nanoparticles in food biosensing
DESCRIPTION
NANOPARTICLES IN FOOD BIOSENSING. J.M. Pingarrón* L. Agüí and P. Yáñez-Sedeño Department of Analytical Chemistry. Faculty of Chemistry University Complutense of Madrid 28040-Madrid SPAIN [email protected]. NANOJASP’2010 Barcelona, December 2010. - PowerPoint PPT PresentationTRANSCRIPT
NANOPARTICLES IN FOOD BIOSENSING
J.M. Pingarrón* L. Agüí and P. Yáñez-SedeñoDepartment of Analytical Chemistry. Faculty of Chemistry
University Complutense of Madrid28040-Madrid SPAIN
NANOJASP’2010
Barcelona, December 2010
Preparation of nanostructured electrode surfacesPreparation of nanostructured electrode surfacesPreparation of nanostructured electrode surfacesPreparation of nanostructured electrode surfaces
Advances in sensors technology
Development of several (bio)assay-transductor strategies
Applications of nanotechnology
Research line combining:Research line combining:
- Wide range of approaches- Use or not of biological systems
Products, processes and systems operating in nanometric magnitude
Improved charge transfer reactions
Electrocatalytic ability: lower detection potential
Antifouling capability
SensitivitySensitivity
SelectivitySelectivity
RepeatabilityRepeatability
AdvantagesAdvantages
Nanostructured electrode surfacesNanostructured electrode surfacesNanostructured electrode surfacesNanostructured electrode surfaces
Electrochim. Acta., 53 (2008) 5848
ELECTROCHEMICAL BIOSENSORS BASED ON GOLD NANOPARTICLE-MODIFIED ELECTRODES
Gold nanoparticlesGold nanoparticles
Ability to provide a stable surface for biomolecules immobilization retaining their biological activity
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
Au
AuUseful interfaces for electrocatalysis of redox processes of H2O2 or NADHUseful interfaces for electrocatalysis of redox processes of H2O2 or NADH
ELECTROCHEMICAL BIOSENSORS FOR FOOD ANALYSIS
DEVELOPMENT AND INNOVATIONDEVELOPMENT AND INNOVATION
FOOD SAFETYFOOD SAFETY FOOD QUALITYFOOD QUALITY
EFFICIENT TRACEABILITY SYSTEMSEFFICIENT TRACEABILITY SYSTEMS
Development of detection, analysis and diagnosis Development of detection, analysis and diagnosis
methodsmethods
RapidRapid
SensitiveSensitive
Automated screening Automated screening
AMPEROMETRIC BIOSENSOR FOR HYPOXANTHINE BASED ON IMMOBILIZED XOD ON NANOCRYSTAL GOLD-CARBON PASTE
ELECTRODES
• Hypoxanthine (Hx) is formed as a product of nucleotide Hypoxanthine (Hx) is formed as a product of nucleotide catabolism during the degradation processes in foodstuffs of catabolism during the degradation processes in foodstuffs of animal origin.animal origin.• Hx is accumulated mostly in the animal muscle and its levels Hx is accumulated mostly in the animal muscle and its levels are used as an index of fish and meat freshness in the food are used as an index of fish and meat freshness in the food industryindustry
XODHx + O2 → X + H2O2
XODX + O2 → Uric acid + H2O2
Gold nanoparticle preparation: Electrodeposition from a HAuCl4 solution on the bulk electrode material
Determination of hypoxanthine based on the enzyme reaction catalyzed by XOD
LOD at 0.00 V: 2.2x10LOD at 0.00 V: 2.2x10-7-7 mol L mol L-1-1
KKmmapp app = 18x10= 18x10-6-6 mol L mol L-1-1
Useful lifetime = at least 15 daysUseful 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-Determination of hypoxanthine in sardines and chicken meat using the GA-BSA-XOD-nAu-CPE biosensorCPE biosensor
Sample
Non-spiked 1
2
3
Spiked 1
3
2
Sardines
Added (mg/100g) Found (mg/100g) Recovery (%)
Chicken
Added (mg/100g) Found (mg/100g) Recovery (%)
- 145 -
- 138 -- 152 -
- 225 105
69.6 213 103
- 210 95
- 91.7 -
- 85.0 -- 94.7 -
- 157.2 103
60.7 145.1 99
- 160.2 103
Mean recoveries (Mean recoveries ( = 0.05): = 0.05): 101 101 ± 8 % sardines± 8 % sardines 102±3% chicken meat102±3% chicken meat
Sens. Actuators B. 113 (2006) 272
AMPEROMETRIC BIOSENSOR FOR HYPOXANTHINE BASED ON IMMOBILIZED XOD ON NANOCRYSTAL GOLD-CARBON PASTE
ELECTRODES
Bienzyme amperometric biosensor using gold nanoparticles-modified electrodes
for the determination of inulin in foods
Anal. Biochem., 375 (2008) 345-353
(C6 H10 O5 )n (n=35)
INULIN
Prebiotic ingredient added to functional foodsPrebiotic ingredient added to functional foods
Vegetal origin: chicory root, artichoke
O
H
HO
H
HO
H
O
OHHH
OH
CH2
OH
OH
OHO
O
CH2OH
CH2OH
OH
OH
O
n
DETERMINATION METHODS
HPLC UV, RI, ED
1st enzymebiosensor
1st enzymebiosensor
This work
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 advantagesBiosensor advantages
AuEAuE CystCyst AuAucolcol TTFTTF FDHFDH InulinaseInulinase
BIENZYME BIOSENSOR FOR INULIN
2e2e
PQQPQQ
PQQHPQQH22
FRUCTOSEFRUCTOSE
5-CETO-D-FRUCTOSE5-CETO-D-FRUCTOSE
2TTF2TTF
2TTF2TTF++
INULININULIN
Redox mediator
E = +0.2 VPBS 0.05 M, pH 4.5
Gold nanoparticle preparation: By adding sodium citrate to a boiling HAuCl4 By adding sodium citrate to a boiling HAuCl4 aqueous solution HAuCl4/sodium citrate aqueous solution HAuCl4/sodium citrate Particle size Particle size
Stability
More than 5 monthsStorage conditions:
0.05 M phosphate buffer, pH 4.5, a 4ºC
0 20 40 60 80 100 120 140 1600.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18
0.20
0.22
-3s
+3s
i, A
time, days
BIENZYME BIOSENSOR FOR INULIN
20 nA
100 s
dextrose
inulin additions
glucose lactose maltose saccharose
[INTERFERENT] / [INULIN] = 1[INTERFERENT] / [INULIN] = 1
Interferences
10% Erel
BIENZYME BIOSENSOR FOR INULIN
Sample
Sample preparation by size exclusion SPE
2 μA
t, min
Bio-Gel P-6 (Bio-Rad)
fructosefructose
inulininulin
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 = 6
0
3
6
9
12
15
0 2 4 6 8 10 12 14 16 18 20
t, min
i, nA
COLLOIDAL GOLD-CARBON NANOTUBES COMPOSITE ELECTROCHEMICAL BIOSENSORS
Hybrid nanoparticles/nanotubes materialsBiocompatible materials with important electroanalytical features
Aucoll-CNT-Teflon electrodeAucoll-CNT-Teflon electrode
0 200 400 600 800 1000-20
4
8
12
16
i, A
E, mV
Aucoll-CNTs-Teflon
CNTs-Teflon (30:70)
graphite-Teflon (30:70)
Slope values of the calibration plot over (1.0-5.0)x10-3 M H2O2, at Eapp=+0.5 V
0.0083μA 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
COLLOIDAL GOLD-CARBON NANOTUBES COMPOSITE ELECTROCHEMICAL BIOSENSORS
Analytical characteristics and kinetic parameters for glucose biosensors based on GOx–CNT electrodes
BIOSENSOR Edet, V Linear range, Slope, LOD, Useful KMapp
mM mA/M μM lifetime
30 1day 1.2 33 0.1-8 +0.5 vs Ag/AgClGOx-CNT-Teflon
14..9 3 months 2.6 17 0.05 – 1 +0.5 vs Ag/AgClGOx-Aucoll-CNT-Teflon
2 40
25
50
75
100
Current, %
Days
6 8
COLLOIDAL GOLD-CARBON NANOTUBES COMPOSITE ELECTROCHEMICAL BIOSENSORS
GOx –Aucoll – CNT-Teflon biosensor
GOx – CNT-Teflon biosensor
1.0 x 10-3 M glucose; Eapp=+0.5 V
Suitable electrode material for NADH detection
Suitable for the preparation of dehydrogenase biosensors
Aucoll-CNT-Teflon
electrocatalytic activityEnhanced electrode kinetics
Aucol CNT
NADH amperometric detection
0 0.2 0.4 0.6 0.8 E, V
50
40
30
20
10
i, µA
Aucoll-CNT-Teflon
CNT-Teflon
Aucoll-graphite-Teflon
graphite-Teflon
enhanced currents at less positive potentials
Aucoll-CNT-Teflon
NADH amperometric detection
Aucoll-CNT-Teflon
CNT-Teflon
Aucoll-graphite -Teflon
graphite -Teflon
50 s
10A
50 s
0.5 A
50 s
0.5 A
Eapp.=+0.3 V; NADH, 1.0 x 10-4M
Aucoll-CNT-Teflon
repeatability
t90%= 10-15 s
rapidity
RSD = 3.7 %(n=10)
NADH, 2.0 x 10-4 M
Analytical characteristics
Electrode Edet, V Linear range, mM
Slope, µA/mM
LOD, µM
Reference
TB-CNT +0.021 - 24 15 Lawrence, 2006
DHB-CNT-GCE -0.05 0.005-0.4 1.66 0.1 Retna, 2006
CNT-epoxy +0.55 hasta 1.0 29.7 - Pumera, 2006
CNT-Chit-GCE +0.6 0.005-0.8 8.7 0.5 Tsai, 2007
PVA-CNT-GCE +0.6 hasta 2.0 5.88 20 Tsai, 2007a
MB-CNT-GCE -0.1 hasta 0.5 0.52 4.8 Zhu, 2007
MB-Chit-CNT-GCE -0.14 hasta 0.08 5.9 0.5 Chakraborti, 2007
PDAB-CNT-GCE +0.07 0.002- 4.0 - 0.5 Zeng, 2007
CNT-sol-gel +0.3 hasta 0.65 2.31 12.4 Zhu, 2007a
Aucolll-CNT-Teflon +0.3 0.010-1.0 37.7 3.0 This work
CNT-Teflon +0.3 0.010-1.0 17.3 - This work
NADH DETECTION
Aucoll-CNT-Teflon
Redox mediator
- the highest calibration plot slope value- low detection potential with no mediator
2e
NAD+
NADH
CH3CH2OH
CH3CHO
ADH
ADH-Aucoll-CNT-Teflon
Alcohol dehydrogenase biosensor based on a colloidal gold-carbon nanotubes composite electrode
Alcohol dehydrogenase biosensor based on a colloidal gold-carbon nanotubes composite electrode
ETHANOL
Electrochim. Acta, 53 (2008) 4007-4012
Analytical characteristics ADH-Aucoll-CNT-Teflon
ETHANOL DETERMINATION
ElectrodE Eap, VLinearrange,
mM
Slope, µA/mM
LOD, µM
Reference
ADH-MB-CNT-CPE 0.0 0.05 - 10.0 0.597 5Santos,
2006
ADH-PVA-CNT-GCE +0.7 up to 1.5 0.196 13 Tsai, 2007
ADH-PDDA-CNT-GCE +0.1 0.5-5.0 - 90 Liu, 2007
ADH- Aucoll -CNT-Teflon +0.3 0.02-1.0 2.27 4.7 This work
ADH-CNT-Teflon +0.3 0.10-10.0 1.8 32 This work
Redox mediator
higher slope value even with no mediator
APPLICATION ADH-AucolL-CNT-Teflon
SAMPLEReference material AO6191
4.8 + 0.4 1.0 + 0.1 5.5 +0.3
4.3 + 0.6 <1 5.5
Ethanol concentration, g / 100 ml* Ethanol concentration, g / 100 ml*
Found
Declared
Sample FREE Sample WITH
* mean value + ts / √n (n = 3)
RESULTSsample
a)us stirringb)dilution
CO2
analyticalsolution
glucosinolates
DETERMINATION OF GLUCOSINOLATE DETERMINATION OF GLUCOSINOLATE IN VEGETABLESIN VEGETABLES
Β-thioglucoside-N-hydroxysulfatesFound in cabbage and broccoliIngredient in functional foodsAnticarcinogenic properties
MYR/GOx-Aucoll-CNT-Teflon
2
H2O
2
O2
2eH O
2
O2
H2O
glucose
GOxFAD
FADH
H O
GOxFAD
MYR
Electroanalysis, 21 (2009) 1527
CONCLUSIONS
Gold nanoparticles allows the construction of electrochemical biosensors exhibiting enhanced performances with respect to other designs
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.
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.
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.