scholarly journals Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples

Polymers ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 1497 ◽  
Author(s):  
Ewa Moczko ◽  
Richard Díaz ◽  
Bernabé Rivas ◽  
Camilo García ◽  
Eduardo Pereira ◽  
...  

In 2004, octopamine was added to the list of drugs banned by the world anti-doping agency (WADA) and prohibited in any sport competition. This work aims to develop a new analytical method to detect octopamine in water and human urine samples. We proposed a pseudo-enzyme-linked immunosorbent assay (pseudo-ELISA) by replacing traditional monoclonal antibodies with molecularly imprinted polymer nanoparticles (nanoMIPs). NanoMIPs were synthesised by a solid-phase approach using a persulfate initiated polymerisation in water. Their performance was analysed in pseudo competitive ELISA based on the competition between free octopamine and octopamine-HRP conjugated. The final assay was able to detect octopamine in water within the range 1 nmol·L−1–0.1 mol·L−1 with a detection limit of 0.047 ± 0.00231 µg·mL−1 and in human urine samples within the range 1 nmol·L−1–0.0001 mol·L−1 with a detection limit of 0.059 ± 0.00281 µg·mL−1. In all experiments, nanoMIPs presented high affinity to the target molecules and almost no cross-reactivity with analogues of octopamine such as pseudophedrine or l-Tyrosine. Only slight interference was observed from the human urine matrix. The high affinity and specificity of nanoMIPs and no need to maintain a cold chain logistics makes the nanoMIPs a competitive alternative to antibodies. Furthermore, this work is the first attempt to use nanoMIPs in pseudo-ELISA assays to detect octopamine.

2017 ◽  
Vol 38 (7) ◽  
pp. 1083-1090 ◽  
Author(s):  
Carolina Nasser Boscari ◽  
Giovanna Rodrigues Mazzuia ◽  
Célio Wisniewski ◽  
Keyller Bastos Borges ◽  
Eduardo Costa Figueiredo

2016 ◽  
Vol 8 (20) ◽  
pp. 4075-4085 ◽  
Author(s):  
Ricky Cássio Santos da Silva ◽  
Valdir Mano ◽  
Arnaldo César Pereira ◽  
Eduardo Costa de Figueiredo ◽  
Keyller Bastos Borges

A simple and selective sample preparation technique employing PT-MIP-μ-SPE coupled to HPLC/DAD was developed for the determination of the cis-enantiomers of ketoconazole in human urine samples.


1997 ◽  
Vol 43 (2) ◽  
pp. 363-368 ◽  
Author(s):  
Frédéric Taran ◽  
Yveline Frobert ◽  
Christophe Créminon ◽  
Jacques Grassi ◽  
Didier Olichon ◽  
...  

Abstract A fast competitive enzyme immunoassay (EIA) for measuring homovanillic acid in human urine samples was developed with a monoclonal antibody and acetylcholinesterase as enzyme label. Enzyme detection was performed by an easy colorimetric assay. Monoclonal antibodies were screened on the basis of sensitivity, specificity, and correlation studies. EIA has a detection limit of 0.5 μmol/L, a CV <10% in the 1.25–10 μmol/L range, and intra- and interassay CVs of <10%. Cross-reactivity with vanillylmandelic acid was 0.5% and <8% for other structurally related catecholamine metabolites. Parallelism of the EIA was shown in dilution studies and the correlation with routine HPLC assay in 62 normal and pathologic samples was EIA = 1.492 (HPLC) − 3.46, Sy|x = 47.52, range = 4–1800 μmol/L, r2 = 0.977. Additional data concerning the validity of this assay were provided by HPLC analysis of urinary immunoreactive material.


Sign in / Sign up

Export Citation Format

Share Document