Isolation of Adrenocorticotropic Hormone from Sheep Pituitaries

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Abstract
A protein free from contaminants and which stimulates the adrenal cortex was prepared as follows: sheep pituitaries were ground and extracted with acidified 80% acetone, and extract precipitated in 90% acetone and dried, then extracted with 0.1 M Na2HPO4, precipitated with (NH4)2SO4. The precipitate was dissolved in water and dialyzed until salt free, adjusted to pH 3.0 and saturated NaCl added to 0.54 M. The supernatant was brought to half-saturation with (NH4)2SO4, the precipitate dissolved in water, half of its vol. of NH4OH added and left at room temp. 4 hrs., then solution was brought to 90% acetone, and the ppt. suspended in water and dialyzed. Saturated (NH4)2SO4 was added and ppt. dialyzed and adjusted to pH 3.0, then saturated NaCl soln. was added to 0.54 M. The supernatant was brought to 1.35 M. The ppt. with NaCl was repeated 4 times and final ppt. analyzed. Tests showed that it contained only one component. The hormone was standardized by 2 methods: repair of adrenal cortex of immature [female] rats and maintenance of adrenal cortex in the [male] rat.

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