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NANOPARTICLES IN FOOD BIOSENSING J.M. Pingarrón* L. Agüí and P. Yáñez-Sedeño Department of Analytical Chemistry. Faculty of Chemistry University Complutense.

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Presentation on theme: "NANOPARTICLES IN FOOD BIOSENSING J.M. Pingarrón* L. Agüí and P. Yáñez-Sedeño Department of Analytical Chemistry. Faculty of Chemistry University Complutense."— Presentation transcript:

1 NANOPARTICLES IN FOOD BIOSENSING J.M. Pingarrón* L. Agüí and P. Yáñez-Sedeño Department of Analytical Chemistry. Faculty of Chemistry University Complutense of Madrid Madrid SPAIN NANOJASP’2010 Barcelona, December 2010

2 Preparation of nanostructured electrode surfaces Advances in sensors technology Development of several (bio)assay-transductor strategies Applications of nanotechnology Research line combining: - Wide range of approaches - Use or not of biological systems Products, processes and systems operating in nanometric magnitude

3 Improved charge transfer reactions Electrocatalytic ability: lower detection potential Antifouling capabilitySensitivitySelectivity Repeatability Advantages Nanostructured electrode surfaces

4 Electrochim. Acta., 53 (2008) 5848 ELECTROCHEMICAL BIOSENSORS BASED ON GOLD NANOPARTICLE-MODIFIED ELECTRODES Gold nanoparticles Ability to provide a stable surface for biomolecules immobilization retaining their biological activity Permit direct electron-transfer between redox proteins and bulk electrode materials no need for electron-transfer mediators High surface-to-volume ratio High surface energy Ability to decrease the distance between proteins and metal particles Functioning as electron- conducting pathways between the prosthetic groups and the electrode surface Permit direct electron-transfer between redox proteins and bulk electrode materials no need for electron-transfer mediators High surface-to-volume ratio High surface energy Ability to decrease the distance between proteins and metal particles Functioning as electron- conducting pathways between the prosthetic groups and the electrode surface Au Useful interfaces for electrocatalysis of redox processes of H 2 O 2 or NADH

5 ELECTROCHEMICAL BIOSENSORS FOR FOOD ANALYSIS DEVELOPMENT AND INNOVATION FOOD SAFETY FOOD QUALITY EFFICIENT TRACEABILITY SYSTEMS Development of detection, analysis and diagnosis methods  Rapid  Sensitive  Automated screening

6 AMPEROMETRIC BIOSENSOR FOR HYPOXANTHINE BASED ON IMMOBILIZED XOD ON NANOCRYSTAL GOLD-CARBON PASTE ELECTRODES Hypoxanthine (Hx) is formed as a product of nucleotide catabolism during the degradation processes in foodstuffs of animal origin. Hypoxanthine (Hx) is formed as a product of nucleotide catabolism during the degradation processes in foodstuffs of animal origin. Hx is accumulated mostly in the animal muscle and its levels are used as an index of fish and meat freshness in the food industry Hx is accumulated mostly in the animal muscle and its levels are used as an index of fish and meat freshness in the food industry XOD Hx + O 2 → X + H 2 O 2 XOD X + O 2 → Uric acid + H 2 O 2 Gold nanoparticle preparation: Electrodeposition from a HAuCl 4 solution on the bulk electrode material Determination of hypoxanthine based on the enzyme reaction catalyzed by XOD

7 LOD at 0.00 V: 2.2x10 -7 mol L -1 K m app = 18x10 -6 mol L -1 Useful lifetime = at least 15 days SEM of a GA-BSA-XOD-nAu-CPE biosensor Determination of hypoxanthine in sardines and chicken meat using the GA-BSA-XOD-nAu- CPE biosensor Sample Non-spiked1 2 3 Spiked1 3 2 Sardines Added (mg/100g)Found (mg/100g) Recovery (%) Chicken Added (mg/100g)Found (mg/100g) Recovery (%) Mean recoveries (  = 0.05):101 ± 8 % sardines102±3% chicken meat Sens. Actuators B. 113 (2006) 272 AMPEROMETRIC BIOSENSOR FOR HYPOXANTHINE BASED ON IMMOBILIZED XOD ON NANOCRYSTAL GOLD-CARBON PASTE ELECTRODES

8 Bienzyme amperometric biosensor using gold nanoparticles-modified electrodes for the determination of inulin in foods Anal. Biochem., 375 (2008)

9 (C 6 H 10 O 5 ) n (n=35) INULIN Prebiotic ingredient added to functional foods Vegetal origin: chicory root, artichoke DETERMINATION METHODS HPLC UV, RI, ED 1st enzyme biosensor 1st enzyme biosensor This work

10 INULIN Determination of interest in: - MONITORING OF PROCESSES - inuline extraction - fructose production - QUALITY CONTROL - diethetic and children’s foods - component of dietary fiber FOOD INDUSTRY - ECONOMIC AND LEGISLATIVE - added value for functional foods - ingredients establish prices inherent specificity simplicity rapidity real time analysis Biosensor advantages

11 AuECyst Au col TTF FDH Inulinase BIENZYME BIOSENSOR FOR INULIN 2e PQQ PQQH 2 FRUCTOSE 5-CETO-D-FRUCTOSE 2TTF 2TTF + INULIN INULIN Redox mediator E = +0.2 V PBS 0.05 M, pH 4.5 By adding sodium citrate to a boiling HAuCl4 aqueous solution HAuCl4/sodium citrate  Particle size  Gold nanoparticle preparation: By adding sodium citrate to a boiling HAuCl4 aqueous solution HAuCl4/sodium citrate  Particle size 

12 Stability More than 5 months Storage conditions: 0.05 M phosphate buffer, pH 4.5, a 4ºC s +3s i,  A time, days BIENZYME BIOSENSOR FOR INULIN

13 20 nA 100 s dextrose inulin additions glucoselactosemaltosesaccharose [INTERFERENT] / [INULIN] = 1 Interferences 10% E rel BIENZYME BIOSENSOR FOR INULIN

14 Sample Sample preparation by size exclusion SPE 2 μ A t, min Bio-Gel P-6 (Bio-Rad) fructose inulin BIENZYME BIOSENSOR FOR INULIN Chicory powder (18.5% inulin) 19.5 ± 0.4%, RSD = 2%, n = 6 Prebiotic food “Mas Vital”(2.0% inulin) 1.8 ± 0.1%, RSD = 5%, n = t, min i, nA

15 COLLOIDAL GOLD-CARBON NANOTUBES COMPOSITE ELECTROCHEMICAL BIOSENSORS Hybrid nanoparticles/nanotubes materials Biocompatible materials with important electroanalytical features Au coll -CNT-Teflon electrode Au coll -CNTs-Teflon CNTs-Teflon (30:70) graphite-Teflon (30:70) Slope values of the calibration plot over ( )x10 -3 M H 2 O 2, at E app =+0.5 V μA mM -1 ; 2.1 μA mM -1 ; 4.3 μA mM -1 Other advantages:Much lower noise level Rapidity J. Electroanal. Chem., 603 (2007) 1

16 COLLOIDAL GOLD-CARBON NANOTUBES COMPOSITE ELECTROCHEMICAL BIOSENSORS Analytical characteristics and kinetic parameters for glucose biosensors based on GOx–CNT electrodes BIOSENSOR E det, V Linear range, Slope, LOD, Useful K M app mM mA/M μM lifetime 30 1day vs Ag/AgClGOx-CNT-Teflon months – vs Ag/AgClGOx-Au coll -CNT-Teflon

17 Current, % Days 6 8 COLLOIDAL GOLD-CARBON NANOTUBES COMPOSITE ELECTROCHEMICAL BIOSENSORS GOx –Au coll – CNT-Teflon biosensor GOx – CNT-Teflon biosensor 1.0 x M glucose; E app =+0.5 V

18 Suitable electrode material for NADH detection Suitable for the preparation of dehydrogenase biosensors Au coll -CNT-Teflon electrocatalytic activity Enhanced electrode kinetics Au col CNT

19 NADH amperometric detection E, V i, µA Au coll -CNT-Teflon CNT-Teflon Au coll -graphite-Teflon graphite-Teflon enhanced currents at less positive potentials Au coll -CNT-Teflon

20 NADH amperometric detection Au coll -CNT-Teflon CNT-Teflon Au coll -graphite -Teflon graphite -Teflon 50 s 10  A 50 s 0.5  A 50 s 0.5  A E app. =+0.3 V; NADH, 1.0 x M Au coll -CNT-Teflon repeatability t 90% = s rapidity RSD = 3.7 % (n=10) NADH, 2.0 x M

21 Analytical characteristics ElectrodeE det, VLinear range, mM Slope, µA/mM LOD, µM Reference TB-CNT Lawrence, 2006 DHB-CNT-GCE Retna, 2006 CNT-epoxy+0.55hasta Pumera, 2006 CNT-Chit-GCE Tsai, 2007 PVA-CNT-GCE+0.6hasta Tsai, 2007a MB-CNT-GCE-0.1hasta Zhu, 2007 MB-Chit-CNT-GCE-0.14hasta Chakraborti, 2007 PDAB-CNT-GCE Zeng, 2007 CNT-sol-gel+0.3hasta Zhu, 2007a Au colll -CNT-Teflon This work CNT-Teflon This work NADH DETECTION Au coll -CNT-Teflon Redox mediator - the highest calibration plot slope value - low detection potential with no mediator

22 2e NAD + NADH CH 3 CH 2 OH CH 3 CHO ADH ADH-Au coll -CNT-Teflon Alcohol dehydrogenase biosensor based on a colloidal gold- carbon nanotubes composite electrode ETHANOL Electrochim. Acta, 53 (2008)

23 Analytical characteristics ADH-Au coll -CNT-Teflon ETHANOL DETERMINATION ElectrodEE ap, V Linear range, mM Slope, µA/mM LOD, µM Reference ADH-MB-CNT-CPE Santos, 2006 ADH-PVA-CNT-GCE+0.7up to Tsai, 2007 ADH-PDDA-CNT-GCE Liu, 2007 ADH- Au coll -CNT-Teflon This work ADH-CNT-Teflon This work Redox mediator higher slope value even with no mediator

24 APPLICATION ADH-Au colL -CNT-Teflon SAMPLE Reference material AO <1 5.5 Ethanol concentration, g / 100 ml * Found Declared Sample FREE Sample WITH * mean value + ts / √n (n = 3) RESULTS sample a)us stirring b)dilution CO 2 analytical solution

25 glucosinolates DETERMINATION OF GLUCOSINOLATE IN VEGETABLES Β-thioglucoside-N-hydroxysulfates Found in cabbage and broccoli Ingredient in functional foods Anticarcinogenic properties MYR/GOx-Au coll -CNT-Teflon 2 H 2 O 2 O 2 2e HO 2 O 2 H 2 O glucose GOx FAD FADH HO GOx FAD MYR Electroanalysis, 21 (2009) 1527

26 CONCLUSIONS Gold nanoparticles allows the construction of electrochemical biosensors exhibiting enhanced performances with respect to other designs The unique properties of gold nanoparticles concerning immobilization of biomolecules retaining their biological activity, and as efficient conducting interfaces with electrocatalytic ability makes them a powerful tool to modify electrode materials and to construct robust and sensitive biosensors. They can be powerful analytical tools to be applied to the food industry. Applications in this field comprise the whole food chain, from the primary production to the final distribution to the consumer, which implies an enormous potential of application to food traceability.

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