Separation of pineal extracts by gelfiltration. VI. Isolation and identification from sheep pineals of biopterin; comparison of the isolated compound with some synthetic pteridines and the biological activity in in vitro and in vivo bioassays.
Aqueous extracts of sheep pineal bodies were separated on Sephadex G-25. Two low molecular weight Sephadex G-25 fractions, F2 and F3, were ultrafiltrated through the Amicon membrane UM-2. The UM-2 filtrate was subsequently filtrated through the ultramembrane UM-05 and the UM-05 filtrate was separated on Sephadex G-10 columns. After paper electrophoresis, preparative paper chromatography was carried out. The fluorescent band showing a Rf value identical with synthetic 6-biopterin was eluted; gas liquid chromatography and mass spectrometry of the isolated compound were carried out. The mass spectra of the isolated compound were shown to be identical with synthetic 6-biopterin. The results of the Crithidia fasciculata test and thinlayer chromatography study revealed that the isolated compound is identical with 6-L-erythro-biopterin. The activities of the isolated compound and of synthetic biopterin in in vitro and in vivo bioassays are demonstrated.
D000818:Animals; D001681:Biological Assay; D001708:Biopterins / Q000032:analysis / Q000302:isolation & purification*; D002850:Chromatography, Gel / Q000379:methods; D005453:Fluorescence; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D010870:Pineal Gland / Q000032:analysis*; D011621:Pteridines / Q000302:isolation & purification*; D051381:Rats; D012756:Sheep
D011621:Pteridines; D001708:Biopterins; D007986:Luteinizing Hormone