OWLS APPLICATION NOTES NO-002

LABEL-FREE IMMUNOSENSOR
FOR HERBICIDE TRIFLURALIN DETECTION
using OPTICAL WAVEGUIDE LIGHTMODE SPECTROSCOPY (OWLS) detection
Abstract CH3 CH2 H2C CH2 CH2 CH3
N
Acting as a potent microtubule inhibitor, the dinitroaniline herbicide trifluralin has been used in
O2N NO2
agricultural applications as a selective, pre-emergence herbicide in various plant cultures
including grain, crops, vegetables, fruits and nuts. In recent years, the compound is a subject to
increasing toxicological and environmental concerns. OWLS offered a highly sensitive label-
free method to develop immobilized antigen-BSA conjugate based competitive immunosensor. CF3

Application of OWLS sensors Standard inhibition curve

as competitive immunosensor for the detection of of the immobilized antigen conjugate based
trifluralin competitive trifluralin immunosensor

polyclonal serum + sample 21

Sensor response (a.u.)

1:1
adding mixture, 19
incubation
17

15

antigen-BSA conjugate 13
antigen
poly- or monoclonal antibody
11
1E-10 1E-08 1E-06 1E-04 1E-02 1E+00

Trifluralin concentration (ng/ml)

Surface Chemistry of OWLS sensors
• Amino functionalisation of waveguide surface
by (γ-aminopropyl)triethoxysilane
• Immobilisation of trifluralin-BSA conjugate on Cross-reactivity
the OW LS sensor surface. of dinitroaniline herbicides, haptens and various
synthetic i nterm edi ates in t he com petitive OW LS
Sensitization of the sensor surface with immunosensors:
trifluralin-BSA conjugate Compound CR (%) (OWLS sensor)
Trifluralin 100
Ethalfluralin 6,9
Benfluralin 3,4
280 1000x Ab 2000x Ab
Isopropalin 0,26
trifl-BSA
2,6-Dinitro-4-trifluoromethylaniline 0,09
Sensor response (a. u.)

230 GA
Pendimethalin >0,01
180 2,6-Dinitro-4-trifluoromethyl-1-
>>0,01
130 chlorobenzene
2,6-Dinitroaniline >>0,01
80 Aniline >>0,01
TRIS
30 d.w.
Conclusion
-20 A competitive or binding inhibition (immobilized
15 35 55 75 95
antigen-bas ed) OW LS immunosensor was developed
Time (min)
and optimized for the detection of trifluralin. The
OW LS sensor was opti m ized for m ajor m ethod
parameters, such as sensitizing antigen
Optimization of antiserum dilution c o n c e n t r a t i o n ( 1 0 µg m l - 1 ) a n d a n t i s e r u m d i l u t i o n
Sensor response for different polyclonal serum (1:2000) and offered a standard curve for trifluralin
in the analyte concentration range of 2 x 10-7 to 3 x
d i l u t i o n s wa s i n v e s t i g a t e d .
10-5 ngml-1 in water.

70
References
Sensor response (a.u.) xx

500x
60
1000x 1. Vörös, J. J. Ramsden, G. Csucs, I. Szendrõ, S.M. De Paul,
50
2000x M. Textor, N. D. Spencer “Optical Grating Coupler
40 4000x
Biosensors”
30 8000x
Biomaterials 23 (2002) 3699-3710
20
2. Székács A., Trummer N., Adányi N., Váradi M., Szendrı I.
10 “Development of a non-labeled immunosensor for the
0 herbicide trifluralin via optical waveguide lightmode
0 2 4 6 8 10 spectroscopic detection”, Analytica Chimica Acta 487, 2003,
Time (min) 31-42

3. www.owls-sensors.com

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