Etoposide 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

Etoposide Injection contains NLT 90.0% and NMT 110.0% of the labeled amount of etoposide (C₂₉H₃₂O₁₃) in a sterile solution in a nonaqueous medium intended for dilution with a suitable parenteral vehicle before intravenous infusion.

2 IDENTIFICATION

A.

Diluent: Chloroform and methanol (9:1)

Standard solution: 0.8 mg/ml. of USP Etoposide RS in Diluent

Sample solution: Equivalent to 0.8 mg/mL of etoposide in Diluent from the Injection

Chromatographic system

(See Chromatography (621), Thin-Layer Chromatography)

Mode: TLC

Adsorbent: 0.25-mm layer of chromatographic silica gel mixture

Application volume: 10 µL

Developing solvent system: Chloroform, acetone, alcohol, and water (80:25:2.5:0.5)

Spray reagent: Add 10 ml of sulfuric acid with cooling and stirring to 70 ml. of dehydrated alcohol in a 100-mi, volumetric flask Dilute with debydrated alcohol to volume, and mix

Analysis:

Samples: Standard solution and Sample solution

Allow the chromatogram to develop until the solvent front has moved 17 cm from the origin. Remove the plate, and allow it to air-dry in a fume hood for 5 min. Replace the plate in the tank, and develop again to a distance of 17 cm from the origin. Remove the plate, and air dry it in a fume hood for about 20 min. Spray the plate with the Spray reagent, and heat in a forced-air oven at 120" for about 15 min.

Acceptance criteria: The appearance and R, value of the principal spot from the Sample solution corresponds to that from the Standard solution

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

3 ASSAY

PROCEDURE

Buffer: 2.72 g/L of sodium acetate in water. Adjust with glacial acetic acid to a pH of 4.0.

Mobile phase: Acetonitrile and Buffer (26:74)

System suitability solution: 0.3 mg/mL of USP Etoposide Resolution Mixture RS in Mobile phase

Standard stock solution: 2.0 mg/ml of USP Etoposide RS in acetonitrile

Standard solution: 0.2 mg/ml. of USP Etoposide BS in Mobile phase from the Standard stock solution

Sample solution: Equivalent to 0.2 mg/ml of etoposide in Mobile phase from the Injection

Chromatographic system

(See Chromatograoby (521), System Suitability)

Mode: LC

Detector: UV 254 nm

Column: 3.9-mm × 30-cm; packing L11

Flow rate: 1 mL/min

Injection volume: 20 μL

Run time: NLT 1.5 times the retention time of etoposide

System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 1.35 between the etoposide and a etoposide peaks, System suitability solution

Relative standard deviation: NMT 2.0% for etoposide, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of etoposide  (C₂₉H₃₂O₁₃) in the portion of Injection taken:

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

r_U = peak response of etoposide from the Sample solution

r_S = peak response of etoposidefrom the Standard solution

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

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

Acceptance criteria: 90.0%-110.0%

4 IMPURITIES

ORGANIC IMPURITIES

Buffer: Prepare as directed in the Assay

Solution A: Acetonitrile and Buffer (20:80)

Solution B: Acetonitrile and Buffer (60:40)

Mobile phase: See Table 1

Diluent: Acetonitrile and 0.02 M sodium acetate previously adjusted with acetic acid to a pH of 4.0 (30:70)

Standard stock solution: 2.0 mg/mL of USP Etoposide RS in Diluent

System suitability stock solution: 0.2 mg/mL of a storyiparaben in Diluent

System suitability solution: Transfer 5.0 ml, of the System suitability stock solution and 5.0 ml of the Standard stock solution to a 50-mL volumetric flask, and dilute with Diluent to volume. Transfer 5.0 mL of this solution to a 100-ml volumetric flask, and dilute with Diluent to volume.

Standard solution: 10 μg/mL of USP Etoposide RS from the Standard stock solution in Diluent

Sample solution: Nominally equivalent to 2.0 mg/ml. of etoposide in Diluent

Chromatographic system

(See Chromatography (621). System Suitability)

Mode: LC

Detector: UV 254 nm

Column: 3.9-mm × 15-cm; less than 5-μm packing L11

Flow rate: 1.5 mL/min

Injection volume: 25 μL

Run time: NLT 40 min

System suitability

[Note—Run time is 15 min in isocratic conditions.]

Sample: System suitability solution

Suitability requirements

Resolution: NLT 1.1 between Propylparaben and etoposide

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) x 100

r_U = peak response of each impurity from the Sample solution

r_S = peak response of etoposide from the Standard solution

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

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

Acceptance criteria

Total impurities: NMT 3.0%

5 SPECIFIC TESTS

pH (791)

Sample solution: 5.0 mL. of Injection in 45 ml of water

Acceptance criteria: 3.0-4.0

ALCOHOL DETERMINATION (611), Method (if present): 90.0%-110.0% of the labeled amount of alcohol (C,H,OH), using n-propyl alcohol as th

internal standard

BACTERIAL ENDOтохіна Тест (85)

Sample solution: Dilute the Injection with sterile water to obtain 0.31 mg/ml. of etoposide activity

Acceptance criteria: NMT 2.0 USP Endotoxin Units/mg of etoposide

BENZYL ALCOHOL CONTENT (if present)

Buffer, Mobile phase, System suitability solution, Sample solution, Chromatographic system, and System suitability: Proceed as directe

in the Assay

Standard solution: Transfer 0.75 mL of freshly distilled benzyl alcohol, accurately weighed, to a 50-ml volumetric flask, dissolve in and dil with Mobile phase to volume, and mix. Transfer 1.0 ml of this solution to a 50-ml, volumetric flask, dilute with Mobile phase to volume, an mix.

Analysis

Samples: Sample solution and Standard solution

Calculate the percentage of the labeled amount of bernzyl alcohol in the portion of Injection taken:

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

r_U = peak response of benzyl alcohol from the Sample solution

r_S = peak response of benzyl alcohol from the Standard solution

C_S = concentration of benzyl alcohol in the Standard solution (mg/ml)

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

Acceptance criteria: 90.0%-110.0%

OTHER REQUIREMENTS: It meets the requirements in Injections and Implanted Drug Products (1),

6 ADDITIONAL REQUIREMENTS

Change to read:

PACKAGING AND STORAGE: Preserve in single dose or multiple-dose containers, preferably of Type I glass.

LABELING: Label it to indicate that it must be diluted with suitable parenteral vehicle before intravenous infusion.