Abstract
Application of liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS) for the analysis of cytokinins was examined. The fragmentation of cytokinins was studied using authentic trans-zeatin (t-Z), trans-zeatin riboside (t-ZR), isopentenyl adenine (i6Ade), isopentenyl adenosine (i6Ado), benzyl adenine (BAde), benzyl adenosine (BAdo), and kinetin. These cytokinins were effectively ionized by APCI in aqueous acetonitrile. t-Z, i6Ade, BAde, and kinetin showed prominent quasi-molecular ions of [M + H]+, and ribosylcytokinins clearly showed both [M + H]+ and a characteristic fragment ion ([M + H-ribose]+), giving some information about their structures. The qualitative and semi-quantitative analyses of cytokinins by LC/APCI-MS were validated in combination with enzyme-linked immunosorbent assay (ELISA) through the analysis of t-ZR in the teratoma of Nicotiana tobacum. t-ZR was conclusively identified and a semi-quantitative estimate of its endogenous levels were provided by the combination of LC/APCI-MS and ELISA. The quantified values obtained by LC/ APCI-MS (single ion detection) and ELISA are in close agreement.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Eberle J, Angelik A, Dieter K, Weiler EW (1986) Monoclonal antibodies to plant growth regulators: III. Zeatin riboside and dihydrozeatin riboside. Plant Physiol 81:516–521
Ishikawa K, Kamada H, Yamaguchi I, Takahashi N, Harada H (1988) Morphology and hormone levels of tobacco and carrot tissues transformed by Agrobacterium tume-faciens: I. Auxin and cytokinin contents of cultured tissues transformed with wild-type and mutant Ti plasmids. Plant Cell Physiol 29:461–466
Murofushi N, Yang Y-Y, Yamaguchi I, Schneider G, Kato Y (1992) Liquid chromatography/atmospheric pressure chemical ionization mass spectrometry of gibberellin conjugates. In: Karssen CM, van Loon LC, Vreugdenhil D (eds) Progress in plant growth regulation. Kluwer Academic Publishers, Dordrecht, Boston, London pp 900–904
Palni LMS, Summons RE, Letham DS (1983) Mass spectrometric analysis of cytokinins in plant tissues: V. Identification of the cytokinin complex of Datura innoxia crown gall tissue. Plant Physiol 72:858–863
Sakairi M, Kambara H (1988) Characteristics of a liquid chromatograph/atmospheric pressure ionization mass spectrometer. Anal Chem 60:774–780
Watanabe N, Yokota T, Takahashi N (1982) Transfer RNA, a possible supplier of free cytokinins, ribosyl-cis-zeatin and ribosyl-2-methylthiozeatin: quantitative comparison between free and transfer cytokinins in various tissues of the hop plant. Plant Cell Physiol 23:479–488
Weiler EW, Wieczorek U (1981) Determination of femtomol quantities of gibberellic acid by radioimmunoassay. Planta 152:159–167
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Yang, YY., Yamaguchi, I., Kalo, Y. et al. Qualitative and semi-quantitative analyses of cytokinins using LC/APCI-MS in combination with ELISA. J Plant Growth Regul 12, 21–25 (1993). https://doi.org/10.1007/BF00193674
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00193674