Microdetermination of adrenocortical steroids by double isotope method. 2. Fractional determination of corticosteroids: free and protein-bound cortisol, corticosterone in plasma

Journal Article · Thu Feb 01 04:00:00 EST 1973 · Endocrinol. Jap., v. 20, no. 1, pp. 23-31

DOI:https://doi.org/10.1507/endocrj1954.20.23· OSTI ID:4360818

Kunugi, A; Matsumoto, U; Nagase, Y

Further examinations were made on the double isotope derivative dilution method by the use of thiosemicarbazide-/sup 35/S as the labeled reagent and /sup 3/H-labeled corticosteroid for correcting the loss during extraction and separation procedure, and fractional determination of free and protein-bound cortisol and corticosterone. The DELTA 4-3ketosteroids with similar chemical structure present in plasma, namely, cortisol, corticosterone, aldosterone, deoxycorticosterone, and cortisone, can be separated by repeating thin-layer chromatography at a low temperature of 10 deg C more than four times. Cortisol and corticosterone can be determined correctly and with good precision, relative standard deviation being 1 to 2%. The other three corticosteroids can be determined correctly and with good precision, relative standard deviation being 3 to 6%. It was found that the protein-bound cortisol and corticosterone in plasma are dissociated during solvent extraction, and the total:amount is determined in free form. It was also found that the protein-bound and free cortisol and corticosterone can be separated by gel filtration, and the combination of gel filtration and the present method of determination has made it possible to carry out fractional determination of bound and free forms of cortisol and corticosterone. The quantity per 100 ml of plasma was found to be 1.83 and 0.18 mu g of free cortisol and corticosterone, and 17.45 and 1.75 mu g of protein- bound cortisol and corticosterone, respectively. (auth)

• Research Organization: Tokyo Coll. of Pharmacy
• NSA Number: NSA-29-010557
• OSTI ID: 4360818
• Journal Information: Endocrinol. Jap., v. 20, no. 1, pp. 23-31, Journal Name: Endocrinol. Jap., v. 20, no. 1, pp. 23-31; ISSN ECJPA
• Country of Publication: Japan
• Language: English

Related Subjects

*CORTICOSTERONE-- RADIOMETRIC ANALYSIS
*HYDROCORTISONE-- RADIOMETRIC ANALYSIS
ACCURACY
BLOOD PLASMA
FILTRATION
ISOTOPE DILUTION
N48830* --Life Sciences--New Tracer Techniques--Other
QUANTITATIVE CHEMICAL ANALYSIS
SEMICARBAZIDES
SULFUR 35
THIN-LAYER CHROMATOGRAPHY
TRITIUM COMPOUNDS