REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY FOR THE DETERMINATION OF HAEMORPHIN-LIKE IMMUNOREACTIVITY IN HUMAN CEREBROSPINAL-FLUID

The haemorphins are opioid peptides derived form the blood protein hae moglobin. This study was focused on the detection and determination of haemorphin-like immunoreactivity in human cerebrospinal fluid (CSF) b y reversed-phase HPLC. For this purpose a SMART System, optimized for micropurification, was applied. Prior to application to HPLC, the pept ide fraction of the CSF sample was extracted using a reversed-phase si lica gel cartridge (Sep-Pak C-18). In the HPLC separation, the peptide -like material associated with haemorphin-7 immunoreactivity was recov ered and determined using a UV detector. The tryptophan residue presen t in the haemorphin sequence allowed UV detection at wavelengths (e.g. , 276 nm) where interference with other co-eluting peptides lacking th is residue is minimized. Recorded levels of haemorphin-like immunoreac tivity were compared with those detected by radioimmunoassay.