Principles of Electronic Nanobiosensors

Unit 3: Sensitivity
Lecture 3.7: Amperometric Sensors:
Glucose Sensors I

By Muhammad A. Alam
Professor of Electrical and Computer Engineering
Purdue University
alam@purdue.edu

A. Shakouri nanoHUB-U Fall 2013 19

1
 Three types of sensors
Potentiometric Amperometric Cantilever
Ref. & Aux.
Fluid Gate Electrode
Gate

Charge to current Chemical to current Mass to frequency

2
 A short history of sensors

Virus/bacteria Protein/DNA PCR

pH-meter Glucose Genome

(potentiometric) (amperometric) sequencer

Vacuum tube MOS IC

3
3
A glucose sensor

Counter/
Ref. Electrode

Working
electrode

Enzyme coating

Alam, Principles of Nanobiosensors, 2013 4

 Basics of a amperometric sensor

Glucose
H2O2
Ag/AgCl

O2
Pt O2
Gluconic acid
2e 2e

2H+

enzyme:
Glucose oxidase
+

350 mV 5
Alam, Principles of Nanobiosensors, 2013
 Glucose sensing

(1) C6 H12O6 + O2 

→   C6 H12O7 + H 2O2
GOx

enzyme:
Gluconic acid
Glucose oxidase

( 2 ) H 2O2 
→ O2 + 2 H
Pt Anode( 350 mV ) +
+ 2e −

( 3) AgCl + e −
→ Ag + Cl
Ag Ref. 0 −

( 4 ) H
4 +
+ O2 
→ 2H 2O2 − 4e
Aux Re f . −

Alam, Principles of Nanobiosensors, 2013 6

 Why amperometric sensors?
• Obviates the problem of screening
• Electro-chemical reactions provide specificity
• A broad range of modern point-of-care sensors
belong to this group (glucose sensor, genome
sequencer)

• Still constrained by the limits of settling time

• Multiple-electrode setup difficult to miniaturize
• New diffusion limits related to reaction products
• Parasitic reactions may compromise selectivity

Alam, Principles of Nanobiosensors, 2013 7

 Outline

• Introduction to amperometric biosensing

– Spontaneous redox reaction vs. electrolysis
– Importance of reference electrodes
– Basic H2O2 based scheme for Glucose
sensing
• Conclusion
• Appendix: Butler-Volmer equation

Alam, Principles of Nanobiosensors, 2013 8

 Let us start with the cell on the left

Glucose
H2O2
Ag/AgCl

O2
Pt O2
2e Gluconic acid
2e

2H+

350 mV 9
Alam, Principles of Nanobiosensors, 2013
 Spontaneous reaction
Zn → Zn 2+ + 2e − (oxidation at anode)
Cu 2+ + 2e − → Cu (reduction in solution)
Zn( s ) + Cu 2+ ( aq ) → Zn 2+ ( aq ) + Cu

Zn

Heat

CuS04

Alam, Principles of Nanobiosensors, 2013 10

Spontaneous reaction and the driving force

Zn → Zn 2+ + 2e −
Zn

Oxidation Potential
0.763

Heat

-0.337
Cu2S04
Cu 2+ + 2e − → Cu

Potential difference drives electron transfer

Alam, Principles of Nanobiosensors, 2013 11
 Driving a motor by chemical energy
Zn( s ) + Cu 2+ ( aq ) → Zn 2+ ( aq ) + Cu

Zn → Zn 2+ + 2e − (oxidation at anode)
Cu 2+ + 2e − → Cu (reduction in cathode)

Zn 2e Cu

2 2e

Salt bridge

Alam, Principles of Nanobiosensors, 2013 12

 Forced oxidation-reduction

Oxidation Potential
Oxidation Potential

Oxidation Potential

difficult redox New molecule Biased

Alam, Principles of Nanobiosensors, 2013 13

Electrolysis: forced oxidation-reduction
350 mV
Pt 2e + Ag

H 2O2 → 2 H + + O2 + 2e − (oxidation at anode)

Alam, Principles of Nanobiosensors, 2013 14

 Outline

• Introduction to amperometric biosensing

– Spontaneous redox reaction vs. electrolysis
– Importance of reference electrodes
– Basic H2O2 based scheme for Glucose
sensing
• Conclusion
• Appendix: Butler-Volmer equation

Alam, Principles of Nanobiosensors, 2013 15

 The issue of reference electrode
Ammeter

Voltmeter
Ag/AgCl
Reference
Electrode Working Electrode

We wish to control the

… but to do that, we need a potential of this working
second electrode in the electrode…
solution to complete the circuit

Ref – Notes by Andrea Mardegan

16
Without a Reference Electrode …

NHE:
Normal Hydrogen electrode

Voltage does not divide symmetrically

17
 The purpose of reference electrode

Ref – Notes by Andrea Mardegan

18
A three electrode cell

Ref – Notes by Andrea Mardegan 19

 Outline

• Introduction to amperometric biosensing

– Spontaneous redox reaction vs. electrolysis
– Importance of reference electrodes
– Basic H2O2 based scheme for Glucose
sensing
• Conclusion
• Appendix: Butler-Volmer equation

Alam, Principles of Nanobiosensors, 2013 20

Current proportional to H2O2 concentration

R
O

−
R  → O + ne
kf
kb

I (
Ae q k f ρ H 2O2 − k=
b ρ O2 )
I Ae q k f ρ s ,R − kb ρ s ,O ( )
Butler-Volmer equation
Alam, Principles of Nanobiosensors, 2013 21
Validation of H2O2 response

I ∝ Ae I∝ρ
Alam, Principles of Nanobiosensors, 2013 22
 Conclusion

• A amperometric sensor responds linearly to

the reactants to be measured and
exponentially to the applied voltage.
• Three electrode configuration is ideal, but may
not be practical.
• A broad range of biologically relevant products
may be measured by amperometric sensors.

Alam, Principles of Nanobiosensors, 2013 23

Appendix: Derivation of Butler-Volmer Equation

−
R  → O + ne
kf
kb

Oxidation Potential
(1−α ) nf(E − E0 )
k f = k0, f e f ≡ F RT
−α n f(E − E0 )
kb = k0,b e

⇒I (
qA k0, f ρ s , R e(1−α ) nf(E − E0 ) − k0,b ρ s ,O e −α n f(E − E0 ) )
Alam, Principles of Nanobiosensors, 2013 24
Appendix: Derivation of Butler-Volmer
Equation
I (
qA k0, f ρ s , R e(1−α ) nf(E − E0 ) − k0,b ρ s ,O e −α n f(E − E0 ) )
Consider, the special case in which the
interface is at equilibrium with a
solution in which ρ O = ρ R
* *
R

In that situation, E = E0 O

k0, f ρ O = k0,b ρ R
* *

Alam, Principles of Nanobiosensors, 2013 25

Appendix: Derivation of Butler-Volmer
Equation
⇒ k0, f =
k0,b
⇒I (
qA k0, f ρ s , R e(1−α ) nf(E − E0 ) − k0,b ρ s ,O e −α n f(E − E0 ) )
In general, if k0, f ≠ k0,b , we can define a
equilibrium potential such that
At E = E eq , I = 0
k BT  k0,b ρ s ,O  f ≡ F RT
⇒ Eeq =E0 + ln  
nq  k0, f ρ s ,R 

Alam, Principles of Nanobiosensors, 2013 26

 Appendix: Derivation of Butler-Volmer
Equation
Let η= E − Eeq
qA ( k0, f ρ s , R ) ( k0,b ρ s ,O ) ( )
α 1−α (1−α ) nf η −α n f η
⇒I e − e
= io ( e(1−α ) nf η − e −α n f η )
qA ( k0, f ρ s , R ) (k ρ s ,O )
α 1−α
= =
io qAk o 0,b

Alam, Principles of Nanobiosensors, 2013 27

 References
• Dill, Ken A., and Sarina Bromberg. Molecular driving forces:
statistical thermodynamics in chemistry and biology. Taylor &
Francis, 2003.
• Brown, Theodore Lawrence. Chemistry: the central science.
Pearson Education, 2009.Van Hal, R. E. G., J. C. T. Eijkel, and P.
Bergveld.
• Janata, Jiˇí. "Introduction to Sensors." Principles of Chemical
Sensors. Springer US, 2009. 1-11.
• S. M. Sze, Semiconductor Sensors, Wiley.
• Zhang, Xueji, Huangxian Ju, and Joseph Wang,
eds. Electrochemical sensors, biosensors and their biomedical
applications. Access Online via Elsevier, 2011.

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