ELISA kit applicable to residue analysis of terrachlor

A pentachloronitrobenzene and residue analysis technology, which is applied in the direction of analyzing materials, biological testing, and material inspection products, can solve problems such as environmental pollution, high requirements for instrumentation, and difficulty in adapting to requirements, achieving less time-consuming, The effect of simple pre-processing

Inactive Publication Date: 2009-09-09
CHINA AGRI UNIV
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  • Abstract
  • Description
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Problems solved by technology

[0003] Conventional detection methods for pentachloronitrobenzene include gas chromatography, high performance liquid chromatography, etc. Applying these physical and chemical analysis techniques to analyze trace amounts of pentachloronitrobenzene residues in environmental, biological, food and other samples requires a relatively high degree of instrumentation. High, and need to go through complicated pretreatment processes such as separation, extraction, purification, derivation, etc., the analysis speed is slow, the cost is high, and the pretreatment process requires the use of a large amount of organic solvents, which has caused new environmental pollution
With the rapid increase in the number of samples to be tested, especially the samples that require rapid on-site detection, traditional pesticide residue analysis methods are difficult to meet the requirements. Therefore, there is an urgent need to develop and apply high-efficiency rapid pesticide residue analysis techniques

Method used

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  • ELISA kit applicable to residue analysis of terrachlor
  • ELISA kit applicable to residue analysis of terrachlor
  • ELISA kit applicable to residue analysis of terrachlor

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Embodiment 1 Kit operation and result calculation

[0019] After the samples to be tested were pretreated, they were made to volume with PBST for later use. Unpack the vacuum packaging bag and take out the microtiter plate, and equilibrate at room temperature for 5 minutes for later use. Prepare 0ng / mL, 10ng / mL, 50ng / mL, 100ng / mL, 500ng / mL, 2000ng / mL pentachloronitrobenzene standard solution, add 50μL standard sample or processed sample to each well, standard sample and Make 4 replicates of the sample, add 50 μL of diluted antibody, and incubate at 37°C for 30 minutes; pour out the liquid in the well, wash 5 times with diluted PBST, invert the plate on absorbent paper and pat dry; add 1: 100 μL of enzyme-labeled goat anti-rabbit secondary antibody diluted at 1000, incubate at 37°C for 30 minutes; pour out the liquid in the well, wash the plate 5 times with PBST, and pat dry; take equal volumes of liquid A and liquid B and mix, add to each well 100 μL, develop color at...

Embodiment 2

[0021] The synthesis of embodiment 2 haptens

[0022] Weigh 2g pentachlorophenol (pentachlorophenol, PCP) and 1gK 2 CO 3 Dissolve in 30mL acetone, add in the reactor, under the protection of nitrogen, add 1.14g bromopropionic acid, heat and reflux for 2 hours, then add 0.5g bromopropionic acid, keep the above conditions unchanged and react overnight (reaction The generated gas is passed into the NaOH absorption bottle for absorption). The organic solvent was evaporated under reduced pressure, the remaining substance was separated with concentrated hydrochloric acid and dichloromethane, the inorganic phase was discarded, and anhydrous Na 2 SO 4 After drying, the organic solvent was evaporated under reduced pressure, and purified by silica gel column chromatography to obtain the crude product of the desired hapten. Yield 50%.

Embodiment 3 5

[0023] Example 3 Pentachloronitrobenzene polyclonal antibody preparation

[0024] Conjugate the hapten with bovine serum albumin by the active ester method, dissolve 2mg of the conjugate in 1mL of normal saline, mix it with 1mL of complete Freund's adjuvant, inject it into the thigh of New Zealand big-eared white rabbit after fully emulsifying, and then inject it into the thigh of New Zealand white rabbit every other day Immunization was boosted once every two weeks, and incomplete Freund's adjuvant was used to mix with the immunogen. The immunization site was subcutaneous on the back of the neck. From the third immunization, blood was collected from the rabbit ear vein one week after each immunization to test the serum titer. A total of 5 times of immunization, the whole blood was collected from the rabbit carotid artery one week after the last immunization, and the rabbit antiserum was crudely extracted by 35% saturated ammonium sulfate salting out method, and finally purifie...

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Abstract

The invention provides an ELISA kit applicable to residue analysis of terrachlor, comprising an ELISA plate coated by terrachlor coating antigens, concentrated cleaning solution, a terrachlor standard, terrachlor polyclonal antibodies, ELIAS secondary antibodies, developing solution and stop solution. The terrachlor with known concentration is detected, and a standard curve is drawn, thus the concentration of the terrachlor to be detected can be calculated. The ELISA kit has the advantages of accurately and sensitively detecting terrachlor residues in environment, being capable of detecting a large amount of samples with simple preprocess procedures and least time consumption, having much lower sample-detecting cost than traditional detecting methods.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay (ELISA) kit, in particular to an enzyme-linked immunosorbent assay kit suitable for the analysis of pentachloronitrobenzene residues. Background technique [0002] Pentachloronitrobenzene (PCNB) is mainly used as a seed dressing agent and soil disinfectant to prevent and control cotton seedling diseases, wheat smut and various diseases of vegetables and fruit trees. It is widely used in the cultivation process of Chinese herbal medicine, especially as a medicament for preventing and treating ginseng diseases at home and abroad. Pentachloronitrobenzene mainly damages the cardiovascular system, central nervous system, liver, kidney and hematopoietic system. my country's mandatory national standards stipulate that the maximum residue limits of pentachloronitrobenzene in vegetables and raw grains are 0.2 and 0.1 mg / kg respectively, and the European Union began to ban pentachloronitrobenzene in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/53
Inventor 李季许艇邵晓龙
Owner CHINA AGRI UNIV
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