Epirubicin Hydrochloride

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 contains NLT 97.0% and NMT 102.0% of epirubicin hydrochloride (C₂₇H₂₉NO₁₁ · HCl), calculated on the anhydrous and solvent-free basis.

2 IDENTIFICATION

Change to read:

A. Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197M (CN 1-May-2020)

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

C. Identification Tests—General, Chloride〈191〉

Solution A: Nitric acid and water (1:1)

Sample solution: 10 mg/mL in Solution A

Acceptance criteria: Meets the requirements

3 ASSAY

Procedure

Allow the System suitability solution, Standard solution, and Sample solution to stand for 3 h before use.

Solution A: Dilute 10 mL of phosphoric acid with water to 100 mL.

Solution B:

Dissolve 3.7 g of sodium lauryl sulfate in 950 mL of water. To the resulting solution, add 28 mL of Solution A, and dilute with water to 1000mL.

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

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

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

Sample solution: 1 mg/mL of Epirubicin Hydrochloride in Mobile phase

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: About 3.5 times the retention time of the epirubicin peak

System suitability

Sample: System suitability solution

Suitability requirements

Resolution: NLT 2.0 between doxorubicin and epirubicin

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of epirubicin hydrochloride (C₂₇H₂₉NO₁₁ · HCl) in the portion of Epirubicin Hydrochloride 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 the Standard solution (mg/mL)

C_U = concentration of the Sample solution (mg/mL)

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

Acceptance criteria: 97.0%–102.0% on the anhydrous and solvent-free basis

4 IMPURITIES

Organic Impurities

Allow the System suitability solution, Sample solution, and Standard solution to stand for 3 h before use.

Mobile phase, System suitability solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay.

Standard solution: 0.01 mg/mL of USP Epirubicin Hydrochloride RS in Mobile phase

Peak identification solution: Dissolve 10 mg of USP Doxorubicin Hydrochloride RS in 10 mL of a mixture of water and phosphoric acid (1:1).

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

Analysis

Samples: Sample solution, Standard solution, and Peak identification solution

[Note—Use the Peak identification solution to identify the doxorubicinone peak.]

Calculate the percentage of each impurity in the portion of Epirubicin Hydrochloride 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 = 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 hydrochloride.

^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 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}.

Limit of Acetone

Analysis: See Residual Solvents 〈467〉.

Acceptance criteria: NMT 1.5%

5 SPECIFIC TESTS

Water Determination, Method Ic〈921〉: NMT 4.0%

pH 〈791〉

Sample solution: 5 mg/mL

Acceptance criteria: 4.0–5.5

Bacterial Endotoxins Test 〈85〉: NMT 1.1 USP Endotoxin Units/mg, where the label states that Epirubicin Hydrochloride is sterile or must be subjected to further processing during the preparation of injectable dosage forms.

Sterility Tests 〈71〉: Meets the requirements where the label states that Epirubicin Hydrochloride is sterile.

6 ADDITIONAL REQUIREMENTS

Packaging and Storage: Store in airtight containers, protected from light. Store as per labeling instructions. Possible storage conditions could include the following: Store at a temperature between 2° and 8°. Store at room temperature. If the substance is sterile, store in a sterile, airtight, tamper-proof container.

USP Reference Standards 〈11〉

USP Doxorubicin Hydrochloride RS

USP Epirubicin Hydrochloride RS