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Enzyme-linked immunosorbent assay kit for detecting quinclorac and its application

A technology for quinclorac and quinclorac is applied in the field of enzyme-linked immunosorbent assay kits for detecting quinclorac, and can solve the problems that it is difficult to meet the rapid detection of a large number of samples and on-site samples, the processing process is complicated, and the cost is high, To achieve the effect of convenient and easy inspection method, low pre-processing requirements and high accuracy

Active Publication Date: 2017-07-21
ZHENGZHOU TOBACCO RES INST OF CNTC +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Currently commonly used detection methods for quinclorac are mainly instrumental methods, such as high performance liquid chromatography and liquid chromatography tandem mass spectrometry. These analytical methods require expensive instruments and specialized technicians, and the sample pretreatment process is complicated and expensive. , time-consuming, difficult to meet the needs of a large number of samples and rapid detection of on-site samples

Method used

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  • Enzyme-linked immunosorbent assay kit for detecting quinclorac and its application
  • Enzyme-linked immunosorbent assay kit for detecting quinclorac and its application
  • Enzyme-linked immunosorbent assay kit for detecting quinclorac and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 Preparation of kit components

[0027] 1. Preparation of quinclorac hapten

[0028] Take 0.43 g bromoacetyl chloride, add 10 mL chloroform to dissolve, add 30.28 g AlCl 3 Stir, react at room temperature for 2 h, add dropwise 20 mL of a pyridine solution containing 1.0 g of quinclorac, and react at 50 °C for 4 h. Stop the reaction, spin evaporate, remove the organic solvent, add water to dissolve, add dilute hydrochloric acid to adjust pH = 5, extract with ethyl acetate, dry with anhydrous sodium sulfate, concentrate to obtain a red oil, put it on a silica gel column, petroleum ether / ethyl acetate ( v / v, 1 / 1), eluted and separated to obtain 1.38 g of bromoacetylated quinclorac hapten product, with a yield of 92%.

[0029] The above haptens were identified by H NMR spectroscopy, 1 H NMR (CDCl 3 , 300 MHZ) δ: 4.353 (2, 2H), 8.330 (7, 1H, d, J=3.832), 8.812 (9, 1H, dd, J=3.832, J=1.700), 8.703 (15, 1H, d , J=1.700) The chemical shift δ=4.35 is the resonanc...

Embodiment 2

[0044] Example 2 The formation of an enzyme-linked immunosorbent assay kit for detecting quinclorac

[0045] Set up the enzyme-linked immunosorbent assay kit that detects quinclorac to include the following components:

[0046] (1) A microtiter plate coated with quinclorac-conjugated antigen;

[0047] (2) 6 bottles of quinclorac standard solution, the concentrations are 0 µg / L, 0.5 µg / L, 1.5 µg / L, 4.5 µg / L, 13.5 µg / L, 40.5 µg / L;

[0048] (3) Working solution of quinclorac monoclonal antibody;

[0049] (4) Anti-quinclorac antibody labeled with horseradish peroxidase;

[0050] (5) The substrate chromogenic solution is composed of liquid A and liquid B, liquid A is carbamide peroxide, and liquid B is tetramethylbenzidine;

[0051] (6) The stop solution is 2 mol / L sulfuric acid;

[0052] (7) The washing solution has a pH value of 7.4, contains 0.5%~1.0% Tween-20, 0.01‰~0.03‰ sodium azide preservative, and 0.1~0.3 mol / L phosphate buffer;

[0053] (8) The complex solution is a ...

Embodiment 3

[0054] Example 3 Detection of quinclorac in soil

[0055] 1. Sample pretreatment

[0056] Weigh 1.0±0.05 g of the homogenized soil sample into a 10 mL polystyrene centrifuge tube, add 5 mL of distilled water, oscillate with an oscillator for 5 min, mix well, and centrifuge at 3000 g for 5 min at room temperature (20-25°C); Take 50 mL of the supernatant to a 2 mL polystyrene centrifuge tube, add 950 mL of sample reconstitution working solution, vortex for 2 min, mix well, and take 50 mL for analysis.

[0057] 2. Detection with kit

[0058] Add 50 mL of standard / sample to the corresponding microwells, then add 50 mL / well of antibody working solution, shake gently to mix, cover the plate with a cover film and place it in a light-proof environment at 25°C for 30 min. Carefully uncover the cover plate membrane, shake off the liquid in the well, add 250 mL / well of washing working solution, wash thoroughly 4-5 times with an interval of 10 s, pour off the washing liquid in the well,...

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Abstract

The invention provides an enzyme-linked immunosorbent assay kit for detecting quinclorac. The enzyme-linked immunosorbent assay kit comprises an ELISA plate coated with a coating antigen, a quinclorac standard substance solution, a quinclorac antibody, an enzyme-labeled second antibody, a substrate developing solution, a stop solution, a scrubbing solution and reconstitution fluid, wherein the coating antigen is a quinclorac coupling antigen, and the enzyme-labeled second antibody is an enzyme-labeled quinclorac antiantibody. The invention further discloses a method for detecting quinclorac with the enzyme-linked immunosorbent assay kit. The method includes the steps that firstly, sample pretreatment is carried out; then, detection is carried out with the kit; finally, a detection result is analyzed. The enzyme-linked immunosorbent assay kit can be used for detecting the content of quinclorac in a soil sample and is easy and convenient to operate, low in cost, high in sensitivity, capable of being monitored on site and suitable for screening of a large number of samples.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent detection technology, in particular to an enzyme-linked immunosorbent assay kit for detecting quinclorac and its application, which can qualitatively and quantitatively detect the residual amount of quinclorac in soil. Background technique [0002] Quinclorac is a new type of quinolinic acid hormone-type herbicide developed by BASF in Germany, which has the characteristics of strong selectivity and long-lasting effect. Quinclorac has obvious selectivity between annual weeds and gramineous crops, and can be used to control weeds in rice fields before and after seedlings or direct seeding rice fields. It is one of the main herbicide varieties in rice fields in my country. Because quinclorac is acidic and degrades slowly in acidic soil, it is easy to cause serious damage to sensitive crops, especially in rice and tobacco rotation areas, which can cause abnormal growth of tobacco. With the increase i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/535G01N33/577
CPCG01N33/535G01N33/577
Inventor 陈黎范子彦李行杨吕松潘立宁胡斌唐纲岭刘惠民贾芳芳崔华鹏樊美娟
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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