, Volume 43, Issue 7–8, pp 419–425 | Cite as

Simultaneous determination of tetrahydro-β-carbolines and β-carbolines

  • H. Tsuchiya
  • M. Sato
  • H. Hayashi
  • H. Kato
  • H. Kureshiro
  • T. Hayashi


Reversed-phase ion-pair HPLC with fluorimetric detectors connected in series was studied for the simultaneous determination of two tetrahydro-β-carbolines and two β-carbolines. After additon of internal standards the samples were reacted with fluorescamine, and then subjected to serial extractions to remove their precursor (tryptamine) from the analytical system. This treatment not only suppressed the artefactual formation of tetrahydro-β-carbolines and β-carbolines during analysis, but also effectively purified them. Under optimum conditions, using trifluoroacetic acid as counter ion, all analytes were separated within 14 min and without major interfering peaks. The quantitative ranges were 0.25–80.0 ng mL−1 for both tetrahydro-β-carbolines and 0.1–30.0 ng mL−1 for both β-carbolines. Replicate spiking experiments showed that recovery from most of the samples tested was over 90% and the relative standard deviation ranged from 0.7 to 10.4% within and between assays. The proposed method was applicable to various materials such as soy sauce, vinegar, ketchup, tabasco, beer, wine, sake, whisky, brandy, cows' milk, coffee, cocoa, cheese, cigarette smoke and urine.

Key Words

Column liquid chromatography Fluorimetric detection Tetrahydro-β-carbolines β-Carbolines 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    M. M. Airaksinen, I. Kari, Med. Biol.59, 190 (1981).Google Scholar
  2. [2]
    S. Y. H. Tse, I.-T. Mak, B.F. Dickens, Biochem. Pharmacol.42, 459 (1991).Google Scholar
  3. [3]
    M. B. Given, G. L. Longenecker, Alcohol Drug Res.7, 383 (1987).Google Scholar
  4. [4]
    J. R. F. Allen, B. R. Holmstedt, Phytochemistry19, 1573 (1980).Google Scholar
  5. [5]
    C. Melchior, M. A. Collins, CRC Crit. Rev. Toxicol.9, 313 (1982).Google Scholar
  6. [6]
    H. Rommelspacher, T. May, R. Susilo, Planta Med.57 (Suppl. 1), S85 (1991).Google Scholar
  7. [7]
    M. M. Airaksinen, I. Kari, Med. Biol.59, 21 (1981).Google Scholar
  8. [8]
    H. Tsuchiya, K. Yamada, H. Todoriki, T. Hayashi, J. Nutr. Biochem., in press.Google Scholar
  9. [9]
    H. Rommelspacher, S. Strauss, J. Lindemann, FEBS Lett.109, 209 (1980).Google Scholar
  10. [10]
    K. Matsubara, S. Fukushima, A. Akane, K. Hama, Y. Fukui, Alcohol Alcoholism21, 339 (1986).Google Scholar
  11. [11]
    T. R. Bosin, C. A. Jarvis, J. Chromatogr.341, 287 (1985).Google Scholar
  12. [12]
    M. J. Schouten, J. Bruinvels, Anal. Biochem.147, 401 (1985).Google Scholar
  13. [13]
    J. Moncrieff, J. Chromatogr.496, 269 (1989).Google Scholar
  14. [14]
    J. Adachi, Y. Mizoi, T. Naito, K. Yamamoto, A. Fujiwara, I. Ninomiya, J. Chromatogr.538, 331 (1991).Google Scholar
  15. [15]
    H. Tsuchiya, S. Ohtani, K. Yamada, N. Takagi, H. Todoriki, T. Hayashi, J. Pharm. Sci.83, 415 (1994).Google Scholar
  16. [16]
    H. Tsuchiya, S. Ohtani, K. Yamada, M. Akagiri, N. Takagi, M. Sato, Analyst119, 1413 (1994).Google Scholar
  17. [17]
    H. Tsuchiya, S. Ohtani, N. Takagi, T. Hayashi, Biomed. Chromatogr.3, 157 (1989).Google Scholar
  18. [18]
    B. J. Compton, W. C. Purdy, Anal. Chim. Acta119, 349 (1980).Google Scholar
  19. [19]
    H. Tsuchiya, S. Ohtani, K. Yamada, K. Tajima, M. Sato, Biochem. Pharmacol.50, 2109 (1995).Google Scholar
  20. [20]
    T. R. Bosin, B. Holmstedt, A. Lundman, O. Beck, Prog. Clin. Biol. Res.90, 15 (1982).Google Scholar
  21. [21]
    T. R. Bosin, B. Holmstedt, A. Lumdman, O. Beck, Anal. Biochem.128, 287 (1983).Google Scholar
  22. [22]
    H. Tsuchiya, K. Yamada, H. Kato, H. Hayashi, T. Miyazaki, T. Hayashi, Phytochem. Anal.6, 297 (1995).Google Scholar
  23. [23]
    H. Tsuchiya, K. Yamada, S. Ohtani, N. Takagi, H. Todoriki, T. Hayashi, J. Neurosci. Meth.62, 37 (1995).Google Scholar
  24. [24]
    J. Gynther, S. P. Lapinjoki, M. M. Airaksinen, P. Peura, Biochem. Pharmacol.35, 2671 (1986).Google Scholar
  25. [25]
    R. Susilo, H. Rommelspacher, Naunyn-Schmiedeberg's Arch. Pharmacol.337, 566 (1988).Google Scholar
  26. [26]
    P. Peura, J. V. Johnson, R. A. Yost, K. F. Faull, J. Neurochem.52, 847 (1989).Google Scholar
  27. [27]
    H. Tsuchiya, S. Ohtani, K. Yamada, N. Takagi, T. Hayashi, J. Pharm. Biomed. Anal.12, 1547 (1994).Google Scholar
  28. [28]
    A. Rehwald, O. Sticher, B. Meier, Phytochem. Anal.6, 96 (1995).Google Scholar
  29. [29]
    T. R. Bosin, K. F. Faull, J. Chromatogr.428, 229 (1988).Google Scholar
  30. [30]
    T. R. Bosin, K. F. Faull, Alcohl. Clin. Exp. Res.12, 679 (1988).Google Scholar
  31. [31]
    O. Beck, B. Holmstedt, Food, Cosmet. Toxicol.19, 173 (1981).Google Scholar
  32. [32]
    H. Rommelspacher, T. May, B. Salewski, Eur. J. Pharmacol.252, 51 (1994).Google Scholar
  33. [33]
    J. Adachi, Y. Mizoi, T. Naito, Y. Ogawa, Y. Uetani, I. Ninomiya, J. Nutr.121, 646 (1991).Google Scholar
  34. [34]
    E. H. Poindexter, Jr., R. D. Carpenter, Phytochemistry1, 215 (1962).Google Scholar
  35. [35]
    H. Rommelspacher, H. Damm, L. Schmidt, G. Schmidt, Psychopharmacology87, 64 (1985).Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1996

Authors and Affiliations

  • H. Tsuchiya
    • 1
  • M. Sato
    • 2
  • H. Hayashi
    • 1
  • H. Kato
    • 1
  • H. Kureshiro
    • 2
  • T. Hayashi
    • 3
  1. 1.Department of Dental PharmacologyAsahi University School of DentistryGifuJapan
  2. 2.Department of Oral MicrobiologyAsahi University School of DentistryGifuJapan
  3. 3.Grelan Pharmaceutical Company Ltd.Hamura, TokyoJapan

Personalised recommendations