Analysis of endogenous neuropeptides by reversed-phase high-performance chromatography and mass spectrometry

Data are presented on the use of high performance liquid chromatography (HPLC) to separate and mass spectrometry (MS) to detect opioid neuropeptides in human tissues and fluids. Reversed phase HPLC (RP-HPLC) has been used in conjunction with a variety of post-column HPLC detector systems, including radioimmunoassay (RIA), radioreceptorassay (RRA), MS, and tandem mass spectrometry (MS/MS). Tandem mass spectrometry connects the protonated molecule ion, (M+H)+, of a peptide with a unique amino acid sequence-determining fragment ion, and therefore provides the highest level of molecular specificity for the detection of a peptide. Tandem MS has also been used to quantify opioid peptides in human pituitary tissues.