Gemcitabine 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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C₉H₁₁F₂N₃O₄·HCI 299.66

Cytidine, 2'-deoxy-2',2'-difluoro-, monohydrochloride;

2'-Deoxy-2,2'-difluorocytidine monohydrochloride (β-isomer) CAS RN^®: 122111-03-9; UNII: U347PV74IL.

1 DEFINITION

Gemcitabine Hydrochloride contains NLT 97.5% and NMT 101.5% of gemcitabine hydrochloride (C₉H₁₁F₂N₃O₄·HCI), calculated on the as-is basis.

[CAUTION-Gemcitabine Hydrochloride is a potent cytotoxic agent. Great care should be taken to prevent inhaling particles and exposing the skin to it.]

2 IDENTIFICATION

A. SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197A or 197K.

Meets the requirements

B. IDENTIFICATION TESTS-GENERAL (191), Chemical Identification Tests, Chloride: Meets the requirements

C. 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

3.1 PROCEDURE

Mobile phase: 13.8 g of monobasic sodium phosphate and 2.5 mL of phosphoric acid in 1 L of water. [NOTE-The pH of this solution is 2.4-2.6.]

System suitability solution: Transfer 10 mg of Gemcitabine Hydrochloride to a small vial, add 4 mL of 168 mg/mL of potassium hydroxide in methanol, cap tightly, and sonicate. Heat at 55° for 6-16 h, allow to cool, and transfer the contents to a 100-mL volumetric flask with successive washes of 1% phosphoric acid. Dilute with 1% (v/v) phosphoric acid to volume. [NOTE-This solution contains about 0.02 mg/mL of gemcitabine α-anomer.]

Standard solution: 0.1 mg/mL of USP Gemcitabine Hydrochloride RS in water

Sample solution: 0.1 mg/mL of Gemcitabine Hydrochloride in water

3.2 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 275 nm

Column: 4.6-mm × 25-cm; 5-µm packing L7

Flow rate: 1.2 mL/min

Injection volume: 20 µL

3.3 System suitability

Samples: System suitability solution and Standard solution

[NOTE-The relative retention times for gemcitabine α-anomer and gemcitabine are about 0.5 and 1.0, respectively.]

Suitability requirements

Resolution: NLT 8.0 between gemcitabine a-anomer and gemcitabine, System suitability solution

Tailing factor: NMT 1.5 for the gemcitabine peak, System suitability solution

Relative standard deviation: NMT 1.0%, Standard solution

3.4 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of gemcitabine hydrochloride (C₉H₁₁F₂N₃O₄·HCI) in the portion of Gemcitabine Hydrochloride taken:

Result = (r_U/r_S) x (C_S/C_U) x 100

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

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

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

Acceptance criteria: 97.5%-101.5% on the as-is basis

4 IMPURITIES

4.1 RESIDUE ON IGNITION (281)

NMT 0.1%

Change to read:

4.2 ORGANIC IMPURITIES

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

Solution A: Use the Mobile phase, prepared as directed in the Assay.

Solution B: Methanol

Mobile phase: See Table 1.

Table 1

Standard solution: 0.002 mg/mL each of USP Gemcitabine Hydrochloride RS and USP Cytosine RS in water

Sample solution: 2 mg/mL of Gemcitabine Hydrochloride in water

System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 8.0 between gemcitabine a-anomer and gemcitabine, System suitability solution

Tailing factor: NMT 1.5 for the gemcitabine peak, System suitability solution

Relative standard deviation: NMT 2.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of cytosine in the portion of Gemcitabine Hydrochloride taken:

Result = (r_U/r_S) x (C_S/C_U) x 100

r_U = peak response of cytosine from the Sample solution

r_S = peak response of cytosine from the Standard solution

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

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

Calculate the percentage of each impurity other than cytosine in the portion of Gemcitabine Hydrochloride taken:

Result = (r_U/r_S) x (C_S/C_U) x 100

r_U = peak response of each impurity from the Sample solution

r_S = peak response of gemcitabine from the Standard solution

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

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

Acceptance criteria: See Table 2. The reporting threshold is 0.02%.

Table 2

^a 4-Amino-1-(2-deoxy-2,2-difluoro-α-D-erythro-pentofuranosyl)-2(1H)-pyrimidinone.

5 SPECIFIC TESTS

5.1 OPTICAL ROTATION (781S), Procedures, Specific Rotation

Sample solution: 10 mg/mL

Acceptance criteria: +43° to +50° at 20°

5.2 PH (791)

Sample solution: 10 mg/mL

Acceptance criteria: 2.0-3.0

5.3 STERILITY TESTS (71), Test for Sterility of the Product to Be Examined, Membrane Filtration

Where the label states that Gemcitabine Hydrochloride is sterile, it meets the requirements.

5.4 BACTERIAL ENDOTOXINS TEST (85)

Where the label states that Gemcitabine Hydrochloride is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.05 USP Endotoxin Units/mg of gemcitabine.

6 ADDITIONAL REQUIREMENTS

6.1 PACKAGING AND STORAGE

Preserve in tight containers.

6.2 LABELING

Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms.

Change to read:

6.3 USP REFERENCE STANDARDS (11)

USP Cytosine RS

Cytosine. (ERR 1-Aug-2021)

C₄H₅N₃O 111.10

USP Gemcitabine Hydrochloride RS