J. Lewis, J. Morley, R. Venn
1993
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Quality indicators
Journal
Journal of chromatography
Abstract
A method is described for the determination of sub-picomole amounts of Lys-Lys-Gly-Glu [the C-terminal tetrapeptide sequence of human beta-endorphin, referred to as melanotropin potentiating factor (MPF)], a putative neurotrophic agent. Attempts to raise antibodies to the peptide were not successful and we have therefore developed a method based on the fluorescence of its 9-fluorenylmethyl chloroformate (FMOC) derivative which provides a sensitivity comparable to that of radioimmunoassay. Standard solutions, cerebrospinal fluid or central nervous tissue extracts are first treated with FMOC-Cl. The resulting mixture of FMOC-peptides is then subjected to high-performance liquid chromatography (HPLC) and quantified using a fluorescence monitor. By this procedure, MPF and related peptides can be analysed from one sample in a single HPLC run. The method was also applied to determine the rate of release into a phosphate-buffered saline medium of a metabolically stable analogue of MPF from a slow-release formulation of the compound.