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Amino Acids

, Volume 40, Issue 4, pp 1185–1193 | Cite as

Determination of trace amino acids in human serum by a selective and sensitive pre-column derivatization method using HPLC-FLD-MS/MS and derivatization optimization by response surface methodology

  • Guoliang Li
  • Yanyan Cui
  • Jinmao YouEmail author
  • Xianen Zhao
  • Zhiwei Sun
  • Lian Xia
  • Yourui Suo
  • Xiao Wang
Original Article

Abstract

Analysis of trace amino acids (AA) in physiological fluids has received more attention, because the analysis of these compounds could provide fundamental and important information for medical, biological, and clinical researches. More accurate method for the determination of those compounds is highly desirable and valuable. In the present study, we developed a selective and sensitive method for trace AA determination in biological samples using 2-[2-(7H-dibenzo [a,g]carbazol-7-yl)-ethoxy] ethyl chloroformate (DBCEC) as labeling reagent by HPLC-FLD-MS/MS. Response surface methodology (RSM) was first employed to optimize the derivatization reaction between DBCEC and AA. Compared with traditional single-factor design, RSM was capable of lessening laborious, time and reagents consumption. The complete derivatization can be achieved within 6.3 min at room temperature. In conjunction with a gradient elution, a baseline resolution of 20 AA containing acidic, neutral, and basic AA was achieved on a reversed-phase Hypersil BDS C18 column. This method showed excellent reproducibility and correlation coefficient, and offered the exciting detection limits of 0.19–1.17 fmol/μL. The developed method was successfully applied to determinate AA in human serum. The sensitive and prognostic index of serum AA for liver diseases has also been discussed.

Keywords

Trace amino acids analysis Human serum HPLC-FLD-MS/MS Pre-column derivatization Response surface methodology 

Notes

Acknowledgments

This work was supported by the National Science Foundation of China (No. 20075016) and supported by 100 Talents Programme of The Chinese Academy of Sciences (No. 328). The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Guoliang Li
    • 1
    • 2
  • Yanyan Cui
    • 5
  • Jinmao You
    • 1
    • 3
    Email author
  • Xianen Zhao
    • 4
  • Zhiwei Sun
    • 1
    • 2
  • Lian Xia
    • 1
    • 2
    • 3
  • Yourui Suo
    • 1
  • Xiao Wang
    • 4
  1. 1.Northwest Plateau Institute of BiologyChinese Academy of SciencesXiningPeople’s Republic of China
  2. 2.Graduate School of the Chinese Academy of SciencesBeijingPeople’s Republic of China
  3. 3.College of Chemistry ScienceQufu Normal UniversityJiningPeople’s Republic of China
  4. 4.Shandong Analysis and Test Center, Shandong Academy of SciencesJinanPeople’s Republic of China
  5. 5.88 Military Hospital of ChinaTaianPeople’s Republic of China

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