, Volume 72, Issue 7–8, pp 641–649 | Cite as

LC-Fluorescence Detection Analysis of Amino Acids from Stellera chamaejasme L. Using 2-[2-(Dibenzocarbazole)-ethoxy] Ethyl Chloroformate as Labeling Reagent

  • Zhiwei Sun
  • Jinmao You
  • Cuihua Song


A pre-column derivatization method for the sensitive determination of amino acids using the tagging reagent 2-[2-(dibenzocarbazole)-ethoxy] ethyl chloroformate (DBCEC) followed by liquid chromatography with fluorescence detection has been developed. Identification of DBCEC-amino acids derivatives was by liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS–MS). DBCEC can easily and quickly label amino acids, and derivatives are stable enough to be efficiently analyzed by LC. Separation of the derivatized amino acids had been optimized on Hypersil BDS C18 column. A perfect baseline separation for 20 amino acid derivatives was achieved with a ternary gradient elution program. The chromophore of dibenzocarbazole group, which comprise a large rigid planar structure with p–π conjugation system, resulted in a sensitive fluorescence detection for amino acid derivatives. The derivatized amino acids were detected with fluorescence detector with excitation maximum and emission maximum at 300 and 390 nm, respectively. Excellent linear responses were observed with coefficients of >0.9993, and detection limits were in the range of 0.78–5.13 fmol (signal-to-noise ratio of 3). The mean accuracy ranged from 83.4 to 98.7% for fluorescence detection. The mean inter-day precision for all standards was <4.2% of the expected concentration. Therefore, the proposed method was a highly sensitive and specific method for the quantitative analysis of amino acids from biological and natural environmental samples.


Column liquid chromatography Fluorescence detection Electrospray mass spectrometry Amino acids Stellera chamaejasme L. 



This work was supported by the National Science Foundation of China (No. 20075016) and supported by 100 Talents Program of the Chinese Academy of Sciences (No. 328).


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Copyright information

© Vieweg+Teubner Verlag | Springer Fachmedien Wiesbaden GmbH 2010

Authors and Affiliations

  1. 1.Northwest Plateau Institute of BiologyChinese Academy of ScienceXiningPeople’s Republic of China
  2. 2.Shandong Provincial Key Laboratory of Life-Organic AnalysisQufu Normal UniversityQufuPeople’s Republic of China
  3. 3.Graduate University of the Chinese Academy of ScienceBeijingPeople’s Republic of China

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