High-Performance Liquid Chromatographic Determination of Peptides Released by Tryptic Degradation from Opioid Peptide Precursors in Rat Brain

概要

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A high-performance liquid chromatographic method involving postcolumn fluorescence derivatization is described for the quantification of five fragment peptides (methionine-enkephalin-Thr-Ser-Glu-Lys, methionine-enkephalin-Lys, methionine-enkephalin-Arg, leucine-enkephalin-Lys and leucine-enkephalin-Arg) released by tryptic digestion from the opioid peptide precursors (proopiomeranocortin and proenkephalins A and B) in rat brain tissues. The tissue proteins containing the precursors are hydrolyzed with trypsin to the fragment peptides. The peptides are separated on an Asahipak ODP-50 column and on-line detected fluorometrically by using hydroxylamine, cobalt(II) and borate buffer reagents. The detection limits (S/N=3) for the peptides are 0.7-2.8 pmol per 100 μl injected. The distribution of the precursors in the brain tissues was also discussed on the basis of the determined values of the fragment peptides.
公益社団法人日本薬学会の論文
1991-09-25