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Levine [1949] has shown that a 'spreading factor', or mucolytic activity, is present in large amounts in the thyroid gland of rats and guinea-pigs and that the enzymic activity varies according to the physiological state of the gland. These findings made it desirable to identify the substrate, and to decide whether its concentration in the colloid also varied in different states of activity. A solution to the problem became possible with the development by Hotchkiss [1948] of a procedure for visualizing polysaccharides. A complicating factor, which was also investigated, is that purified thyroglobulin is rich in carbohydrates, of which glucosamine is the chief component [Brand, Kassell & Heidelberger, 1939; Salter, 1940]. As expected, this protein gave a positive spot test with the periodic acid-leucofuchsin method and its presence in colloid would suggest that at least two groups of reacting substances might be found there. Two such substances have now been