Graphitized carbon nanofiber-Pt nanoparticle hybrids as sensitive tool for preparation of screen printing biosensors. Detection of lactate in wines and ciders.

This work describes the fabrication of a new lactate biosensor. The strategy is based on the use of a novel hybrid nanomaterial for amperometric biosensors i.e. platinum nanoparticles (PtNps) supported on graphitized carbon nanofibers (PtNps/GCNF) prepared by chemical reduction of the Pt precursor at GCNF surfaces. The biosensors were constructed by covalent immobilization of lactate oxidase (LOx) onto screen printed carbon electrodes (SPCEs) modified with PtNps (PtNps/GCNF-SPCEs) using polyethyleneimine (PEI) and glutaraldehyde (GA). Experimental variables concerning both the biosensor design and the detection process were investigated for an optimal analytical performance. Lactate biosensors show good reproducibility (RSD 4.9%, n=10) and sensitivity (41,302±546) µA/Mcm(2), with a good limit of detection (6.9µM). Covalent immobilization of the enzyme allows the reuse of the biosensor for several measurements, converting them in a cheap alternative to the solid electrodes. The long-term stability of the biosensors was also evaluated. 90% of the signal was kept after 3months of storage at room temperature (RT), while 95% was retained after 18months at -20°C. These results demonstrate that the method provides sensitive electrochemical lactate biosensors where the stability of the enzymatic activity can be preserved for a long period of time in adequate storage conditions.

Disposable amperometric biosensor based on lactate oxidase immobilised on platinum nanoparticle-decorated carbon nanofiber and poly(diallyldimethylammonium chloride) films.

A novel biosensor for lactate has been developed, using screen-printed carbon electrodes (SPCE) and lactate oxidase (LOx). The active surface of the electrodes was modified using a dispersion of platinum nanoparticle decorated carbon nanofibers (PtNp-CNF) in poly(diallyldimethylammonium) chloride (PDDA) solution. In this way, sensitive, disposable, low cost and reliable hydrogen peroxide sensors were obtained. The immobilisation of LOx on top of these PtNp-CNF-PDDA/SPCEs resulted in amperometric biosensors with high operational stability. The sensitivity of the optimised lactate biosensor was 36.8 (mA/Mcm(2)) with a linear range of 25-1500 µM. The limit of detection was 11 µM (S/N=3). Reproducibility, selectivity and storage stability were also evaluated. Additionally, the stability of the biosensor was also predicted by a model based on thermal degradation. Finally, lactate in sweat and blood samples was determined in a sport test using LOx/PtNp-CNF-PDDA/SPCEs and commercial biosensors respectively. Based on these data, the validity of the sweat lactate for the determination of the lactate threshold is discussed.

Nanostructured disposable impedimetric sensors as tools for specific biomolecular interactions: sensitive recognition of concanavalin A.

The development of sensors to detect specific weak biological interactions is still today a challenging topic. Characteristics of carbohydrate-protein (lectin) interactions include high specificity and low affinity. This work describes the development of nanostructured impedimetric sensors for the detection of concanavalin A (Con A) binding to immobilized thiolated carbohydrate derivatives (D-mannose or D-glucose) onto screen-printed carbon electrodes (SPCEs) modified with gold nanoparticles. Thiolated D-galactose derivative was employed as negative control to evaluate the selectivity of the proposed methodology. After binding the thiolated carbohydrate to the nanostructured SPCEs, different functionalized thiols were employed to form mixed self-assembled monolayers (SAM). Electrochemical impedance spectroscopy (EIS) was employed as a technique to evaluate the binding of Con A to selected carbohydrates through the increase of electron transfer resistance of the ferri/ferrocyanide redox probe at the differently SAM modified electrodes. Different variables of the assay protocol were studied in order to optimize the sensor performance. Selective Con A determinations were only achieved by the formation of mixed SAMs with adequate functionalized thiols. Important differences were obtained depending on the chain lengths and functional groups of these thiols. For the 3-mercapto-1-propanesulfonate mixed SAMs, the electron transfer resistance varied linearly with the Con A concentration in the 2.2-40.0 µg mL(-1) range for D-mannose and D-glucose modified sensors. Low detection limits (0.099 and 0.078 pmol) and good reproducibility (6.9 and 6.1%, n=10) were obtained for the D-glucose and D-mannose modified sensors, respectively, without any amplification strategy.