Document Type : Research paper


1 Department of Horticulture, Faculty of Agriculture, University of Zanjan, Zanjan, Iran

2 Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran, Iran

3 Department of Photochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran, Iran


Licorice (Glycyrrhiza glabra L.) as a strategic and highly valuable medicinal plant in Iran with numerous beneficial pharmaceutical properties contributes substantially to Iranian herbs exports. In the present study, a variation on the phytochemical and antioxidant activity of 25 populations of valuable and profitable medicinal plant of G. glabra was investigated. The climate variables and soil properties were evaluated in various habitates of G. glabra. Total phenol (Folin–Ciocalteu method), total flavonoid (Aluminum Chloride method), anthocyanin (pH difference method) and antioxidant activity (DPPH method) were studied. Glycyrrhizic acid, glabridin, liquiritigenin and liquiritin content of root were evaluated by liquid chromatography. The content of major components in G. glabra varied in different regions. Outstanding quantitative variability of glycyrrhizic acid, glabridin, liquiritin and liquiritigenin content were observed in roots of licorice from different regions of Iran. Glycyrrhizic acid content was varied from 93.89 (mg/g dry weight) for the Sepidan population to 20.68 (mg/g dry weight) for the Ahar population. The maximum and minimum amounts of glabridin were recorded in Kashmar and Bajgah populations, respectively. The highest levels of liquritin and liquiritigenin were in the population of Kazerun and Yasuj, respectively, and the lowest was observed in Kashmar. Also, the highest antioxidant activity (the lowest IC50) for licorice root was associated with Semirom population. In conclusion, The populations with high amount of each active ingredient in licorice root and those with considerable antioxidant activity can be exploited depending on the purpose of breeding and cultivation.


Ahmadi Hosseini S.M, Kazme Souri M, Farhadi N, Moghaddam M, Omidbaigi R. 2014. Changes in Glycyrrhizin Content of Iranian licorice (Glycyrrhiza glabra L.) Affected by Different Root Diameter and Ecological Conditions. Agricultural Communications 2, 27-33.
2. Allison L, Bollen W, Moodie C. 1965. Total carbon. In: Black CA (ed.). Methods of Soil Analysis. Part 2. Chemical and Microbiological Properties. Agron Monogr no. 9, Soil Science Society of America, Madison, WI.
3. Asif H, Aftab A, Mohd M, Shah A.K, Afnan G.H, Firoz A. 2015. Quantitative Analysis of Total Phenolic, Flavonoid Contents and HPTLC Fingerprinting for Standardization of Glycyrrhiza glabra Linn. Roots. herbal-medicine 1, 1-9.
4. Becerro, M.A, Paul, V.J. 2004. Effects of depth and light on secondary metabolites production and cyanobacterial symbionts of the sponge Dysideagranulose. Marine Ecology Progress Series, 280, 115-28.
5. Blois M.S. 1958. Antioxidant determinations by the use of a stable free radical. Nature 181, 1199-1200.
6. Bremner J.M, Mulvaney C. 1982. Nitrogen-total. In: Page AL, Miller RH (eds.) Methods of Soil Analysis: Part 2. Chemical and Microbiological Properties. 2nd ed. Agron Monogr 9. ASA and SSSA, Madison, WI.
7. Cinatl J, Morgenstern B, Bauer G, Chandra P, Rabenau H, Doerr H. 2003. Glycyrrhizin, an
398 Int. J. Hort. Sci. Technol; Vol. 7, No. 4; December 2020
active component of liquorice roots, and replication of SARS-associated coronavirus. Lancet 361, 2045–2046.
8. Dehpour A, Zolfaghari M, Samadian T, Kobarfard F, Faizi M, Assari M. 1995. Antiulcer activities of liquorice and its derivatives in experimental gastric lesion induced by ibuprofen in rats. International Journal of Pharmaceutics 119, 133–138.
9. Deka K, Borthakur S.K, Tanti B. 2017. Distribution and population dynamics of Calamus nambariensis Becc. An endemic and threatened cane of Assam. Annals of Plant Sciences 6, 1829-1834.
10. Eghlima G.h, Hadian J, Motallbi Azar A.R. 2018. Survey on Diversity of Morphological and Biological Production Traits of Satureja rechingeri Jamzad Clones in Dezfool Climate. Plant Products 40:4, 41-53.
11. Fenwick G.K, Lutomski J, Nieman C. 1990. Licorice, Glycyrrhiza glabra L. composition, uses and analysis. Food Chemistry 38, 43-57.
12. Fu B, Liu J, Li H, Li L, Lee F.S, Wang X. 2005. The application of macroporous resins in the separation of licorice flavonoids and glycyrrhizic acid. Journal of Chromatography a 1089, 18–24.
13. Fujisawa Y, Sakamoto M, Matsushita M, Fujita T, Nishioka K. 2000. Glycyrrhizin inhibits the lytic pathway of complement—possible mechanism of its anti‐inflammatory effect on liver cells in viral hepatitis—. Microbiol. Immunol 44, 799–804.
14. Fukai T, Satoh K, Nomura T, Sakagami H. 2003. Preliminary evaluation of antinephritis and radical scavenging activities of glabridin from Glycyrrhiza glabra. Fitoterapia 74, 624-629.
15. Gairola S, Sharma C.M, Rana C.S, Ghildiyal S.K, Suyal S. 2010. Phytodiversity (Angiosperms and Gymnosperms) in Mandal-Chopta forest of Garhwal Himalaya, Uttarakhand, India. Natural Sciences 8, 1–17.
16. Gee G, Bauder J. 1979. Particle size analysis by hydrometer: a simplified method for routine textural analysis and a sensitivity test of measurement parameters. Soil Science Society of America Journal 43, 1004-1007.
17. Gobbo-Neto L, Guaratini T, Pessoa C, Moraes M, Costa-Lotufo L, Vieira R.F, Colepicolo P, Lopes N.P. 2010. Differential metabolic and biological profiles of Lychnophora ericoides from different localities in Brazilian “Campos
rupestres. Brazilian Chemical Society 21(4), 750- 9.
18. Guannan L, Dejan N, van Breemen R.B. 2016. Identification and Chemical Standardization of Licorice Raw Materials and Dietary Supplements Using UHPLC-MS/MS. Journal of Agricultural and Food Chemistry 64, 8062−8070.
19. Hadian J, Mirjalili M.H, Ganjpoor N. 2011. Morphological and phytochemical characterization of natural population of Satureja khuzestanica. Chemistry & Biodiversity 8, 1-15.
20. Haraguchi H, Yoshida N, Ishikawa H, Tamura Y, Mizutani K, Kinoshita T. 2000. Protection of mitochondrial functions against oxidative stresses by isoflavans from Glycyrrhiza glabra. Journal of Pharmacy and Pharmacology 52, 219-223.
21. Hayashi H, Hattori S, Inoue K, Khodzhimatov O, Ashurmetov O, Ito M, Honda G. 2003. Field survey of Glycyrrhiza plants in Central Asia (3). Chemical characterization of G. glabra collected in Uzbekistan. Chemical and Pharmaceutical Bulletin 51, 1338-1340.
22. Hayashi H, Shibano M, Kusano G, Yamamoto H, Ikeshiro Y. 1998. A field survey of Glycyrrhiza glabra L in Sicily and Spain. Nature Medicine 52, 259–264.
23. Hosseini S.M.A, Souri M.K, Farhadi N, Moghadam M, Omidbeigi R. 2014. Changes in glycyrrhizin content of Iranian licorice (Glycyrrhiza glabra L.) affected by different root diameter and ecological conditions. Agricultural Communications 2, 27-33.
24. Husain A, Ahmad A, Mujeeb M, Khan S.A, Alghamdi A.G, Anwar F. 2015. Quantitative analysis of total phenolic, flavonoid contents and HPTLC fingerprinting for standardization of Glycyrrhiza glabra Linn. Roots. Journal of Herbal Medicine 1, 1-9.
25. Jovancevic M, Balijagic J, Menkovic N, Savikin K, Zdunic G, Jankovic T, Dekic-Ivankovic M. 2011. Analysis of Phenolic compounds in wild populations of bilberry (Vaccinium myrtillus) from Montenegro. Journal of Medicinal Plants Research 5(6), 910- 4.
26. Karami Z, Mirzaei H, Emam-Djomeh Z, Sadeghi Mahoonak A.R, Khomeiri M, 2013. Effect of harvest time on antioxidant activity of Glycyrrhiza glabra root extract and evaluation of its antibacterial activity. International Food Research Journal 20, 2951-2957.
Investigation of Phytochemical Variability, Antioxidant … 399
27. Khanahmadi M, NaghdiBadi H, Akhondzadeh S, Khalighi–Sigaroodi F, Mehrafarin A, Shahriari S, Hajiaghaee R, 2013. A Review on Medicinal Plant of Glycyrrhiza glabra L. Journal of Medicinal Plants 2, 1-12.
28. Khym J.X. 1974. Analytical Ion-Exchange Procedures in Chemistry and Biology: Theory, Equipment, Techniques. Prentice-Hall, NJ.
29. Kim Y.W, Ki S.H, Lee J.R, Lee S.J, Kim C.W, Kim S.C, Kim S.G. 2006. Liquiritigenin, an aglycone of liquiritin in Glycyrrhizae radix, prevents acute liver injuries in rats induced by acetaminophen with or without buthionine sulfoximine. Chemico-Biological Interactions 161, 125-138.
30. Ko Y.H, Kwon S.H, Hwang J.Y, Kim K.I, Seo J.Y, Nguyen T.L, Lee S.Y, Kim H.C, Jang C.G. 2018. The memory-enhancing effects of liquiritigenin by activation of NMDA receptors and the CREB signaling pathway in mice. Biomolecules & Therapeutics 26, 109-114.
31. Kovalenko P.G, Antonjuk V.P, Maliuta S.S. 2004. Secondary metabolites synthesis in transformed cells of Glycyrrhiza glabra L. and Potentilla alba L. as producents of radioprotective compounds. Ukrainica Bioorganica Acta 1, 13-22.
32. Kuang Y, Li B, Fan J, Qiao X, Ye M, 2018. Antitussive and expectorant activities of licorice and its major compounds. Bioorganic & Medicinal Chemistry 26, 278-284.
33. Mareshige K, Hayashi S, Shibata T, Yamamoto Y, Sekizaki H. 2011. Variation of glycyrrhizin and liquiritin contents within a population of 5-year-old licorice (Glycyrrhiza uralensis) plants cultivated under the same conditions. Biological and Pharmaceutical Bulletin 34(8), 1334-1337.
34. Mareshige K, Shigeki H, Toshiro S, Yutaka Y, Haruo S, 2011. Variation of Glycyrrhizin and Liquiritin Contents within a Population of 5-Year-Old Licorice (Glycyrrhiza uralensis) Plants Cultivated under the Same Conditions. Biological and Pharmaceutical Bulletin 34, 1334-1337.
35. Medina-Holguin B.A.L, Micheletto S, Holguin F.O, Rodriguez J, O’connell MA. 2007. Environmental influences on essential oils in roots of Anemopsis californica. Horticultural Science 42, 1578–1583.
36. Nelson D, Sommers L.E. 1982. Total carbon, organic carbon, and organic matter. Methods of soil analysis. Part 2. Chemical and
microbiological properties (methodsofsoilan) 2, 539-579.
37. Oloumi H, Hassibi N. 2011. Study the correlation between some climate parameters and the content of phenolic compounds in roots of Glycyrrhiza glabra. Journal of Medicinal Plants Research 5, 6011-6016.
38. Olukoga A, Donaldson D. 1998. Historical perspectives on health. The history of liquorice: the plant, its extract, cultivation, and commercialisation and etymology. Journal of the Royal Society of Health 118, 300-304.
39. Omidbeigi R. 2006. Processing and production of medicinal plants. Razavi Publications, Mashhad. In Persian.
40. Park S.M, Ki S.H, Han N.R, Cho I.J, Ku S.K, Kim S.C, Zhao R.J, Kim Y.W. 2015. Tacrine, an oral acetylcholinesterase inhibitor, induced hepatic oxidative damage, which was blocked by liquiritigenin through GSK3-beta inhibition. Biological and Pharmaceutical Bulletin 38, 184-192.
41. Parvaiz M, Hussain K, Khalid S, Hussnain N, Iram N. 2014. A Review: Medicinal Importance of Glycyrrhiza glabra L. (Fabaceae Family). Global Journal of Pharmacology 8, 8-13.
42. Peluso M.R. 2006. Flavonoids attenuate cardiovascular disease, inhibit phosphodiesterase, and modulate lipid homeostasis in adipose tissue and liver. Experimental Biology and Medicine 231(8), 1287 1299.
43. Pourmorad F, Hosseinimehr SJ, Shahabimajd N. 2006. Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants. African journal of biotechnology 5(11), 1142-1145.
44. Sanja V, Filip S, Izabella S, Istvan Z, Imre O, Suzana J.S. 2018. Chemical composition, antioxidant and anticancer activity of licorice from Fruska Gora locality. Industrial Crops and Products 112, 217–224.
45. Saraf B.D, Inam F, Deo S.S. 2013. Antimicrobial and Antioxidant Activities of Methanol Extract Roots of Glycyrrhiza glabra and HPLC analysis. International Journal of Pharmacy and Pharmaceutical Sciences 5, 157-160.
46. Scherf A, Treutwein J, Kleeberg H, Schmitt A, 2012. Efficacy of leaf extract fractions of Glycyrrhiza glabra L. against downy mildew of cucumber (Pseudoperonospora cubensis). European Journal of Plant Pathology 134, 755–762.
400 Int. J. Hort. Sci. Technol; Vol. 7, No. 4; December 2020
47. Simmler C, Pauli G.F, Chen S.N. 2013. Phytochemistry and biological properties of glabridin. Fitoterapia 90, 160-184.
48. Siracusa L, Saija A, Cristani M, Cimino F, D’Arrigo M, Trombetta D, Rao F, Ruberto G. 2011. Phytocomplexes from liquorice (Glycyrrhiza glabra L.) leaves – chemical characterization and evaluation of their antioxidant, anti-genotoxic and anti-inflammatory activity. Fitoterapia 82, 546–556.
49. Song W, Qiao X, Chen K, Wang Y, Ji S. 2017. Biosynthesis Based Quantitative Analysis of 151 Secondary Metabolites of Licorice to differentiate Medicinal Glycyrrhiza Species and Their Hybrids. Analytical Chemistry 89, 3146-3153.
50. Sultana S, Haque A, Hamid K, Urmi K, Roy S. 2010. Antimicrobial, cytotoxic and antioxidant activity of methanolic extract of Glycyrrhiza glabra. Agriculture and Biology Journal of North America 1, 957–960.
51. Tamir S, Eizenberg M, Somjen, D, Stern N, Shelach R, Kaye A, Vaya J. 2000. Estrogenic and antiproliferative properties of glabridin from licorice in human breast cancer cells. Cancer Research 60, 5704–5709.
52. Tupe R.S, Kemse N.G, Khaire A.A, 2013. Evaluation of antioxidant potentials and total phenolic contents of selected Indian herbs powder extracts. International Food Research Journal 20, 1053-1063.
53. Visavadiya N.P, Narasimhacharya A. 2006. Hypocholesterolaemic and antioxidant effects of Glycyrrhiza glabra (Linn) in rats. Molecular Nutrition and Food Research 23, 1080–1086.
54. Wang L, Yang R, Yuan B, Liu Y, Liu C. 2015. The antiviral and antimicrobial activities of licorice, a widely-used Chinese herb. Acta Pharmaceutica Sinica B 5, 310–315.
55. Yang R, Wang L.Q, Yuan B.C, Liu Y. 2015. The pharmacological activities of licorice. Planta Medica 81, 1654–1669.
56. Zhang J, Xu B, Li M. 2011. Relationships between the bioactive compound content and environmental variables in Glycyrrhiza uralensis populations in different habitats of North China. Journal experimental botany 80, 161-166.
57. Zhang Q, Ye M, 2009. Chemical analysis of the Chinese herbal medicine Gan-Cao (licorice). Journal of Chromatography A 1216, 1954–1969.
58. Zhang Y, He Y, Yu H, Ma F, Wu J, Zhang X. 2015. Liquiritigenin protects rats from carbon etrachloride Induced Hepatic Injury through PGC-1α Pathway. Evidence-Based Complementary and Alternative Medicine, 649-568.
59. Zheng H, Li Y, Wang Y, Zhao H, Zhang J, Chai H, Tang T, Yue J, Guo A.M, Yang J. 2014. Downregulation of COX-2 and CYP 4A signaling by isoliquiritigenin inhibits human breast cancer metastasis through preventing anoikis resistance, migration and invasion. Toxicology and Applied Pharmacology 280, 10–20.
60. Zhou C.M. 2006. Cultivation techniques for Glycyrrhiza uralensis. Xinjiang Farmland Reclamation Science and Technology 15, 14-15.