Chromatography derivatization

This chapter will focus on topic 3, which is normally regarded to be chiral derivatization chromatography, but will also cover other topics that might be considered when applying derivatization techniques. The goal for the separation of the race-mates may be their analysis or their preparation. Both topics will be covered in this chapter. 

One of the most useful applications of chiral derivatization chromatography is the quantification of free amino acid enantiomers. Using this indirect method, it is possible to quantify very small amounts of enantiomeric amino acids in parallel and in highly complex natural matrices. While direct determination of free amino acids is in itself not trivial, direct methods often fail completely when the enantiomeric ratio of amino acid from protein hydrolysis must be monitored in complex matrices. 

Fig. 7-1. General reaction scheme of chiral derivatization chromatography.

Glutaraldehyde also can be used to create aldehydes on amine-containing polymers. The use of this reagent in derivatizing chromatography supports and other soluble polymers is well known (Hermanson et al., 1992). 

Schulte, M. Chiral derivatization chromatography, in Chiral Separation Techniques, Subramanian, G. (Ed.), Wiley-VCH, Weinheim, 2001... 

Inoue, T. Umemura, H. Yoshida, H. Todoroki, K. Nohta, H. Yamaguchi, M. Sensitive determination of oxytocin-related compounds by liquid chromatography with pre-column excimer fluorescence derivatization. Chromatography 2009, 30(Suppl. 1), 17-18. 

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