Fludarabine Phosphate for 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

Fludarabine Phosphate for Injection contains NLT 95.0% and NMT 105.0% of the labeled amount of fludarabine phosphate (C₁₀H₁₃FN₅O₇P).

[Caution—Fludarabine Phosphate is potentially cytotoxic. Great care should be taken to prevent inhaling particles and exposing the skin to it.]

2 IDENTIFICATION

Change to read:

A. Spectroscopic Identification Tests 〈197〉, Ultraviolet-Visible Spectroscopy: 197U _{(CN 1-May-2020)}

Sample solution: 27 µg/mL in 0.1 M hydrochloric acid

Acceptance criteria: Meets the requirements

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

Solution A: 10 mM of monobasic potassium phosphate

Mobile phase: Methanol and Solution A (6:94)

Standard solution: 0.02 mg/mL of USP Fludarabine Phosphate RS in Mobile phase

Sample stock solution: 1 mg/mL of udarabine phosphate in Mobile phase prepared as follows. Inject 2.0 mL of Mobile phase into each of ve vials of Fludarabine Phosphate for Injection. Transfer the contents of the vials into a 250-mL volumetric ask, using Mobile phase rinses. Dilute with Mobile phase to volume.

Sample solution: 0.02 mg/mL of udarabine phosphate in Mobile phase, from Sample stock solution

3.1 Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 260 nm

Column: 4.6-mm × 15-cm; 5-µm packing L1

Flow rate: 1.0 mL/min

Injection volume: 10 µL

3.2 System suitability

Sample: Standard solution

Suitability requirements

Relative standard deviation: NMT 2.0%

3.3 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of fludarabine phosphate (C₁₀H₁₃FN₅O₇P) in the portion of Fludarabine Phosphate for Injection taken:

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

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

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

C_U = nominal concentration of fludarabine phosphate in the Sample solution (mg/mL)

Acceptance criteria: 95.0%–105.0%

4 PERFORMANCE TESTS

Uniformity of Dosage Units 〈905〉: Meets the requirements

5 IMPURITIES

5.1 Organic Impurities Procedure 1: Early-Eluting Impurities

Mobile phase, Standard solution, and Chromatographic system: Proceed as directed in the Assay.

System suitability solution: 10 mg of Fludarabine Phosphate in 10 mL of 0.1 N hydrochloric acid. Heat the solution at 80° in a water bath for 15 min.

Sensitivity solution: 0.5 µg/mL of USP Fludarabine Phosphate RS in Mobile phase, from the Standard solution

Sample solution: Use Sample stock solution as directed in the Assay.

5.1.1 System suitability

Samples: Standard solution, System suitability solution, and Sensitivity solution

Suitability requirements

Resolution: NLT 2.0 between the iso-ara-guanine monophosphate and isoguanine peaks, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution

Signal-to-noise ratio: NLT 10, Sensitivity solution

5.1.2 Analysis

Sample: Sample solution

Calculate the percentage of each early-eluting impurity in the portion of Fludarabine Phosphate for Injection taken:

Result = (r_U /r_S ) × (1/F₁) × 100

r_U = peak response of each individual impurity from the Sample solution

r_S = peak response of fludarabine phosphate from the Sample solution

F₁ = relative response factor for each individual impurity (see Table 1)

Acceptance criteria: See Table 1.

Table 1

a This is a process impurity and controlled in the drug substance monograph.

5.2 Organic Impurities Procedure 2: Late-Eluting Impurities

Solvent A: 10 mM monobasic potassium phosphate

Mobile phase: Methanol and Solvent A (1:4)

Standard solution and Chromatographic system: Proceed as directed in the Assay.

Sensitivity solution and Sample solution: Prepare as directed in Organic Impurities Procedure 1: Early-Eluting Impurities.

5.2.1 System suitability

Samples: Standard solution and Sensitivity solution

Suitability requirements

Tailing factor: NMT 2.0, Standard solution

Relative standard deviation: NMT 2.0%, Standard solution

Signal-to-noise ratio: NLT 10, Sensitivity solution

5.2.2 Analysis

Sample: Sample solution

Calculate the percentage of each late-eluting impurity in the portion of Fludarabine Phosphate for Injection taken:

Result = (r_U /r_S × (1/F₂) × 100

r_U = peak response of each individual impurity from the Sample solution

r_S = peak response of fludarabine phosphate from the Sample solution

F₂ = relative response factor for each individual impurity (see Table 2)

Acceptance criteria: See Table 2.

Table 2

a This is a process impurity and controlled in the drug substance monograph.

b The sum of all degradation products found in Organic Impurities Procedure 1 and Organic Impurities Procedure 2.

6 SPECIFIC TESTS

Bacterial Endotoxins Test 〈85〉: NMT 7.7 USP Endotoxin Units/mg of fludarabine phosphate

pH 〈791〉: 7.2–8.2

Sterility Tests 〈71〉: Meets the requirements when tested as directed in Test for Sterility of the Product to Be Examined, Membrane Filtration

Water Determination, Method I〈921〉: NMT 5.0%

Constituted Solution: At the time of use, it meets the requirements for Injections and Implanted Drug Products 〈1〉, Specic Tests, Completeness and clarity of solutions.

7 ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve as described in Packaging and Storage Requirements 〈659〉, Injection Packaging, Packaging for constitution, between 2° and 30°, or at controlled room temperature.

USP Reference Standards 〈11〉

USP Fludarabine Phosphate RS