Planta Med 2010; 76(7): 729-733
DOI: 10.1055/s-0029-1240690
Analytical Studies
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

Determination of Flavonoids in Licorice Using Acid Hydrolysis and Reversed-Phase HPLC and Evaluation of the Chemical Quality of Cultivated Licorice

Makio Shibano1 , Kazuo Ozaki2 , Hitoshi Watanabe2 , Akinori Tabata1 , Masahiko Taniguchi1 , Kimiye Baba1
  • 1Osaka University of Pharmaceutical Sciences, Nasahara, Takatsuki, Osaka, Japan
  • 2Kyoto Herbal Garden, Takeda Pharmaceutical Company, Ltd., Ichijoji, Sakyoku, Kyoto, Japan
Further Information

Publication History

received Sept. 3, 2009 revised Nov. 10, 2009

accepted Nov. 13, 2009

Publication Date:
14 December 2009 (online)

Abstract

Licorice contains flavonoids and triterpenoids as the major bioactive components. Most of the flavonoids are the glycosidic form of liquiritigenin (LIQ), isoliquiritigenin (ISO) and formononetin (FOR). A reversed-phase HPLC method for the quantification of LIQ, ISO and FOR in licorice was developed. This method does not measure each glycoside but measures the aglycones using acid hydrolysis. All calibration curves showed good linear regression (r > 0.9998). The method showed good precision for intraday (RSD < 2.14 %) and interday (RSD < 0.51 %) assays. The limit of detection was 0.031 µg for LIQ, 0.011 µg for ISO and 0.006 µg for FOR. The limit of quantification was 0.31 µg for LIQ, 0.11 µg for ISO and 0.06 µg for FOR. The flavonoid contents along with the glycyrrhizin content of cultivated licorice from seedling plants in Japan and commercial wild licorice were investigated. This new method could be extremely useful for evaluating the quality of licorice.

References

  • 1 Yamamoto Y, Tani T. Growth and glycyrrhizin contents in Glycyrrhiza uralensis roots cultivated for four years in eastern Nei-Meng-gu of China.  J Tradit Med. 2002;  19 87-92
  • 2 Chin Y W, Jung H A, Liu Y, Su B N, Castoro J A, Keller W J, Pereira M A, Kinghorn A D. Anti-oxidant constituents of the roots and stolons of licorice (Glycyrrhiza glabra).  J Agric Food Chem. 2007;  55 4691-4697
  • 3 Kwon G T, Cho H J, Chung W Y, Park K K, Moon A, Park J H Y. Isoliquiritigenin inhibits migration and invasion of prostate cancer cells: possible mediation by decreased JNK/AP-1 signaling.  J Nutr Biochem. 2009;  20 663-676
  • 4 Jayaprakasam B, Doddaga S, Wang R, Holmes D, Goldfarb J, Li X M. Licorice flavonoids inhibit eotaxin-1 secretion by human fetal lung fibroblasts in vitro.  J Agric Food Chem. 2009;  57 820-825
  • 5 Ma J, Fu N Y, Pang D B, Wu W Y, Xu A L. Apoptosis induced by isoliquiritigenin in human gastric cancer MGC-803 cells.  Planta Med. 2001;  67 754-757
  • 6 Kumar S, Sharma A, Madan B, Singhal V, Ghosh B. Isoliquiritigenin inhibits IκB kinase activity and ROS generation to block TNF-α induced expression of cell adhesion molecules on human endothelial cells.  Biochem Pharmacol. 2007;  73 1602-1612
  • 7 Wang Y C, Yang Y S. Simultaneous quantification of flavonoids and triterpenoids in licorice using HPLC.  J Chromatogr B Anal Technol Biomed Life Sci. 2007;  850 392-399
  • 8 Li H, Chen B T, Liu L, Liu Q. Simultaneous determination of six compounds in licorice and related Chinese herbal preparations.  Chromatographia. 2009;  69 229-235
  • 9 Hayashi H, Inoue K, Ozaki K, Watanabe H. Comparative analysis of ten strains of Glycyrrhiza uralensis cultivated in Japan.  Biol Pharm Bull. 2005;  28 1113-1116
  • 10 Shibano M, Matsumoto Y, Kusano G, Shibata T. Researches of Glycyrrhiza species grown at medicinal plant gardens in Japan and basic studies for selection of pharmaceutically fine races. 1. Comparative studies by HPLC patterns and constituents of aerial parts.  Nat Med. 1996;  50 273-283
  • 11 Yamamoto Y, Tani T. Evaluation of glycyrrhizin variation in Gancao collected from 1986 to 2000 at drug market in Japan and discrimination of Daitou-Gancao and Dongbei-Gancao.  J Tradit Med. 2001;  18 197-202

Makio Shibano

Osaka University of Pharmaceutical Sciences

4–20–1 Nasahara

Takatsuki

569–1094 Osaka

Japan

Phone: + 81 7 26 90 10 73

Fax: + 81 7 26 90 10 05

Email: shibano@gly.oups.ac.jp

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