Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples
Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples
Authors
Moczko, Ewa
Diaz, Richard
Rivas, Bernabe
Garcia, Camilo
Pereira, Eduardo
Piletsky, Sergey
Caceres, Cesar
Diaz, Richard
Rivas, Bernabe
Garcia, Camilo
Pereira, Eduardo
Piletsky, Sergey
Caceres, Cesar
Authors
Date
Datos de publicaciĆ³n:
POLYMERS,Vol.11,,2019
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Abstract
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 mu g.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 mu g.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.