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
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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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