Buchanan on Glutathione

Buchanan reported On January 9, 1929, after many years of speculation and four and one half years of work, I arrived at the point in my investigations where it seemed reasonable to believe that pernicious anemia was due to a yeast (6). He concluded that the active principle of liver extracts has apparently every chemical resemblance and physiological action of oxidized glutathione. Although Fleming (37) later declared that glutathione, mostly in oxidized form, enters into the composition of the antianemic factor, Koser in 1936 (72) denied the importance of glutathione in this respect. 

In 1935, Erdos (32) reported on the preparation of active material from liver and on certain of its properties. Finely hashed liver was first ground with quartz powder and then extracted with dilute H2SO4 at different temperatures. The extracts, concentrated at 60-70° C. until the dry wei t constituted about 60 per cent, were then mixed with about ten fold volumes of H2O, wanned to 50°, cooled, filtered, brought to pH 7.5-8 wdth NaHCQj, 

Analysis (of that portion of the powder which is water-soluble). 

Erdos assayed his material for activity by noting its influence on tiie anemia produced in dogs and rabbits by the administration of phenyl-hydrazine (33). In the normal animal, the administration of phenyl-hydrazine hydrochloride (16 mg. per kg. body Aveight) lowered the red cell count 25 to 40 per cent in 48 to 72 hours, after Avhich time, spontaneous remission occurred. HoAvever, in those animals Avhich received liver preparations along Avith the phenylhydrazine, the ma, imal decrease of red cells Avas reached only after 96 to 120 hours. 

In 1942, Erdos modified his previous method of isolation. Finely ground liver AA as extracted Avith dilute H2SO4 at different temperatures and the sulfuric acid was removed Avith Ba(OH)2. The concentrated filtrate Avas precipitated with 70% alcohol. The insoluble fraction Ayas removed and the filtrate Avas concentrated and precipitated with AgNOj. The Ag precipitate Avas separated and decomposed with HCl. The silver free filtrate Avas concentrated and precipitated AA ith alcohol. 

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