Fabrication and Characterization of Biomimetic Molecularly Imprinted Polymer for Paranitrophenol Detection

Abhijit Chandra Roy Roll No. 11202061

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Introduction to biosensors

Working Principle

Microelectr onics signal analysis

Major constraints of biosensor

Low stability High price Natural receptors and enzymes very sensitive to their environment. As for example antibodies

Molecular Imprinting Polymers a suitable answer

Types of molecular imprinting

Mainly two types
Covalent MI Due to the covalent bond formation the rebinding is slow This imprinting approach have been found suitable for only a number of compounds like alcohol, aldehydes, ketones, amines and carboxylic acids Non-covalent MI Non covalent imprinting is based on the non-covalent forces such as H-bonding, ion pairing and dipole-dipole interactions

Easier production, faster rebinding of the template, and greater variety of available functional monomers than the covalent approach Mechanism: Antibody like recognition capability
Wulff, G.; Sharhan, A.; Zabrocki, K. Tetrahedron Lett.1973, 14, 4329-4332.

Comparison of Natural and Artificial recognition elements

Natural biomolecules
Low stability

MIPs
Stable to low/high pH, pressure and
temperature (<180 C)

High price of enzymes and receptors

Inexpensive and easy to prepare

Generally poor performance in non-aqueous MIPs can work in organic solvents

media
Different natural biomolecules have their own Due to their the minimal design of operational MIP-based

operational requirements (pH, ionic strength, requirements, temperature, substrate etc.)

multisensors is a relatively easy task

Natural receptors and enzymes exist for only a In principle, MIPs can be prepared for
limited number of important analytes Poor compatibility with practically any compound are fully compatible with

micromachining Polymers

technology and miniaturization

micromachining technology

Polyaniline
Polyaniline is a conducting polymer Chemical structure of polyaniline
n
NH2
HCl Oxidation
NH NH
Cl
-

NH

NH
Cl
-

Main advantages Less expensive in processing Good electrical conductivity Mechanical stability It can be operated at lower applied voltages and temperatures It interacts more favorably with organic compounds Ability to form hydrogen bonding etc

Para-nitrophenol (PNP) as environmental pollutant

Para-nitrophenol (PNP) is one of the harmful toxic pesticides that cause rapid environment contamination It is a carcinogenic , mutagenic , cyto- embryotoxic to mammals PNP irritates eyes, skin, and respiratory tract and might lead to inflammation It exists in soil as well as in waste water
Rosenkranz, H. S et. al. Mutagenesis 1990, 5 (6), 525-527. Massey, I. J. et. at. Environ. Toxicol. Chem. 1994, 13(11), 17431752. Elia, M.C et al. Environ. Mol. Mutagen. 1994, 24 (3) 181191.

Molecularly imprinted polyaniline-polyvinyl sulphonic acid (PANI-PVSA)composite for PNP detection

Working principle

Electrochemical deposition characterization

1.2 1.1 1.0

Voltage (Volts)

0.9 0.8 0.7 0.6 0.5 0 50 100 150 200

(a) (b)
PANI PANI+PVSA PANI+PVSA+PNP

(c)

250

300

350

Time (Sec)
P. D. Gaikwad et al. Bull. Mater. Sci., Vol. 29, No. 4, August 2006

Contact angle measurements

Addition of PNP, the contact angle decreases from 57.4 to 27.1 and is due to increase in hydrophilicity of the polymer surface

Over oxidation and proper rinsing with deionized water of the Para-nitophenol imprinted PANI-PVSA polymer electrode shows increased contact angle of 55.30

Rebinding of PNP results change in contact angle from from 55.30 to 32.80
Contact angle pictures (a) PANI-PVSA/ITO, (b) PNP-PVSAPANI/ITO, (c) PI-PANI-PVSA/ITO and (d) ReI-PANIPVSA/ITO electrodes, respectively

UV studies
343 nm 418 nm
H O

+O

Intensity (a.u.)

The UV studies have been performed to confirm the formation of PNP imprinted PANI-PVSA/ITO film

(i)

(ii)
N+
O (i)

N
O

PNP in aqueous sol. PANI/ITO PANI-PVSA/ITO PNP-PANI-PVSA/ITO PNPI-PNP-PANI-PVSA/ITO

(ii)

838 nm

317 nm

(e) (d) (c) (b)

The incorporation of PNP results the increment of peak intensity at 418 nm and broadening of the peak near 800 nm occurs. The increase in the peak intensity and broadening of peak is due to the interaction of PNP in the polymer matrix.

430 nm

(a)

300

400

500

600

700

800

900

1000

Wavelength (nm)

The UV visible spectra of the electrochemically deposited films

M. Scully, M. C. Petty, A. P. Monkman, Synth. Met . 183 (1993) 5557.

1. 1527, 1314, 1246, 1208, 1121 and 812 cm-1represent the vibrational peaks of the C=C bond of PANI 2. The band seen at 1527cm-1 is attributed to C=C quinoid (Q) ring stretching vibrations 3. The band seen at 685 cm-1 corresponds to C-S stretching vibrations. The peaks at 1034 and 1111.87 cm-1 are related to SO2 symmetric stretching vibrations peak at 1295 cm-1 related to antisymmetric stretch vibrations 4. The vibrational band observed at 1382 cm-1 corresponds to N=O symmetric stretching of aromatic nitro compounds. The strong peak observed at 1517 cm-1 corresponds to the antisymmetric stretching vibration of NO2 group & indicate the incorporation of PNP inside the polyaniline-poly vinyl sulphonic A composite

FTIR studies
PANI-PVSA
250

PANI

PNP-PANI-PVSA

1295
(b)

Transmittance(T%)

200

685
1517
50

1527

(a) (c)

1382
0 500 1000 1500 2000 2500 3000 3500 4000

4500

-1 Wavelength(cm )(cm-1) Wave number

Figure 3 FTIR spectra of (a) PANI/ITO, (b) PANIPVSA/ITO, (c) PANI-PVSA-PNP/ITO, electrodes
C. Dhand, S. K. Arya, S. P. Singh, B. P. Singh, M. Datta, B. D. Malhotra, Carbon 46 (2008) 1727.

SEM studies
(a)
200 nm

(b) b)

200 nm

200 nm

Hindrance

Scanning Electron Micrographs of (a) PANI-PVSA/ITO( b)PNP-PANI-PVSA/ITO

Electrochemical studies
The PANI-PVSA/ITO electrode shows anodic and cathodic peaks at +0.06V and -0.5V. After the incorporation of PNP in the polymer matrix the anodic and cathodic peaks are shifted to higher voltage (0.07V &0.44V). This shifting of peaks may be attributed to the interaction of PNP in the polymer matrix.
The peak current ratio [Ipa / Ipc] have been calculated and shows the value in the order PNP-PANI-PVSA/ITO (0.896) > PANI-PVSA/ITO (0.892)> PANI-/ITO (0.690) and indicating uniform facile charge transfer kinetics
0.0002 PANI PANI+PVSA+PNP PANI+PVSA

(c) (b) (a)

0.0001

Current (A)

0.0000

-0.0001

-0.0002

-0.0003 -0.8 -0.6 -0.4 -0.2 0.0 0.2 0.4 0.6 0.8

Voltages (Volts)
Cyclic voltametric curve of (a) PANI/ITO electrode, (b) PANI-PVSA/ITO electrode, (c) PANI-PVSAPNP/ITO, electrodes

DPV studies
3.5x10
-5

Significant enhancement in the magnitude of amperometric peak current of the PNP- PANI-PVSA/ITO electrode with respect to that of the NIP- PANIPVSA/ITO (NIPnon imprinted polymer) electrode

3.0x10

-5

(b)
Current (A)

2.5x10

-5

2.0x10

-5

(a)
1.5x10
-5

1.0x10

-5

-0.4

-0.2

0.0

0.2

0.4

0.6

Voltage (mV)

Differential pulse voltametric curves PVSA/ITO and PNP- PANI-PVSA/ITO

NIP-

PANI-

P H study
P -6
H

P -7
1.14 1.12

P - 7.4

P -8

The amperometric response measurement of the MIP electrode in the PH range of 6 to 8 reveal that this molecularly imprinted PANI electrode shows excellent amperometric response at neutral PH

0.00012

Current (Amp)

0.00010
Current (Amp)

1.10 1.08 1.06 1.04 1.02 1.00 0.98

0.00008

0.00006

0.96 6.0 6.5 7.0 Voltage (Volt) 7.5 8.0

0.00004 0.0 0.2 0.4 0.6 0.8 1.0 1.2 Voltage (Volt)

Influence of PH on the molecularly imprinted PIPANI-PVSA/ITOelectrode has been examined with different pH buffer solutions

Sensing studies
This PNP selective molecularly imprinted PANI-PVSA electrode shows low detection limit of 0.001 mM having value of sensitivity of 1.5x10-3 mM with correlation coefficient of 0.9961. It shows good reusability and excellent fabrication stability.
0.000045 0.000040 0.000035

Current (A)

0 mM 0.012mM 0.024mM 0.036mM 0.048mM

41 40 39 38

Current

37 36 35 34 33 32 0.00 0.01 0.02 0.03 0.04 0.05

0.000030 0.000025 0.000020 0.000015 0.000010 0.000005 -0.2 0.0 0.2

PNP Concenration (mM)

0.4

0.6

0.8

Voltage (V)
Effect of concentration of PNP on the DPV response of PIPANI-PVSA/ITO electrodes

Conclusion
Artificial complimentary molecular site based electrochemical sensor is very simple, cost effective and easily doable The polymer composite exhibit increase electrical conductivity, this result not only helps to fabricate our sensor, it may give a variety of technological applications It has been shown that this PNP selective molecularly imprinted PANI-PVSA electrode has detection limit of ~0.001 mM of PNP, a sensitivity of 1.5x10-3 AmM-1

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