J Integr Plant Biol. ›› 2010, Vol. 52 ›› Issue (10): 925-932.DOI: 10.1111/j.1744-7909.2010.00989.x

• Research Articles • Previous Articles     Next Articles

Quantitative Analysis of Cytokinins in Plants by High Performance Liquid Chromatography: Electronspray Ionization Ion Trap Mass Spectrometry

Weiqi Chen1, Ying Gai1, Shichang Liu1, Renxiao Wang2 and Xiangning Jiang1,*   

  1. 1College of Life Sciences and Biotechnology, Beijing Forestry University, The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of Chinese Forestry Administration, National Engineering Laboratory for Tree Breeding, Beijing 100083, China
    2Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
  • Received:2010-05-21 Accepted:2010-07-28 Published:2010-10-01
  • About author:*Corresponding author Tel(Fax): +86 10 6233 8063; E-mail: jiangxn@bjfu.edu.cn

Abstract:

The present paper introduces a highly sensitive and selective method for simultaneous quantification of 12 cytokinins (free form and their conjugates). The method includes a protocol of extraction with methanol/water/formic acid (15/4/1, v/v/v) to the micro-scale samples, pre-purification with solid phase extraction (SPE) cartridges of the extracts, separation with a high performance liquid chromatography (HPLC) and detection by an electrospray ionization ion trap mass spectrometry (ESI-Ion trap-MS) system in a consecutive ion monitoring (CRM) mode at the three stage fragmentation of mass spectrometry (MS3). The lowest detection level of the cytokinins of the method reaches 0.1–2.0 pg with a very wide range of linear regression from 1–512 pg, at the coefficient factors of 0.98–0.99. The feasibility of this method has been proven in the application of the method to the analysis of the trace-amount contents of cytokinins in the micro-scale samples of various types of plant materials, such as aerial parts of rice and poplar leaves etc. 12 endogenous cytokinins had been identified and quantified in the plant tissues, with an acceptable relatively higher recovery rate from 40% to 70%.

Chen W, Gai Y, Liu S, Wang R, Jiang X (2010) Quantitative analysis of cytokinins in plants by high performance liquid chromatography: electronspray ionization ion trap mass spectrometry. J. Integr. Plant Biol. 52(10), 925–932.

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