Provided by Beijing YinglaikeODS-100VReference to the 2015 edition of the Chinese Pharmacopoeia for the determination of glycyrrhizin content in chromatographic columns

liquiritinAlso known as glycyrrhizin. It is an important monomeric active ingredient in licorice flavonoids, with various pharmacological effects such as antioxidant and anti HIV.

On May 3, 2020, Beijing Daily reported that the team led by Xie Zhengwei from Peking University and the military team led by Qin Zhengfeng collaborated to publish a research paper in bioRxiv (an artificial intelligence system showed that glycyrrhizin inhibits SARS-CoV-2 by mimicking type I interferon). This research paper analyzed the changes in cellular transcriptome after the action of glycyrrhizin, revealed the potential antiviral mechanism of glycyrrhizin, and evaluated its safety at the mouse level, laying the foundation for its pharmacological development.

This application refers to the 2015 edition of the Chinese Pharmacopoeia and uses high-performance liquid chromatographyC18 chromatographic column TSKgel ODS-100V 5 μ m, 4.6 mm I.D. × 15 cm，Separate and detect glycyrrhizin. The analysis results indicate that glycyrrhizin and glycyrrhetinic acid are well separated, and the column efficiency can meet the requirement of more than 5000 specified in the pharmacopoeia.

sample preparation

Preparation of reference solution: Take appropriate amounts of glycyrrhizin reference substance and ammonium glycyrrhizinate reference substance, accurately weigh them, and add 70% ethanol to make 1 mL of solution containing 20 μ g glycyrrhizin and 0.2 mg ammonium glycyrrhizinate, respectively. This is the result (glycyrrhizic acid weight=ammonium glycyrrhizinate weight/1.0207).

Analysis conditions

Chromatography column: TSKgel ODS-100V (4.6mm I.D. × 15cm, 5 μ m)

Mobile phase: A: CH3CN, B: 0.05% H3PO4

Gradient: 0 min(19%A)→ 8 min(19%A)→ 35 min(50%A)→ 36 min(100%A)→ 40 min(19%A)

Flow rate: 1.0 mL/min

Column temperature: 40 ℃

Detector parameters: UV@237nm

Sample: Glycyrrhizin (20 μ g/mL, 70% ethanol), Ammonium Glycyrrhetinic Acid (0.2mg/mL, 70% ethanol)

Injection volume: 10 μ L

Analysis results

Chromatogram of Glycyrrhizin Separation

1. Glycyrrhizin (TP=19400>5000) 2. Glycyrrhetinic acid