(BIo)analytical microsystems based on the use of nanoparticles. Microreactors for the synthesis of nanoparticles

Resumen

Demand for new automated analytical procedures and instrumentation for the continuous monitoring of (bio)chemical parameters, such as those affecting water quality, gathers special importance due to the significant consequences that the use or the consumption of (bio)chemically contaminated water might have. µ- Total Analysis Systems or Lab-on-a-Chip devices are increasingly employed with this purpose owing to the high integration and automation of these devices that allow, through miniaturization, the possibility of performing in situ measurements. Besides, the use of nanoparticles with analytical purposes has demonstrated to improve the sensitivity and detection limits of optical methods. Within this general context, the present work is focused on the development of automated analytical microsystems to perform colorimetric or fluorimetric analyses, based on the use of nanoparticles as optical labels, for the rapid detection of water pollutants or organisms, such as heavy metals or bacteria. However, the use of nanoparticles with identical physical characteristics is a must in order to obtain reliable and reproducible analytical measures. Therefore, the first part of this work has been addressed to the development of microreactors for the synthesis of metallic, semiconductor and carbon nanoparticles. Seven different ceramic microreactors have been developed, in which the influence of the design (dimensions and configuration of the channels) and the hydrodynamic (flow rates and dosage volumes) parameters have been studied. Moreover, a miniaturized optical detection system has been implemented for the on-line monitoring of the synthesis; and a thermal module has been also developed to reach and accurately control temperatures up to 300 ºC for when required. It is important to highlight that all prototypes operate automatically, which simplifies the synthesis and improves the reproducibility of the obtained nanoparticles. The second part focuses on the development of analytical microsystems based on the use of the nanoparticles previously synthesized for water quality analysis. Two different prototypes have been constructed and tested. The first described allows the monitoring of mercuric ion in water. The system is based on the selective recognition of the analyte by an ionophore (a thiourea derivative), attached onto gold nanoparticles surface. The metal-ionophore interaction generates a change on the surface plasmon resonance band of the nanoparticles, resulting in a quantifiable optical signal, which is on-line registered by a miniaturized optical detection system integrated in the microfluidic platform. Once optimized, the device is capable to automatically detect up to 11 ppb of mercuric ion. Finally, a prototype for the specific determination of Escherichia coli in water has been developed. An oligonucleotide sandwich assay is performed in the microfluidic system. On the one hand, magnetic beads are employed as substrate support of the assay, which allow simplifying and improving the different steps of the procedure. On the other hand, and as a first approximation to the use of fluorescent nanoparticles within microsystems, ß -galactosidase enzyme is used as probe label. This change responds to the necessity of improving the optical detection system (while maintaining its portability and low cost), due to the low sensitivity observed when fluorescent nanoparticles were used as labels, which at the moment is not feasible. The enzyme generates a coloured product (o-nitrophenol) with the addition of the substrate, which is registered through the implemented miniaturized optical system. Once optimized, the device can detect up to 1 ppb of the target oligonucleotide in only 20 minutes. The presented results demonstrate the great potential of automated analytical microsystems based on the use of nanoparticles for the monitoring of water quality. Similarly, the suitability of microreactors for the synthesis of nanoparticles has been well proved.