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Chemiluminescence detection kit for sulfametoxydiazine and preparation method thereof

A sulfonamide-to-methoxine, chemiluminescence detection technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve the problem of easy inactivation of enzyme markers, low sensitivity, instrument operation Cumbersome and other problems, to achieve the effect of stable kit, high sensitivity and good specificity

Inactive Publication Date: 2018-01-09
太原瑞盛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the liquid chromatography analysis method lacks highly sensitive detectors, and the instrument operation is cumbersome, and the pretreatment is complicated and time-consuming.
The enzyme-linked immunosorbent method uses horseradish peroxidase or alkaline phosphate markers. The enzyme markers are easily inactivated, and the chromogenic substrates are easily decomposed when exposed to light. Inaccurate test results
[0004] CN 201852831U (2011.06) discloses an ELISA detection kit for sulfadimethoxine. The kit uses an enzyme-labeled secondary antibody, which mainly uses enzymes to catalyze the color development of TEM substrates. Reaction time, pH, disinfectant, etc. will all affect enzyme activity. There is a large impact, resulting in poor stability of the reagent
And its sensitivity is low, and the operation process is cumbersome
[0005] CN 201016977 Y (2008.02) used the colloidal gold method to detect the residual amount of sulfa-methoxine in honey and milk products. The final detection result of the kit was characterized by the color visible to the naked eye, and the error was large, and the operation was cumbersome, and the process was relatively complicated. More, more prone to errors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: The formation of kit 1 and the preparation of its specific components

[0021] 1. Assembly of the kit

[0022] A chemiluminescent detection kit for sulfamethoxine, which contains the following components:

[0023] acridinium ester-labeled sulfamethoxine antigen;

[0024] Carboxyl magnetic beads coupled with sulfa-methoxine antibody;

[0025] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0026] Sulfamethoxine series standard solutions, the standard concentrations are: 0 μg / L, 0.01 μg / L, 0.1 μg / L, 1.0 μg / L, 10.0 μg / L, 20.0 μg / L, the buffer solution is Tris-HCl containing 0.5-5.0% BSA and 0.1-0.5% PC300;

[0027] The cleaning solution is specifically a Tris-HCl solution with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl and 0.05% Tween-20 at a concentration of 0.15 mol / L.

[0028] 2. Preparation of magnetic bead-conjugated antibodies

[0029] (1) Take 1 mg of carboxyl magnetic particles in...

Embodiment 2

[0054] Embodiment 2: the formation of kit 2 and the preparation of its components

[0055] 1. Assembly of the kit

[0056] A chemiluminescent detection kit for sulfamethoxine, which contains the following components:

[0057] Acridinate-labeled sulfa-methoxine antibody;

[0058] Carboxyl magnetic beads coupled with sulfa-methoxine antigen;

[0059] Chemiluminescence pre-excitation solution A and chemiluminescence excitation solution B;

[0060] Sulfamethoxine series standard solution, the standard concentrations are: 0 μg / L, 0.01 μg / L, 0.1 μg / L, 1.0 μg / L, 10.0 μg / L, 20.0 μg / L, the buffer solution is Tris-HCl containing 0.5-5.0% BSA and 0.1-0.5% PC300;

[0061] The cleaning solution is specifically a Tris-HCl solution with a pH of 7.2 and a concentration of 25 mmol / L, which contains NaCl and 0.05% Tween-20 at a concentration of 0.15 mol / L.

[0062] 2. Preparation of magnetic bead-conjugated antigen

[0063] (1) Take 1 mg of carboxyl magnetic particles in a 0.5 mL centrifu...

Embodiment 3

[0075] Embodiment 3: the detection of kit

[0076] 1. Sample pretreatment

[0077] (1) Obtaining the milk test sample solution: Take 150 μL of fresh milk in a 500 μL centrifuge tube, centrifuge at 4°C for 10 min (3000 r / min), and discard the upper layer of fat. Pipette 25 μL of the centrifuged milk sample into a clean glass test tube, and add 950 μL of 0.02 mol / L phosphate buffer for dilution.

[0078] (2) Take 1 g of honey and add it to a 25 mL centrifuge tube, add 2 mL of 0.5 mol / L hydrochloric acid solution, oscillate with an oscillator until the honey is completely dissolved, warm it in hot water at 55-75°C for several minutes, add 5 mL of Sodium hydroxide with a concentration of 0.2 mol / L, adjust the pH range to 4.0-6.0, shake and mix. Add 8 mL of ethyl acetate, shake up and down for 5 min, and then centrifuge for 10 min (3000 r / min). Take 5 mL of the organic solvent in the upper layer and put it in a clean glass tube, dry it in the air, add 0.5 mL of 0.02 mol / L phosph...

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Abstract

The invention discloses a chemiluminescence detection kit for sulfamethoxine and a preparation method thereof. The kit includes: magnetic particles coupled with sulfa-methoxine antigen or antibody, acridinium ester markers, sulfa-methoxine series standards, chemiluminescence pre-excitation solution A, chemiluminescence excitation solution B, cleaning solution . Compared with the existing technology for detecting sulfa-methoxine, the kit has the advantages of high sensitivity, high signal-to-noise ratio and rapid detection. Its analytical sensitivity can reach 0.030μg / L.

Description

technical field [0001] The invention relates to the technical field of food detection, in particular to a chemiluminescent detection kit for sulfamethoxine in food and a preparation method thereof. Background technique [0002] Sulfamethoxine, also known as N-(5-methoxy-2-pyrimidinyl)-4-aminobenzenesulfonamide. A sulfonamide drug with antibacterial properties. In veterinary medicine, it is mainly used to prevent or treat bacterial infectious diseases. It is usually mixed into the feed and used by animals as an additive. Once sulfamethoxine is ingested in the body, it takes a long time for metabolism. The blood concentration of sulfamethoxine should not exceed 200 μg / mL. When people eat animal-derived foods containing sulfa-methoxine for a long time, if the concentration exceeds this concentration, the incidence of adverse reactions will increase, causing chronic poisoning, inducing renal failure, and even leading to cancer. occur. In addition, the long-term presence of ...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/532G01N33/543
Inventor 胡雪婷常燕刘丽青曹晶杜爱铭徐兵
Owner 太原瑞盛生物科技有限公司
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