Derivatization and fluorescence detection of amino acids and peptides with 9-fluorenylmethyl chloroformate on the surface of a solid adsorbent.
An approach that exploits the surface of a solid adsorbent is proposed for precolumn FMOC derivatization of amino acids and peptides. Amino acids (Ser, Glu, GABA, Val, Phe, Lys) and two neuropeptides (substance P and Leuenkephalin) were adsorbed on alkaline silica gel cartridges. After drying, they were reacted with 9-fluorenyl-methyl chloroformate (FMOC-Cl) in toluene. After washing off the excess FMOC-Cl with ethyl acetate, the derivatives were eluted with aqueous eluant. The eluates were separated and detected by means of HPLC with fluorescence detection. Compared with the traditional derivatization in the liquid phase, the extent of formation of byproducts of FMOC-Cl with water was greatly decreased, and the excess FMOC-Cl was eliminated completely.
Shangguan
D
D
Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing, People's Republic of China.
Zhao
Y
Y
Han
H
H
Zhao
R
R
Liu
G
G
D000596:Amino Acids / Q000032:analysis*; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004743:Enkephalin, Leucine / Q000032:analysis / Q000737:chemistry; D005449:Fluorenes; D018698:Glutamic Acid / Q000032:analysis; D007202:Indicators and Reagents; D008239:Lysine / Q000032:analysis; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D010649:Phenylalanine / Q000032:analysis; D015203:Reproducibility of Results; D012694:Serine / Q000032:analysis; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013373:Substance P / Q000032:analysis / Q000737:chemistry; D014633:Valine / Q000032:analysis; D005680:gamma-Aminobutyric Acid / Q000032:analysis
D000596:Amino Acids; D005449:Fluorenes; D007202:Indicators and Reagents; D010455:Peptides; D013373:Substance P; D018698:Glutamic Acid; D012694:Serine; D010649:Phenylalanine; D005680:gamma-Aminobutyric Acid; D004743:Enkephalin, Leucine; C054007:1-(9-fluorenyl)methyl chloroformate; D014633:Valine; D008239:Lysine