Analysis and Enantioseparation of Amino Acids by Liquid Chromatography

  • Poonam Malik
  • Ravi BhushanEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2030)


Enantioseparation studies of proteinogenic, non-proteinogenic, and dansyl amino acids are described herein by using liquid chromatographic techniques, i.e., HPLC and TLC. A researcher who wants to perform amino acid (AA) analysis or separate enantiomers of AAs by HPLC or TLC can follow the method. Figures included represent the actual experiments.

Synthesis and application of chiral derivatizing reagents (CDRs) based on cyanuric chloride (CC) and difluorodinitrobenzene (DFDNB) have been described for AA analysis and enantioseparation by indirect approach. The methods represent pre-column derivatization of AAs and represent a good and less expensive substitute of AA analyzer. The application of commercial “Chiralplate” and use of erythromycin and l-tartaric acid have been described as chiral selector either as impregnating reagent in the stationary phase or as an additive in the mobile phase for direct enantioseparation by TLC. Application of the homemade TLC plates has also been described; the methods are successful in obtaining the native enantiomer as well.

Key words

Amino acids Enantiomeric separation Amino acid analysis RP-HPLC TLC Erythromycin l-Tartaric acid Cyanuric chloride DFDNB 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of ChemistryIndian Institute of Technology RoorkeeRoorkeeIndia

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