection

Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 254 nm. When this procedure is used for Identi

cation test B, use a diode array detector set at 200–400 nm.

Column: 4.6-mm × 25-cm; 5-μm packing L1

Flow rate: 1 mL/min

Injection volume: 10 μL

System suitability

Sample: Standard solution

Suitability requirements

Relative standard deviation: NMT 2.0%

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of epirubicin hydrochloride (C₂₇H₂₉NO₁₁ · HCl) in the portion of Injection taken:

Result = (r_U/r_S) × (C_S/C_U) × P × 100

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

C_S = concentration of USP Epirubicin Hydrochloride RS in the Standard solution (mg/mL)

C_U = nominal concentration of epirubicin hydrochloride in the Sample solution (mg/mL)

P = potency of epirubicin hydrochloride in USP Epirubicin Hydrochloride RS (mg/mg)

Acceptance criteria: 90.0%–110.0%

4 IMPURITIES

Organic Impurities

Solution A: Dissolve 3.7 g of sodium lauryl sulfate in 950 mL of water. To the resulting solution add 28 mL of phosphoric acid and dilute with water to 1 L.

Solution B: Dilute 28 mL of phosphoric acid with water to 1 L.

Mobile phase: Acetonitrile, methanol, and Solution A (29:17:54)

Diluent: Acetonitrile, methanol, and Solution B (29:17:27)

System suitability solution: 0.1 mg/mL each of USP Epirubicin Hydrochloride RS and USP Doxorubicin Hydrochloride RS in Mobile phase

Peak identification solution: Dissolve 10 mg of USP Doxorubicin Hydrochloride RS in a mixture of 5 mL of water and 5 mL of phosphoric acid.

Allow to stand for 30 min. Adjust with 2 M sodium hydroxide to a pH of 2.6. Add 15 mL of acetonitrile and 10 mL of methanol, and mix.

Standard stock solution: 0.5 mg/mL of USP Epirubicin Hydrochloride RS in Mobile phase

Standard solution: 0.01 mg/mL of USP Epirubicin Hydrochloride RS from Standard stock solution in Diluent

Sample solution: Nominally 1 mg/mL of epirubicin hydrochloride from Injection in Diluent. Store at room temperature and use within 4 h.

Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 254 nm

Column: 4.6-mm × 25-cm; 5-μm packing L13

Column temperature: 35°

Flow rate: 2.5 mL/min

Injection volume: 10 μL

Run time

For the Standard solution: About 2 times the retention time of the epirubicin peak

For the System suitability solution, Peak identification solution, and Sample solution: About 4.5 times the retention time of the epirubicin peak

System suitability

Samples: System suitability solution, Peak identi fication solution, and Standard solution. [Note—Use the Peak identification solution to identify the doxorubicinone peak.]

Suitability requirements

Resolution: NLT 2.0 between epirubicin and doxorubicin, System suitability solution

Relative standard deviation: NMT 5.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of each impurity in the portion of Injection taken:

Result = (r_U/r_S) × (C_S/C_U) × P × (1/F) ×100

r_U = peak response of each impurity from the Sample solution

r_S = peak response of epirubicin from the Standard solution

C_S = concentration of USP Epirubicin Hydrochloride RS in the Standard solution (mg/mL)

C_U = nominal concentration of epirubicin hydrochloride in the Sample solution (mg/mL)

P = potency of epirubicin hydrochloride in USP Epirubicin Hydrochloride RS (mg/mg)

F = relative response factor (see Table 1)

Acceptance criteria: See Table 1. The reporting threshold is 0.05% of the area of the epirubicin peak in the Standard solution.

Table 1

^a (8S,10S)-6,8,10,11-Tetrahydroxy-8-(hydroxyacetyl)-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione.

^b (8S,10S)-8-Acetyl-6,8,10,11-tetrahydroxy-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione.

^c Dihydrodaunorubicin; (8S,10S)-10-[(3-Amino-2,3,6-trideoxy-α-l-lyxo-hexopyranosyl)oxy]-6,8,11-trihydroxy-8-(1-hydroxyethyl)-1-methoxy-7,8,9,10-tetrahydrotetracene-5,12-dione.

^d (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.

^e These impurities do not have individual limits; they are included in total impurities.

^f 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}.

5 SPECIFIC TESTS

pH 〈791〉: 2.5–3.5

Sterility Tests 〈71〉: Meets the requirements

Bacterial Endotoxins Test 〈85〉: NMT 1.61 USP Endotoxin Units/mg of epirubicin

Other Requirements: It meets the requirements in Injections and Implanted Drug Products 〈1〉.

6 ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve in light-resistant containers. Store in a refrigerator.

USP Reference Standards 〈11〉

USP Doxorubicin Hydrochloride RS

USP Epirubicin Hydrochloride RS