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. 2005 Nov 28;17(1):25–31. doi: 10.1002/pca.877

Simultaneous HPLC analysis, with isocratic elution, of glycyrrhizin and glycyrrhetic acid in liquorice roots and confectionery products

Cesare Sabbioni 1, Anna Ferranti 1, Francesca Bugamelli 1, Giorgio Cantelli Forti 2, Maria Augusta Raggi 1,
PMCID: PMC7167644  PMID: 16454473

Abstract

Glycyrrhizin (1), the main active principle of Glycyrrhiza glabra (liquorice) roots, is extensively used in herbal medicines, in pharmaceutical preparations and confectionery products. A feasible and reliable method which allows the simultaneous analysis of 1 and its aglycone, 18β‐glycyrrhetic acid (2), by means of an isocratic HPLC procedure is described. The system uses a C8 column as the stationary phase, and a mixture of acetonitrile, methanol, water and glacial acetic acid as the mobile phase. Good linearity was found in the concentration ranges 1–50 and 0.05–2.50 µg/mL for 1 and 2, respectively. A simple and rapid sample pre‐treatment, based on the extraction of the two analytes with a mixture of water and ethanol, was developed for the examination of liquorice confectionery products and root samples. The HPLC method was shown to be appropriate, in terms of precision and feasibility, for the quality control of the analytes in these matrices. Copyright © 2005 John Wiley & Sons, Ltd.

Keywords: Liquid chromatography, glycyrrhizin, glycyrrhetic acid, confectionery products, liquorice root, Glycyrrhiza glabra

REFERENCES

  1. Baltina LA. 2003. Chemical modification of glycyrrhizic acid as a route to new bioactive compounds for medicine. Curr Med Chem 10: 155– 171. [DOI] [PubMed] [Google Scholar]
  2. Bernardi M, D'Intino PE, Trevisani F, Cantelli‐Forti G, Raggi MA, Turchetto E, Gasbarrini G. 1994. Effects of prolonged ingestion of graded doses of licorice by healthy volunteers. Life Sci 55: 863– 872. [DOI] [PubMed] [Google Scholar]
  3. Cantelli Forti G, Maffei F, Hrelia P, Bugamelli F, Bernardi M, D'Intino P, Maranesi M, Raggi MA. 1994. Interaction of licorice on glycyrrhizin pharmacokinetics. Environ Health Perspect 102: 65– 69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cinatl J, Morgenstern B, Bauer G, Chandra P, Rabenau H, Doerr HW. 2003. Glycyrrhizin, an active component of licorice roots, and replication of SARS‐associated coronavirus. Lancet 361: 2045– 2046 [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Collinge A, Hermesse B, Noirfalise A. 1985. Study and determination by high‐performance liquid chromatography of glycyrrhizin in food products. J Food Chem Biotechnol 40: 143– 145. [Google Scholar]
  6. De Groot G, Koops R, Hogendoorn EA, Goewie CE, Savelkoul TJ, Van Vloten P. 1988. Improvement of selectivity and sensitivity by column switching in the determination of glycyrrhizin and glycyrrhetic acid in human plasma by high‐performance liquid chromatography. J Chromatogr 456: 71– 81. [DOI] [PubMed] [Google Scholar]
  7. Doll R, Hill ID, Hutton C, Underwood DJ. 1962. Clinical trial of a triterpenoid liquorice compound in gastric and duodenal ulcer. Lancet 2: 793– 796. [DOI] [PubMed] [Google Scholar]
  8. European Pharmacopoeia. 2005. 5th edn. Council of Europe: Strasbourg; 1920– 1921. [Google Scholar]
  9. Fenwick GR, Lutomski J, Nieman C. 1990. Liquorice, Glycyrrhiza glabra L. Composition, uses and analysis. Food Chem 38: 119– 143. [Google Scholar]
  10. Francischetti IM, Monteiro RQ, Guimaraes JA, Francischetti B. 1997. Identification of glycyrrhizin as a thrombin inhibitor. Biochem Biophys Res Commun 9: 259– 263. [DOI] [PubMed] [Google Scholar]
  11. Frattini C, Bicchi C, Barettini C, Nano GM. 1977. Volatile flavor components of licorice. J Agric Food Chem 25: 1238– 1241. [Google Scholar]
  12. Fujioka T, Kondou T, Fukuhara A, Tounou S, Mine M, Mataki N, Hanada K, Ozaka M, Mitani K, Nakaya T, Iwai T, Miyakawa H. 2003. Efficacy of a glycyrrhizin suppository for the treatment of chronic hepatitis C: a pilot study. Hepatol Res 26: 10– 14. [DOI] [PubMed] [Google Scholar]
  13. Hermesse B, Collinge A, Noirfalise A. 1986. Glycyrrhizin levels in some food products. Arch Belges 44: 60– 67. [PubMed] [Google Scholar]
  14. Hurst W, McJeffrey KJ, Michael, MRA . 1983. High‐performance liquid chromatographic determination of glycyrrhizin in licorice products. J Agric Food Chem 31: 387– 389. [Google Scholar]
  15. Ichikawa T, Ishida S, Sakiya Y, Yoko A, Yoshinobu U. 1984. Highperformance liquid chromatographic determination of glycyrrhizin and glycyrrhetinic acid in biological materials Chem Pharm Bull 32: 3734– 3738. [DOI] [PubMed] [Google Scholar]
  16. Lauren DR, Jensen DJ, Douglas JA, Follet JM. 2001. Efficient method for determining the glycyrrhizin content of fresh and dried roots, and root extracts, of Glycyrrhiza species. Phytochem Anal 12: 332– 335. [DOI] [PubMed] [Google Scholar]
  17. Liu J, Manheimer E, Tsutani K, Gluud C. 2003. Medicinal herbs for hepatitis C virus infection: a Cochrane hepatobiliary systematic review of randomized trials. Am J Gastroenterol 98: 538– 544. [DOI] [PubMed] [Google Scholar]
  18. Mizutani K, Kuramoto T, Tamura Y, Ohtake N, Doi S, Nakaura M, Tanaka O. 1994. Sweetness of glycyrrhetic acid 3‐Oβ‐d‐monoglucuronide and the related glycosides Biosci Biotechnol Biochem 58: 554– 555. [DOI] [PubMed] [Google Scholar]
  19. Okamura N, Maki T, Miyauchi H, Shimoe M, Yokono S, Yoshitomi H, Yagi A. 2001. Simultaneous determination of glycyrrhizin, glycyrrhetic acid and glycyrrhetic acid mono‐glucuronide in Shakuyaku‐kanzo‐to incubated with rat feces by semi‐micro high‐performance liquid chromatography. Biol Pharm Bull 24: 1161– 1164. [DOI] [PubMed] [Google Scholar]
  20. Ong ES, Len SM. 2003. Pressurized hot water extraction of berberine, baicalein and glycyrrhizin in medicinal plants. Anal Chim Acta 482: 81– 89. [Google Scholar]
  21. Raggi MA, Maffei F, Bugamelli F, Cantelli Forti G. 1994a. Bioavailability of glycyrrhizin and licorice extract in rat and human plasma as detected by a HLPC method. Pharmazie 49: 269– 272. [PubMed] [Google Scholar]
  22. Raggi MA, Bugamelli F, Nobile L, Schiavone P, Cantelli Forti G. 1994b. HPLC determination of glycyrrhizin and glycyrrhetic acid in biological fluids, after licorice extract administration to humans and rats. Boll Chim Farm 133: 704– 708. [PubMed] [Google Scholar]
  23. Sharaf A, Gomaa N, El‐Gamal MHA. 1975. Glycyrrhetic acid as an active estrogenic substance separated from Glycyrrhiza glabra (licorice). Egypt J Pharm Sci 16: 245– 251. [Google Scholar]
  24. Spinks EA, Fenwick GR. 1990. The determination of glycyrrhizin in selected UK licorice products. Food Add Contam 7: 769– 771. [DOI] [PubMed] [Google Scholar]
  25. Ukiya M, Akihisa T, Tokuda H, Suzuki H, Mukainaka T, Ichiishi E, Yasukawa K, Kasahara Y, Nishino H. 2002. Constituents of Compositae plants III. Anti‐tumor promoting effects and cytotoxic activity against human cancer cell lines of triterpene diols and triols from edible chrysanthemum flowers. Cancer Lett 177: 7– 12. [DOI] [PubMed] [Google Scholar]
  26. United States Pharmacopeia. 2005. 28th edn. United States Pharmacopeial Convention: Rockville, MD; 2748– 2751. [Google Scholar]
  27. Van Rossum TG, Vulto AG, De Man RA, Brouwer JT, Schalm SW. 1998. Glycyrrhizin as a potential treatment for chronic hepatitis C. Aliment Pharmac Ther 12: 199– 205. [DOI] [PubMed] [Google Scholar]
  28. Yamamura Y, Kawakami J, Santa T, Kotaki H, Uchino K, Sawada Y, Iga T. 1991. Selective high‐performance liquid chromatographic method for the determination of glycyrrhizin and glycyrrhetic acid‐3‐O‐glucuronide in biological fluids: application of ion‐pair extraction and fluorescence labelling agent. J Chromatogr 567: 151– 160. [DOI] [PubMed] [Google Scholar]

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