, Volume 63, Supplement 13, pp S111–S114 | Cite as

HPLC Analysis of Alizarin and Purpurin Produced by Rubia tinctorum L. Hairy Root Cultures

  • P. Bányai
  • I. N. Kuzovkina
  • L. Kursinszki
  • É. Szőke


We have determined the quantities of the anthraquinones alizarin and purpurin, with especial regard to their effective antigenotoxic activity, in genetically transformed hairy root cultures of Rubia tinctorum L. Hairy roots were cultured on solid and in liquid Gamborg B5 and 1/2 NMS media in a shaking cabinet and in a bioreactor. Methanolic extracts of lyophilized hairy roots were hydrolysed, and then purified by solid phase extraction with good recovery. A new HPLC method was developed for the determination of alizarin and purpurin. The analysis was performed on a Luna C8 RP column using a 45:55 (v/v) mixture of acetonitrile:20 mM ammonium formate-formic acid buffer (pH 3.00) as eluent. Peaks were identified by addition of standards and by diode-array detection. External standardization allowed the determination of alizarin and purpurin with good sensitivity and reliability. The maximum purpurin content was observed in cultures cultivated on solid B5 medium (5.94 mg g−1). The highest alizarin content was measured in cultures cultivated on solid 1/2 NMS medium (2.14 mg g−1).


Column liquid chromatography Rubia tinctorum L Hairy root culture Alizarin Purpurin 


  1. Marec F, Kollárová I, Jegorov A (2001) Planta Med 67:127–131CrossRefGoogle Scholar
  2. Kawasaki Y, Goda Y, Noguchi H, Yamada T (1994) Chem Pharm Bull 42 (9):1971–1973Google Scholar
  3. Kawasaki Y, Goda Y, Yoshihiro K (1992) Chem Pharm Bull 40 (6):1504–1509Google Scholar
  4. Hao JN, Huang MP, Lee H (1995) Mutat Res 328:183–191Google Scholar
  5. Marczylo TH, Hayatsu T, Arimoto-Kobayashi S, Tada M, Fujita K, Kamataki T, Nakayama K, Hayatsu H (1999) Mutat Res 444 (2):451–461Google Scholar
  6. Marczylo TH, Arimoto-Kobayashi S, Hayatsu H (2000) Mutagenesis 15 (3):223–228Google Scholar
  7. Takahashi E, Marczylo TH, Watanabe T, Nagai S, Hayatsu H, Negishi T (2001) Mutat Res 480-481:139–145Google Scholar
  8. Tóth ZA, Raatikainen O, Naaranlahti T, Auriola S (1993) J Chromatogr 630:423–428CrossRefGoogle Scholar
  9. Krizsán K, Szókán Gy, Tóth ZA, Hollósy F, László M, Khlafulla A (1996) J Liq Chrom & Rel Technol 19 (14):2295–2314Google Scholar
  10. Eichinger D, Adelbert B, Zenk MH, Eisenreich W (1999) J Am Chem Soc 121 (33):7469–7475Google Scholar
  11. Xu XR, Li HB, Gu JD, Paeng KJ (2004) Chromatographia 59 (11-12):745–747Google Scholar
  12. Derksen GCH, van Beek TA, de Groot Æ, Capelle A (1998) J Chromatogr A 816:277–281CrossRefGoogle Scholar
  13. Derksen GCH, Niederländer HAG, van Beek TA (2002) J Chromatogr A 978:119–127CrossRefGoogle Scholar
  14. Derksen GCH, Naayer M, van Beek TA, Capelle A, Haaksman IK, van Doren HA, de Groot Æ (2003) Phytochem Anal 14:137–144CrossRefGoogle Scholar
  15. Derksen GCH, Lelyveld GP, van Beek TA, Capelle A, de Groot Æ (2004) Phytochem Anal 15:397–406CrossRefGoogle Scholar
  16. van der Heijden R, Verpoorte R, Hoekstra SS, Hoge JHC (1994) Plant Physiol Biochem 32 (3):399–404Google Scholar
  17. Kino-oka M, Mine K, Taya M, Tone S, Ichi T (1994) J Ferment Bioeng 77:103–106CrossRefGoogle Scholar
  18. Sato K, Yamazaki T, Okuyama E, Yoshihira K, Shimomura K (1991) Phytochemistry 30:1507–1509CrossRefGoogle Scholar
  19. Murashige T, Skoog F (1962) Physiol. Plant. 15:473–497Google Scholar
  20. Kuzovkina IN, Mantrova OV, Al'terman IE, Yakimov SA (1996) Rus J Plant Physiol 43 (2):291–298Google Scholar
  21. Mantrova OV, Dunaeva MV, Kuzovkina IN, Schneider B, Müller-Uri F (1999) Rus J Plant Physiol 46 (2):248–251Google Scholar
  22. Gamborg OK, Miller RA, Ojima K (1968) Exp Cell Res 50:151–158CrossRefGoogle Scholar
  23. Bálványos I, Kursinszki L, Bányai P, Szőke É (2004) Chromatographia 60:235–238Google Scholar

Copyright information

© Friedr. Vieweg & Sohn/GWV Fachverlage GmbH 2006

Authors and Affiliations

  • P. Bányai
    • 1
  • I. N. Kuzovkina
    • 2
  • L. Kursinszki
    • 1
  • É. Szőke
    • 1
  1. 1.Department of PharmacognosySemmelweis UniversityBudapestHungary
  2. 2.Institute of Plant PhysiologyRussian Academy of SciencesMoscowRussia

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