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Enzyme linked immunosorbent reagent casing for detecting furantoin metabolite and uses thereof

A furanotazone and metabolite technology, applied in the field of enzyme-linked immunosorbent assay kits, can solve the problems of complex instruments and equipment, unsuitable for on-site monitoring, screening of a large number of samples, and tedious processes, so as to overcome technical problems and reduce the lower limit of sample detection. , the effect of improving the detection sensitivity

Inactive Publication Date: 2007-08-08
BEIJING WANGER KANGTAI BIOTECH +1
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0003] The most common methods used to detect furaltadone metabolites are LC-UV, LC-MS and LC-MS / MS, which are not suitable for on-site monitoring and screening of a large number of samples due to complex instruments and cumbersome processes

Method used

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  • Enzyme linked immunosorbent reagent casing for detecting furantoin metabolite and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1 Preparation of kit components

[0067] 1. Antigen Synthesis

[0068] a. Synthesis of furaltadone metabolite derivative hapten

[0069] The furaltadone metabolite and m-carboxybenzaldehyde are reacted in water for derivatization to obtain the furaltadone metabolite derivative hapten.

[0070] Preparation process of furaltadone metabolite derivative hapten:

[0071] Dissolve 2000mg of furaltadone metabolite in 20ml of pure water, dissolve 1500mg of m-carboxybenzaldehyde in 30ml of water, add it to the furaltadone metabolite solution and react at room temperature for 48 hours to obtain a light yellow precipitate which is the furaltadone metabolite Derivative haptens. Repeatedly washing with 50ml of water for 5-6 times, drying to obtain the furaltadone metabolite derivative hapten.

[0072] b. Preparation of immunogenic furaltadone metabolite derivative conjugated antigen

[0073] The immunogen is obtained by coupling the furaltadone metabolite derivative ...

Embodiment 2

[0096] Example 2 Construction of an enzyme-linked immunosorbent assay kit for detecting furaltadone metabolites

[0097] An enzyme-linked immunosorbent assay kit for detecting furaltadone metabolites was constructed to include the following components:

[0098] (1) Enzyme plate coated with anti-furaltadone metabolite antigen;

[0099] (2) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0100] (3) Monoclonal antibody to furaltadone metabolite;

[0101] (4) 6 bottles of furaltadone standard solution, the concentrations were 0 μg / L, 0.1 μg / L, 0.3 μg / L, 0.9 μg / L, 2.7 μg / L, 8.1 μg / L;

[0102] (5) The substrate chromogenic solution is composed of A liquid and B liquid, the substrate chromogenic liquid A liquid is carbamide peroxide, and the substrate chromogenic liquid B liquid is tetramethylbenzidine;

[0103] (6) The stop solution is 2mol / L hydrochloric acid;

[0104] (7) The concentrated washing solution is 0.01M, pH7.6, containing 0.8% Tween 20 and 1‰ so...

Embodiment 3

[0106] Example 3 Detection of furaltadone metabolite residues in samples

[0107] 1. Sample pretreatment

[0108] Animal tissues (chicken, pork, fish and shrimp)

[0109] Take 1±0.05g tissue sample homogenate, add 4ml distilled water, 0.5ml 1M HCl and 100μl 10mM 2-nitrobenzaldehyde, shake well; incubate overnight at 37°C (about 16h); add 5ml 0.1M K 2 HPO 4 , 0.4ml 1M NaOH and 5ml ethyl acetate, shake vigorously for 30s; centrifuge at room temperature (20~25℃ / 68~77) over 3000g for 10min; take out 2.5ml ethyl acetate and blow it with nitrogen at 50℃ Dry or evaporate to dryness with a rotary evaporator; dissolve the dry matter with 1ml of n-hexane, and mix well with 1ml of the diluted complex solution; centrifuge at room temperature (20~25℃ / 68~77) over 3000g for 10min; use 50μl of the lower layer liquid for analysis.

[0110] 2. Detection with kit

[0111] Add 50 μl of a series of standard solutions or sample solutions to the microwells of the microtiter plate coated with ...

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Abstract

The invention provides a detecting furanketone metabolite ELISA kit, which contains: an ELISA plate covered by the original coated, enzyme marker, furanketone metabolite specific antibody or furanketone metabolite derivative antibody, furanketone metabolite standard solution or furanketone metabolite derivative standard solution, the substrate color solution, the termination solution, the condensed washing solution, the condensed complex solution. The invention also provides a method to apply the above ELISA kit detecting furanketone metabolite, which includes steps: firstly, sample pre-treatment, and then using the kit for testing, and finally analyzing testing results. The invention is to provide the furanketone metabolite residues in the ELISA kit for detection of animal derived foods, such as chicken, pork, fish, shrimp, milk, honey, egg, and other samples, and the detection method is simple, low cost, high sensitivity, and it can monitor on the scene and suitable for screening large number of samples.

Description

technical field [0001] The invention relates to an enzyme-linked immunoimmunoassay kit for detecting residues of furaltadone metabolites in samples such as chicken, pork, fish and shrimp, milk, honey and eggs in animal-derived foods and a detection method using the kit . Background technique [0002] Nitrofuran drugs have been widely used as growth-promoting additives for poultry, aquatic products and pigs because of their excellent antibacterial and pharmacokinetic properties. However, in the course of long-term experimental research, it was found that both nitrofuran drugs and metabolites can cause cancer and gene mutation in experimental animals, so these drugs are prohibited from being used in treatment and feed. Furaltadone was banned in 1995. [0003] The most common methods used to detect furaltadone metabolites are LC-UV, LC-MS and LC-MS / MS, which are not suitable for on-site monitoring and screening of a large number of samples due to complex instruments and cumbe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/566G01N33/535
Inventor 沈建忠何方洋冯才茂吴小平万宇平冯才伟汪善良赵正苗余厚美刘玉梅罗贵昆刘平陈炜琳丁双阳张素霞江海洋
Owner BEIJING WANGER KANGTAI BIOTECH
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