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Method for detecting content of fenthion

A technology for fenthion content, which is applied in the field of detection of fenthion content, can solve the problems that EMIT sensitivity assessment has not yet revealed

Active Publication Date: 2008-04-02
MYONGJI UNIV IND & ACAD COOPERATION FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the assessment of bridging group structure on EMIT sensitivity has not yet been revealed

Method used

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  • Method for detecting content of fenthion
  • Method for detecting content of fenthion
  • Method for detecting content of fenthion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of G6PDH-hapten conjugates

[0019] (1) Chemicals

[0020] Glucose-6-phosphate dehydrogenase (G6PDH), glucose-6-phosphate (G6P), β-nicotinamide adenine dinucleotide (NAD) from Leucinostoc mensenteroiders + ), oxaloacetic acid (OAA), and reduced NAD + Form (ie NADH), which was obtained from Sigma (St. Louis, MO), and fenthion (fenthion) was purchased from Dr. Ehrenstorfer (Augsburg, Germany). Coupling buffers for enzyme-hapten conjugates were 0.10M sodium bicarbonate buffer, pH 9.0. The working assay buffer for G6PDH-hapten conjugation is 0.05M tris(hydroxymethyl)aminomethane-hydrochloric acid buffer (tris(hydroxyl)aminomethane-hydrochloricacid buffer), pH 7.8 , containing 0.1M NaCl, 0.01% (w / v) NaN 3 , and 0.3% (w / v) gelatin (gelatin). All buffers were prepared using deionized water, each experiment was repeated three times and the median values ​​are shown.

[0021] (2) Synthesis of hapten and production of antibodies

[0022] Please refer...

Embodiment 2

[0033] Example 2: Determination of enzyme activity and maximum percent inhibition of G6PDH-hapten conjugates.

[0034] (1) Instrument

[0035] Enzyme activity was determined by a Gilford-Stasaar-III spectrophotometer (spectrophotometer), which was equipped with a vacuum-operated sampling system (vacuum-operated sampling system) and temperature-controlled cuvette (temperature-controlled cuvette) (during the whole experiment maintained at 30°C). This spectrophotometer is connected to Syva CP-5000 EMIT ClinicalProcessor (Mountain View, CA), a kind of clinical processor, is used for setting the record of interval reading (reading intervals) and absorbance value (absorbance values) automatically, utilizes Pierce company ( Rockford, IL) produced a microdialyzer system (microdialyzer system 500), used for dialyzer-hapten conjugates.

[0036] (2) Determination of enzyme activity

[0037] The activity of the G6PDH-hapten conjugate, which was detected by adding 100 μl of NAD + (32.5...

Embodiment 3

[0050] Example 3: Kinetic studies on inhibition by G6PDH-hapten conjugates.

[0051] (1) Combined kinetic research

[0052] The binding rate between the 6PDH-hapten conjugate and the anti-hapten antibody was determined using different incubation times (incubation time), and the detection solution contained 100 μl of the appropriately diluted conjugate (1 / 100 dilutions; 8.96×10 -7 concentration of M), and 100 μl of antibody solution (diluted 1 / 1250) were mixed and incubated for 0 to 30 minutes. After each incubation period, the enzymatic activity of the conjugates was determined by addition of substrate, as described above, and kinetic curves were developed by plotting the rate of inhibition versus the position of the incubation period.

[0053] (2) The influence of the concentration of different antibodies and G6PDH-hapten conjugates for inhibition.

[0054] Using an Aliquots type aliquot injector (100 μl), the antibody solution containing different antibodies with differe...

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Abstract

The invention relates to a method for examining the fenthion content, especially by an enzyme linked immunoanalysis test, comprising following steps: (A) mixing an antibody (anti-fenthion) in a reaction solution and glucose-6-phosphatedehydrogenase fenthionconjugate; (B) test and sampling; (C) adding the NAD+ and glucose-6-phosphate to the mixing solution; and (D) examining the growth speed of the NADH by a spectrometer at 340 nm.

Description

technical field [0001] The present invention relates to a method for detecting the content of fenthion. The steps include: (A) mixing an anti-fenthion antibody and glucose-6-phosphate dehydrogenase into a reaction solution (glucose-6-phosphate dehydrogenase, G6PDH) fenthion conjugate (fenthion conjugate); (B) test sampling; (C) NAD + and glucose-6-phosphate dehydrogenase (glucose-6-phosphate dehydrogenase, G6PDH) into the mixed solution; and (D) using a spectrometer to detect the growth rate of NADH at 340nm. Background technique [0002] Due to the widespread use of insecticides, the pollution of the environment and food has become increasingly serious. Among the many types of insecticides, fenthion [O, O-dimethyl O-[3-methyl-4(formazol Thio)phenyl]phosphorothioate]([O,O-dimethyl O-[3-methyl-4-(methylthio)phenyl]phosphorothioate]) is a kind of organophosphorus insecticide (organophosphorus) commonly used in the world insecticide), which is used in different types of culti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/25C12Q1/00
Inventor 李相喜
Owner MYONGJI UNIV IND & ACAD COOPERATION FOUND
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