Use of Radioisotopes in Steroid Methodology
Quantitative analyses of organic compounds in a mixture of biologic origin are extremely difficult or inconvenient if the compounds to be analyzed are present in trace amounts and are similar to other components of the mixture. Thus, it frequently is necessary to use labeled (radioactive) indications in the analytical assay so that quantitative isolation of the compound of interest is not required. C14- and tritium-labeled steroids can be used to measure quantitatively, by the isotope dilution procedure, the amount of a particular steroid present in a mixture when the steroid to be analyzed cannot be quantitatively separated from the mixture, or when the analytical problem cannot be readily solved by other means. With isotope dilution procedures, quantitative isolation of a compound is not necessary for accurate analyses. The only requirement is a sufficiently pure sample for assay of specific activity. Previous publications have described the methods for isotope dilution analyses of various steroids using C14-labeled steroids [1, 2]. These methods depend on the measured dilution of the isotopically labeled steroid by a nonlabeled variant of the same steroid, and consist of three general types: (a) direct isotope dilution; (b) reverse isotope dilution; and (c) double isotope dilution.
KeywordsCortisol Dichloromethane Anhydride Aldosterone Tritium
Unable to display preview. Download preview PDF.
- (1).Peterson, R. E„ Karrer, A., and Guerra, S. L. “Evaluation of the Silber-Porter Procedure for the Determination of Plasma Hydrocortisone, ” A.al. Chem. 29, 144 (1957).Google Scholar
- (2).Peterson, R. E. “The Identification of Corticosterone in Human Plasma and Its Assay by Isotope Dilution,” J. Biol. Chem. 225, 25, (1957).Google Scholar
- (7).Peterson, R. E., Kliman, B., and Bollier, M. E. “A Physical Method for Determination of Steroids in Biological Extracts,” Abstracts of the 131st Meeting of the Am. Chem. Soc., Miami, Florida, April 7–12, 1957, 59C.Google Scholar
- (8).Kliman, B. and Peterson, R. E. “Double Isotope Derivative Assay of Aldosterone in Biological Fluids,” “Extracts.” J. Biol. Chem. 235, 1639 (1960).Google Scholar
- (10).Peterson, R. E. “The Miscible Pool and Turnover Rate of Adrenocortical Steroids in Man,” Rec. Frog, in Hormone Res. 15, 231 (1959)Google Scholar
- (11).Okita, G.T., 3Kabara. J. J., Richardson, F., and Le Roy, G. V, G.T., 3Kabara. J. J., Richardson, F., and Le Roy, G. V. “Assaying Compounds Containing H and C,” Nucleonics 15, 111 (1957).Google Scholar