High-performance liquid chromatographic determination of substance p-like arginine-containing peptide in rat brain by on-line post-column fluorescence derivatization with benzoin

摘要: A high-performance liquid chromatographic method with fluorescence detection is described for the determination of substance P, one neuropeptides, in hypothalamus tissue rat brain. The based on on-line post-column derivatization selective arginine-containing peptides. endogenous P-like peptide extracted from and [d-Phe11]-neurotensin as an internal standard were separated various interfering substances a reversed-phase column (TSKgel ODS-120T) by gradient elution acetonitrile-phosphate buffer (pH 2.3). peptides eluate then automatically converted into fluorescent derivatives reaction benzoin. Arginine-containing fragments produced enzyme P fraction trypsin also detected, identification peptide. was sensitive enough to permit quantitative at concentration low 580 fmol/mg protein brain homogenate. values 9.45±1.50 pmol/mg (six determinations).