A quasi-reagentless point-of-care test for nitrite and unaffected by oxygen and cyanide.

The ubiquitous nitrite is a major analyte in the management of human health and environmental risks. The current analytical methods are complex techniques that do not fulfil the need for simple, robust and low-cost tools for on-site monitoring. Electrochemical reductase-based biosensors are presented as a powerful alternative, due to their good analytical performance and miniaturization potential. However, their real-world application is limited by the need of anoxic working conditions, and the standard oxygen removal strategies are incompatible with point-of-care measurements. Instead, a bienzymatic oxygen scavenger system comprising glucose oxidase and catalase can be used to promote anoxic conditions in aired environments. Herein, carbon screen-printed electrodes were modified with cytochrome c nitrite reductase together with glucose oxidase and catalase, so that nitrite cathodic detection could be performed by cyclic voltammetry under ambient air. The resulting biosensor displayed good linear response to the analyte (2-200 µM, sensitivity of 326 ± 5 mA M-1 cm-2 at -0.8 V; 0.8-150 µM, sensitivity of 511 ± 11 mA M-1 cm-2 at -0.5 V), while being free from oxygen interference and stable up to 1 month. Furthermore, the biosensor's catalytic response was unaffected by the presence of cyanide, a well-known inhibitor of heme-enzymes.

Ensuring Food Integrity by Metrology and FAIR Data Principles.

Food integrity is a general term for sound, nutritive, healthy, tasty, safe, authentic, traceable, as well as ethically, safely, environment-friendly, and sustainably produced foods. In order to verify these properties, analytical methods with a higher degree of accuracy, sensitivity, standardization and harmonization and a harmonized system for their application in analytical laboratories are required. In this view, metrology offers the opportunity to achieve these goals. In this perspective article the current global challenges in food analysis and the principles of metrology to fill these gaps are presented. Therefore, the pan-European project METROFOOD-RI within the framework of the European Strategy Forum on Research Infrastructures (ESFRI) was developed to establish a strategy to allow reliable and comparable analytical measurements in foods along the whole process line starting from primary producers until consumers and to make all data findable, accessible, interoperable, and re-usable according to the FAIR data principles. The initiative currently consists of 48 partners from 18 European Countries and concluded its "Early Phase" as research infrastructure by organizing its future structure and presenting a proof of concept by preparing, distributing and comprehensively analyzing three candidate Reference Materials (rice grain, rice flour, and oyster tissue) and establishing a system how to compile, process, and store the generated data and how to exchange, compare them and make them accessible in data bases.

Construction of effective disposable biosensors for point of care testing of nitrite.

In this paper we aim to demonstrate, as a proof-of-concept, the feasibility of the mass production of effective point of care tests for nitrite quantification in environmental, food and clinical samples. Following our previous work on the development of third generation electrochemical biosensors based on the ammonia forming nitrite reductase (ccNiR), herein we reduced the size of the electrodes' system to a miniaturized format, solved the problem of oxygen interference and performed simple quantification assays in real samples. In particular, carbon paste screen printed electrodes (SPE) were coated with a ccNiR/carbon ink composite homogenized in organic solvents and cured at low temperatures. The biocompatibility of these chemical and thermal treatments was evaluated by cyclic voltammetry showing that the catalytic performance was higher with the combination acetone and a 40°C curing temperature. The successful incorporation of the protein in the carbon ink/solvent composite, while remaining catalytically competent, attests for ccNiR's robustness and suitability for application in screen printed based biosensors. Because the direct electrochemical reduction of molecular oxygen occurs when electroanalytical measurements are performed at the negative potentials required to activate ccNiR (ca.-0.4V vs Ag/AgCl), an oxygen scavenging system based on the coupling of glucose oxidase and catalase activities was successfully used. This enabled the quantification of nitrite in different samples (milk, water, plasma and urine) in a straightforward way and with small error (1-6%). The sensitivity of the biosensor towards nitrite reduction under optimized conditions was 0.55 A M(-1) cm(-2) with a linear response range 0.7-370 µM.

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.