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 dis...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 HPLC with 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 side\|products 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. FITC was also not suitable for this method because of its strong polarity.展开更多
文摘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 HPLC with 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 side\|products 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. FITC was also not suitable for this method because of its strong polarity.
