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Studies on the Biosensor for Detection of Methyl Parathion
Chouhan, Raghuraj Singh (2010) Studies on the Biosensor for Detection of Methyl Parathion. PhD thesis, University of Mysore.
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Abstract
Methyl Parathion (MP), an organophosphorus pesticide holds concern due to its extensive use in agricultural practices. MP being highly toxic at trace levels to human beings requires detection at high sensitivity, which is a challenging task. The sensitive detection of MP by immunosensor methods have been delineated in this thesis. Initial approach was emphasized on the synthesis of MP hapten. Synthesized MP hapten was conjugated to a carrier protein molecule and immunized in chicken to produce anti-MP IgY antibodies. Antibodies were purified in bulk using affinity column chromatographic technique. Further, antibodies were tested for cross reactivity with structurally related OP pesticides. MP-HRP conjugate, which have been used as a labeled marker for immunosensor applications, was stabilized with 0.1% BSA. Furthermore, chemiluminescence (CL) parameters were standardized for MP-HRP conjugate by different biochemical methods. Immunosensor detection of MP requires efficient immobilization of antibodies and dissociation of MP from the immuno-reactor column. Study revealed that Gly-HCl (pH-2.4) in combination with 1% methanol was the best dissociating buffer for reusability of the antibody immobilized in immuno-reactor column. The CL and charge coupled device (CCD) based method of detections were standardized for the sensitive detection of MP in water and different fruit beverages. With CL techniques the detection limit was 0.005 ng/mL and with CCD it was 0.01 ng/mL. Quantum dot based MP detection showed the credibility of the method to detect at 0.1 ng/mL. The visual colloidal gold based dipstick technique was also employed for the qualitative detection of MP and detection limit was 0.1 ng/mL. Validation of all these methods over 22 existing ELISA was carried out in present investigation. The results indicated the superiority of developed immuno-sensors against conventional techniques for the sensitive detection of MP in water and fruit beverages.
| Item Type: | Thesis (PhD) |
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| Uncontrolled Keywords: | Methyl Parathion (MP), organophosphorus pesticide, immuno-sensors |
| Subjects: | 500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 26 Pesticide Chemistry 600 Technology > 05 Chemical engineering > 01 Biotechnology and Bioengineering |
| Divisions: | Fermentation Technology and Bioengineering |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 15 May 2012 06:25 |
| Last Modified: | 15 May 2012 06:25 |
| URI: | http://ir.cftri.res.in/id/eprint/10752 |
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