A DOUBLE ISOTOPE DERIVATIVE METHOD FOR THE DETERMINATION OF ALDOSTERONE IN PERIPHERAL PLASMA

1961 ◽  
Vol 37 (4) ◽  
pp. 541-558 ◽  
Author(s):  
Ejgil Bojesen ◽  
Hans Degn
Keyword(s):  
Isotope Dilution ◽  
Half Life ◽  
Acid Anhydride ◽  
Hydroxyl Group ◽  
Aldosterone Level ◽  
Peripheral Plasma ◽  
Derivative Method ◽  
Double Isotope

ABSTRACT The details of a method for the determination of aldosterone at the level of 1 ng or more are described. A combination of the isotope derivative principle and the isotope dilution principle, using 35S and 131I labelled piodobenzenesulfonic acid anhydride (pipsan) as the labelled reagents, has been employed. A quantitative or almost quantitative and reproducible esterification of the 21-hydroxyl group of aldosterone and an 80% recovery of the steroid added to plasma can be achieved by the procedure described. The blank value has been determined by the analysis of epripheral plasma of adrenalectomized dogs and was found to be indistinguishable from zero and therefore less than 3 ng per 100 ml of plasma. In salt depleted anaesthetized dogs the aldosterone level was 20–30 ng per 100 ml. The concentration increased considerably during adrenalectomy and decreased with a »half life« of 20–30 minutes from the time this was completed.

PROCEDURE FOR DETERMINATION OF ALDOSTERONE IN HUMAN PERIPHERAL PLASMA BY DOUBLE-ISOTOPE DERIVATIVE ASSAY, AND ITS APPLICATION FOR MEASUREMENT OF SECRETORY RATE AND URINARY EXCRETION

1967 ◽  
Vol 45 (12) ◽  
pp. 1919-1936 ◽  
Author(s):  
Wojciech Nowaczynski ◽  
Jack Silah ◽  
Jacques Genest
Keyword(s):  
Acetic Anhydride ◽  
Urinary Excretion ◽  
Specific Activity ◽  
High Specific Activity ◽  
Plasma Aldosterone ◽  
Chromatographic Purification ◽  
Secretory Rate ◽  
Peripheral Plasma ◽  
Double Isotope

This is a report of a procedure for the determination of peripheral plasma aldosterone, in which 14C-labeled aldosterone serves as the marker, and the acetylating agent is relatively inexpensive tritium-labeled acetic anhydride of high specific activity. A new method for the chromatographic purification of aldosterone and aldosterone diacetate is presented. This procedure provides a better separation of these two compounds from the unlabeled and labeled impurities.

PLASMA ALDOSTERONE DETERMINATION WITH 35S-p-TOLUENE-SULPHONIC ANHYDRIDE

1972 ◽  
Vol 70 (3) ◽  
pp. 552-566 ◽  
Author(s):  
D. Scholer ◽  
A. M. Riondel ◽  
E. L. Manning
Keyword(s):  
Plasma Level ◽  
Thin Layer ◽  
Water Samples ◽  
Regression Line ◽  
Mean Value ◽  
Plasma Aldosterone ◽  
Peripheral Plasma ◽  
Derivative Method ◽  
Replicate Measurements ◽  
Double Isotope

ABSTRACT The double isotope derivative method for plasma aldosterone, (Bojesen & Thuneberg 1967), has been applied to human peripheral plasma. 35S-p-toluene-sulphonic anhydride was used as the labelled reagent (100–150 mCi/mEq.) and 3H-aldosterone (30 Ci/mm; 0.05 ng added to each sample) as indicator. The initial derivative (aldosterone-21-35S-tosylester) was transformed to its 11,18-γ-lactone and finally to its 3-dinitrophenylhydrazone. Purification was achieved by 1 paper chromatography and 4 thin-layer chromatographies. The overall-recovery was 10 ± 2.7 (sd)%, n = 134. The non-specific blank was 0.053 ± 0.044 (sd), when determined on water samples, and 0.068 ± 0.034 (sd) ng/5 ml, when determined on adrenalectomised human plasmas (non-specific water blank of the same series: 0.064±0.037 (sd) ng). The addition of a known amount of aldosterone to a plasma resulted in a linear relationship over the range examined: y = (0.947 ± 0.013) × + 0.696, P = 0.000, and aldosterone from 1.0 to 5.0 ng was measured with an accuracy of 95%. Replicate measurements of different volumes of the same plasma from 1.0 to 7.5 ml showed a calculated regression line of y = (0.148 ± 0.005) × −0.01, P = 0.002. Replicate measurements of the same plasma (n = 8) gave a mean value of 0.14±0.01 (sd) ng, a range between 0.13 and 0.15 ng and a coefficient of variation of 7%. Thus, it was possible to determine a peripheral plasma level of 7 ng/100 ml in 2 ml plasma and a level of 3.5 ng/100 ml in 4 ml plasma.

Comparison of Results by Four Different Procedures for Determination of 17-Hydroxycorticosteroids

1973 ◽  
Vol 19 (7) ◽  
pp. 748-752 ◽  
Author(s):  
Joseph Bruton ◽  
Ting-Kai Li ◽  
Gerald D Smith
Keyword(s):  
Protein Binding ◽  
Binding Assay ◽  
Assay Procedure ◽  
Protein Binding Assay ◽  
Normal Individuals ◽  
Derivative Method ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding ◽  
Double Isotope

Abstract A Porter and Silber procedure, a fluorometric procedure, a double-isotope derivative procedure, and a competitive protein-binding assay procedure were used to determine 17-hydroxycorticosteroids in plasma from normal individuals and from patients with endocrinopathies. Results from each of these procedures were intercompared. Results of the fluorometric procedure and the competitive protein-binding assay compared favorably with results obtained by the more elaborate and difficult double-isotope derivative method. The Porter and Silber method is less specific. This study should be a useful aid in comparing values with those of other laboratories and in selecting the most advantageous method for use by a particular laboratory.

BESTIMMUNG VON TESTOSTERON IM PERIPHEREN BLUT VON SCHWEIN UND RIND MIT EINER DOPPELISOTOPEN-DERIVAT-VERDÜNNUNGSMETHODE

1970 ◽  
Vol 64 (2) ◽  
pp. 377-384 ◽  
Author(s):  
B. Hoffmann ◽  
R. Claus ◽  
H. Karg
Keyword(s):  
Isotope Dilution ◽  
Peripheral Blood ◽  
Derivative Method ◽  
Double Isotope

ABSTRACT Using a double-isotope-dilution derivative method combined with a cristallisation technique testosterone was measured in pig and bovine peripheral blood. In the male pig the values varied from 0.94 to 1.61 μg per 100 ml plasma and were 7- resp. 11-times higher than in castrated and female animals. Values increased with age. The concentration measured in a bull was 0.59 μg per 100 ml plasma.

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