Enhanced Electron Transfer Efficiency of Fructose Dehydrogenase onto Roll-to-Roll Thermal Stamped Laser-Patterned Reduced Graphene Oxide Films.

Herein, a strategy to stamp laser-produced reduced graphene oxide (rGO) onto flexible polymers using only office-grade tools, namely, roll-to-roll thermal stamping, is proposed, proving for the first time its effectiveness for direct bioelectrocatalysis. This straightforward, scalable, and low-cost approach allows us to overcome the limits of the integration of laser-induced rGO-films in bioanalytical devices. Laser-produced rGO has been thermally stamped (TS) onto different polymeric substrates (PET, PVC, and EVA) using a simple roll-laminator; the obtained TS-rGO films have been compared with the native rGO (untransferred) via morphochemical and electrochemical characterization. Particularly, the direct electron transfer (DET) reaction between fructose dehydrogenase (FDH) and TS-rGO transducers has been investigated, with respect to the influence of the amount of enzyme on the catalytic process. Remarkable differences have been observed among TS-rGO transducers; PET proved to be the elective substrate to support the transfer of the laser-induced rGO, allowing the preservation of the morphochemical features of the native material and returning a reduced capacitive current. Noteworthily, TS-rGOs ensure superior electrocatalysis using a very low amount of FDH units (15 mU). Eventually, TS-rGO-based third-generation complete enzymatic biosensors were fabricated via low-cost benchtop technologies. TS-rGOPET exhibited bioanalytical performances superior to the native rGO, allowing a sensitive (0.0289 µA cm-2 µM-1) and reproducible (RSD = 3%, n = 3) d-fructose determination at the nanomolar level (LOD = 0.2 µM). TS-rGO exploitability as a point-of-need device was proved via the monitoring of d-fructose during banana (Musa acuminata) postharvest ripening, returning accurate (recoveries 110-90%; relative error -13/+1%) and reproducible (RSD ≤ 7%; n = 3) data.

Impedimetric immunosensor for microalbuminuria based on a WS2/Au water-phase assembled nanocomposite.

An electrochemical impedimetric biosensor for human serum albumin (HSA) determination is proposed. The biosensor is based on water-phase assembled nanocomposites made of 2D WS2 nanoflakes and Au nanoparticles (AuNPs). The WS2 has been produced using a liquid-phase exfoliation strategy assisted by sodium cholate, obtaining a water-stable suspension that allowed the straightforward decoration with AuNPs directly in the aqueous phase. The resulting WS2/Au nanocomposite has been characterized by atomic force microscopy and Raman spectroscopy and, then, employed to modify screen-printed electrodes. Good electron-transfer features have been achieved. An electrochemical immunosensing platform has been assembled exploiting cysteamine-glutaraldehyde covalent chemistry for antibody (Ab) immobilization. The resulting immunosensor exhibited good sensitivity for HSA detection (LOD = 2 ng mL-1), with extended linear range (0.005 - 100 µg mL-1), providing a useful analytical tool for HSA determination in urine at relevant clinical ranges for microalbuminuria screening. The HSA quantification in human urine samples resulted in recoveries from 91.8 to 112.4% and was also reproducible (RSD < 7.5%, n = 3), with marked selectivity. This nanocomposite, thanks to the reliable performance and the ease of the assembling strategy, is a promising alternative for electrochemical immunosensing of health relevant markers.

Lab-made flexible third-generation fructose biosensors based on 0D-nanostructured transducers.

Herein, we report a scalable benchtop electrode fabrication method to produce highly sensitive and flexible third-generation fructose dehydrogenase amperometric biosensors based on water-dispersed 0D-nanomaterials. The electrochemical platform was fabricated via Stencil-Printing (StPE) and insulated via xurography. Carbon black (CB) and mesoporous carbon (MS) were employed as 0D-nanomaterials promoting an efficient direct electron transfer (DET) between fructose dehydrogenase (FDH) and the transducer. Both nanomaterials were prepared in water-phase via a sonochemical approach. The nano-StPE exhibited enhanced electrocatalytic currents compared to conventional commercial electrodes. The enzymatic sensors were exploited for the determination of D-fructose in model solutions and various food and biological samples. StPE-CB and StPE-MS integrated biosensors showed appreciable sensitivity (∼150 µA cm-2 mM-1) with µmolar limit of detection (0.35 and 0.16 µM, respectively) and extended linear range (2-500 and 1-250 µM, respectively); the selectivity of the biosensors, ensured by the low working overpotential (+0.15 V), has been also demonstrated. Good accuracy (recoveries between 95 and 116%) and reproducibility (RSD ≤8.6%) were achieved for food and urine samples. The proposed approach because of manufacturing versatility and the electro-catalytic features of the water-nanostructured 0D-NMs opens new paths for affordable and customizable FDH-based bioelectronics.

Rapid pretreatment-free evaluation of antioxidant capacity in extra virgin olive oil using a laser-nanodecorated electrochemical lab-on-strip.

An electroanalytical lab-on-a-strip device for the direct extra-virgin olive oil (EVOO) antioxidant capacity evaluation is proposed. The lab-made device is composed of a CO2 laser nanodecorated sensor combined with a cutter-plotter molded paper-strip designed for EVOOs sampling and extraction. Satisfactory performance towards the most representative o-diphenols of EVOOs i.e., hydroxytyrosol (HY) and oleuropein (OL) were achieved; good sensitivity (LODHY = 2 µM; LODOL = 0.6 µM), extended linear ranges (HY: 10-250 µM; OL: 2.5-50 µM) and outstanding reproducibility (RSD < 5%, n = 3) were obtained in rectified oil. The device was challenged for the extraction-free analysis of 15 different EVOO samples, with satisfactory recoveries (90-94%; RSD < 5%, n = 3) and correlation with classical photometric assays (r ≥ 0.91). The proposed device includes all analysis steps, needs 4 µL of sample, and returns reliable results in 2 min, resulting portable and usable with a smartphone.

Nanofibrillar biochar from industrial waste as hosting network for transition metal dichalcogenides. Novel sustainable 1D/2D nanocomposites for electrochemical sensing.

Industrial wastes have become elective sustainable sources to obtain materials for electronic/electroanalytical purposes; on the other hand, easy and green strategies to include semiconductor 2D graphene-like materials in conductive networks are highly required. In this work, 1D/2D nanocomposites (NCs) based on nanofibrillar biochar (BH) from paper industry waste and transition metal dichalcogenides (TMDs: MoS2, WS2, MoSe2, and WSe2), were prepared in water via liquid phase exfoliation (LPE) using sodium cholate as bioderived surfactant. The TMD amount in the NCs has been carefully optimized, searching for the best compromise between electron transfer ability and electroanalytical performances. Four different water-dispersed BH-TMD NCs have been selected and comprehensively studied from the electrochemical point of view and morphologically characterized. The BH-TMDs potentiality have been demonstrated in model solutions and real samples towards different analytes of biological and agri-food interest. The most performing NCs have been selected and used for the simultaneous determination of the neurotransmitters dopamine (DP) and serotonin (SR), and the flavonoids quercetin (QR) and rutin (RT), obtaining good linearity (R2 ≥ 0.9956) with limits of detection ranging from 10 to 200 nM. Reproducible quantitative recovery values (90-112%, RSD ≤6%, n = 3) were obtained analyzing simultaneously DP and SR in synthetic biological fluid and drugs, and QR and RT in food supplements, proving the usability of the proposed materials for real analyses. This work proves that BH-nanofibers act as a sustainable conductive hosting network for 2D-TMDs, allowing full exploit their electroanalytical potential. The proposed BH-TMD NCs represent a sustainable, affordable, and captivating opportunity for the electrochemical and (bio)sensoristic field.

Carbon Black Functionalized with Naturally Occurring Compounds in Water Phase for Electrochemical Sensing of Antioxidant Compounds.

A new sustainable route to nanodispersed and functionalized carbon black in water phase (W-CB) is proposed. The sonochemical strategy exploits ultrasounds to disaggregate the CB, while two selected functional naturally derived compounds, sodium cholate (SC) and rosmarinic acid (RA), act as stabilizing agents ensuring dispersibility in water adhering onto the CB nanoparticles' surface. Strategically, the CB-RA compound is used to drive the AuNPs self-assembling at room temperature, resulting in a CB surface that is nanodecorated; further, this is achieved without the need for additional reagents. Electrochemical sensors based on the proposed nanomaterials are realized and characterized both morphologically and electrochemically. The W-CBs' electroanalytical potential is proved in the anodic and cathodic window using caffeic acid (CF) and hydroquinone (HQ), two antioxidant compounds that are significant for food and the environment. For both antioxidants, repeatable (RSD ≤ 3.3%; n = 10) and reproducible (RSD ≤ 3.8%; n = 3) electroanalysis results were obtained, achieving nanomolar detection limits (CF: 29 nM; HQ: 44 nM). CF and HQ are successfully determined in food and environmental samples (recoveries 97-113%), and also in the presence of other phenolic classes and HQ structural isomers. The water dispersibility of the proposed materials can be an opportunity for (bio) sensor fabrication and sustainable device realization.

Modular graphene mediator film-based electrochemical pocket device for chlorpyrifos determination.

In this work, a redox-graphene (Rx-Gr) film with electron-mediating ability has been integrated into a modular flexible pocket device, giving rise to a reusable biosensing platform. The Rx-Gr has been obtained in water from graphite taking advantage of catechin, a redox-antioxidant, able to assist the sonochemical layered-material exfoliation, conferring electron mediating feature. A film composed exclusively of Rx-Gr has been transferred via thermal rolling onto a flexible PET-support that was used as the biosensor base. The biosensing platform, composed of office-grade materials, was then fabricated using a cutter-plotter and assembled by thermal lamination; an interchangeable paper-based strip was used to host the enzymatic reaction and drive the capillary flow. An acetylcholinesterase-based inhibition assay has been optimized onboard the pocket device to determine chlorpyriphos, a widespread environmental pesticide. The proposed set-up allows the determination of chlorpyriphos at low overpotential (0.2 V) with satisfactory sensitivity (LOD = 0.2 ppb), thanks to the straightforward electroactivity of the Rx-Gr film towards thiocholine (enzymatic product). The modular design allows 5 consecutive complete inhibition assays (control + inhibition measure) retaining the performance (RSD = 5.4%; n = 5). The coupling of bench-top technologies and a new functional graphene film resulted in the development of a cost-effective, reusable, transportable, and within everyone's reach biosensing platform.

(+)-Catechin-assisted graphene production by sonochemical exfoliation in water. A new redox-active nanomaterial for electromediated sensing.

A new green and effective sonochemical liquid-phase exfoliation (LPE) is proposed wherein a flavonoid compound, catechin (CT), promotes the formation of conductive, redox-active, water-phase stable graphene nanoflakes (GF). To maximize the GF-CT redox activity, the CT concentration and sonication time have been studied, and the best performing nanomaterial-fraction selected. Physicochemical and electrochemical methods have been employed to characterize the morphological, structural, and electrochemical features of the GF-CT nanoflakes. The obtained GF intercalated with CT exhibits fully reversible electrochemistry (ΔEp = 28 mV, ipa/ipc = ⁓1) because of the catecholic adducts. GF-CT-integrated electrochemistry was generated directly during LPE of graphite, with no need of graphene oxide production, nor activation steps, electropolymerization, or ex-post functionalization. The GF-CT electro-mediator ability has been proven towards hydrazine (HY) and ß-nicotinamide adenine dinucleotide (NADH) by simply drop-casting the redox-material onto screen-printed electrodes. GF-CT-based electrodes by using amperometry exhibited high sensitivity and extended linear ranges (HY: LOD = 0.1 µM, L.R. 0.5-150 µM; NADH: LOD = 0.6 µM, L.R. 2.5-200 µM) at low overpotential (+ 0.15 V) with no electrode fouling. The GF-CT electrodes are performing significantly better than commercial graphite electrodes and graphene nanoflakes exfoliated with a conventional surfactant, such as sodium cholate. Recoveries of 94-107% with RSD ≤ 8% (n = 3) for determination of HY and NADH in environmental and biological samples were achieved, proving the material functionality also in challenging analytical media. The presented GF-CT is a new functional redox-active material obtainable with a single-pot sustainable strategy, exhibiting standout properties particularly prone to (bio)sensors and cutting-edge device development.

Class-selective voltammetric determination of hydroxycinnamic acids structural analogs using a WS2/catechin-capped AuNPs/carbon black-based nanocomposite sensor.

A high-performance screen-printed electrode (SPE) based nanocomposite sensor integrating tungsten disulfide (WS2) flakes decorated with catechin-capped gold nanoparticles (AuNP-CT) and carbon black (CB) has been developed. The excellent antifouling properties of WS2 decorated with AuNP-CT into a high conductivity network of CB results in high selectivity, sensitivity, and reproducibility for the simultaneous determination of hydroxycinnamic acid (hCN) structural analogs: caffeic (CF), sinapic (SP), and p-coumaric acids (CM). Using differential pulse voltammetry (DPV), the target hCNs resulted in three well-resolved oxidation peaks at SPE-CB-WS2/AuNP-CT sensor. Excellent antifouling performance (RSD ip,a ≤ 3%, n = 15 for three analytes' simultaneous measure) and low detection limits (CF 0.10 µmol L-1; SP, 0.40 µmol L-1; CM, 0.40 µmol L-1) are obtained despite the analyzed compounds having a high passivation tendency towards carbon-based sensors. The SPE-CB-WS2/AuNP-CT sensor was successfully applied to determine CF, SP, and CM in food samples with good precision (RSD ≤ 4%, n = 3) and recoveries (86-109%; RSD ≤ 5%, n = 3). The proposed sensor is the first example exploiting the simultaneous determination of these compounds in food samples. Given its excellent electrochemical performance, low cost, disposability, and ease of use, this SPE-CB-WS2/AuNP-CT nanocomposite sensor represents a powerful candidate for the realization of electrochemical devices for the determination of (bio)compounds with high passivation tendency. Graphical abstract.