method of growth and metabolism contributes to the low molecular weight ainee n isolation from blood.

摘要

1,188,656. Anabolic substance from haemolysed blood W. HELMBOLD. 20 April, 1967 [20 April, 1966], No. 18146/67. Heading A5B. [Also in Division C1] A substance having growth promoting and anabolic activity is prepared by fractionating a haemolysate of the blood cells from the blood of young animals, by one or more of: ion exchange, absorption chromatography preparative high voltage electrophoresis, high voltage electrodialysis, gel filtration, preparative thin-layer chromatography and counter current distribution, and isolating the substance from the fraction having an ultra-violet absorption maximum at 245-246mu. The starting material is preferably spray-dried haemolysed blood, and the fractionation is preferably preceded by ultra filtration, e.g. through a high pressure membrane filter permeable to molecules up to mol. wt. of 10,000, to filter out high molecular weight impurities. Suitable chromatographic agents are dextran or cellulose based anion exchangers, or anion exchange synthetic resins or hydroxy-apatite. Counter current distribution may be effected in propanol or butanol, and thin layer chromatography is performed on silica gel with a butanol/acetic acid/water system. The substance is a water soluble anionic compound having a molecular weight of less than 700, and is not a peptide, sugar, lipid, nucleoside or nucleotide. It has the same electrophoretic mobility as serum albumin at pH 8.6; an Rf-value of 0.93-0.96 in butanol/acetic acid/water 0:1:1 on silica gel; and the following U.V. absorption maxima: 245-246 mÁ at neutral pH; 250-260 mn (255.5 mÁ at pH 2.0; and 280-281mÁ at pH 12.0. Compositions comprising the substance together with a carrier are used for pharmaceutical application or for stimulating the growth of crops (see Division C1). Pharmaceutical compositions suitably contain the active substance in such a concentration that, after 20 fold dilution with water, they have an extinction coefficient of at least 0.6 at 246 mÁ.