Ammonium Glycyrrhizate

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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C₄₂H₆₂O₁₆ . NH₃                          839.97 (ERR 1-Jul-2022)

Monoammonium glycyrrhizinate;

Glycyrrhizic acid ammonium salt;

α-D-Glucopyranosiduronic acid, (3β,20β)-20-carboxy-11-oxo-30-norolean-12-en-3-yl 2-O-β-D-glucopyranuronosyl-, ammonium salt (1:1);

α-D-Glucopyranosiduronic acid, (3β,20β)-20-carboxy-11-oxo-30-norolean-12-en-3-yl 2-O-β-D-glucopyranuronosyl-, monoammonium salt CAS

RN: 53956-04-0.

1 DEFINITION

Ammonium Glycyrrhizate is a mixture of ammonium 18α- and 18β-glycyrrhizate (ammonium salt of (20β)-3β-[[2-0-(β-D-glucopyranosyluronic acid)-α-D-glucopyranosyluronic acid oxy]-11-oxoolean-12-en-29-oic acid), and the 18β-isomer is the main component. It contains NLT 78.0% and NMT 102.0% of ammonium 18α- and 18β-glycyrrhizate, on the anhydrous basis.

2 IDENTIFICATION

A. The retention times of the peaks of 18α- and 18β-glycyrrhizic acid from the Sample solution correspond to those from the System suitability solution, as obtained in the Content of Ammonium 18α- and 18β-Glycyrrhizate. [NOTE-The peak of 18α-glycyrrhizic acid could be absent in the Sample solution.]

B. IDENTIFICATION TESTS-GENERAL, Ammonium (191)

Acceptance criteria: Meets the requirements

3 ASSAY

Change to read:

3.1 CONTENT OF AMMONIUM 18α AND 18β-GLYCYRRHIZATE

3.1.1 Mobile phase

Acetonitrile, glacial acetic acid, and water (38:1:61)

3.1.2 Standard solution

0.5 mg/mL of USP Glycyrrhizic Acid RS in Mobile phase

3.1.3 System suitability solution

0.5 mg/mL of USP Ammonium Glycyrrhizate RS in Mobile phase

3.1.4 Sample solution

0.5 mg/mL of Ammonium Glycyrrhizate in Mobile phase

3.1.5 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 254 nm

Column: 3.9-mm x 30-cm analytical column; 5-10 µm packing L1

Flow rate: 2.0 mL/min

Injection volume: 10 µL

3.1.6 System suitability

Samples: System suitability solution and Standard solution

[NOTE-The relative retention times of 18β-glycyrrhizic acid and 18α-glycyrrhizic acid are about 1.0 and 1.2, respectively, System suitability solution.]

Suitability requirements

• Resolution: NLT 2.0 between the peaks due to 18β-glycyrrhizic acid and 18α-glycyrrhizic acid, System suitability solution
• Relative standard deviation: NMT 2.0%, Standard solution

3.1.7 Analysis

3.1.7.1 Samples

Standard solution, System suitability solution, and Sample solution

Determine the peak areas for each isomer (18α-glycyrrhizic acid or 18β-glycyrrhizic acid).

Calculate the percentage of ammonium 18α-glycyrrhizate (or ammonium 18β-glycyrrhizate) in the portion of Ammonium Glycyrrhizate

taken:

Result = (r_u/r_s) × (C_s/C_u) × (M_W(Salt)/M_W(Acid)) × 100

r_u = peak area of the 18α-glycyrrhizic acid (or 18β-glycyrrhizic acid) in the Sample solution

r_s = peak area of the 18β-glycyrrhizic acid in the Standard solution

C_s  = concentration of the USP Glycyrrhizic Acid RS in the Standard solution (mg/mL)

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

M_W(Salt) = molecular weight of ammonium glycyrrhizate, 839.97 (ERR 1-Jul-2022) g/mol

M_W(Acid) = molecular weight of glycyrrhizic acid, 822.94 (ERR 1-Jul-2022) g/mol

3.1.7.2 Acceptance criteria

The total percentage of ammonium 18α-glycyrrhizate and ammonium 18β-glycyrrhizate is 78.0%-102.0%, and the percentage of ammonium 18α-glycyrrhizate is NMT 13.0%, on the anhydrous basis.

4 IMPURITIES

4.1 RESIDUE ON IGNITION (281)

Sample: 1.0 g

Acceptance criteria: NMT 0.5%

4.2 LIMIT OF ORGANIC IMPURITIES

Mobile phase, System suitability solution, and Chromatographic system: Proceed as directed in Content of Ammonium 18α- and 188-

Glycyrrhizate.

Sample solution: 1.0 mg/mL of Ammonium Glycyrrhizate in the Mobile phase

Reference solution A: 0.05 mg/mL of Ammonium Glycyrrhizate in the Mobile phase, prepared from the Sample solution

Reference solution B: 0.057 mg/mL of Ammonium Glycyrrhizate in the Mobile phase, prepared from the Sample solution

System suitability

Sample: System suitability solution

[NOTE-The relative retention times for 24-hydroxyglycyrrhizinic acid, 18ẞ-glycyrrhizic acid, and 18α-glycyrrhizic acid are about 0.7, 1.0 and 1.2, respectively.]

Suitability requirements

Resolution: NLT 2.0 between the peaks due to 18ẞ-glycyrrhizic acid and 18α-glycyrrhizic acid

Analysis

Samples: System suitability solution, Reference solution A, Reference solution B, and Sample solution

Acceptance criteria: See Table 1.

a: (4β,20β)-3β-[[2-0-(B-D-Glucopyranosyluronic acid)-α-D-glucopyranosyluronic acid)oxy]-23-hydroxy-11-oxoolean-12-en-29-oic acid.

5 SPECIFIC TESTS

Change to read:

5.1 OPTICAL ROTATION, Specific Rotation (781S) (ERR-1-JUL-2022)

Sample solution: 10.0 mg/mL of Ammonium Glycyrrhizate in 50% ethanol

Acceptance criteria: +49.0° (ERR 1-Jul-2022) to +55.0" (ERR 1-Jul-2022) on the anhydrous basis

5.2 WATER DETERMINATION, Method la(921).

Sample: 0.25 g

5.3 Acceptance criteria: NMT 6.0%

6 ADDITIONAL REQUIREMENTS

6.1 PACKAGING AND STORAGE

Preserve in tight containers, and store in a cool, dry place.

6.2 USP REFERENCE STANDARDS (11)

USP Ammonium Glycyrrhizate RS

JSP Glycyrrhizic Acid RS.