Summary
A reversed-phase high-performance liquid chromatography procedure combined with radioimmunoassay (rp-HPLC-RIA) is presented. It makes possible the quantitative determination of the neuropeptides,β-endorphin and substance P, in small volumes of cerebrospinal fluid (CSF). The concentration-dependence of the recovery of the method is demonstrated and the effect of free silanol groups of the stationary phase on the recovery is discussed. CSF was shown to contain 9.6±1.1fmolβ h-endorphin and 12.0±0.9 fmol substance P per ml.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Emrich HM, Nassett L, Gramsch C, John S (1983) Heroin addiction: betaendorphin immunoreactivity in plasma increases during withdrawal. Pharmakopsychiatry 16: 93–96
Emson PC, Arregui A, Clement-Jones V, Sandberg BEB, Rossor M (1980) Regional distribution of methionine-enkephalin and substance P-like immunoreactivity in normal human brain and in Huntingtons disease. Brain Res 199: 147–180
Guillemin R, Ling N, Vargo T (1977) Radioimmunoassays forα-endorphin andβ-endorphin. Biochem Biophys Res Commun 77: 361–366
Hermann K, Lang RE, Unger T, Bayer C, Ganten D (1984) Combined HPLC-RIA for the characterization and quantitation measurement of neuropeptides. J Chromatogr 312: 273–284
Hosobuchi Y, Emson PC, Iversen LL (1982) Elevated cerebrospinal fluid substance P in arachnoiditis is reduced by systemic administration of morphine. In: Costa E, Trabucchi M (eds) Regulatory peptides: from molecular biology to function. Raven Press, New York, pp 497–500
Hunter WM, Greenwood FC (1962) Preparation of iodine-131 labeled human growth hormone of high specific activity. Nature 194: 495–496
Javoy-Agid F, Ruberg M, Pique L, Bertagna X, Taquet H, Studler JM, Cesselin F, Epelbaum J, Agid Y (1984) Biochemistry of the hypothalamus in Parkinsons disease. Neurology 34: 672–675
Jolkkonen JT, Soininen HS, Riekkinen PJ (1984)β-endorphin-like-immunoreactivity in cerebrospinal fluid of patients with alzheimer disease. Acta Neurol Scand 69: 234–235
Laatikainen TJ, Salminen KR, Sulkava RO, Leppäluoto JP (1985) Determination ofβ-endorphin andβ-lipotropin in human cerebrospinal fluid. Clin Chim Acta 145: 325–329
Lewis RV, Stein S, Udenfriend S (1979) Separation of opioid peptides utilizing high-performance liquid chromatography. Int J Peptide Prot Res 13: 493–497
Loeber JG, Verhoff J, Burbach JPH, Witter A (1979) Combination of highperformance liquid chromatography and radioimmunoassay is a powerful tool for the specific and quantitative determination of endorphins and related peptides. Biochem Biophys Res Commun 86: 1288–1295
Loeber JG, Verhoff J (1981) HPLC and RIA for the specific and quantitative determination of endorphins and related peptides. Meth Enzymol 73: 261–268
McDermott JR, Smith AJ, Biggins JA, Chyad Al-Noaemi M, Edwardson JA (1981) Characterization and determination of neuropeptides by HPLC and RIA. J Chromatogr 222: 371–379
Morris HR, Etienne AT, Dell A, Albuquerke R (1980) A rapid and specific method for the high resolution purification and characterization of neuropeptides. J Neurochem 34: 574–582
Nakao K, Nakai Y, Matsubara S, Konishi T, Nishitana H, Imura H (1980) Immunoreactiveβ-endorphin in human cerebrospinal fluid. J Clin Endocrinol Metab 50: 230–233
Nutt JG (1983) Substance P in human cerebrospinal fluid. In: Wood JH (ed) Neurobiology of CSF II. Plenum, New York, pp 67–75
Pezzoli G, Panerai AE, Di Giulio A, Longo A, Passerini D, Carenzi A (1984) Methionine-enkephalin, substance P, and homovanillic acid in the CSF of parkinsonian patients. Neurology 34: 516–519
Rachlin JR, Fessler RG, Duda EE, Brown FD, Fang V, Mullan S (1984)β- endorphin-like-immunoreactivity increases in human cerebrospinal fluid following routine metrizamid myelography. J Neurosurg 60: 800–802
Savoldi F, Mazzella GL, Facchinetti F, Nappi G, Petraglia F, Sinforiani E, Parrini D, Genazzani AR (1983)β-endorphin,β-lipotropin and adrenocorticotropic hormone levels in cerebrospinal fluid, and brain damage in chronic alcoholics. Eur Neurol 22: 265–271
Smyth DG (1984) Chromatography of peptides related toβ-endorphin. Anal Biochem 136: 127–135
Tenovuo O, Rinne UK, Viljanen MK (1984) Substance P immunoreactivity in the post mortem Parkinsonian brain. Brain Res 303: 113–116
Wardlaw SL, Schlachter LB, Tindall GT, Frantz AG (1982)β-endorphin in cerebrospinal fluid after total hypophysectomy. Clin Res 30: 278 A
Wiedemann K, Teschemacher H (1983)β-endorphin immunoreactive materials in human plasma determined by a multiple radioimmunoassay system. Life Sci 33 [suppl] 1: 89–92
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Henning, K., Wallasch, T.M. & Przuntek, H. Determination of β-endorphin and substance P in small volumes of human cerebrospinal fluid. J. Neural Transmission 67, 105–112 (1986). https://doi.org/10.1007/BF01243363
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01243363