OWLS APPLICATION NOTES NO-002

LABEL-FREE IMMUNOSENSOR FOR HERBICIDE TRIFLURALIN DETECTION
using OPTICAL WAVEGUIDE LIGHTMODE SPECTROSCOPY (OWLS) detection

Abstract
Acting as a potent microtubule inhibitor, the dinitroaniline herbicide trifluralin has been used in 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 labelfree method to develop immobilized antigen-BSA conjugate based competitive immunosensor.

CH3 CH2 H2C N O2N

CH2 CH2 CH3 NO2

CF3

Application of OWLS sensors
as competitive immunosensor for the detection of trifluralin
polyclonal serum + sample 1:1 adding mixture, incubation

Standard inhibition curve
of the immobilized antigen conjugate based competitive trifluralin immunosensor
21

Sensor response (a.u.)

19 17 15 13 11 1E-10 1E-08 1E-06 1E-04 1E-02 1E+00

antigen-BSA conjugate antigen poly- or monoclonal antibody

Surface Chemistry of OWLS sensors
• Amino functionalisation of waveguide surface by (γ-aminopropyl)triethoxysilane Immobilisation of trifluralin-BSA conjugate on the OW LS sensor surface.

Trifluralin concentration (ng/ml)

Cross-reactivity
of dinitroaniline herbicides, haptens and various synthetic i nterm edi ates in t he com petitive OW LS immunosensors: Compound Trifluralin Ethalfluralin Benfluralin Isopropalin 2,6-Dinitro-4-trifluoromethylaniline Pendimethalin 2,6-Dinitro-4-trifluoromethyl-1chlorobenzene 2,6-Dinitroaniline Aniline CR (%) (OWLS sensor) 100 6,9 3,4 0,26 0,09 >0,01 >>0,01 >>0,01 >>0,01

Sensitization of the sensor surface with trifluralin-BSA conjugate

280
Sensor response (a. u.)
trifl-BSA

1000x Ab GA

2000x Ab

230 180 130 80 30 -20 15

TRIS d.w.

Conclusion
55
Time (min)

35

75

95

Optimization of antiserum dilution
Sensor response for different polyclonal serum d i l u t i o n s wa s i n v e s t i g a t e d .

A competitive or binding inhibition (immobilized antigen-bas ed) OW LS immunosensor was developed 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 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 (1:2000) and offered a standard curve for trifluralin in the analyte concentration range of 2 x 10-7 to 3 x 10-5 ngml-1 in water.

Sensor response (a.u.) xx

70 60 50 40 30 20 10 0 0

References
500x 1000x 2000x 4000x 8000x

1. Vörös, J. J. Ramsden, G. Csucs, I. Szendrõ, S.M. De Paul,
M. Textor, N. D. Spencer “Optical Grating Coupler Biosensors” Biomaterials 23 (2002) 3699-3710

2. Székács A., Trummer N., Adányi N., Váradi M., Szendrı I.
2 4 6 8 10

Time (min)

“Development of a non-labeled immunosensor for the herbicide trifluralin via optical waveguide lightmode spectroscopic detection”, Analytica Chimica Acta 487, 2003, 31-42

3. www.owls-sensors.com

MicroVacuum Ltd. H-1147, Kerékgyártó u. 10., Budapest, Hungary
Web: http://www.owls-sensors.com/; E-mail: info@owls-sensors.com

Phone: + 36 1 252 1991; +36 1 467 0108

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