Direct solid-phase enzymoimmunoassay of testosterone in saliva.

1989 ◽  
Vol 35 (10) ◽  
pp. 2044-2047 ◽  
Author(s):  
K Howard ◽  
M Kane ◽  
A Madden ◽  
J P Gosling ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Medical Applications ◽  
Microtiter Plates ◽  
Large Numbers ◽  
Coefficients Of Variation ◽  
Serial Sampling ◽  
Direct Solid

Abstract This competitive, solid-phase enzymoimmunoassay for testosterone in saliva is carried out on microtiter plates and involves no chromatographic or extraction steps. With an overnight incubation the detection limit of the assay is 230 fg per well (16.1 pmol/L). There was a good correlation (correlation coefficient 0.95) between testosterone concentrations measured with and without prior extraction of the saliva samples. Repeated assay of three control saliva samples containing a range of testosterone concentrations (200-1000 pmol/L) gave within- and between-assay coefficients of variation of 5.5-13.2%. The analytical procedure is simple and closely resembles already published procedures for the determination of progesterone and estrone (with extraction) in saliva. One person can assay 200 samples in 24 h and the assay is suitable for reproductive and sports medical applications, particularly for projects involving serial sampling and yielding large numbers of samples.

Solid-phase enzymoimmunoassay of estrone in saliva.

1989 ◽  
Vol 35 (4) ◽  
pp. 569-572 ◽  
Author(s):  
J Folan ◽  
J P Gosling ◽  
M F Finn ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Diethyl Ether ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Ovarian Activity ◽  
Microtiter Plates ◽  
Coefficients Of Variation ◽  
Serial Sampling ◽  
High Concentrations ◽  
Working Day

Abstract In this solid-phase enzymoimmunoassay for estrone in saliva, microtiter plates are used after extraction of the sample with diethyl ether. No chromatographic step is involved. The detection limit of the assay is 450 fg per well (33 pmol/L). Intra- and interassay coefficients of variation for the assay of low, medium, and high concentrations of estrone in saliva were respectively 4.2, 12.7; 5.2, 8.7; and 2.7, 5.8%. Using this assay, we found a highly significant correlation (P less than 0.001) between estrone concentrations in time-matched serum and saliva samples. The analytical procedure is rapid and relatively simple. One person can assay 50-60 saliva samples during a normal working day. We conclude that the assay is very suitable, even in small laboratories, for saliva estrone measurements, which, in facilitating serial sampling, enables dynamic observations of estrone concentrations and ovarian activity to be more easily made.

Solid-phase enzymoimmunoassay of estrone in serum.

1988 ◽  
Vol 34 (9) ◽  
pp. 1843-1846 ◽  
Author(s):  
J Folan ◽  
J P Gosling ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Correlation Coefficient ◽  
Diethyl Ether ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Serum Samples ◽  
Microtiter Plates ◽  
Coefficients Of Variation ◽  
High Concentrations ◽  
Working Day

Abstract This rapid solid-phase enzymoimmunoassay for estrone in serum or plasma is done on microtiter plates after the serum is extracted with diethyl ether. No chromatographic or centrifugation steps are involved. The detection limit of the assay is 380 fg per well (28 pmol/L). Intra- and interassay coefficients of variation for the assay of low, medium, and high concentrations of estrone in plasma were respectively 4.4, 9.3; 2.3, 9.1; and 2.0, 6.3 percent. There was a good correlation (correlation coefficient 0.95) between estrone concentrations measured with this assay and with a commercial radioimmunoassay. The analytical procedure is simple, and one person can assay 80 serum samples per working day. We conclude that the assay is very suitable for serum estrone measurements and is more convenient than published radioimmunoassays.

Direct chemiluminescence immunoassay of estradiol in saliva

1990 ◽  
Vol 36 (12) ◽  
pp. 2036-2041 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
J Bouve ◽  
D Leyseele ◽  
D VandeKerckhove
Keyword(s):  
Detection Limit ◽  
Incubation Time ◽  
Solid Phase ◽  
Chemiluminescence Immunoassay ◽  
Microtiter Plates ◽  
Analytical Recovery ◽  
Plate Reader ◽  
Steroid Binding ◽  
Method R ◽  
Direct Solid

Abstract A sensitive and simple direct solid-phase chemiluminescence immunoassay is described for estradiol in saliva. In this assay, a second antibody is bound to the wells of microtiter plates. Either buffer with standards or saliva (100 microL) is incubated in these wells with monoclonal anti-estradiol antibody and with estradiol-isoluminol conjugate. Incubation time is 2 h. Chemiluminescence of the bound fraction is measured in a manually operated luminometer (Biocounter). The assay has a detection limit of 3.8 pmol/L; analytical recovery of added estradiol is 96.8% (SD 7.0%); within- and between-assay CVs range between 2.5% and 12.7%. Forty unknown saliva samples can be assayed and results calculated within 4.5 h. Results of a slightly modified procedure-with black microtiter plates and a prototype of an automated plate reader (Luminoskan)--compare well with those of the described method (r = 0.97). Because steroid-binding globulins have been found in saliva, the effect of displacing agents on the results of the direct chemiluminescence assay is described.

scholarly journals Determination of Citrinin in Barley by Indirect and Direct Enzyme Immunoassay

1996 ◽  
Vol 79 (6) ◽  
pp. 1325-1329 ◽  
Author(s):  
David Abramson ◽  
Ewald Usleber ◽  
Erwin Martlbauer
Keyword(s):  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Polyclonal Antibodies ◽  
Detection Limits ◽  
Keyhole Limpet Hemocyanin ◽  
Buffer Solutions ◽  
Microtiter Plates ◽  
Coefficients Of Variation ◽  
Optimum Extraction

Abstract Polyclonal antibodies against the mycotoxin citrinin were raised in rabbits after immunization with citrinin conjugated to keyhole limpet hemocyanin. The antibodies were used in a competitive indirect enzyme immunoassay (EIA) with citrinin coupled to glucose oxidase as the solid-phase antigen for coating microtiter plates. Detection limits of this indirect EIA for citrinin ranged from 0.4 to 0.8 ng/mL for buffer solutions. Recoveries of citrinin added to ground barley at 100–2000 ng/g ranged from 105 to 112%, with coefficients of variation between 4.5 and 12%. A direct competitive EIA also was established, with citrinin coupled to horseradish peroxidase as the labeled antigen. Detection limits of this direct EIA for citrinin ranged from 2 to 4 ng/mL for buffer solutions. Recoveries of citrinin added to ground barley at 500–2000 ng/g ranged from 108 to 111%, with coefficients of variation between 8.4 and 26.9%. In naturally contaminated barley samples assayed with the indirect EIA, optimum extraction of citrinin was obtained in 30 min, and only one extraction was necessary to recover 72–76% of the analyte.

Determination of Thiabendazole Residues in Pears by Solid-Phase Spectrofluorimetry

1994 ◽  
Vol 77 (6) ◽  
pp. 1651-1653 ◽  
Author(s):  
Luis Fermin Cápitan-Vallvey ◽  
Ramiro Avidad ◽  
Jose Luis Vilchez
Keyword(s):  
Acetic Acid ◽  
Detection Limit ◽  
Fluorescence Intensity ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Solid Support ◽  
Relative Fluorescence Intensity ◽  
Buffer Solution ◽  
Silica Cell

Abstract An analytical procedure for determination of thiabendazole (TBZ) residues in pears is described. The method involves extracting the chemical from the chopped fruit with buffer solution (acetic acid–acetate, pH 4.6), use of Sephadex G-15 dextrantype gel as a solid support, and determination of TBZ by solid-phase spectrof luorimetry (SPF). The relative fluorescence intensity of the Sephadex G- 15 gel–TBZ system, packed in a 1 mm-thickness silica cell, was measured directly at λex = 303 nm and λem = 350 nm with a solid-phase attachment. The applicable concentration range was 5.0–20.0 ppb with a detection limit of 0.5 ppb. Recoveries were from 98.7 to 102.0% when 15.0 ppb of TBZ was added to the fruit.

Determination of Residual Acrylamide Monomer in Medical Polyacrylamide Hydrogel by HPLC-SPE

2005 ◽  
Vol 288-289 ◽  
pp. 405-408
Author(s):  
Zi Yi Wan ◽  
Ting Fei Xi ◽  
P. Zhao ◽  
Y. Sun ◽  
Z.G. Feng
Keyword(s):  
Detection Limit ◽  
Solid Phase ◽  
Polyacrylamide Hydrogel ◽  
Simple Method ◽  
Lower Detection Limit ◽  
Response Range ◽  
Coefficients Of Variation ◽  
Lower Detection ◽  
Linear Response Range

The polyacrylamide hydrogel (PAMG) has been used in cosmetology in China, Ukraine and Russia since 1990s. Because the monomer acrylamide(AM) used to produce PAMG has been implicated as a potential mutagen and reproductive toxicant[1,2], it is important to accurately determine the amount of residual AM monomer in the PAMG. In this study, a quick, practical and simple method to determine AM is presented with respect to the hydrogel. AM is analysed quantitatively by ODS-3 column with ultraviolet (UV) absorbance detector. AM is separated from interferential component with an aqueous solution of 0.9%NaCl (NS) adjusted at pH~3.7 using hydrochloric acid and then detected at a UV wavelength of 210 nm. The results show that ODS-3 is effective approach for quantifying AM concentrations in PAMG. This method has a lower detection limit of 0.003µg/ml and a linear response range of 0.003 and 0.9 µg/ml (depending on the range required for analysis). Precision studies give coefficients of variation of <3.2%(n=5) for 0.003µg/ml. The recoveries for this method are greater than 90%. When AM content in PAMG is lower than the detection limit of this method, SPE (solid phase extraction) could be used to concentrate AM. In the case, C18 cartridge is used. And the recoveries are about 70% for SPE when AM concentration is lower than ppb.

Direct solid-phase chemiluminescence immunoassay for salivary progesterone.

1986 ◽  
Vol 32 (5) ◽  
pp. 763-767 ◽  
Author(s):  
J De Boever ◽  
F Kohen ◽  
D Vandekerckhove
Keyword(s):  
Detection Limit ◽  
Luteal Phase ◽  
Solid Phase ◽  
Free Ligand ◽  
Follicular Phase ◽  
Chemiluminescence Immunoassay ◽  
Coefficients Of Variation ◽  
Analytical Recovery ◽  
Unstimulated Saliva ◽  
Direct Solid

Abstract A simple, direct chemiluminescence immunoassay for progesterone in mixed, unstimulated saliva is described. We use purified polyclonal anti-progesterone antibodies covalently coupled to polyacrylamide beads and progesterone-11 alpha-hemisuccinyl-aminobutylethyl isoluminol as the chemiluminescent ligand marker. Bound and free ligand are separated by simple centrifugation. The detection limit of the assay is 1.5 pg per tube (38 pmol/L). Intra- and interassay coefficients of variation for low, medium, and high progesterone concentrations are 7.9, 6.8, 8.8% and 9.3, 6.9, 8.5%, respectively. Analytical recovery of added progesterone is 99.6%. Mean +/- SD progesterone concentrations (pmol/L) in saliva are 178 +/- 46 in the follicular phase, 313 +/- 90 in the periovulatory phase, and 658 +/- 166 in the luteal phase of the menstrual cycle. Correlation between progesterone concentrations in serum (assayed by RIA after extraction) and in saliva is good (r = 0.88, p less than 0.001, n = 96). The assay is simple, fast (4 h, including 1.5 h of incubation time), and reliable.

scholarly journals Preconcentration of Copper with Solid Phase Extraction and its Determination by Flame Atomic Absorption Spectrometry

2008 ◽  
Vol 5 (4) ◽  
pp. 878-883 ◽  
Author(s):  
Ayob Parchehbaf Jadid ◽  
Habibollah Eskandari
Keyword(s):  
Detection Limit ◽  
Solid Phase Extraction ◽  
Solid Phase ◽  
Copper Ions ◽  
Absorption Spectrometry ◽  
Phase Extraction ◽  
Flame Atomic Absorption ◽  
Low Levels ◽  
Determination Of Copper

A new method for the solid phase extraction (SPE) and determination of copper ions at low levels is presented. Extraction percent and the effects of some factors were evaluated. The detection limit was in the range of 2.26 µg·L-1. This procedure has been successfully applied to determination of copper in water samples.

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

Sign in / Sign up

Export Citation Format

Share Document