Enzyme linked immunosorbent assay kit detecting sodium pentachlorophenate and application of enzyme linked immunosorbent assay kit

An enzyme-linked immunosorbent reagent, sodium pentachlorophenate technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high capital and personnel investment costs, water pollution, etc., and achieve simple pre-treatment process, high accuracy, The effect of high sensitivity

Inactive Publication Date: 2015-11-04
BEIJING KWINBON BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the 1950s and 1960s, my country once used sodium pentachlorophenate in large quantities to kill the host snails of schistosomiasis, and the effect was obvious. However, sodium pentachlorophenate was relatively stable and polluted water bodies.
[0003] GB 29708-2013 "National Food Safety Standard Determina...

Method used

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  • Enzyme linked immunosorbent assay kit detecting sodium pentachlorophenate and application of enzyme linked immunosorbent assay kit
  • Enzyme linked immunosorbent assay kit detecting sodium pentachlorophenate and application of enzyme linked immunosorbent assay kit
  • Enzyme linked immunosorbent assay kit detecting sodium pentachlorophenate and application of enzyme linked immunosorbent assay kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The preparation of embodiment 1 kit components

[0028] 1. Preparation of sodium pentachlorophenate hapten

[0029] 1.0g of sodium pentachlorophenate and ethyl 4 bromobutyrate in DMF, under the catalysis of anhydrous potassium carbonate and sodium iodide, react to generate pentachlorophenol-4 bromo-ethyl butyrate product, which is extracted with ethyl acetate , washed with water, purified and separated by column chromatography to obtain an intermediate product, which was heated and reacted in an aqueous solution of potassium hydroxide, hydrolyzed to generate a hapten, extracted with ethyl acetate, and recrystallized from dichloromethane to obtain a hapten product.

[0030] Get above-mentioned product and measure through proton nuclear magnetic resonance spectrum, such as figure 2 As shown, the chemical shift at 4.0ppm is the methylene signal peak adjacent to the phenolic oxygen, the two groups at 2.0ppm are the methylene signal peak, and the carboxyl hydrogen signal p...

Embodiment 2

[0045] Embodiment 2 detects the formation of the ELISA kit of sodium pentachlorophenate

[0046] Set up the enzyme-linked immunosorbent assay kit that detects pentachlorophenate sodium, make it comprise following components:

[0047] (1) a microtiter plate coated with sodium pentachlorophenate-coupled antigen;

[0048] (2) 6 bottles of sodium pentachlorophenate standard solution, the concentrations are 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 μg / L, 4.05 μg / L;

[0049] (3) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0050] (4) Antibody specific to sodium pentachlorophenate;

[0051] (5) Substrate chromogenic solution is made up of A liquid and B liquid, and A liquid is carbamide peroxide, and B liquid is tetramethylbenzidine;

[0052] (6) The stop solution is 2mol / L sulfuric acid;

[0053] (7) The washing solution has a pH value of 7.4, contains 0.5% to 1.0% Tween-20, 0.01‰ to 0.03‰ sodium azide preservative, and 0.1 to 0.3mol / L phosphate buffe...

Embodiment 4 5

[0064] The determination test of embodiment 4 sodium pentachlorophenate technical parameter

[0065] 1. Kit sensitivity and detection limit

[0066] Determine the sensitivity of the kit according to conventional methods, the range of the standard curve is 0.05-4.05 μg / L, IC 50 (50% inhibitory concentration) floating range is 0.30~0.40 μ g / L; 20 samples are detected, find out the concentration corresponding to each percentage absorbance value from the standard curve, add 3 to the average value of 20 samples concentrations Times standard deviation represents the detection limit, and the results show that the detection limit of this method for shrimp, fish, pork and water samples is 0.75 μg / kg.

[0067] 2. Sample precision and accuracy test

[0068] The recovery rate was used as the accuracy evaluation index, and the relative standard deviation (RSD%) of the test results of repeated determination of a certain concentration sample was used as the precision evaluation index. The...

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Abstract

The invention provides an enzyme linked immunosorbent assay kit detecting sodium pentachlorophenate. The enzyme linked immunosorbent assay kit detecting sodium pentachlorophenate comprises an elisa plate coated with coating antigens, a sodium pentachlorophenate standard product solution, HRP-conjugated antibodies, sodium pentachlorophenate specific antibodies, a substrate color developing solution, a stop solution, washing liquid and reconstruction fluid. The coating antigens are sodium pentachlorophenate coupling antigens. The HRP-conjugated antibodies are goat-anti-mouse antibodies labeled by enzyme. The invention further discloses a method applying the enzyme linked immunosorbent assay kit to detect sodium pentachlorophenate. The method includes the steps that firstly, samples are preprocessed; then; the kit is used for detection; finally, a detection result is analyzed. The enzyme linked immunosorbent assay kit can be used for detecting the content of sodium pentachlorophenate in shrimp mince, fish flesh, pork and water samples and is easy and convenient to operate, low in cost, high in sensitivity, capable of performing on-site monitoring and suitable for screening a large number of samples.

Description

technical field [0001] The invention relates to enzyme-linked immunosorbent detection technology, in particular to an enzyme-linked immunosorbent assay kit for detecting sodium pentachlorophenate, which can qualitatively and quantitatively detect the residual amount of sodium pentachlorophenate in shrimp meat, fish meat, pork and water samples . Background technique [0002] PentachlorophenoI-Na (PCP-Na for short) is the sodium salt of pentachlorophene, which belongs to organochlorine pesticides. It is usually white needle-like or scale-like crystals, volatile, and has a distinct pungent smell. It is easily soluble in water, with a solubility of 20.8% at 4°C. Its aqueous solution has a strong phenolic odor, is stable in nature, and is not easy to decompose under direct sunlight. Sodium pentachlorophenate is widely used in the world as an efficient and cheap insecticide, antibacterial agent, preservative and herbicide. In the 1950s and 1960s, my country used a large amount ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543
CPCG01N33/543G01N33/535G01N2430/00
Inventor 罗晓琴万宇平何方洋贾芳芳彭鸽杨秀贤王建霞张猛
Owner BEIJING KWINBON BIOTECH
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