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Biomedical subjects

Fernando Rubio

Publications and source records attributed to Fernando Rubio.

3 recordsLinked to original sources

Evaluation of commercial immunoassays for the detection of estrogens in water by comparison with high-performance liquid chromatography tandem mass spectrometry HPLC-MS/MS (QqQ).

In this work four different commercially available enzyme-linked immunosorbent assays (ELISA) (from Japan EnviroChemicals, Ltd., Tokyo, Japan) were evaluated in terms of performance for the rapid screening of estrogens in different water matrices, including natural and spiked samples from urban wastewater, river water and ground water. All four test kits are based on monoclonal antibodies. The compounds detected by these immunoassays are (1) 17-beta-estradiol, (2) estrone, (3) 17-alpha-ethynyl estradiol and (4) estrogens in general, with high recognition properties for 17-beta-estradiol, estrone and estriol. Standards were prepared in water containing 10% (v/v) methanol. The IC50 (corresponding to the 50% of the effective concentration) values, the dynamic ranges, and the limits of detection of the ELISA kits were 0.060-0.304 microg/L, 0.05-5 microg/L and 0.05 microg/L, respectively. All samples were extracted by solid-phase extraction (SPE) beforehand, and the evaluation was carried out by comparing the results obtained by ELISA with those obtained by HPLC-MS/MS using a triple quadrupole (QqQ) instrument. In addition, two different solid-phase extraction procedures were carried out and compared. Except for moderate overestimation in the results observed with the ELISA kits in the analysis of complex wastewater samples, the results obtained using all of the tested techniques were generally in very good agreement.

Chromatography, High Pressure Liquid↗

Glyphosate immunosensor. Application for water and soil analysis.

A fully automated immunosensor for the herbicide glyphosate has been developed on the basis of the immunocomplex capture assay protocol. The sensor carries out on-line analyte derivatization prior to the assay and uses a selective anti-glyphosate serum, a glyphosate peroxidase enzyme tracer, and fluorescent detection. Under optimal conditions, the detection limit achieved is 0.021 microg/L with an analysis rate of 25 min per assay, autonomy of more than 48 h, and sensor reusability >500 analytical cycles. The immunosensor is able to discriminate structurally related molecules, such as aminomethylphosphonic acid, the main metabolite of glyphosate, and other related herbicides, such as glufosinate and glyphosine. Interferences from naturally occurring species (anions, cations, and humic substances) and their elimination were also studied. The immunosensor has been successfully applied to water and soil sample analysis, with good recoveries at levels lower than 1 microg/L. Results obtained with the immunosensor correlate well with data from a magnetic particle ELISA and LC/LC/MS chromatographic method.

Glycine↗

Comparison of a direct ELISA and an HPLC method for glyphosate determinations in water.

A competitive direct enzyme-linked immunosorbent assay (ELISA) and high-pressure liquid chromatographic (HPLC) methods were compared in terms of accuracy and precision for the detection and quantification of glyphosate-spiked Nanopure, tap, and river waters. The ELISA had a detection limit of 0.6 ng mL(-)(1) and a linear working range of 1-25 ng mL(-)(1), whereas the HPLC method had a detection limit of 50 ng mL(-)(1) and a linear working range of 100-10000 ng mL(-)(l). No statistically significant differences (95% confidence interval) were found between the ELISA and HPLC analysis of the three water matrixes. The coefficients of variation obtained with the ELISA in tap water were between 10 and 19%, whereas the coefficients of variation for the HPLC analysis were between 7 and 15%. The use of ELISA for the analysis of glyphosate in water is a cost-effective and reliable method capable of meeting water quality guidelines established for Europe and North America.

Chromatography, High Pressure Liquid↗