关键词: 固相荧光衍生, FMOC-Cl, 碱性硅胶, 氨基酸, 多肽

Abstract: A solid phase derivatization method of amino acids and peptides was described. Samples were added on alkaline silica gel cartridge and dried. The derivatization was carried out by passing the derivatization regent dissolved in anhydrous solvent through the cartridge slowly, then the cartridge was washed with a selected solvent, finally, the derivatives were eluted. The eluates were separated and detected by means of HPLCwith fluorescence detector. Three derivatization reagents, 9-fluorenylmethyl chloroformate(FMOC-Cl), 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole(NBD-Cl) and fluorescein isothiocyanate(FITC) were investigated with this method. The results showed that the method was suitable for derivatization of amino acids and some of peptides with FMOC-Cl because of its highly reacting activity and low polarity, the sideproducts of the FMOC-Cl with water were greatly decreased, the excess FMOC-Cl was eliminated completely, and the silica gel support did not significantly affect the reaction rate and yield for amino acids. NBD-Cl was not suitable for this method because of the low yield of derivatization. FITCwas also not suitable for this method because of its strong polarity.

Key words: Solid phase fluorescent derivatization, FMOC-Cl, Alkaline silica gel, Amino acids, Peptides