A COST REDUCTIVE METHOD FOR THE RADIOIMMUNOASSAY OF PLASMA ANDROSTENEDIONE. A COMPARISON OF ANTISERA AND THE EFFECTS OF CHROMATOGRAPHY

1974 ◽  
Vol 76 (3) ◽  
pp. 597-607 ◽  
Author(s):  
John F. Hennam ◽  
William P. Collins
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Significant Difference ◽  
Assay Tube ◽  
Before And After ◽  
Process Effects ◽  
The Cost ◽  
Potential Use

ABSTRACT A method is described and evaluated for the determination of androstenedione (A) in peripheral venous plasma. The potential use of antisera to A-6-carboxymethyl thioether-bovine serum albumin and A-11α-succinyl-bovine serum albumin has been evaluated. The same plasma samples have been analysed before and after chromatography on a micro column of Sephadex LH-20. A dye, azobenzene is used to locate the fraction containing androstenedione. The results show that there is no significant difference in the values of apparent A using either antisera (overall mean 3.2 ng/ml plasma from men and 2.8 ng/ml from women). After chromatography these values are reduced by 62 and 42% respectively to 1.2 and 1.6 ng/100 ml. A new method using a mixture of ammonium and calcium sulphates is described for the separation of steroid bound to antibody. The precipitate is then resuspended and the amount of radioactivity determined, directly in the assay tube, by liquid scintillation counting. This process effects a 68 % reduction in the cost of the assay of each plasma sample.

Radioimmunoassay for free and conjugated urinary metanephrine.

1977 ◽  
Vol 23 (7) ◽  
pp. 1264-1267 ◽  
Author(s):  
R W Lam ◽  
R Artal ◽  
D A Fisher
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Incubation Time ◽  
Bovine Serum ◽  
Normal Subjects ◽  
Specific Radioimmunoassay ◽  
Coefficients Of Variation ◽  
Assay Tube ◽  
Before And After ◽  
Urine Specimens

Abstract We describe a convenient and specific radioimmunoassay for urinary metanephrine, in which we used an antiserum generated in rabbits by injecting with synephrine conjugated to bovine serum albumin as described by Grota and Brown [Endocrinology 98, 615 (1976)]. The antiserum is specific for metanephrine and epinephrine. Epinephrine cross reaction is minimized to 1% by adding 2 ng of epinephrine to each assay tube. Incubation time is 12 h. The sensitivity of the assay is 40 pg. Within-and between-assay coefficients of variation are 7.0 and 9.7%, respectively. We assayed first-voided morning urine and 24-h urine specimens from 15 normal subjects before and after acid hydrolysis, to assess total and free metanephrine concentrations. Mean (+/-SD) total and free urinary metanephrine excretion were 17 +/- 11 and 3.5 +/- 1.0 micrograms/24 h in these subjects; corresponding values for the morning specimens were 13.4 +/- 8.3 and 3.5 +/- 1.5 micrograms/liter, respectively.

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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