An improved method for the extraction of corticosterone from cell homogenates and subcellular fractions of the rat adrenal cortex.
An improved technique is described for the extraction and analysis of corticosterone (11 beta,21-dihydroxy-4-pregnene-3,20-dione) from homogenates and subcellular fractions of the rat adrenal cortex. Factors influencing complete extraction of corticosterone were the nature of the organic solvent system and the concentration of the tissue being extracted. The continued activity of steroidogenic enzymes during subcellular fractionation was presented by 0.1 mM 1-benzylimidazole. For optimum extraction, homogenates were diluted 1:12 (v/v) in 0.25 M sucrose, containing 0.1 M potassium hydroxide. Dilute homogenate was mixed with absolute ethanol (1:10, v/v) and extracted three times with diethyl ether (1:5, v/v). Following extraction, corticosterone in each sample was isolated by thin-layer chromatography (TLC), quantitated by radioimmunoassay (RIA), and corrected by measuring the recovery of added H3 corticosterone. With these procedures, 90-100% of corticosterone found in extracts of adrenal homogenates was recovered in extracts of subcellular fractions of the homogenates.
D000302:Adrenal Cortex / Q000032:analysis* / Q000187:drug effects; D000616:Aminoglutethimide / Q000494:pharmacology; D000818:Animals; D003345:Corticosterone / Q000302:isolation & purification*; D000431:Ethanol; D007093:Imidazoles / Q000494:pharmacology; D008297:Male; D008797:Metyrapone / Q000494:pharmacology; D051381:Rats; D012966:Sodium Cyanide / Q000494:pharmacology; D012997:Solvents; D013347:Subcellular Fractions / Q000032:analysis
D007093:Imidazoles; D012997:Solvents; D000616:Aminoglutethimide; D000431:Ethanol; C017062:1-benzylimidazole; D012966:Sodium Cyanide; D003345:Corticosterone; D008797:Metyrapone