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 haemoglobin. This study was focused on the detection and determination of haemorphin-like immunoreactivity in human cerebrospinal fluid (CSF) by reversed-phase HPLC. For this purpose a SMART System, optimized for micropurification, was applied. Prior to application to HPLC, the peptide fraction of the CSF sample was extracted using a reversed-phase silica gel cartridge (Sep-Pak C18). In the HPLC separation, the peptide-like material associated with haemorphin-7 immunoreactivity was recovered and determined using a UV detector. The tryptophan residue present in the haemorphin sequence allowed UV detection at wavelengths (e.g., 276 nm) where interference with other co-eluting peptides lacking this residue is minimized. Recorded levels of haemorphin-like immunoreactivity were compared with those detected by radioimmunoassay.
Sanderson
K
K
Department of Pharmaceutical Biosciences, Uppsala University, Sweden.
Thörnwall
M
M
Nyberg
G
G
Glämsta
E L
EL
Nyberg
F
F
D000595:Amino Acid Sequence; D002543:Cerebral Hemorrhage / Q000134:cerebrospinal fluid; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006408:Hematoma, Subdural / Q000134:cerebrospinal fluid; D006454:Hemoglobins / Q000134:cerebrospinal fluid* / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D011863:Radioimmunoassay; D013345:Subarachnoid Hemorrhage / Q000134:cerebrospinal fluid
D006454:Hemoglobins