RADIOIMMUNOASSAYS OF TETRAHYDROCORTISONE AND TETRAHYDROCORTISOL IN HUMAN URINE

1977 ◽  
Vol 86 (2) ◽  
pp. 369-379 ◽  
Author(s):  
H. Will ◽  
R. Aderjan ◽  
Th. Winkler ◽  
B. Penke ◽  
P. Vecsei
Keyword(s):  
New Zealand ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Secretion Rate ◽  
Cortisol Secretion ◽  
Cross Reactions ◽  
New Zealand Rabbits

ABSTRACT A simple radioimmunoassay for the determination of tetrahydrocortisone (THE)1), and tetrahydrocortisol (THF) as well as their 21-glucosiduronate conjugates is described. White New Zealand rabbits were immunized with THF- and THE-20-oxime-bovine serum-albumin conjugates. The resulting antisera proved to be very specific. Both allo-THE and allo-THF gave minimal cross-reactions; however, THE- and THF-21-glucosiduronate bind with the corresponding antibodies at a level of 100 %. The following types of estimations were introduced: a) Estimation of the sum of unconjugated THF and THF-21-glucosiduronate in highly diluted unprocessed human urine. b) A similar estimation of THE and THF-21-glucosiduronate as in a). c) Estimation of THF after β-glucuronidase treatment and dilution without further procedures. d) A similar estimation for THE as in c). THF and THE values correlate significantly with known parameters that characterize the cortisol production, such as cortisol secretion rate and the urinary corticoid (C21-α-ketolic-steroid) levels.

RADIOIMMUNOASSAY OF TETRAHYDROCORTICOSTERONE (THB) IN HUMAN URINE

1978 ◽  
Vol 88 (1) ◽  
pp. 139-148 ◽  
Author(s):  
K.-H. Kohl ◽  
K.-H. Gless ◽  
S. Abdelhamid ◽  
B. Penke ◽  
P. Vecsei
Keyword(s):  
New Zealand ◽  
Bovine Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Normal Values ◽  
Chromatographic System ◽  
Acth Stimulation ◽  
Cross Reactions ◽  
Dexamethasone Suppression ◽  
New Zealand Rabbits

ABSTRACT Antisera against tetrahydrocorticosterone (THB) (one of the metabolites of the glucocorticoid corticosterone) were raised in white New Zealand rabbits. A 3α,5β-THB-20-oxime-bovine serum albumin complex was used as antigen. The resulting titers were 1/300–1/19 000. The cross-reactions of the best antiserum were: THDOC 2.25%, THB-21-glucuronide 39%, and THF-21-glucuronide 4.8%. Other steroids and steroid metabolites have given negligible cross-reactions, e. g. allo-(3α,5α-)-THB 0.53 %, tetrahydro-11-dehydro-corticosterone (THA) 0.92%, allo-THA 0%, and THB-3-glucuronide 0 %. Two methods were introduced: a) for screening an estimation of THB-glucuronide, mostly THB-21-glucuronide ("THB-gluc."), in unprocessed urine after simple dilution b) specific estimation of THB using one paper chromatographic system. The values estimated by method a) showed a highly significant correlation (r = 0.9654, P < 0.01) with the values obtained by technique b). Normal values (method b) measured in subjects hospitalized under standardized conditions were: 21.8–177.6; mean 89.1 μg/24 h. THB levels and "THB-gluc." levels react to ACTH stimulation and dexamethasone suppression and thus we propose that the methods are useful for the diagnosis of hyper- and hypocorticosterone production states.

scholarly journals An enzyme-linked immunoassay for the collagen cross-link pyridinoline

1982 ◽  
Vol 207 (3) ◽  
pp. 617-620 ◽  
Author(s):  
S P Robins
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Human Urine ◽  
Bovine Serum ◽  
Urine Samples ◽  
Cross Link ◽  
Human Urine Samples

An inhibition immunoassay method for the determination of pyridinoline was developed with the use of microtitre plates coated with a pyridinoline-gelatin conjugate and rabbit antisera directed against pyridinoline linked to bovine serum albumin. The sensitivity of the assay is about 2pmol of pyridinoline, and the presence of related pyridinium and lysine-derived compounds does not significantly interfere with the procedure. Its application to tissue and human urine samples is described.

scholarly journals Electrochemical characterization and determination of carbamazepine as pharmaceutical standard and tablet content on gold electrode

2014 ◽  
Vol 68 (2) ◽  
pp. 207-212 ◽  
Author(s):  
Nemanja Trisovic ◽  
Bojan Bozic ◽  
Slobodan Petrovic ◽  
Svetlana Tadic ◽  
Milka Avramov-Ivic
Keyword(s):  
Cyclic Voltammetry ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Phosphate Buffer ◽  
Bovine Serum ◽  
Gold Electrode ◽  
Anticonvulsant Drug ◽  
Voltammetric Techniques ◽  
Anodic Behaviour

The anodic behaviour of carbamazepine (CBZ), an anticonvulsant drug, has been studied on gold electrode in 0.1 mol dm-3 phosphate buffer of pH 7.0 by using cyclic voltammetry. It has been found that the value of the oxidative current of pure CBZ at +0.90 V is a linear function of the concentration in a range from 1.0?10-7 to 1.0?10?4 mol dm?3. The detection of CBZ in the concentration of 1.0?10-8 mol dm-3 is among the lowest that have been reported for this drug using voltammetric techniques. CBZ as a content of tablet Galepsine? has been quantitatively determined. It has also been demonstrated that the modification of gold electrode with bovine serum albumin (BSA) results in a decrease of the oxidative peak current due to the binding of the drug to BSA.

A solid-phase enzymeimmunoassay for the determination of progesterone in bovine, porcine and ovine plasma

1995 ◽  
Vol 75 (4) ◽  
pp. 525-529 ◽  
Author(s):  
R. P. Del Vecchio ◽  
W. D. Sutherland ◽  
M. L. Connor
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Petroleum Ether ◽  
Bovine Serum ◽  
Solid Phase ◽  
Plasma Samples ◽  
Assay Sensitivity ◽  
Coefficients Of Variation ◽  
Rabbit Igg

The purpose of this project was to develop a valid quantitative enzymeimmunoassay (EIA) for progesterone in blood plasma of cattle, pigs and sheep. Rabbit anti-progesterone, mouse monoclonal anti-rabbit IgG, authentic progesterone, and acetylcholine esterase bound covalently to progesterone were the principal reagents used to develop the EIA. Ninety-six well microliter plates were coated with mouse monoclonal anti-rabbit IgG and saturated with bovine serum albumin before use. Rabbit anti-progesterone was diluted to a working dilution of 1:2.0 × 106. Standard curves were linear and ranged from 1.56 to 400 pg of progesterone per well which allowed for the measurement of 0.03125 to 8.0 ng mL−1. Assay sensitivity averaged 1.56 pg well−1. Progesterone was extracted from plasma samples with petroleum ether. Plasma samples (n = 3 or 4 from each species) with unknown amounts of progesterone that were extracted and serially diluted with EIA buffer did not deviate from parallelism with progesterone standard curves in buffer. The correlation between EIA and radioimmunoassay (RIA) measurements of progesterone in the same plasma samples was high (P < 0.0001) for all three species (r = 0.96 for bovine; r = 0.96 for porcine; r = 0.94 for ovine). The regression of EIA data on RIA data produced the following equations:[Formula: see text]The intra- and inter-assay coefficients of variation were 5.4 and 10.6% for bovine, 5.8 and 11.0% for porcine and, 6.1 and 12.3% for ovine, respectively. These data show that this EIA is a valid and reliable memod for quantitating progesterone in extracts of bovine, porcine and ovine plasma. Key words: Enzymeimmunoassay, progesterone, plasma, bovine, porcine, ovine

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