Gemcitabine 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

Gemcitabine for Injection contains an amount of gemcitabine hydrochloride equivalent to NLT 95% and NMT 105% of the labeled amount of gemcitabine (C₉H₁₁F₂N₃O₄).

[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), Ultraviolet-Visible Spectroscopy: 197U

Medium: 0.14 M phosphate buffer with a pH of 2.5 prepared as follows. Add 13.8 g of monobasic sodium phosphate and 2.5 mL of phosphoric acid to 1 L of water.

Sample solution: 16 µg/mL of gemcitabine in Medium

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

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 a-anomer.]

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

Sample solution: Equivalent to 0.1 mg/mL of gemcitabine in water from Gemcitabine for Injection prepared as follows. Reconstitute a suitable number of vials with an appropriate amount of water, based on the labeled amount of gemcitabine.

3.2 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 275 nm

Column: 4.6-mm x 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 a-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 the labeled amount of gemcitabine (C₉H₁₁F₂N₃O₄) in the portion of Gemcitabine for Injection taken:

Result = (r_U/r_S) x (C_S/C_U) x (M_r1/M_r2) 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 = nominal concentration of gemcitabine in the Sample solution (mg/mL)

M_r1 = molecular weight of gemcitabine, 263.20

M_r2 = molecular weight of gemcitabine hydrochloride, 299.66

Acceptance criteria: 95%-105%

4 PERFORMANCE TESTS

UNIFORMITY OF DOSAGE UNITS, Weight Variation (905): Meets the requirements

5 IMPURITIES

Change to read:

5.1 ORGANIC IMPURITIES

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

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

Solution B: Methanol

Mobile phase: See Table 1.

Table 1

Standard solution: 2 µg/mL each of USP Gemcitabine Hydrochloride RS and USP Cytosine RS in water

Sample solution: Equivalent to 2 mg/mL of gemcitabine in water prepared by reconstituting the vial with an appropriate amount of water, based on the labeled amount of gemcitabine

5.2 System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 8.0 between gemcitabine α-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

5.3 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of cytosine, expressed as a percentage of gemcitabine hydrochloride (C₉H₁₁F₂N₃O₄·HCI), in the portion of Gemcitabine for Injection taken:

Result = (r_U/r_S) x (C_S/C_U) x (M_r1/M_r2) 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 = nominal concentration of gemcitabine in the Sample solution (mg/mL)

M_r1 = molecular weight of gemcitabine, 263.20

M_r2 = molecular weight of gemcitabine hydrochloride, 299.66

Calculate the percentage of each impurity other than cytosine, expressed as a percentage of gemcitabine hydrochloride (C₉H₁₁F₂N₃O₄·HCI), in the portion of Gemcitabine for Injection taken:

Result = (r_U/r_S) x (C_S/C_U) x (M_r1/M_r2) x 100

r_U = peak response for 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 = nominal concentration of gemcitabine in the Sample solution (mg/mL)

M_r1 = molecular weight of gemcitabine, 263.20

M_r2 = molecular weight of gemcitabine hydrochloride, 299.66

Acceptance criteria: See Table 2. Disregard any impurity peaks less than 0.02%.

Table 2

^a 2'-Deoxy-2',2'-difluorocytidine (α-isomer).

6 SPECIFIC TESTS

6.1 PARTICULATE MATTER IN INJECTIONS (788)

It meets the requirements for small-volume injections.

6.2 PH (791)

Sample solution: 40 mg/mL of gemcitabine in 0.9% sodium chloride solution

Acceptance criteria: 2.7-3.3

6.3 CLARITY OF SOLUTION

Sample solution: Dissolve it in the solvent and at the concentration recommended in the labeling.

Analysis: Determine the turbidity by ratio turbidimetry within 15 min of reconstitution, corrected for a diluent blank (see Nephelometry and Turbidimetry (855)).

Acceptance criteria: NMT 10 NTU

6.4 BACTERIAL ENDOTOXINS TEST (85)

It contains NMT 0.05 USP Endotoxin Unit/mg of gemcitabine.

6.5 STERILITY TESTS (71)

It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration.

7 ADDITIONAL REQUIREMENTS

7.1 PACKAGING AND STORAGE

Preserve as described in Packaging and Storage Requirements (659), Injection Packaging, Packaging for constitution. Store at controlled room temperature. Do not refrigerate after reconstitution.

Change to read:

7.2 USP REFERENCE STANDARDS (11)

USP Cytosine RS

Cytosine (ERR 1-Aug-2021)

C₄H₅N₃O 111.10

USP Gemcitabine Hydrochloride RS