Glucagon 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

Glucagon for Injection is a sterile lyophilized mixture of the hydrochloride of glucagon with one or more suitable buffering and stabilizing agents. It contains NLT 65% and NMT 110% of the labeled amount of glucagon.

2 IDENTIFICATION

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

B. GLUCAGON BIOIDENTITY TESTS (123): Meets the requirements

3 ASSAY

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3.1 PROCEDURE

Solution A: Dissolve 16.3 g of monobasic potassium phosphate in 750 mL of water, adjust with phosphoric acid to a pH of 2.7 (±0.05), add water to 800 mL, add 200 mL of acetonitrile, and degas.

Solution B: Prepare a degassed solution of acetonitrile and water (4:6).

Mobile phase: See Table 1. [NOTE-The ratio of Solution A to Solution B can be adjusted to obtain a retention time of about 21 min for the main peak.]

Table 1

^a The end time of the isocratic elution can be adjusted so that the gradient begins after the fourth desamido peak elutes (relative retention time about 1.4). The rest of the program is then adjusted accordingly with this offset.

System suitability solution: Reconstitute a vial of USP Glucagon (Human) RSA (USP 1-Aug-2022) in 0.01 N hydrochloric acid to obtain a solution with a concentration of about 0.5 mg/mL. Let stand at 50° for 48 h. At least 7% total of all four desamido glucagons should be present in the solution.

Standard solution: Reconstitute a vial of USP Glucagon (Human) RSA (USP 1-Aug-2022) in 0.01 N hydrochloric acid to obtain a solution with a concentration of about 0.5 mg/mL.

Sample solution: Dissolve the substance to be examined in 0.01 N hydrochloric acid in order to obtain a concentration of 0.5 mg/mL of glucagon.

3.2 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 214 nm

Column: 3-mm x 15-cm; 3-µm or less packing L1

Column temperature: 45°

Flow rate: 0.5 mL/min

Injection volume: 15 µL

3.3 System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 1.5 between the main peak and the first eluting desamido peak. Four peaks eluting after the glucagon peak that correspond to the desamido glucagons are clearly visible, System suitability solution.

Tailing factor: NMT 1.8 for the glucagon peak, Standard solution

Relative standard deviation: NMT 2.0%, Standard solution

3.4 Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of glucagon (C₁₅₃H₂₂₅N₄₃O₄₉S) in the portion of Glucagon for Injection 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 the Standard solution (mg/mL)

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

Acceptance criteria: 65%-110%

4 PERFORMANCE TESTS

UNIFORMITY OF DOSAGE UNITS (905): Meets the requirements

5 IMPURITIES

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ORGANIC IMPURITIES

Solution A, Solution B, Mobile phase, System suitability solution, Standard solution, Chromatographic system, and System suitability: Proceed as directed in the Assay.

Sample solution: Dissolve the substance to be examined in 0.01 N hydrochloric acid (USP 1-Aug-2022) in order to obtain a concentration of 0.5 mg/mL of glucagon.

Analysis

Sample: Sample solution

Calculate separately the percentage of each impurity in the portion of Glucagon for Injection taken:

Result = (r_U/r_T) x 100

r_U = peak response for each impurity

r_T = sum of all the peak responses

Acceptance criteria: NMT 14% total of all four desamido glucagons is found, and NMT 31% of total impurities and related compounds is found.

6 SPECIFIC TESTS

6.1 WATER DETERMINATION (921), Method I, Method Ic

NMT 4.0%

6.2 PH AND CLARITY OF SOLUTION

Dissolve it in the solvent and in the concentration recommended in the labeling: the pH of the solution is between 1.7 and 3.5, and the solution is clear.

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6.3 BACTERIAL ENDOTOXINS TEST (85)

Meets the requirements (USP 1-Aug-2022)

6.4 STERILITY TESTS (71)

Meets the requirements

6.5 CONSTITUTED SOLUTION

At the time of use, it meets the requirements for Injections and Implanted Drug Products (1), Product Quality Tests Common to Parenteral Dosage Forms, Specific Tests, Completeness and Clarity of Solutions.

7 ADDITIONAL REQUIREMENTS

7.1 PACKAGING AND STORAGE

Preserve as described in Packaging and Storage Requirements (659), Injection Packaging, Packaging for Constitution.

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7.2 LABELING

The labeling states that the material is synthetic or of recombinant DNA origin. (USP 1-Aug-2022)

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7.3 USP REFERENCE STANDARDS (11)

USP Glucagon (Human) RSA (USP 1-Aug-2022)